DOCSLIB.ORG

Detection of Secreted Antimicrobial Peptides Isolated from Cell-Free Culture Supernatant of Paenibacillus Alvei AN5

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Detection of Secreted Antimicrobial Peptides Isolated from Cell-Free Culture Supernatant of Paenibacillus Alvei AN5 View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Crossref J Ind Microbiol Biotechnol (2013) 40:571–579 DOI 10.1007/s10295-013-1259-5 BIOTECHNOLOGY METHODS Detection of secreted antimicrobial peptides isolated from cell-free culture supernatant of Paenibacillus alvei AN5 Bassam Alkotaini • Nurina Anuar • Abdul Amir Hassan Kadhum • Asmahani Azira Abdu Sani Received: 21 December 2012 / Accepted: 4 March 2013 / Published online: 19 March 2013 Ó The Author(s) 2013. This article is published with open access at Springerlink.com Abstract An antimicrobial substance produced by the Abbreviations Paenibacillus alvei strain AN5 was detected in fermenta- NB Nutrient broth tion broth. Subsequently, cell-free culture supernatant NA Nutrient agar (CFCS) was obtained by medium centrifugation and fil- TSB Tryptic soy broth tration, and its antimicrobial activity was tested. This TSA Tryptic soy agar showed a broad inhibitory spectrum against both Gram- CFCS Cell free culture supernatant positive and -negative bacterial strains. The CFCS was then AU Arbitrary unit purified and subjected to SDS-PAGE and infrared spec- troscopy, which indicated the proteinaceous nature of the antimicrobial compound. Some de novo sequencing using Introduction an automatic Q-TOF premier system determined the amino acid sequence of the purified antimicrobial peptide as Y-S- Due to the increasing numbers of resistant pathogenic K-S-L-P-L-S-V-L-N-P (1,316 Da). The novel peptide was bacteria and side effects caused by existing antibiotics, new designated as peptide AN5-1. Its mode of action was antimicrobial compounds with effective properties are bactericidal, inducing cell lysis in E. coli ATCC 29522 and needed [9]. Antimicrobial peptides are attracting increasing S. aureus, and non-cell lysis in both S. marcescens and B. interest as a potential substitute. The importance of anti- cereus ATCC 14579. Peptide AN5-1 displayed stability at microbial peptides has been appreciated in the last 20 years a wide range of pH values (2–12) and remained active after due to pathogenic bacterial resistance against existing exposure to high temperatures (100 °C). It also maintained antibiotics [24]. its antimicrobial activity after incubation with chemicals Antimicrobial peptides are small proteins occurring in such as SDS, urea and EDTA. living organisms and are produced as defense molecules against pathogens such as bacteria. Therefore, it is con- Keywords Paenibacillus alvei AN5 Á Antimicrobial sidered the first line of defense in invaded eukaryotic and peptide AN5-1 Á LC/ESI–MS Á Action mode Á Stability prokaryotic cells [24]. Their mode of action varies between peptides. Three factors play a major role in determining the mode of action: the net positive charge on the surface, its 3-D amphipathic structure and the selective disruption B. Alkotaini Á N. Anuar (&) Á A. A. H. Kadhum points on the target cell membrane [17]. These actions Department of Chemical and Process Engineering, Faculty were studied by Hallock et al. [10] based on solid-state of Engineering and Built Environment, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia NMR, which showed an interaction between antimicrobial e-mail: [email protected] peptides and membranes, leading to membrane disruption. However, in terms of medical and food applications, anti- A. A. A. Sani microbial peptides have no toxic effects on mammalian Proteomic Laboratory, Malaysia Genome Institute, Ministry of Science, Technology and Innovation, cells [20], making them a promising candidate for further 43000 Jalan Bangi, Selangor, Malaysia studies. 123 572 J Ind Microbiol Biotechnol (2013) 40:571–579 In 1925, the first antimicrobial peptide was discovered Table 1 Antimicrobial activity of cell free culture supernatant from Gram-negative bacteria, Escherichia coli, known (CFCS) from P. alvei AN5 and the MICs of the synthesized antimi- ascolicins. This was followed by the discovery of nisin in crobial peptide AN5-1 against test strains 1928, which was an antimicrobial peptide produced by the Bacterial test strains Inhibition zones MICs (lgml-1) Gram-positive Streptococcus lactis (formerly known as of CFCS of AN5-1 Lactococcus lactis)[12]. Since then, several Gram-positive Staphylococcus aureusa ??? 64 bacteria, in particular soil bacteria bacilli, have been Streptococcus agalactiae –– reported to produce antimicrobial agents such as bacitracin, ATCC 12386 lichenin and cerein, which are produced by Bacillus sub- Staphylococcus –– tilis, Bacillus licheniformis and Bacillus cereus, respec- epidermidis tively [7, 19, 23]. Production of antimicrobial peptides Lactobacillus plantarum ??? 32 is considered an unstable bacterial characteristic because ATCC 8014 it changes according to growth conditions. A tradeoff Lactococcus lactis ATCC ? [128 11454 between metabolism and gene expression affects antimi- crobial peptide biosynthesis during bacterial growth [26]. Bacillus subtilis ??? 32 Paenibacillus alvei are Gram-positive, rod-shaped, Bacillus cereus ATCC ?? 64 14579 motile, spore-forming and catalase-positive bacteria [22]. Bacillus anthracis ? 128 The first report of antimicrobial peptide production from Bacillus licheniformis ? [128 this bacteria was by Anandaraj et al. [1], who isolated a Bacillus flexusa ???? 32 strain from fermented tomato fruit and detected two anti- MRSA – – microbial peptides, Paenibacillin P and Paenibacillin N. Escherichia coli ATCC ??? 8 Other species of Paenibacillus have been reported as 25922 antimicrobial peptide producers too. For example, poly- Escherichia coli ??? 8 myxins, LI-F complex, saltavalin and gatavalin are pro- Salmonella enteritidis ???? 4 duced by different strains of Paenibacillus polymyxa [25]. ATCC 13076 This study aimed to isolate an antimicrobial peptide from Pseudomonas aeruginosa –– the cell-free culture supernatant (CFCS) of P. alvei AN5, Enterobacter spp. ??? 16 screen its antimicrobial activity and identify the amino acid Serratia marcescensa ???? 32 sequence of the antimicrobial peptide. –, no inhibition zones; ?, zone diameter 1–11 mm; ??, zone diameter 12–16 mm; ???, zone diameter 16–21 mm. ???? zone Materials and methods diameter C22 mm a Test strains are incubated in NA Microorganisms Paenibacillus alvei AN5 with a 16S rRNA gene accession of 20 g l-1 glucose, 2 g l-1 yeast extract, 5.0 g l-1 -1 -1 number of JQ868768 at GenBank was obtained from the L-glutamic acid, 2 g l KH2PO4, 0.16 mg l CuSO4, -1 -1 -1 Biochemical Engineering Laboratory, Faculty of Engi- 0.5 g l MgSO4Á7H2O, 0.15 mg l FeSO4, 0.5 g l -1 -1 -1 neering and Built Environment, Universiti Kebangsaan KCl, 4 g l NaCl, 1 g l (NH4)NO2, 1gl NH4SO4 -1 Malaysia. It was grown in nutrient broth (NB) (Oxoid and 45.0 mg l MnSO4ÁH2O, pH 7. The sample was limited, UK) at 30 °C, under agitation at 150 rpm for 24 h. incubated at 30 °C, under agitation at 150 rpm for 24 h. The test strains used in this study (Table 1) were obtained The CFCS was obtained at 2-h intervals by culture cen- from the American Type Culture Collection (ATCC) and trifugation at 9,000 rpm for 20 min followed by 0.22-lm Biochemical Engineering Laboratory, Faculty of Engi- membrane filtration. The CFCS was heated up to 65 °C for neering and Built Environment, UKM. All bacterial strains 20 min for protease inactivation. The antimicrobial sub- were grown in tryptic soy broth (TSB) (Difco, France) and stance production during the P. alvei AN5 growth cycle NB at 37 °C, under agitation at 150 rpm for 24 h. was determined by measuring total activity in arbitrary units (AU), defined as the reciprocal of the highest dilution Production of the antimicrobial compound of CFCS yielding clear inhibition zones against the indi- cator microorganism [19]. In this study, Escherichia coli Paenibacillus alvei AN5 was grown in NB at 30 °C for ATCC 29522 was used as the indicator strain during 24 h. The resulting culture was used to inoculate a 2 l flask purification and characterization of the antimicrobial containing 500 ml of modified Landy medium consisting compound. 123 J Ind Microbiol Biotechnol (2013) 40:571–579 573 Determination of antimicrobial activity Identification of the antimicrobial peptide Antimicrobial activity was assessed by agar well diffusion The purified sample was further analyzed by integrated assays as described [6] with some modifications. Briefly, nano liquid chromatography electrospray ionization mass 100 ll of each test strain (OD600 = 0.1) was mixed with spectroscopy (LC/ESI–MS) (Waters, UK). Nano ultra 20 ml of TSA or NA before being solidified and cast in a performance liquid chromatography (UPLC) separation Petri dish. A total of three circular wells were cut with was performed with a nano ACQUITY@ UPLC system, diameters of 6 mm and filled with 100 ll of CFCS equipped with a Symmetry C18 5 lm, 20 mm 9 180 lm obtained after 20 h of cultivation. Plates were incubated at pre-column and BEH C18 1.7 lm, 20 cm 9 75 lm, ana- 4 °C for 8 h to allow absorption and were further incubated lytical reversed-phase column. Following the manufac- at 37 °C for 24 h. Tests were carried out in triplicate and turer’s instructions, 500 ng of sample was initially loaded averages of inhibition zones were reported. and transferred with 0.1 % of aqueous formic acid solution to the pre-column at a 15 ll min-1 flow rate for 3 min. The Purification of the antimicrobial compound mobile phase consisted of 0.1 % formic acid in water (Solvent A) and 0.1 % formic acid in acetonitrile (Solvent Cell-free culture supernatant was subjected to ultra-filtra- B). The sample was separated with a gradient of 1–40 % of tion using 5, 10 and 30 kDa molecular weight cut-offs solvent B in solvent A for 90 min at a flow rate of (MWCO) (Sigma Aldrich, Germany). Four fractions were 3 ll min-1, followed by a 3-min rinse with 85 % of sol- obtained and tested against the indicator strain.
Load more

Recommended publications
  • Laboratory and Field Performance of Some Soil Bacteria Used As Seed Treatments on Meloidogyne Incognita in Chickpea
    08 Khan_143 4-01-2013 17:33 Pagina 143 Nematol. medit. (2012), 40: 143-151 143 LABORATORY AND FIELD PERFORMANCE OF SOME SOIL BACTERIA USED AS SEED TREATMENTS ON MELOIDOGYNE INCOGNITA IN CHICKPEA M.R. Khan*, M.M. Khan, M.A. Anwer and Z. Haque Department of Plant Protection, Aligarh Muslim University, 202002, India Received: 26 May 2012; Accepted: 27 September 2012. Summary. Experiments were conducted under in vitro and field conditions to assess the efficacy of the soil bacteria Bacillus sub- tilis, Pseudomonas fluorescens, P. stutzeri and Paenibacillus polymyxa for controlling the root knot nematode, Meloidogyne incogni- ta, in chickpea, Cicer arietinum, in India. The bacterial strains tested solubilized phosphorous under in vitro and soil conditions and produced indole acetic acid, ammonia and hydrogen cyanide in vitro. Both pure culture and culture filtrates of the bacteria reduced egg hatching and increased juvenile mortality of the nematode. Under field conditions, seed treatment (at 5 ml/kg seed) with cultures containing 1012 colony forming units/ml of P. fluorescens and P. stutzerisignificantly increased yield and root nodula- tion of chickpea. Inoculation with 2000 juveniles of M. incognita/spot (plant) caused severe root galling and decreased the yield of chickpea by 24%. Treatment with P. fluorescens suppressed gall formation, and treatment with P. fluorescensor B. subtilis sup- pressed reproduction and soil populations of M. incognita. However, the suppressive effects of the two bacteria on the nematode were less than that of fenamiphos. In nematodes infested plots, only treatments with P. fluorescens increased the yield (14%) com- pared to fenamiphos, being 31% above the untreated nematode control.
    [Show full text]
  • Paenibacillus Polymyxa: Antibiotics, Hydrolytic Enzymes and Hazard Assessment
    002_OfferedReview_419 13-11-2008 14:35 Pagina 419 Journal of Plant Pathology (2008), 90 (3), 419-430 Edizioni ETS Pisa, 2008 419 OFFERED REVIEW PAENIBACILLUS POLYMYXA: ANTIBIOTICS, HYDROLYTIC ENZYMES AND HAZARD ASSESSMENT W. Raza, W. Yang and Q-R. Shen 1 College of Resource and Environmental Sciences, Nanjing Agriculture University, Nanjing, 210095, Jiangsu Province, P.R. China SUMMARY pressing several plant diseases and promoting plant growth (Benedict and Langlykke, 1947; Ryu and Park, Certain Paenibacillus polymyxa strains that associate 1997). These strains have been isolated from the rhizos- with many plant species have been used effectively in phere of a variety of crops like wheat (Triticum aes- the control of plant pathogenic fungi and bacteria. In tivum), barley (Hordeum gramineae) (Lindberg and this article we review the possible mechanism of action Granhall, 1984), white clover (Trifolium repens), peren- by which P. polymyxa promotes plant growth and sup- nial ryegrass (Lolium perenne), crested wheatgrass presses some plant diseases. Furthermore we present an (Agropyron cristatum) (Holl et al., 1988), lodgepole pine updated summary of antibiotics, autolysis, hydrolytic (Pinus contorta latifolia) (Holl and Chanway, 1992), and autolytic enzymes and levanase produced by this Douglas fir (Pseudotsuga menziesii) (Shishido et al., bacterium. Some hazards and mild pathogenic effects 1996), green bean (Phaseolus vulgaris) (Petersen et al., are also reported, but these appear to be strain-specific 1996) and garlic (Allium sativum ) (Kajimura and Kane- and negligible. The association between plants and P. da, 1996). P. polymyxa has been successfully used to con- polymyxa seems to be specific and to involve co-adapta- trol Botrytis cinerea, the causal agent of grey mould, in tion processes.
    [Show full text]
  • Bacterial Extracellular Polymeric Substances
    A Seminar Paper on Bacterial Extracellular Polymeric Substances: Characteristics and Bioremoval of Heavy Metals Course Title: Seminar Course Code: ENS 598 Term: Summer, 2020 Submitted To: Course Instructors Major Professor Dr. A. K. M. Aminul Islam Dr. Md. Manjurul Haque Professor Professor Department of Environmental Dr. Md. Mizanur Rahman Science Professor BSMRAU, Gazipur Dr. Dinesh Chandra Shaha Associate Professor Dr. Md. Sanaullah Biswas Associate Professor BSMRAU, Gazipur Submitted By: Md. Mohiminul Haque Mithun Reg. No. 15-05-3509 MS Student Term: Summer, 2020 Department of Environmental Science BANGABANDHU SHEIKH MUJIBUR RAHMAN AGRICULTURAL UNIVERSITY GAZIPUR-1706 1 ABSTRACT Extracellular polymeric substances (EPS) of microbial origin are a fancy mixture of biopolymers having polysaccharides, proteins, nucleic acids, uronic acids, humic substances, lipids, etc. Bacterial secretions, cell lysates and adsorption of organic constituents from the environment result in EPS formation in a wide variety of free-living bacteria as well as microbial aggregates like biofilms, bioflocs and biogranules. EPS could be loosely attached to the cell surface or bacteria may be embedded in EPS. Regulated by the organic and inorganic constituents of the microenvironment compositional variation exists amongst EPS extracted from pure bacterial cultures and heterogeneous microbial communities. EPS function mainly works as cell-to-cell aggregation, adhesion to substratum, formation of flocs, protection from dessication and resistance to harmful exogenous materials. Additionaly exopolymers fuction biosorbing agents by accumulating nutrients from the encircling environment and also play an important role in biosorption of heavy metals. EPS produced by Bacillus sp. reported for the removal of copper, lead and zinc from different solutions. Some other EPS produced bacterial strain like Pseudomonas sp.
    [Show full text]
  • Application of a Bacterial Extracellular Polymeric Substance in Heavy Metal Adsorption in a Co-Contaminated Aqueous System
    Brazilian Journal of Microbiology (2008) 39:780-786 ISSN 1517-8382 APPLICATION OF A BACTERIAL EXTRACELLULAR POLYMERIC SUBSTANCE IN HEAVY METAL ADSORPTION IN A CO-CONTAMINATED AQUEOUS SYSTEM Paula Salles de Oliveira Martins*; Narcisa Furtado de Almeida; Selma Gomes Ferreira Leite Departamento de Engenharia Bioquímica, Centro de Tecnologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brasil Submitted: November 30, 2007; Returned to authors for corrections: March 18, 2008; Approved: November 02, 2008. ABSTRACT The application of a bacterial extracellular polymeric substance (EPS) in the bioremediation of heavy metals (Cd, Zn and Cu) by a microbial consortium in a hydrocarbon co-contaminated aqueous system was studied. At the low concentrations used in this work (1.00 ppm of each metal), it was not observed an inhibitory effect on the cellular growing. In the other hand, the application of the EPS lead to a lower concentration of the free heavy metals in solution, once a great part of them is adsorbed in the polymeric matrix (87.12% of Cd; 19.82% of Zn; and 37.64% of Cu), when compared to what is adsorbed or internalized by biomass (5.35% of Cd; 47.35% of Zn; and 24.93% of Cu). It was noted an increase of 24% in the consumption of ethylbenzene, among the gasoline components that were quantified, in the small interval of time evaluated (30 hours). Our results suggest that, if the experiments were conducted in a larger interval of time, it would possibly be noted a higher effect in the degradation of gasoline compounds. Still, considering the low concentrations that were evaluated, it is possible that a real system could be bioremediated by natural attenuation process, demonstrated by the low effect of those levels of contaminants and co-contaminants over the naturally present microbial consortium.
    [Show full text]
  • Paenibacillus Polymyxa
    Liu et al. BMC Microbiology (2021) 21:70 https://doi.org/10.1186/s12866-021-02132-2 RESEARCH ARTICLE Open Access Interactional mechanisms of Paenibacillus polymyxa SC2 and pepper (Capsicum annuum L.) suggested by transcriptomics Hu Liu, Yufei Li, Ke Ge, Binghai Du, Kai Liu, Chengqiang Wang* and Yanqin Ding* Abstract Background: Paenibacillus polymyxa SC2, a bacterium isolated from the rhizosphere soil of pepper (Capsicum annuum L.), promotes growth and biocontrol of pepper. However, the mechanisms of interaction between P. polymyxa SC2 and pepper have not yet been elucidated. This study aimed to investigate the interactional relationship of P. polymyxa SC2 and pepper using transcriptomics. Results: P. polymyxa SC2 promotes growth of pepper stems and leaves in pot experiments in the greenhouse. Under interaction conditions, peppers stimulate the expression of genes related to quorum sensing, chemotaxis, and biofilm formation in P. polymyxa SC2. Peppers induced the expression of polymyxin and fusaricidin biosynthesis genes in P. polymyxa SC2, and these genes were up-regulated 2.93- to 6.13-fold and 2.77- to 7.88-fold, respectively. Under the stimulation of medium which has been used to culture pepper, the bacteriostatic diameter of P. polymyxa SC2 against Xanthomonas citri increased significantly. Concurrently, under the stimulation of P. polymyxa SC2, expression of transcription factor genes WRKY2 and WRKY40 in pepper was up-regulated 1.17-fold and 3.5-fold, respectively. Conclusions: Through the interaction with pepper, the ability of P. polymyxa SC2 to inhibit pathogens was enhanced. P. polymyxa SC2 also induces systemic resistance in pepper by stimulating expression of corresponding transcription regulators.
    [Show full text]
  • Paenibacillus Polymyxa Initiates Biocontrol Against Crown Rot Disease W.M
    Journal of Applied Microbiology ISSN 1364-5072 ORIGINAL ARTICLE Colonization of peanut roots by biofilm-forming Paenibacillus polymyxa initiates biocontrol against crown rot disease W.M. Haggag1 and S. Timmusk2 1 Department of Plant Pathology, National Research Center, Dokki, Cairo, Egypt 2 Department of Biology, University of Waterloo, Waterloo, Canada Keywords Abstract Aspergillus niger, biofilm, colonization, Paenibacillus polymyxa, population dynamics. Aim: To investigate the role of biofilm-forming Paenibacillus polymyxa strains in controlling crown root rot disease. Correspondence Methods and Results: Two plant growth-promoting P. polymyxa strains were S. Timmusk, Department of Forest Mycology isolated from the peanut rhizosphere, from Aspergillus niger-suppressive soils. and Pathology, SLU, Box 7026, SE-750 07 The strains were tested, under greenhouse and field conditions for inhibition Uppsala, Sweden. E-mail: of the crown root rot pathogen of the peanut, as well as for biofilm formation [email protected] in the peanut rhizosphere. The strains’ colonization and biofilm formation 2007 ⁄ 0343: received 5 March 2007, revised were further studied on roots of the model plant Arabidopsis thaliana and with 24 July 2007 and accepted 5 September solid surface assays. Their crown root rot inhibition performance was studied 2007 in field and pot experiments. The strains’ ability to form biofilms in gnotobi- otic and soil systems was studied employing scanning electron microscope. doi:10.1111/j.1365-2672.2007.03611.x Conclusion: Both strains were able to suppress the pathogen but the superior biofilm former offers significantly better protection against crown rot. Significance and Impact of the Study: The study highlights the importance of efficient rhizosphere colonization and biofilm formation in biocontrol.
    [Show full text]
  • Discovery of a Paenibacillus Isolate for Biocontrol of Black Rot in Brassicas
    Lincoln University Digital Thesis Copyright Statement The digital copy of this thesis is protected by the Copyright Act 1994 (New Zealand). This thesis may be consulted by you, provided you comply with the provisions of the Act and the following conditions of use: you will use the copy only for the purposes of research or private study you will recognise the author's right to be identified as the author of the thesis and due acknowledgement will be made to the author where appropriate you will obtain the author's permission before publishing any material from the thesis. Discovery of a Paenibacillus isolate for biocontrol of black rot in brassicas A thesis submitted in partial fulfilment of the requirements for the Degree of Doctor of Philosophy at Lincoln University by Hoda Ghazalibiglar Lincoln University 2014 DECLARATION This dissertation/thesis (please circle one) is submitted in partial fulfilment of the requirements for the Lincoln University Degree of ________________________________________ The regulations for the degree are set out in the Lincoln University Calendar and are elaborated in a practice manual known as House Rules for the Study of Doctor of Philosophy or Masters Degrees at Lincoln University. Supervisor’s Declaration I confirm that, to the best of my knowledge: • the research was carried out and the dissertation was prepared under my direct supervision; • except where otherwise approved by the Academic Administration Committee of Lincoln University, the research was conducted in accordance with the degree regulations and house rules; • the dissertation/thesis (please circle one)represents the original research work of the candidate; • the contribution made to the research by me, by other members of the supervisory team, by other members of staff of the University and by others was consistent with normal supervisory practice.
    [Show full text]
  • Paenibacillaceae Cover
    The Family Paenibacillaceae Strain Catalog and Reference • BGSC • Daniel R. Zeigler, Director The Family Paenibacillaceae Bacillus Genetic Stock Center Catalog of Strains Part 5 Daniel R. Zeigler, Ph.D. BGSC Director © 2013 Daniel R. Zeigler Bacillus Genetic Stock Center 484 West Twelfth Avenue Biological Sciences 556 Columbus OH 43210 USA www.bgsc.org The Bacillus Genetic Stock Center is supported in part by a grant from the National Sciences Foundation, Award Number: DBI-1349029 The author disclaims any conflict of interest. Description or mention of instrumentation, software, or other products in this book does not imply endorsement by the author or by the Ohio State University. Cover: Paenibacillus dendritiformus colony pattern formation. Color added for effect. Image courtesy of Eshel Ben Jacob. TABLE OF CONTENTS Table of Contents .......................................................................................................................................................... 1 Welcome to the Bacillus Genetic Stock Center ............................................................................................................. 2 What is the Bacillus Genetic Stock Center? ............................................................................................................... 2 What kinds of cultures are available from the BGSC? ............................................................................................... 2 What you can do to help the BGSC ...........................................................................................................................
    [Show full text]
  • Genome Snapshot of Paenibacillus Polymyxa ATCC 842T
    J. Microbiol. Biotechnol. (2006), 16(10), 1650–1655 Genome Snapshot of Paenibacillus polymyxa ATCC 842T JEONG, HAEYOUNG, JIHYUN F. KIM, YON-KYOUNG PARK, SEONG-BIN KIM, CHANGHOON KIM†, AND SEUNG-HWAN PARK* Laboratory of Microbial Genomics, Systems Microbiology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), P.O. Box 115, Yuseong, Daejeon 305-600, Korea Received: May 11, 2006 Accepted: June 22, 2006 Abstract Bacteria belonging to the genus Paenibacillus are rhizosphere and soil, and their useful traits have been facultatively anaerobic endospore formers and are attracting analyzed [3, 6-8, 10, 15, 22, 26]. At present, the genus growing ecological and agricultural interest, yet their genome Paenibacillus consists of 84 species (NCBI Taxonomy information is very limited. The present study surveyed the Homepage at http://www.ncbi.nlm.nih.gov/Taxonomy/ genomic features of P. polymyxa ATCC 842 T using pulse-field taxonomyhome.html, February 2006). gel electrophoresis of restriction fragments and sample genome Nonetheless, despite the growing interest in Paenibacillus, sequencing of 1,747 reads (approximately 17.5% coverage of its genomic information is very scarce. Most of the completely the genome). Putative functions were assigned to more than sequenced organisms currently belong to the Bacillaceae 60% of the sequences. Functional classification of the sequences family, in particular to the Bacillus genus, whereas data on showed a similar pattern to that of B. subtilis. Sequence Paenibacillaceae sequences is limited even at the draft analysis suggests nitrogen fixation and antibiotic production level. P. polymyxa, the type species of the genus Paenibacillus, by P. polymyxa ATCC 842 T, which may explain its plant is also of great ecological and agricultural importance, owing growth-promoting effects.
    [Show full text]
  • Fusaricidin Produced by Paenibacillus Polymyxa WLY78 Induces Systemic Resistance Against Fusarium Wilt of Cucumber
    International Journal of Molecular Sciences Article Fusaricidin Produced by Paenibacillus polymyxa WLY78 Induces Systemic Resistance against Fusarium Wilt of Cucumber Yunlong Li and Sanfeng Chen * State Key Laboratory of Agrobiotechnology and College of Biological Sciences, China Agricultural University, Beijing 100094, China; [email protected] * Correspondence: [email protected]; Tel.: +86-10-6273-1551 Received: 7 July 2019; Accepted: 17 October 2019; Published: 22 October 2019 Abstract: Cucumber is an important vegetable crop in China. Fusarium wilt is a soil-borne disease that can significantly reduce cucumber yields. Paenibacillus polymyxa WLY78 can strongly inhibit Fusarium oxysporum f. sp. Cucumerium, which causes Fusarium wilt disease. In this study, we screened the genome of WLY78 and found eight potential antibiotic biosynthesis gene clusters. Mutation analysis showed that among the eight clusters, the fusaricidin synthesis (fus) gene cluster is involved in inhibiting the Fusarium genus, Verticillium albo-atrum, Monilia persoon, Alternaria mali, Botrytis cinereal, and Aspergillus niger. Further mutation analysis revealed that with the exception of fusTE, the seven genes fusG, fusF, fusE, fusD, fusC, fusB, and fusA within the fus cluster were all involved in inhibiting fungi. This is the first time that demonstrated that fusTE was not essential. We first report the inhibitory mode of fusaricidin to inhibit spore germination and disrupt hyphal membranes. A biocontrol assay demonstrated that fusaricidin played a major role in controlling Fusarium wilt disease. Additionally, qRT-PCR demonstrated that fusaricidin could induce systemic resistance via salicylic acid (SA) signal against Fusarium wilt of cucumber. WLY78 is the first reported strain to both produce fusaricidin and fix nitrogen.
    [Show full text]
  • Butanediol Production by Paenibacillus Polymyxa DSM 365
    Process development and metabolic engineering to enhance 2,3- butanediol production by Paenibacillus polymyxa DSM 365 DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Christopher Chukwudi Okonkwo Graduate Program in Animal Sciences The Ohio State University 2017 Dissertation Committee: Thaddeus C. Ezeji, Advisor Ramesh Selvaraj Katrina Cornish Ana Alonso Copyrighted by Christopher Chukwudi Okonkwo 2017 Abstract 2,3-Butanediol (2,3-BD) is a platform chemical with vast industrial applications; particularly for its use in the production of 1,3-butadiene (1,3-BD), the monomer from which synthetic rubber is manufactured. Currently, 2,3-BD production is by chemical synthesis using petroleum-derived feedstocks such as propylene, acetylene, butene and butane. Microbial 2,3-BD fermentation is aimed at producing 2,3-BD renewably, and potentially reduce dependency on finite petroleum-derived feedstocks. However, fermentative production of 2,3-BD is hampered by (1) cost of food-based substrates; (2) low 2,3-BD titer, yield and productivity during 2,3-BD fermentation stemming from formation of competing products such as exopolysaccharides (EPS), ethanol, lactic, formic and acetic acids; and (3) high cost of 2,3-BD purification, due partly to additional purification steps necessary to remove 2,3-BD co-products especially EPS prior to 2,3- BD recovery. The objectives of this study were conceived to examine use of process design, alternative substrates and metabolic engineering, to enhance 2,3-BD production. Chapter 3 (objective 1) focused on identification of key fermentation parameters that influence 2,3-BD fermentation by Paenibacillus polymyxa and optimization of them for maximum 2,3-BD production.
    [Show full text]
  • Involvement of Growth-Promoting Rhizobacterium Paenibacillus Polymyxa in Root Rot of Stored Korean Ginseng
    J. Microbiol. Biotechnol. (2003), 13(6), 881–891 Involvement of Growth-Promoting Rhizobacterium Paenibacillus polymyxa in Root Rot of Stored Korean Ginseng JEON, YONG HO, SUNG PAE CHANG, INGYU HWANG, AND YOUNG HO KIM School of Agricultural Biotechnology & Center for Plant Molecular Genetics and Breeding Research, Seoul National University, Seoul 151-742, Korea Received: May 19, 2003 Accepted: August 8, 2003 Abstract Paenibacillus polymyxa is a plant growth- Key words: Ginseng, biolog, gas chromatography of fatty promoting rhizobacterium (PGPR) which can be used for acid methyl esters, Paenibacillus polymyxa, 16S rDNA, PCR biological control of plant diseases. Several bacterial strains were isolated from rotten roots of Korean ginseng (Panax ginseng C. A. Meyer) that were in storage. These strains were Biological control of plant disease using microorganisms identified as P. p o l y m y x a , based on a RAPD analysis using has long been an effective alternative to chemical control, a P. polymyxa-specific primer, cultural and physiological especially since the former is more environmentally- characteristics, an analysis utilizing the Biolog system, gas friendly and safer for humans than the latter. A number of chromatography of fatty acid methyl esters (GC-FAME), and antagonistic microorganisms, including fungi and bacteria, the 16S rDNA sequence analysis. These strains were found to have been developed as biocontrol agents [7]. Microbial cause the rot in stored ginseng roots. Twenty-six P. polymyxa pesticides registered with the EPA in the United States strains, including twenty GBR strains, were phylogenetically include bacteria belonging to the Agrobacterium, Bacillus, classified into two groups according to the ERIC and BOX- Pseudomonas, and Streptomyces genera, and fungi belonging PCR analyses and 16S rDNA sequencing, and the resulting to the Ampelomyces, Candida, Coniothyrium, and Trichoderma groupings systematized to the degrees of virulence of each genera.
    [Show full text]