Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832

International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 3 Number 2 (2014) pp. 801-832 http://www.ijcmas.com

Review Article

Actinomycetes: Source, Identification, and Their Applications

Mukesh Sharma*, Pinki Dangi and Meenakshi Choudhary

Department of Biotechnology, Jaipur Institute of Biotechnology, Maharaj Vinayak Global University, Jaipur (Rajasthan) India *Corresponding author

ABSTRACT

The taxonomic and ecological positions of antibiotic producing actinomycetes are well recognized for their metabolic flexibility, commonly accompanied by the production of primary and secondary metabolites of economic significance. Various approaches including classical, chemo taxonomical, numerical taxonomic

and molecular have been routinely employed for the identification of K e yw or ds actinomycetes. The metabolic perspective of actinomycetes not only provides an

interesting area for research but also offers the possibility of commercialization of Actinomycetes; the metabolites generated in the process. Enzymes such as amylase, lipase, and antibiotic; cellulases produced from actinomycetes play an important role in food, bioremediation fermentation, textile and paper industries. Certain enzymes used as therapeutic en zymes; agents in human cancer, mostly in acute lymphoblastic leukemia. Actinomycetes metabolic . are useful in cancer treatment, bioremediation and it produces some valuable antibiotics such as novobiocin, amphotericin , vancomycin, neomycin, gentamycin, chloramphenicol, tetracycline, erythromycin, nystatin, etc. Actinomycetes are also

used as plant growth promoting agents (help to produce plant growth hormone Indole-3-acetic acid), biocontrol tools, biopesticide agents, antifungal compounds, and biocorrosion and as a source of agroactive compounds. Therefore, actinomycetes play a significant role in the production of various antimicrobial agents and other industrially important substances such as enzymes. The potential of actinomycetes in the discovery of novel compounds with activity against microorganisms has been realized, and hence opens exciting avenues in the field of

biotechnology and biomedical research.

Introduction

During 1914 to 1939, Selman A. growth. In 1940, he was able to isolate an Waksman had been consistently effective T.B. antibiotic, actinomycin and systematically screening soil and for this he got success in 1944, with the fungi to find an antibiotic for tuberculosis. discovery of Spectromycin.For all this In 1939, he discovers the effect of certain work in 1952, he got the Noble prize in fungi specially actinomycetes on bacterial physiological & medicine.

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Actinomycetes are filamentous Gram- (aclarubicin, daunomycin and positive bacteria, characterized by a doxorubicin), peptides (bleomycin and complex life cycle belonging to the actinomycin D), aureolic acids phylum , which represents (mithramycin), enediynes one of the largest taxonomic units among (neocarzinostatin), antimetabolites the 18 major lineages currently recognized (pentostatin), carzinophilin, mitomycins, within the Domain Bacteria (Ventura et al. etc (Newman and Cragg 2007; Olano et 2007). Actinobacteria are widely al., 2009). However, the search for novel distributed in both terrestrial and aquatic drugs is still a priority goal for cancer ecosystems, mainly in soil, where they therapy.The rapid development of play an essential role in recycling resistance to multiple chemotherapeutic refractory biomaterials by decomposing drugs and their undesirable side effects has complex mixtures of polymers in dead increased demand for novel antitumor plants, animals and fungal materials. They drugs that are active against fewer side are also important in soil biodegradation effects with untreatable tumors, and with and humus formation as they recycle the the greater therapeutic efficiency (Demain nutrients associated with recalcitrant and Sanchez 2009). polymers, such as chitin, keratin, and lignocelluloses, (Goodfellow and Williams Progress has been made recently on drug 1983, McCarthy and Williams 1992, Stach discovery from actinomycetes by using and Bull 2005) this produces several high-throughput fermentation and volatile substances like geosmin screening, combinatorial biosynthesis and responsible of the characteristic “wet earth mining genomes for cryptic pathways, to odor” (Wilkins 1996) and exhibit diverse generate new secondary metabolites physiological and metabolic properties, for related to existing pharmacophores (Baltz example the manufacture of extracellular 2008). The isolation of marine enzymes (McCarthy and Williams 1992, actinomycetes has been a great source of Schrempf 2001). new compounds and their isolation all around the world from deepest sediments The bioactive secondary metabolites to the shallow costal sediments from the produced by microorganisms is reported to Mariana Trench, demonstrates that be around 23,000 of which 10,000 are actinomycetes are ever-present in marine produced by actinomycetes,thus sediments, but at lower numbers than in representing 45% of all bioactive soil (Ghanem et al. 2000, Zheng et al. microbial metabolites discovered (Berdy 2000, Fiedler et al. 2005, Maldonado et al. 2005). Among actinomycetes, 2009). Marine microorganisms encompass approximately 7,600 compounds are a complex and diverse assemblage of produced by species (Berdy microscopic life forms, of which it is 2005). Several of these secondary estimated that only 1% has been cultured metabolites are potent antibiotics. As a or identified (Bernan et al. 2004). In result of which streptomycetes have addition, marine actinomycetes have been become the primary antibiotic-producing found in symbiosis with different marine organisms exploited by the pharmaceutical invertebrates, especially sponges (Piel industry (Berdy 2005). Members of this 2004, Kim and Fuerst 2006). Marine group are producers of clinically useful actinomycetes have attracted great antitumor drugs such as anthracyclines attention since they have developed unique

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Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832 metabolic and physiological capabilities Presumably this resemblance results partly that not only ensure survival in extreme from adaptation to the same habitat. habitats, but also offer the prospective to Studies of the fine structure of produce compounds with antitumor and actinomycetes spores during germination other interesting pharmacological have been confined to the genera activities that would not be observed in Streptomyces (Kalakoutswl and Agre terrestrial microorganisms (Blunt et al. 1973). The latter genus forms endospores 2006, Mayer et al. 2007, Williams 2009, which behave in a similar way to those of Blunt et al. 2009, Fenical et al. 2002), Bacillus, a new wall layer being perhaps because of their close synthesized inside the cortex of the spore relationships with marine eukaryotic and extending to form the germ-tube wall. organisms including mammals (Baltz 2008, Piel 2004). In the Streptomyces species studied, the spores had a two-layered wall and the However, one of the main problems inner one extended to form the germ-tube associated with marine actinomycetes is wall. It is not clear if this layer is newly the difficulty often found in their culture, synthesized during germination or if it is because of specific necessities like sea salt formed by reorganization of wall material while in some cases these microorganisms existing in the dormant spore. Ultra are obligate halophiles (Tsueng et al. structural changes during the germination 2008). There are a number of reports on of fungal spores have been studied more techniques and approaches for accessing extensively. Most fungi fall into one of previously uncultured soil actinomycetes two groups: (i) those in which the germ- and the biosynthesis gene clusters they tube wall is formed from a layer which is harbor (Janssen et al. 2002, donadio et al. synthesized de now within the existing 2002). In the case of marine actinomycetes spore wall; (ii) those in which the germ- these studies are only beginning, several tube wall is formed by the extension of a attempts to optimize their isolation and wall layer already present in the dormant growth from several sources (Piel 2004, spore (Bartnicki-Garcsi 1968). Some Bull and Stach 2007, Bull et al. 2005) as conflicting results have been obtained and well as the improvement of the closely related species have been reported fermentation process for the production of to fall into different groups (Khan 1975). specific compounds (Tsueng et al. 2008, This may be partly due to the use of Lam et al. 2007, Selvin et al. 2009) and different fixatives, potassium the development of tools to facilitate the permanganate giving inferior results to genetic manipulation of the isolated those obtained with osmium tetroxide or biosynthesis gene clusters (Moore et al. aldehydes (Borderd and Trincia 1970). 2005). Marked changes in spore wall layers can also be induced by hydration during Structure of Actinomycetes specimen preparation (Florancee et al. 1972). The actinomycetes (sing. actinomycete) are a large group of aerobic, high G-C When grown on an agar-surface, the percentage gram-positive bacteria that actinomycetes branch forming a network form branching filaments or hyphae and of hyphae growing both on the surface and asexual spores. These bacteria closely under-surface of the agar. The on-the- resemble fungi in overall morphology. surface hyphae are called aerial hyphae 803

Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832 and the under-surface hyphae are called Molecular Approach substrate hyphae. The most influential approaches to Septa normally divide the hyphae into are through the study of nucleic long cells (20 mm and longer) possessing acids as a result of this square measure many bacterial chromosomes (nucleoids). either direct cistron merchandise or the These are the aerial hyphae that extend genes themselves and comparisons of above the substratum and reproduce nucleic acids yield goodly data true asexually. Most actinomycetes are non- connexion. motile. When motality is present, it is confined to flagellated spores. Molecular science, which has each classification and identification, has its Cell Wall Composition origin within the early supermolecule

The composition of cell wall in crossbreeding studies, however has actinomycetes varies greatly among achieved a new standing following the different groups and is of considerable introduction of supermolecule sequencing taxonomic significance. Four major cell techniques (O‟Donnell et al. 1993). wall types are distinguished in these Importance of phyletic studies supported filamentous bacteria on the basis of the 16S rDNA sequences is increasing within three features of peptidoglycan the science of bacterium and composition and structure. These features actinomycetes (Yokota 1997). Sequences are (i) diaminopimelic acid isomer on of 16S rDNA have provided tetrapeptide side chain position 3, (ii) actinomycetologists with a phyletic tree sugar content of peptidoglycan, and (iii) that enables the investigation of evolution the presence of glycine in interpeptide of actinomycetes and conjointly provides bridges. As is evident in, characteristic the premise for identification. sugar patterns are present only in cell wall types II-IV of those actinomycetes with meso-diaminopimelic acid. Analysis of the 16S rDNA begins by analytic DNA (Hapwood et al. 1985) and Isolation of actinomycetes amplifying the gene coding for 16S rRNA exploitation the enzyme chain reaction For the isolation of actinomycetes, various (Siva Kumar 2001. The refined DNA methods can be performed on the basis of fragments are directly sequenced. The different sources and media. Samples were sequencing reactions are performed collected from different ecological exploitation DNA sequencer so as to work habitats. Further characterization can be out the order during which the bases are performed to study the different strains of organized at intervals the length of sample actinomycetes. (Xu et al. 1999) and a computer is then used for finding out the sequence for Identification identification exploitationphyletic analysis procedures. Though, analysis of 16S Various approaches for the identification rDNA generally allows us to identify the of actinomycetes square measure given in organism‟s upto the genus level only. brief below:

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Table.1 Different sources and media for isolation of actinomycetes.

SOURCE MEDIA REFERENCES FROM SOIL: Forest Soil Starch-casein medium Kuster & Williams(1964) Humus Layer of Forest Soil (a)Humic acid-vitamin agar Cho et al, 1994 (b)Starch casein nitrate agar(SCS) Hayakawa et al, 1987a

(c)Hair hydrolysate vitamin Hayakawa et al, 1987b agar(HHVA)

(d)Bennet‟s agar(BA) Seong C.N., 1992 Corn Field, Cow Barn yard, Forest (a)Arginine-glycerol Porter et al, 1960 salt(AGS)medium (b)Chitin medium Lingappa & Lockwood, 1961 (c)Modified Benedict‟s medium Porter,Wilhelm & Tresner, 1960

(d)Soybean meal-glucose medium Tsao,Leben & Keitt, 1960

(e)Gauze‟s agar medium Rehacek, 1959 (f)Czapek‟s agar medium Waksman, 1961 (g)Egg albumen medium Waksman, 1961 (h)Glucose-asparagine medium Waksman, 1961

(i)Glycerol-asparaginate agar 2 Waksman, 1961

Lake Soil Chitin agar S.C. HSU & J.L. Lockwood, 1975

Soil Coal-vitamin agar Wakisaka et al, 1982 Antartic Soil Mineral salt(MS) medium Kosmachev (1954) Mitidja plain (Algeria) Yeast extract-malt extract agar Shirling & Gottlieb, 1966

Marine Soil Starch casein nitrate(SCN) agar Ravel J, Amorso (1998) medium FROM WATER: Stream Sediments & Lake muds (a)Chitin agar media Lingappa & Lockwood (1961,1962)

(b)M3 agar medium Jones, 1949 (c)Benett‟s medium Jones, 1949 Marine Sediments (a)Starch-casein agar A.Grein & S.P. Meyers, 1958 (b)Asparagine agar A.Grein & S.P. Meyers, 1958 (c)Glycerol-glycine agar Lindenbein, 1952 Marine Sediments(South China) (a)AIM medium J.L. You et al

FROM OTHER SOURCES: FROM ROOT & STEM SAMPLES OF FOUR PLANTS: Cinnamomum zeylanicum,Zingiber Starch yeast casein Zin et al, 2007 spectabile,Elettariopsis curtisii, agar(SYCA),Actinomycetes Isolation Labisia pumila agar (AIA), Humic Acid vitamin gellan gum (HVG),Tap water yeast extract agar (TWYE), Coal –vitamin agar (CVA) Mangroove Sediments Asparagine-glucose agar medium Shirling & Gottlieb, 1966

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Procedure: Various steps for the Isolation and characterization of actinomycetes were performed which are mentioned below:

Collection of samples from different habitats (According to table no.1)

Enrichment of samples using different treatments

Incubation

Sample were culture on different media (according to table no.1)

Isolate was characterized by Morphological, Physiological, Biochemical and Molecular methods.

Strains of actinomycetes isolated

Morphological Physiological Biochemical methods: Molecular methods: methods: methods: •Catalase production •RFLP using any one (a)Macroscopic • Range of pH for •Urease production of genomic DNA methods growth •Hydrogen sulfide •RAPD •Cover slip culture • Optimum production •PFGE •Nitrate reduction •ARDRA (b)Microscopic temperature for growth methods •Starch hydrolysis •Use of genus specific •Slide culture method •Salinity •Gelatin liquefaction primers •Methyl red test •Vogues-proskauer test •Indole production •Citrate utilization

•Casein hydrolysis

;:

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Chemotaxonomical Approach Classical Approach

Chemotaxonomy is that the study of Classical approaches for classification chemical variation in organisms and also build use of physiological, morphological, the use of chemical characters within the and biochemical characters. The classical classification and identification. It‟s one in methodology delineated within the all the precious strategies to spot the identification key by Nonomura genera of actinomycetes. Studies of (Nonomura 1974) and Bergey‟s Manual of Cummins and Harris (Cummins and Harris Determinative Bacteriology (Buchanan 1956) established that actinomycetes have and Gibbons 1974) is very much useful in a cytomembrane composition comparable the identification of streptomycetes. These to that of gram-positive bacterium, and characteristics are normally utilized in conjointly indicated that the chemical employed in taxonomy of streptomycetes composition of the cytomembrane may for several years. They are quite helpful in furnish sensible strategies of routine identification. These are discussed differentiating numerous varieties of below: actinomycetes. This can be due to the actual fact that chemical components of 1. Aerial Mass Color the organisms that satisfy the subsequent conditions have important which means in The colour of the mature sporulating aerial science. mycelium is recorded in an exceedingly straightforward method (White, grey, red, They must be distributed universally green, blue and violet). Once the aerial among the microorganisms studied; and, mass color falls between two colors series, both the colors are recorded. If the aerial The parts ought to be homologous among mass color of a strain to be studied shows the strains at intervals a taxonomic group, intermediate tints, then also, both the whereas important variations exist colors series are noted. between the taxa to be differentiated. 2. Melanoid Pigments Presence of Diaminopimelic Acid (DAP) isomers is one in all the foremost The grouping is formed on the assembly of necessary cell-wall properties of gram- melanoid pigments (i.e.light-green brown, positive bacterium and actinomycetes. brown black or distinct brown, pigment Most bacterium has a characteristic wall changed by alternative colours) on the envelope, composed of peptidoglycan. The medium. The strains are grouped as 2, 6‐ Diaminopimelic Acid (DAP) is melanoid pigment created (+) and not cosmopolitan as a key amino acid and its created (‐). optical isomers. The systematic significance lies largely within the key 3. Reverse Side Pigments amino acid with two amino bases, and determination of the key amino acid is The strains were divided into two groups, typically adequate for characterization. If consistent with their ability to provide DAP is present,bacterium typically contain characteristic pigments on the reverse one of the isomers, the LL‐form or the aspect of the colony, namely, distinctive meso-form, largely settled within the (+) and not distinctive or none (‐). In peptidoglycan. 807

Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832 case, a color with low saturation like Numerical Taxonomic Approach yellowness, olive or yellowish brown occurs, it is included in the latter group Numerical taxonomy involves examining (‐). several strains for a large number of characters prior to assigning the test 4. Soluble Pigments organism to a cluster based on shared options. The numerically defined taxa are The strains are divided into two groups by polythetic; therefore, no single property is their ability to provide soluble pigments either indispensable or adequate to entitle apart from melanin: particularly, produced an organism for membership of a group. (+) and not produced (‐). The color is Once classification has been achieved, recorded (orange, red, green, violet, blue cluster‐specific or predictive characters and yellow). is chosen for identification (Williams et al. 1983). Numerical taxonomy was initially 5. Spore Chain Morphology applied to Streptomyces (Silvestri et al. 1962). The numerical taxonomic study of With relevancy to spore chains, the strains the genus Streptomyces by Williams et al. are sorted into „sections‟. The species (1983) involves determination of 139 unit belonging to the genus Streptomyces are characters for 394 type cultures of divided into three sections (Shirling and Streptomyces; clusters were outlined at Gottlieb 1966), particularly rectiflexibiles 77.5% or 81% Ssm and 63% Sj similarity (RF), retinaculiaperti (RA) and Spirales levels, and also the former co-effieient is (S). Once a strain forms two types of spore being employed to outline the clusters. His chains, both are noted (e.g. SRA). study includes 23 major, 20 minor and 25 single member clusters. 6. Reproductive Structure Surface The numerical classification of the genus Spore morphology and its surface options Streptomyces by Kampfer et al. (Kampfer ought to be determined under the scanning et al. 1991) involves determination of 329 electron microscope. The cross hatched physiological tests. His study includes 15 cultures arranged for observation under the major clusters, 34 minor clusters and 40 light microscope can be used for this single member clusters which are defined purpose. at 81.5% similarity level Ssm using the simple matching coefficient (Sokal and The electron grid ought to be cleaned and Michener 1958) and 59.6 to 64.6% adhesive tape should be placed on the similarity level Sj using Jaccard surface of the grid. The mature spores of coefficient (Sneath 1957). Thus, numerical the strain ought to be rigorously placed on taxonomy provides us with a useful the surface of the adhesive tape and gold framework for Streptomyces taxonomy, as coating should be applied for half an hour well as identification of species. and also the specimen is examined under the electron microscope at completely Role of Streptomycetes different magnifications. The reproductive structure silhouettes are characterized as Streptomyces is the largest genus of spiny, smooth, warty and hairy. Actinobacteria and the type genus of the family (Kampfer et al.

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1991). Over 500 species of Streptomyces first carbapenem beta-actam antibiotic). A bacteria have been described by Euzeby number of the antibiotics produced by (Euzeby 2008). Streptomycetes have Streptomyces have proven to be too toxic genomes with high GC-content and these for use as antibiotics in humans, other than are gram-positive (Madigan and Martinko because of their toxicity towards cells 2005). Found predominantly in soil and (specifically dividing cells) they have been decaying vegetation, mainly reinvented as chemotherapy drugs. We are streptomycetes produce spores and are talking drugs like: actinomycin-D (the noted for their distinct “earthy” odor original), bleomycin (glycopeptide made which results from production of a volatile by S. verticullus), mitomycin (aziridine metabolite, geosmin. Streptomycetes are made by S. lavendulae) and plicamycin characterized by a complex secondary (made by S.plicatus) (Birnbaum et al. metabolism. They make over two-thirds of 1985). the clinically useful antibiotics of natural origin (e.g. neomycin, chloramphenicol) The Research of microbial metabolites

(Kieser et al. 2000). A variety of actinomycetales, first of all the filamentous fungi and Streptomyces Streptomyces-derived antifungals tend to species, and to a lesser extent several be macrolide polyenes (large ring structure bacterial species are the most important with lots of conjugated carbon-carbon producers both in respect of versatility, double bonds) and include such illustrious diversity, and numbers of structures of the members as: nystatin (the first produced metabolites. The frequency and Actinobacteria-sourced human antifungal, significance of these major types of made by S. noursei), amphotericin B microbes as producers of bioactive (made by S. nodosus, originally isolated metabolites had varied significantly during from a sample of Venezuelan soil) and the last decades. At the start of the natamycin (made by S. natalensis). There antibiotic era the fungal (penicillin, are a friggin‟ tonne of Steptomyces- Griseofulvin) and bacterial (Gramicidin) derived antibiotics used specifically as species were in the forefront of the antibacterial agents. These include interest, but after the detection of Streptomycin by S. griseus, neomycin and streptomycin and afterward kanamycin, respectively produced by S. cholramphenicol, tetracyclines and fradiae and S. kanamyceticus. Other macrolides the attention turned to the antibacterial antibiotics of note include: species of Streptomyces. erythromycin (a macrolide that often subs for penicillin when people be allergic to it, In the fifties and sixties the majority (70%) made by S. erythraea), tetracycline ( a of antibiotics was discovered from these longstanding acne drug that makes you species. The most characteristic and a little light-sensitive, made by S. rimosus), bit surprising feature of the recent years chloramphenicol (cheap, effective, but can just is this declining representation of the cause aplastic anemia, made by S. formerly exhaustively investigated venezuelae), vancomycin (a relatively Actinomycetes. They contribute to among ginormous glycopeptide that can turn all microbial products present in only 30- people red, made by S. orientalis) and 35%, in contrast to the 75-80% share from thienamycin (made by S. cattleya, the sixties to the eighties. The most modified by us to make imipenem, the frequent producers, the Streptomyces

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Table.2 In Table the numbers of actinomycetales species, including the all rare actinos, known to produce bioactive metabolites, are summarized.

Actinomycetales species No. Actinomycetales species No. Streptomycetaceae: Thermomonosporaceae: Streptomyces 8000 Actinomadura 345 Streptoverlicillium 258 Saccharothrix 68 Kitasatosporia 37 Microbispora 54 Chainia 30 Actinosynnema 51 Microellobosporia 11 Nocardiopsis 41 Nocardioides 9 Microtetraspora/Nonomuria 26/21 Micromonosporaceae: (Actinoplanetes) Thermomonospora 19 Micromonospora 740 Micropolyspora/Faenia 13/3 Actinoplanes 248 Thermoactinomyces 14 Dactylosporangium 58 Thermopolyspora 1 Ampullariella 9 Thermoactinopolyspora 1 Glycomyces 2 Mycobacteriaceae: (Actinobacteria) Catenuloplanes 3 Nocardia 357 Catellatospora 1 Mycobacterium 57 Pseudonocardiaceae: Arthrobacter 25 Saccharopolyspora 131 Brevibacterium 17 Amycalotopsis/Nocardia 120/357 Proactinomyces 14 Kibdellosporangium 34 Rhodococcus 13 Pseudonocardia 27 Other (unclassified) species: Amycolata 12 Actinosporangium 30 Saccharomonospora 2 Microellobosporia 11 Actinopolyspora 1 Frankia 7 Streptosporangiaceae: (Maduromycetes) Westerdykella 6 Streptosporangium 79 Kitasatoa 5 Streptoalloteichus 48 Synnenomyces 4 Spirillospora 11 Sebekia 3 Planobispora 10 Elaktomyces 3 Kutzneria 4 Excelsospora 3 Planomonospora 2 Waksmania 3 Alkalomyces 1 Catellatospora 1 Erythrosporangium 1 Streptoplanospora 1 Microechinospora 1 Salinospora 1

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Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832 species produce 7600 compounds (74% of importance since they possess a capacity all actinomycetales), although the atypical to produce and secrete a variety of Actinomycetes represent 26%, altogether extracellular hydrolytic enzymes (Saadoun 2500 compounds. The representation of et al. 2007, Tan et al. 2009). Many rare action products in 1970 was only 5%. actinomycetes have been isolated from In the group Nocardia, Streptoverticillium, various natural sources, as well as in plant Micromonospora, Streptosporangium, tissues and rhizospheric soil. Biological Actinoplanes, Saccharopolyspora and functions of actinomycetes mainly depend Actinomadura, species are the most on sources from which the bacteria are frequent producers; each produces several isolated. Microbial alkaline proteases for hundreds of antibiotics. manufacturing uses are produced mostly from Streptomyces and Bacillus. Enzyme production from Actinomycetes Actinomycetes, particularly Streptomycetes are known to secrete Marine actinomycetes physiological, multiple proteases in culture medium (Sharmin 2005). biochemical and molecular characteristics such as 16SrRNA and terrestrial Actinomycetes have been revealed to be actinomycetes a great difference, followed an excellent resource for L-asparaginase. by metabolic pathway is also different A range of actinomycetes, mainly those from terrestrial actinomycetes, which isolated from soils such as Streptomyces produced a variety of biologically active griseus, S. karnatakensis, S. albidoflavus enzymes. and Nocardia sp. have abilities to produce L-asparaginase enzyme (DeJong 1972, Actinomycetes secrete amylases to the Narayana et al. 2007, Mostafa and Salama outside of the cells to carry out 1979). The production of L-asparaginase extracellular digestion. α amylase starch has been studied in Serratia marcescens degrading amylolytic enzymes is of great (Khan et al. 1970), Erwinia carotovora significance in biotechnological (Maladkar et al. 2000), Enterobacter applications such as food industry, aerogenes (Mukherjee et al. 2000), fermentation and textile to paper industries Pseudomonas aeruginosa (Abdel-Fattah (Pandey et al. 2000). Actinomycetes are and Olama 2002), Bacillus subtilis (Fisher one of the known cellulose producers and Wray 2002) and Saccharomyces (Jang and Chenks 2003, Arunachalam et cerevisiae (Ferrara et al. 2006). Microbial al. 2010). Cellulases are a collection of L-asparaginase has been generally used as hydrolytic enzymes which hydrolyze the a therapeutic agent in the cure of certain glucosidic bonds of cellulose and related human cancers, mostly in acute cello-digosaccharide derivatives (Ito lymphoblastic leukemia (Gallagher et al. 1997). Lipase is produced from a variety 1989, Verma et al. 2007). The roots and of actinomycetes, bacteria, and fungi rhizomes of several Thai medicinal plants (Kulkarni and Gadre 2002). Lipases have such as lemon grass (Cymbopogon broad applications in the detergent citratus), ginger (Zingiber officinale) have industries, foodstuff, oleochemical, long been used in Thai traditional diagnostic settings and also in industries of medicine for stomach ache and asthma pharmaceutical fields (Schmid et al. 1998). treatment (Wutthithamavet 1997). Actinomycetes are of enormous Rhizosphere soil of these plants may be an

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Fig.1 Applications of enzymes produced from actinomycetes.

attractive actinomycetes source, able of bioremediation. They are significantly producing novel secondary metabolites. well adapted to survival in harsh Catalase, Chitinase and Urease enzymes environments. Some are able to grow at also produce from actinomycetes. elevated temperatures (>50°C) and are essential to the composting method. Applications of Actinomycetes Human Health Importance Ecological Importance Antibiotics Actinomycetes are abundant in soil, and are responsible for much of the digestion Actinomycetes are produced many of resistant carbohydrates such as chitin antibiotics, that are best recognized and and cellulose. They are liable for the most valuable. These antibiotics include pleasant odor of freshly turned soil. amphotericin, nystatin, chloramphenicol, Several actinomycetes and other gentamycin, erythromycin, vancomycin, actinobacteria are renowned as degraders tetracycline, novobiocin, neomycin, etc. of toxic materials and are used in

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Fig no.2 Applications of Actinomycetes

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In these antibiotics some are targeted farmers (especially dairy farmers) handle bacterial ribosome‟s and are used in stored hay in winter and early spring. The treating respiratory infections, for example same fungi that cause molding of hay are in treating the Legionnaires‟ disease used common inhabitants of soil, and have tetracycline and erythromycin. additionally been documented to colonize Vancomycin antibiotic are attacks on ventilation systems, garments dryers, deadly organisms such as methicillin- refrigerator drip pans, and any other site resistant staphylococcus aureus (MRSA) that combines heat, cellulosic or other (multiply drug resistant) and bacterial cell carbohydrate material, and water. walls. Rifamycins are useful for treating Common species include leprosy and tuberculosis, these targets Thermoactinomyces vulgaris bacterial RNA polymerase. Amphotericin Saccharopolyspora rectivirgula is one of the minority antibiotics that Thermoactinomyces viridis and others. attack fugal membranes. These antibiotics usually do not influence human cells and Volatile Organic Compounds (VOCs) for that reason have fewer side effects. On the other hand actinomycetes metabolites The odor of freshly turned soil is that the for example adriamycin, prevent DNA results of geosmin, a volatile organic replication, because of this it is used in compound made by actinomycetes. treating the cancer, although rapamymycin Geosmin is additionally made by some is used to repress the immune system to cyanobacteria and produces an earthy taste facilitate organ transplants. in drinking water. Some fungi also produce geosmin, which might impart an equivalent earthy taste to wine made of Infections moldy grapes. In general, folks realize the geosmin odor pleasant in soil. However, Members of the actinomycetes genus are one indoor air research group is usual commensal members of oral cavities investigating the possibility that exposure in human. They can be able to cause to geosmin is related to building-related serious infections when they attack on symptoms. The data at present is too tissue through breaks in the oral mucosa. restricted for conclusions. However, in the The disease is becoming less universal, but future, assortment of samples which will in the USA it is still present, mainly in reveal these organisms could be suggested. inner immunity. Additionally Nocardia species may also be involved. Actinomycetes as Antifungals

Urauchimycins be a Member of antimycin Hypersensitivity pneumonitis (HP) class, a set of well-identified antifungals. Antimycins act by inhibiting the electron Thermophilic actinomycetes are the most flow in the mitochondrial respiratory chain common cause of HP. Farmer's lung (Barrow et al. 1997). Antimycins have disease is HP resulting from exposure to been identified in Streptomyces isolated hay that has become colonized with from the integument of attine ants thermophilic actinomycetes, which (Schoenian et al. 2011, Seipke et al. 2011, produce an abundance of airborne spores. Seipke et al. 2012). Schoenian and Clouds of these spores are released when colleagues (Schoenian et al. 2011) Identify

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Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832 the well-know antimycins A1–A4 in 50% appealing and essential to establish the of the actinobacteria identified as toxicity obtainable by urauchimycin B, to Streptomyces isolated from workers of evaluate whether it can be used as an several Acromyrmex species. Compounds antifungal agent for humans and animals. of this class may have an effective role in In addition, assessment of the isolated the attine ant-microbe association. Another compound against Candida species as antifungal compound broadly Disperse in opposed to commercially offer antifungal Streptomyces related to attine ants is agents should be performed to prove the candicidin (Haeder et al. 2009, Schoenian potential of this relatively unexplored et al. 2011, Seipke et al. 2011, Seipke et antifungal. Actinobacteria of attine ants al. 2012). Urauchimycins A and B were are capable to create antifungal isolated from Streptomyces sp. From a compounds active against other fungal marine sponge Ni-80 was isolated. species and not only against the specific

In 2006, two new urauchimycins were fungal parasite Escovopsis. The few represented: urauchimycin C, isolated current studies that focused on the from Streptomyces sp. B1751 from marine chemical characterization of bioactive sediment, and urauchimycin D, isolated compounds formed by Actinobacteria from Streptomyces sp. AdM21 from soil associated with attine ants support the (Yao et al. 2006). In the study by Imamura potential isolation of novel molecules with and coworkers (Imamura et al. 1993), the biological activity (Oh et al. 2009, Barke urauchimycins A and B repressed the et al. 2010, Carr et al. 2012, Haeder et al. morphological differentiation of C. 2009, Schoenian et al. 2011). Thus, an albicans equal to a concentration of 10 µg exploration program of isolation of mL−1. Urauchimycins C and D showed no bioactive molecules from actinobacteria inhibitory action against C. albicans, from attine ants definitely will result in the Mucor miehei, and bacteria (Yao et al. discovery of novel compounds with 2006). activity against microorganisms that are potentially pathogenic to humans. Urauchimycin B showed inhibitory activity against all Candida strains Actinomycetes for Extracellular evaluated, showing MIC like to those Peroxidase Activity provided by nystatin. Urauchimycin B showed a wide spectrum of activity Peroxidases catalyze the peroxide- against Candida spp. with MIC values dependent oxidation of a range of equal to the nystatin antifungal, which inorganic and organic compounds and are indicates the potential for medical use. widely distributed throughout plants, Antimycins were used from many years animals, and microorganisms. They are for the cure of human infections, but due primarily intracellular enzymes with vital to its mechanism of action and allied side roles in cellular processes (Everse et al. effects, its use in the treatment of human 1990), but extracellular peroxidases disease was discontinued (Barrow et al. concerned within the degradation of 1997). However, with the urgent need for complex organic compounds have new antifungal agents that complement or conjointly been described. The white-rot alternate for the insufficient products basidiomycete Phanerochaete obtainable in the marketplace, it is chrysosporium secretes a complex array of

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Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832 peroxidases throughout secondary high-level extracellular peroxidase metabolism, and their characterization and producers, and in these strains peroxidase role in lignocellulose degradation are well activity was conjointly determined with documented (Gold and Alic 1993). The alternative assay systems. The objective production of extracellular peroxidases by was to evaluate the reliability and actinomycete bacteria has been described applicability of a range of available assays (Godden et al. 1992, Ramachandra et al. for the determination of actinomycete 1988, Winter et al. 1991), but evidence for peroxidase activity. the involvement of this peroxidase activity within the degradation of lignin is ill Actinomycetes as source of Agroactive defined (Godden et al 1992, Spiker et al. compounds 1992). The intracellular peroxidases of streptomycetes have conjointly been Actinomycetes have the most fruitful specifically studied in relevance their role source of microorganisms for all types of within the biosynthesis of halogenated bioactive metabolites, including agroactive antibiotics (Van Pee et al. 1987). type. Over one thousand secondary metabolites from actinomycetes were Extracellular peroxidases would be discovered during 1988-1992. Most of expected to possess improved stability these compounds are produced by various over their intracellular counterparts, species of the genus Streptomyces. In fact, significantly those from thermophiles about 60% of the new insecticides and (Iqbal et al. 1994), and so have potential herbicides reported in the past 5 yr for applications in, as an example, originate from Streptomyces (Tanaka and diagnostic kits. Actinomycetes are a Omura 1993). It is also estimated that as potentially rich source of peroxidases for many as three-quarters of ail introduction into a market that‟s Streptomycetes species are capable of substantial and almost totally dominated antibiotic production (Alexander 1977). by horseradish peroxidase (HRP), which is Actinomycetes produce a variety of both well characterized and extremely antibiotics with diverse chemical active. Peroxide-dependent oxidation of structures such as polyketides, b-lactams luminal by HRP is that the basis of variety and peptides in addition to a variety of of diagnostic immunoassays (Thorpe et al. other secondary metabolites that have 1985) and therefore the enhanced antifungal, anti-tumor and chemiluminescence system for immunosuppressive activities (Behal nonradioactive detection of nucleic acid 2000). hybridization (Stone and Durrant 1991). Screening actinomycetes for extracellular Kasugamycin is a bactericidal and peroxidases is hampered by the necessity fungicidal metabolite discovered in to preconcentrate culture supernatants, but Streptomyces kasugaensis (Umezawa et al. in this paper researchers describe a unique 1965). This antibiotic acts as an inhibitor use of the chemiluminescent assay with of protein biosynthesis in microorganisms the Amerlite analyzer (Amersham PLC, but not in mammals, and its toxicological Bucks, and United Kingdom) to examine a properties are excellent. Hokko Chemical taxonomic range of actinomycetes for Industries developed a production process extracellular peroxidase activity. This to market the systemically active enabled the identification of a group of kasugamycin for control of rice blast

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Pyricularia oryzae Cavara and bacterial 1978), but the fact that this product never Pseudomonas diseases in several crops. appeared in recent publications would indicate, however, that Takeda's efforts to Polyoxin B and D were isolated as develop mildiomycin might not be metabolites of Streptomyces cacaoivar. successful yet. Asoensis in 1965 by (Isono et al. 1965) as a new class of natural fungicides. The The compounds mentioned above are a mode of action of the polyoxins makes few examples of agroactive compounds them very acceptable with regard to isolated from actinomycetes. Microbial environmental considerations. They screening and chemistry procedures have interfere with the fungal cell wall been until recently the main tools to synthesis by specifically inhibiting chitin discover new agroactive compounds. syntheses (Endo and Misato 1969). However, genomic technologies that allow Polyoxin B found application against a the rapid characterization of microbial number of fungal pathogens in fruits, genomes will certainly become the method vegetables and ornamentals. Polyoxin D is of choice for the discovery of new marketed by several companies to control bioactive molecules in the coming years. rice sheath blight caused by Rhizoctonia Furthermore, molecular techniques such as solani Kùhn. combinatorial biosynthesis (Hutchinson 1999) may lead to the discovery of drugs The validamycin family was detected by that cannot be found in nature. Indeed, researchers in 1968 in a greenhouse assay genetic domains, modules and clusters when screening Streptomycetes extracts involved in the microbial biosynthesis of for activity against rice sheath blight. known secondary metabolites can be Validamycin A was found to be a prodrug interchanged and modified to produce which is converted within the fungal cell bioactive products with unique properties. to validoxylamine A, an extremely strong inhibitor of trehalase (Kameda et al. Actinomyccetes as plant growth 1987). This mode of action gives promoting Agents validamycin a favorable biological selectivity because vertebrates do not In attempts to develop commercial depend on the hydrolysis of the biocontrol and plant growth promoting disaccharide trehalase for their products using rhizobacteria, it is metabolism. important to recognize the specific challenges they present. To begin with, the The isolation of the antifungal metabolite interaction between PGPR species and mildiomycin from a culture of their plant symbionts appears to be Streptoverticillium rimofaciens Niida was specific, even within a crop or cultivar reported in 1978, also by Takeda scientists (Chanway et al. 1988, Glick 1995, (Iwasa et al. 1978). Mildiomycin is KIoepper 1996, Lazarovits and Nowak strongly active against several powdery 1997). While a rhizobacterium screened mildews on various crops (Harada and for growth promotion may reveal the Kishi 1978), acting as an inhibitor of the positive effects on one crop, it may have fungal protein biosynthesis (Feduchi et al. no effect, or even retard growth of another 1985). Its low toxicity in vertebrates crop (Gardner et al. 1984, O'Neill et al. would make it an environmentally sound 1992). crop protection agent (Harada and Kishi 817

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Although rhizobacteria may present bacteria, or when PGPR or else facilitates unique challenges to our attempts to plant uptake of soil nutrients. Possibilities harness their beneficial attributes, the include siderophore synthesis, nitrogen prospects for improved agriculture by the fixation, solubilization and phytohormone use of biocontrol-PGPR seem excellent. synthesis, of minerals to make them Advances in our understanding of the available for plant uptake and use (Glick PGPR Systems responsible for plant 1995). growth improvement is a first logical step in opening the way to improving these Some researchers isolate as a seed bacterial strains through genetic treatment of oat, barley, carrot and wheat, engineering, and creating more interest in in order to increase their growth. The their progress for widespread commercial isolate was originally selected for the use for both biocontrol and plant growth biological control of Rhizoctonia solani. promotion. Although the S. griseus isolate did enhance the average dry foliage weight, Despite the well-documented history of tiller number, grain yield, and advanced Streptomyces in biocontrol and head emergence for wheat and oat over the preliminary evidence of their capacity to controls, the differences were not enhance plant growth (Aldesuquy et al. statistically significant. As a seed 1998), Streptomyces species have been treatment for carrot, the isolate was more poorly investigated specifically for their successful. Marketable yields were potential as PGPR. These are amazing as increased over controls by 17% and 15% streptomycetes, usually accounting for an in two separate field trials. Specifically, abundant proportion of the soil microflora, both trials also indicated an increased are particularly effective colonizers of yield of large and very large grade carrots plant root Systems and are able to endure over the controls (Merriman et al. 1974). unfavorable growth conditions by forming Nearly 20 yr later, El-Abyad et al. (El- spores (Alexander 1977). While the Abyad et al. 1993) described the use of beneficial effect of some strains of PGPR three Streptomyces spp. in the control of on particular crops is certain, the bacterial, Fusa rium and Verticillium wilts, mechanisms employed by PGPR are early blight, and bacterial canker of unclear (Glick 1995). PGPR can affect tomato. The isolâtes used were S. pulcher, plant growth in two general ways, either S. canescens, and S. citreofluorescens. As directly or indirectly. Indirect promotion a seed-coating, all three of the strains were occurs when PGPR lessen or prevent the effective at variable levels in controlling harmful effects of one or more deleterious the test pathogens. In addition, tomato microorganisms. This is mainly attained growth was observed to be significantly through biocontrol, or the antagonism of improved with the antagonistic pathogens of soil plant. Specifically, Streptomyces spp. as a seed-coating. colonization or the biosynthesis of antibiotics (Fenton et al. 1992) and other secondary metabolites can prevent The culture filtrates alone of two different pathogen establishment and invasion. Streptomyces spp. (S. olivaceoviridis Direct promotion of plant growth by (Preobrazhenskaya and Ryabova) Pridham PGPR occurs when the plant is supplied et al. and S. rochei Berger et al.) was with a compound that is synthesized by the found to significantly increase the shoot

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Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832 length and shoot fresh mass, respectively, Polonenko 1996) and Europe (Tahvonen of wheat plants. Hormone extraction, 1982a). Many properties related to purification, and bioassay showed that actinomycetes may justify the ability of both species produced substantial amounts sereval of them to act as biocontrol tools. of growth-regulating substances, including Those properties are the ability to colonize auxins, gibberellins, and cytokinins plant surface, the antibiosis against plant (Aldesuquy et al. 1998). This pathogens, the synthesis of particular demonstrated that selected Streptomyces extracellular proteins, and also the spp. produces at least one class of degradation of phytotoxins. compounds that directly influence plant growth. Plant colonization and biocontrol

Direct and indirect interactions between Evidence indicates that actinomycetes are actinomycetes and other nonpathogenic quantitatively and qualitatively vital soil microorganisms also influence plant within the rhizosphere (Barakate et al. growth. For example, some researchers 2002, Crawford et al. 1993, Doumbou et (Mohammadi and Lahdenpera 1992), al. 2001, Miller et al. 1990), where they reported that actinomycetes stimulated the may influence plant growth and defend intensity of mycorrhizal formation and that plant roots against invasion by root resulted in improved plant growth. pathogenic fungi (Lechevalier 1988). However root microorganism interactions Actinomycetes as Biocontrol tools have been extensively studied just for the nitrogenfixing Frankia species (Sardi et al. A prime example of Streptomyces 1992) and a small number of species of the biocontrol agent is Streptomyces genus Streptomyces that are griseoviridis Anderson et al. strain K61. phytopathogens (Loria et al. 1997). This strain, originally isolated from light coloured Sphagnum peat (Tahvonen It is usually assumed that root colonization 1982a, Tahvonen 1982b), has been by introduced bacteria is important for the reported to be antagonistic to a variety of biocontrol of root pathogens which plant pathogens together with Alternaria increasing the population of such an brassicola (Schw.) Wiltsh., Botrytis introduced bacteria on roots should cinerea Pers.:Fr., Fusarium avenaceum enhance disease control (Suslow and (Fr.:Fr.) Sacc, F. culmorum (Wm. G. Schroth 1982]. The key to the idea of root Smith) (Tahvonen 1982a, Tahvonen 1982b colonization is that rootcolonizing bacteria ,Tahvonen and Avikainen 1987). grow on roots in the présence of the Streptomyces griseoviridis strain K61 is indigenous microflora (Schroth and used in root dipping or growth nutrient Hancock 1982) and therefore root treatment of eut flowers, potted plants, colonists are compétitive with soil bacteria greenhouse cucumbers, and varied and fungi. Whereas the tendency to use the alternative vegetables (Mohammadi and term root colonization,alternative terms Lahdenpera 1992). Mycostop™ have been proposed. Rhizosphere (developed by Kemira Oy) is a compétence was used (Schmidt 1979) in biofungicide that contains S. griseoviridis relation to rhizobia, to explain soil as the active ingrédient. This product is microorganisms that show better growth in offered in United States (Cross and response to developing plant roots. In this

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Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832 context, rhizosphere compétent technique ail indicate that S. griseoviridis microorganisms are those that show the colonizes, at least at the seedling stage, classical rhizosphere impact. The term turnip râpe better than carrot root. Since rhizosphere compétence has been the responses of S. griseoviridis to root employed in relation to biological control colonization of two plant species in agents, and Baker (Baker 1991) redefined standard conditions were clearly différent, it because the ability of a microorganism, the mechanism of root colonization should applied by seed treatment, to colonize the be affected by some property that varies rhizosphere of developing roots, a between different plant species. Plant définition that does not differ considerably species are known to produce various from that proposed by Schmidt (Schmidt types and quantities of root exudates (Curl 1979). Many reports hâve used and Truelove 1986), which influence root rhizosphere ability and root colonization colonization (Weller 1988). It‟s potential interchangeably as synonyms (Hozore and that the root exudates of carrot lack some Alexander 1991, Suslow and Schroth characteristics necessary for the 1982). Whereas every définition differs prolifération of S. griseoviridis. The value from the others, there is gênerai agreement of S. griseoviridis seed dressing on barley that root colonization is a vigorous method and spring wheat against foot rot disease involving growth of the introduced was investigated by (Tahvonen et al. 1994) bacteria on or around roots and isn‟t health organization verified that wheat merely a passive chance encounter yields is exaggerated by seed dressirigs between a soil bacterium and a root. more efficiently than those of barley. Researchers consider true colonists to be those bacteria that colonize plant surfaces Proteins involved in biocontrol in compétitive conditions, in natural field soils. Actinomycetes have the capability to produce a wide variety of extracellular A microorganism that colonizes roots is enzymes that permit them to degrade idéal to be used as a biocontrol agent varied biopolymers in soil. The capability against soil-borne diseases (Weller 1988). of actinomycetes to produce extracellular Streptomyces griseoviridis may be a enzymes gained revived attention because example for colonization of plant of their vital role in biocontrol. Especially, rhizosphere by actinomycetes. S. various correlations between fungal griseoviridis is an antagonistic antagonism and bacterial production of microorganism effective in biocontrol of chitinases or glucanases have been noted plant diseases like the the Fusarium wilt of (Fayad et al. 2001, Lim et al. 1991, Valois carnation, the damping-off of Brassica and et al. 1996). Chitin and b-1,3-glucans are also the root rot disease of cucumber major constituents of many fungal cell (Tahvonen and Lahdenpera. 1988). The walls (Sietsma and Wessels 1979), and active root-colonization ability of the various workers have confirmed in vitro biocontrol agent S. griseoviridis was tested lysis of fungal cell walls either by bacterial on turnip râpe and carrot in nonsterile soil chitinases or glucanases alone or by a and by plate test (Kortemaa et al. 1994). mixture of each enzymes (Fiske et al. Plate test and successful root-colonization, 1990, Ordentlich et al. 1988). These root-colonization frequencies and studies have lent support to the hypothesis population densities for sand-tube that these hydrolytic enzymes might

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Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832 contribute to biocontrol efficacy. Several structures like polyketides, p-lactams and attempts to test this hypothesis by means peptides in addition to a variety of of both genetic and molecular approaches alternative secondary metabolites that hâve been undertaken recently. hâve antifungal, anti-tumor and immunosuppressive activities (Behal Many Streptomyces species are 2000). Antibiotics are usually considered lignocellulose decomposers (Chamberlain to be organic compounds of low molecular and Crawford 2000) and are sources of weight produced by microbes. It is antibiotics (Tanaka and Omura 1993). proposed that the production of antibiotics Strains with both the abilities to degrade increases an organism aptitude for survival lignocellulose and antagonize fungal root in the former case by acting as an pathogens should have sensible potential alternative (chemical) défense mechanism for development into a biocontrol product, (Maplestone et al. 1992). Many studies which could be useful to turf grass hâve reported antagonism between growers or managers, to regulate each actinomycetes and a diversity of thatch accumulation and fungal diseases of phytopathogens like Altemaria, Fusarium, turf (Chamberlain and Crawford 2000). Macrophomina, Phytophthora, Pythium, The ability to degrade complex substrates Rhizoctonia, Verticillium (Chattopadhyay could also be an asset in biocontrol. and Nandi 1982, Hussain et al. 1990, Doumbou et al. (Doumbou et al. 1998) Merriman et al. 1974, Valois et al. 1996). showed that actinomycetes degrading thaxtomin A, a phytotoxin created by the Antibiosis as a mechanism of biological plant pathogenic S. scabies, protected control of plant disease has been studied in growing potato plants against common many Systems (Chamberlain and scab. Crawford 2000, El-Abyad et al. 1993, Kortemaa et al. 1994). Gottlieb (Gottlieb Actinomycetal proteins apart from 1976) has reviewed the proof that hydrolases may additionally be concerned antibiotics may be produced by members in biocontrol. For example, Vernekar et al. of soil microflora in their natural (Vernekar et al. 1999) discovered an environment, and function there in an alkaline protease inhibitor (API) as a antagonistic capacity. Various experiments unique category of antifungal proteins hâve verified the difficulty in introducing against phytopathogenic fungi like a new organism into normal soil which Altemaria, Fusarium, and Rhizoctonia. already has an established indigenous The activity of API seems to be related to population and, in contrast, the benefit its ability to inhibit the fungal serine with that organism is introduced into alkaline protease, which is indispensable sterile soil (Gottlieb 1976). Such for their growth. experiments clearly demonstrate that microorganisms in soil are in direct Antibiosis and biocontrol competition, so that, any issue which kills other organisms would certainly be Over one thousand secondary metabolites advantageous to the producer. The from actinomycetes were discovered evidence does not prove that antibiotics throughout the years 1988-1992. are responsible for the competitive Actinomycetes produce a variety of antagonism between species since, as yet, antibiotics with various chemical antibiotics haven‟t been physically isolated from soil. However, there‟s proof 821

Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832 for the analytical detection of antibiotics in Additionally, actinomycetes are important soil;especially, (Zviagintsev et al. 1976) for the production of enzymes, like antibiotic heliomycin is created by chitinase (eg. Streptomyces viridificans), Actinomyces olivocinereus Vinogradova in cellulases (eg. Thermonospora spp.), unsterilised, unsupplemented soil. They peptidases, proteases (Nocardia spp.), used the method of fluorescent Xylanases (Microbispora spp.), ligninases microscopy, utilizing the intrinsic (Nocardia autotrophica), amylases fluorescence of the actinomycete, to (Thermomonospora curvata), sugar examine the development of precursor and isomerases (Actinoplanes missouriensis), synthesis of the antibiotic directly in soil. pectinase, hemicellulase and keratinase (Solans and Vobis 2003). Actinomycetes as Biopesticide Agents To select non-streptomycete As the environmental contamination by actinomycetes by reducing the numbers of toxic chemicals increases, different streptomycete actinomycetes on isolation approaches for controlling pest plates, Streptomyces phages was applied populations became analysis priorities. (Kurtböke et al. 1992, Long and Amphlett These have enclosed biological or 1996). The isolation of Streptomyces ecological management strategies for phages are of sensible importance for a limiting the harmful impacts of pest range of reasons such as the evils they populations, particularly in agriculture cause to fermentation industries (Chater (Nakas and Hagedorn 1990, Canaday 1986), their value for typing 1995, Hokkanen and Lynch 1995). streptomycetes in taxonomic studies (Korn–Wendish and Schneider 1992), Several sorts of microorganisms including their use for the detection and fungi, bacteria, nematodes and viruses that understanding of host controlled are antagonistic to insects are reported as restriction-modification systems (Diaz et methods to biologically control them. al. 1989), their utilization as tools for Actinomycetes play a significant role in genetic exchange and analysis in the biological control of insects through Streptomyces spp. (Herron and Wellington the production of insecticidally active 1990), the study of their general and compounds against the house fly Musca molecular biology (Lomovskaya et al. domestica (Hussain et al. 2002). The 1980) and ecology (Williams et al. 1987). mortality of larval and pupal stages, were terribly high reaching up to 90% after Chitinase is originally an enzyme used by actinomycetes treatments (Hussain et al. insects to degrade the structural 2002). Actinomycetes were effectively polysaccharide “chitin” during the molting used against Culex quinquefasciatus process (Zhang et al. 2002). The largest (Sundarapandian et al. 2002). chitinase activity among bacteria has been determined in species of Streptomyces, Actinomycetes are a vital cluster of Serratia, Vibrio and Bacillus (Reguera and microorganisms, not only as degraders of Leschine 2001). Chitinase enzyme is organic matter within the natural extremely necessary within the biological environment, but also as producers of control of insects (Reguera and Leschine antibiotics and other valuable compounds 2001) and plant pathogenic fungi (El- of commercial interest (Saugar et al.2002, Tarabily et al. 2000, El–Tarabily 2003). Bentley et al. 2002, Basilio et al. 2003). 822

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Species of Streptomyces showed high dominance (Paciorek et al. 2006, multiplicity of chitinase genes Dobbelaere et al. 1999, Bennett et al. (Williamson et al. 2000, Saito et al. 2003), 1998). as in the case of Streptomyces coelicolor and Streptomyces griseus (Itoh et al. Manulis6 observed induced synthesis of 2003). However, screens for antagonism IAA by six diverse Streptomyces species have focused mostly on bacteria, fungi, in the presence of tryptophan and viruses and nematodes (Collier et al. recommended indole-3-acetamide as the 2001). There‟s scarcity of published main pathway, as S. violaceus and S. information with respect to the utilization exfolitus catabolized indole-3- acetamide of actinomycetes significantly, rare non- (IAM), indole-3-lactic acid (ILA), indole- streptomycete actinomycetes, as 3- ethanol (IEt) and indole-3-acetaldehyde biocontrol agents of insect pests. (IAAld) into IAA, besides attainable presence of different pathways for IAA Actinomycetes as production of plant biosynthesis. growth hormone (indole-3-acetic acid) In recent years the tactic of immobilizing Actinomycetes have an extended tradition living cells has gained a large variety of in the analysis of bioactive compounds. applications (D‟Souza et al. 1999, Baianu Several species manufacture a large form et al. 2004). Encapsulation of microbial of secondary metabolites, including anti- cells for soil application provides a variety helminthic compounds, anti-tumour agents of benefits like application to the soil, and majority of identified antibiotics. reduced off-site drifting, and protection of Free-living actinomycetes have cells from environmental stress (Leung et additionally been concerned in the al. 1997, Bashan 1986). Additionally, they improvement of plant growth by possess high cellloading capability, high production of plant growth-producing retention of cell viability, increased rate of substances like auxins and gibberellin-like production of microbial products and also compounds (Persello-Cartieaux et al. act as a reservoir, which releases the 2003, Bloemberg et al. 2001). bacteria at a slow and constant rate (Mahmoud and Rehm 1987). Indole-3-acetic acid (IAA) is the principal form of auxin, which regulates many basic Actinomycetes are known to be durable cellular processes including cell division, organisms and thus appropriate for soil elongation and differentiation. applications. The spores of most actinomycetes endure desiccation and It also leads to decrease in root length and show slightly higher resistance to dry or increase in root hair formation, so wet heat than vegetative cells. enhancing the potential of the plant to Actinomycetes will colonize dry soil absorb soil nutrients. Besides, there are owing to their filamentous nature and exist several developmental processes in which in soil for extended periods as resting auxin plays a role, together with embryo arthrospores that germinate in the and fruit development, organogenesis, occasional presence of exogenous vascular tissue differentiation, root substrates4. So far, the potential of patterning, elongation and tropistic filamentous actinomycetes in encapsulated growth, apical hook formation and apical state for the assembly of IAA has neither

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Int.J.Curr.Microbiol.App.Sci (2014) 3(2): 801-832 been completely examined nor used in activity was seen over a broad range of field conditions to any noticeable extent. pH, and after treatment with several chemicals and heat but not with Proteinase Abiotic soil factors have an effect on the K and trypsin. The AMS, of proteic population dynamics of the inoculant, nature, has publicized to be promising for imposing stresses of varied natures on the use in oil making plants, given its stability cells. They‟ll additionally act indirectly, in the presence of several chemicals and by affecting the activity of the indigenous solvents, and over a broad range of soil microflora (Van Veen et al. 1997). temperature and pH values. Typical environmental stresses faced by the organisms in the soil may include Actinomycetes in Bioremediation salinity, unfavourable soil pH, extremes in temperature, inadequate or excessive soil Petroleum hydrocarbons are widely used moisture, significant metal toxicity and in our daily life as chemical compounds biocides (Slonczewski 2000, McGrath et and fuel. Greater use of result, petroleum al. 1997, Manna et al.2001). has become one of the most common contaminants of large soil surfaces and Actinomycetes in Biocorrosion eventually is considered as a major environmental problem (Sanscartier et al. Corrosion is a principal reason of pipe 2009). failure and high preservation costs in gas pipelines (Zhu et al. 2003). Biocorrosion There are several ways in which is defined as a caustic harm initiated or hydrocarbons degraded in the aggravated by the direct or indirect environment. One mechanism through activities of microorganisms (Zuo 2007). which they can be removed from the A broad range of bacteria is present in environment is Bioremediation. most if not all areas of oil production and Bioremediation is the use of soil microbes have been described from water injection to degrade pollutants to harmless plants, drilling mud, and live reservoir substances (Collin 2001). cores (Feio et al. 2000, Magot et al. 2000, Korenblum et al. 2005, Von Der Weid et Actinomycetes possess many properties al. 2008). that make them good candidates for application in bioremediation of soils Antimicrobial substance (AMS) formed by contaminated with organic pollutants. a Streptomycetes strain having its activity They play an important role in the against an aerobic bacterium B. pumilus recycling of organic carbon and are able to LF-4, and sulfate-reducing bacterium D. degrade complex polymers (Goodfellow alaskensis NCIMB 13491 known to be and Williams 1983). Some reports involved in biofilm formation and indicated that Streptomyces flora could biocorrosion. Strain 235 was identified as play a very important role in degradation belonging to S. lunalinharesii species of hydrocarbons (Radwan et al. 1998, cluster, was initially isolated from a Barabas et al. 2001). Many strains have Brazilian soil. This strain was previously the ability to solubilise lignin and degrade recognized as producer of bioactive lignin-related compounds by producing compounds against phytopathogenic cellulose- and hemicellulose-degrading bacteria and fungi. The antimicrobial enzymes and extracellular peroxidase

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