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Production of Enzymes and Antimicrobial Compounds by Halophilic Antarctic Nocardioides Sp. Grown on Different Carbon Sources

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Production of Enzymes and Antimicrobial Compounds by Halophilic Antarctic Nocardioides Sp. Grown on Different Carbon Sources Production of enzymes and antimicrobial compounds by halophilic Antarctic Nocardioides sp. grown on different carbon sources. Victoria Gesheva, Evgenia Vasileva-Tonkova To cite this version: Victoria Gesheva, Evgenia Vasileva-Tonkova. Production of enzymes and antimicrobial compounds by halophilic Antarctic Nocardioides sp. grown on different carbon sources.. World Journal of Micro- biology and Biotechnology, Springer Verlag, 2012, 28 (5), pp.2069-76. 10.1007/s11274-012-1009-2. pasteur-00748960 HAL Id: pasteur-00748960 https://hal-riip.archives-ouvertes.fr/pasteur-00748960 Submitted on 6 Nov 2013 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Production of enzymes and antimicrobial compounds by halophilic Antarctic Nocardioides sp. grown on different carbon sources Victoria Gesheva & Evgenia Vasileva- Tonkova World Journal of Microbiology and Biotechnology ISSN 0959-3993 Volume 28 Number 5 World J Microbiol Biotechnol (2012) 28:2069-2076 DOI 10.1007/s11274-012-1009-2 1 23 Your article is protected by copyright and all rights are held exclusively by Springer Science+Business Media B.V.. This e-offprint is for personal use only and shall not be self- archived in electronic repositories. If you wish to self-archive your work, please use the accepted author’s version for posting to your own website or your institution’s repository. You may further deposit the accepted author’s version on a funder’s repository at a funder’s request, provided it is not made publicly available until 12 months after publication. 1 23 Author's personal copy World J Microbiol Biotechnol (2012) 28:2069–2076 DOI 10.1007/s11274-012-1009-2 ORIGINAL PAPER Production of enzymes and antimicrobial compounds by halophilic Antarctic Nocardioides sp. grown on different carbon sources Victoria Gesheva • Evgenia Vasileva-Tonkova Received: 7 April 2011 / Accepted: 27 January 2012 / Published online: 7 February 2012 Ó Springer Science+Business Media B.V. 2012 Abstract This study demonstrated the potential of Introduction microbial isolates from Antarctic soils to produce hydro- lytic enzymes by using specific substrates. The results Psychrophilic organisms, in particular bacteria, yeasts, revealed potential of the strains to produce a broad spec- unicellular algae and fungi are able to grow efficiently at trum of hydrolytic enzymes. Strain A-1 isolated from soil sub-zero temperatures and have successfully colonized samples in Casey Station, Wilkes Land, was identified as polar and alpine regions (Gounot and Russell 1999). Nocardioides sp. on the basis of morphological, biochem- In Antarctica, microorganisms have been detected in all ical, physiological observations and also chemotaxonomy habitats such as lakes, ponds, rivers, streams, rocks and analysis. Enzymatic and antimicrobial activities of the soils. The habitats differ one from another with respect to cell-free supernatants were explored after growth of strain nutrients, range of temperature, water activity and other A-1 in mineral salts medium supplemented with different physical–chemical parameters, and the microbial flora carbon sources. It was found that the carbon sources varies from one habitat to another. Prokaryotes dominate in favored the production of a broad spectrum of enzymes as many Antarctic ecosystems and play a major role in food well as compounds with antimicrobial activity against chains, biogeochemical cycles and mineralization of Gram-positive and Gram-negative bacteria, especially pollutants (Nichols et al. 1999). Staphylococcus aureus and Xanthomonas oryzae. Pre- Cold-active microorganisms have developed various liminary analysis showed that the compounds with anti- adaptations enabling them to compensate for the deleteri- microbial activity produced by the strain A-1 are mainly ous effect of low temperatures and to grow in the extreme glycolipids and/or lipopeptides depending on the used environments (Russell 1998; Rampelotto 2010). These carbon source. The results revealed a great potential of the challenges include: reduced enzyme activity; decreased Antarctic Nocardioides sp. strain A-1 for biotechnological, membrane fluidity; altered transport of nutrients and waste biopharmaceutical and biocontrol applications as a source products; decreased rates of transcription, translation and of industrially important enzymes and antimicrobial/anti- cell division; protein cold-denaturation; inappropriate fungal compounds. protein folding, and intracellular ice formation (D’Amico et al. 2006). Keywords Antarctic soils Á Actinomycetes Á Psychrophilic microorganisms mainly bacteria and their Nocardioides Á Psychrophiles Á Hydrolytic enzymes Á enzymes have assumed considerable importance due to Antimicrobial activity biotechnological applications. The catalytic properties of cold-active enzymes as well as their high specific activity at low and moderate temperatures can be extremely useful in different biotechnological fields, for example, in the V. Gesheva Á E. Vasileva-Tonkova (&) detergent and food industries, for the production of fine Bulgarian Academy of Sciences, The Stephan Angeloff Institute chemicals and in bioremediation processes (Gerday et al. of Microbiology, Acad. G. Bonchev Str., Bl. 26, 1113 Sofia, Bulgaria 2000; Huston 2008; Cavicchioli et al. 2011). Cold-adapted e-mail: [email protected] enzymes play a crucial ecological role in recycling the 123 Author's personal copy 2070 World J Microbiol Biotechnol (2012) 28:2069–2076 organic matter in Antarctic environments. Permanent cold Taxonomic characterization of strain A-1 climate requires that extracellular enzymes act efficiently at low temperatures, and on the other hand, the low Strain A-1 was isolated from Casey Station, Budd Coast, concentration of the particulate organic matter requires Wilkes Land (Vasileva-Tonkova and Gesheva 2004). efficient extracellular enzymes to achieve an adequate Mmorphological, biochemical, physiological features, uptake of nutrients. In this regard, selection of samples, utilization of carbon sources, enzyme tests and also assays techniques for isolation of bacterial strains and screening of the cell sugars and amino acids in whole-cell hydroly- for enzymes constitutes an important aspect of studies on sate were studied according to Vasileva-Tonkova and cold-active microorganisms. Furthermore, these microor- Gesheva (2005). Antibiotic formation was observed on ganisms appear to be inexhaustible source of new bioactive MPA after 4–6 days of cultivation at 25°C. Antibiotic metabolites with antibiotic effect against pathogenic activity was determined by agar diffusion method against microbes, and continuous efforts are being made in the the following test cultures: Bacillus subtilis ATCC 6633, research on microbial secondary metabolites with potential Sarcina lutea, Staphylococcus aureus 605P, Escherihia applications (Giudice et al. 2007). coli, Candida tropicalis, and Saccaharomyces cerevisiae. In our previous work, seventeen microbial strains were isolated from soil samples of three regions of Antarctica Growth of strain A-1 on different carbon sources and investigated for their ability to grow on hydrocarbons (Vasileva-Tonkova and Gesheva 2004). Some of the Strain A-1 was cultivated in 200-ml Erlenmeyer flasks isolates were psychrophilic actinomycetes able to produce containing mineral salts medium (MSM) with composition enzymes and glycolipid biosurfactants (Vasileva-Tonkova as described earlier (Vasileva-Tonkova and Gesheva and Gesheva 2004; Gesheva 2009a, 2010). In the present 2004). The medium was supplemented with 2% of different work, the potential capability of the Antarctic isolates to carbon sources; phenanthrene was used in concentration produce hydrolytic enzymes was investigated aiming to 10 g l-1 (Table 4). A control flask without any carbon find sources of industrially relevant enzymes. Isolate source was also prepared. The pH value was adjusted to marked as A-1 was selected, and the effect of different 7.0 ± 0.2 and the media were autoclaved at 121 °C for carbon sources on the production of hydrolytic enzymes 15 min. Culture after 24 h of growth in meat extract-pep- and antimicrobial/antifungal compounds by the strain A-1 tone broth (MPB) was used as an inoculum. Experimental was studied. flasks were incubated statically at 25 ± 1°C until the stationary growth phase. Growth was monitored by mea- suring the whole cell protein increase. Periodically, sam- Materials and methods ples (0.5-ml) were taken from each flask, heated for 10 min with 0.05 ml of 1 N NaOH, and the protein content was Enzymatic activities assay then determined by the method of Bradford (1976) using human serum albumin as a standard. Agar well diffusion assay was used for testing the enzy- matic activities of bacterial cultures and cell-free super- Antimicrobial activity assay natants of strain A-1. The proteolytic, amylolytic, lipolytic, and ribonuclease (RNase) activities were assayed by using Antimicrobial activity of the cell-free supernatants/extracts skim-milk (30% v/v), gelatin (1% w/v), soluble starch obtained after growth of
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