Purification of Chitinase enzymes from Bacillus subtilis bacteria TV-125, investigation of kinetic properties and antifungal activity against Fusarium culmorum

Annals of Clinical Microbiology and Antimicrobials

Research

Merve Senol¹  Neslihan Dikbas¹  Hayrunnisa Nadaroglu²  Recep Kotan³ 

[1] Department of Biotechnology, Faculty of Agriculture, Ataturk University, 25240, Erzurum, Turkey;Department of Food Technology, Erzurum Vocational Training School, Ataturk University, 25240, Erzurum, Turkey;Department of Plant Protect, Faculty of Agriculture, Ataturk University, 25240, Erzurum, Turkey;

关键词: Chitinase;    Purification;    Antifungal activity;    Fusarium culmorum;    Bacillus subtilis;   

DOI  :  10.1186/s12941-014-0035-3

 received in 2014-05-17, accepted in 2014-07-13,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundChitin is the main structural component of cell walls of fungi, exoskeletons of insects and other arthropods and shells of crustaceans. Chitinase enzyme is capable of degrading chitin, and this enzyme can be used as a biological fungicide against phytopathogenic fungi, as well as an insecticide against insect pests.MethodsIn this study, 158 isolates, which were derived from bacteria cultures isolated from leaves and root rhizospheres of certain plants in Turkey, were selected after confirming that they are not phytopathogenic based on the hypersensitivity test performed on tobacco; and antifungal activity test was performed against Fusarium culmorum, which is a pathogenic fungi that cause decomposition of roots of vegetables. Accordingly, chitinase enzyme activity assay was performed on 31 isolates that have an antifungal activity, and among them the isolate of Bacillus subtilis TV-125 was selected, which has demonstrated the highest activity.ResultsChitinase enzyme was purified by using ammonium sulphate and DEAE-sephadex ion exchange chromatography. Ammonium sulphate precipitation of chitinase enzyme from Bacillus subtilis TV-125 isolate was performed at maximum range of 0-20%, and 28.4-fold purification was obtained with a 13.4% of yield. Optimum activity of the purified enzyme was observed at pH 4.0 and at 50°C of temperature. In addition, it was identified that Bacillus subtilis TV-125A isolate retains 42% of its activity at 80°C temperature.ConclusionIn the last phase of the study, chitinase enzyme purified from Bacillus subtilis TV-125A was tested on four fungal agents, although all the results were positive, it was particularly effective on F. culmorum according to the findings.

【 授权许可】

Unknown   
© Senol et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd.