Molecular characterization of alkaline protease of Bacillus amyloliquefaciens SP1 involved in biocontrol of Fusarium oxysporum
| Autor: | C. K. Shirkot, Anjali Chauhan, Abhishek Walia, Shiwani Guleria |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
DNA
Bacterial 0301 basic medicine Bacillus amyloliquefaciens medicine.medical_treatment 030106 microbiology Bacillus subtilis Microbiology 03 medical and health sciences chemistry.chemical_compound Bacterial Proteins Fusarium Endopeptidases Fusarium oxysporum Escherichia coli medicine Protease Inhibitors Amino Acid Sequence Subtilisins Cloning Molecular Peptide sequence Protease Base Sequence biology Nucleic acid sequence Sequence Analysis DNA General Medicine biology.organism_classification Protease inhibitor (biology) 030104 developmental biology Biological Control Agents chemistry Biochemistry PMSF Food Science medicine.drug |
| Zdroj: | International Journal of Food Microbiology. 232:134-143 |
| ISSN: | 0168-1605 |
| DOI: | 10.1016/j.ijfoodmicro.2016.05.030 |
| Popis: | An alkaline protease gene was amplified from genomic DNA of Bacillus amyloliquefaciens SP1 which was involved in effective biocontrol of Fusarium oxysporum. We investigated the antagonistic capacity of protease of B. amyloliquifaciens SP1, under in vitro conditions. The 5.62 fold purified enzyme with specific activity of 607.69 U/mg reported 24.14% growth inhibition of F. oxysporum. However, no antagonistic activity was found after addition of protease inhibitor i.e. PMSF (15 mM) to purified enzyme. An 1149 bp nucleotide sequence of protease gene encoded 382 amino acids of 43 kDa and calculated isoelectric point of 9.29. Analysis of deduced amino acid sequence revealed high homology (86%) with subtilisin E of Bacillus subtilis. The B. amyloliquefaciens SP1 protease gene was expressed in Escherichiax coli BL21. The expressed protease was secreted into culture medium by E. coli and exhibited optimum activity at pH 8.0 and 60 °C. The most reliable three dimensional structure of alkaline protease was determined using Phyre 2 server which was validated on the basis of Ramachandran plot and ERRAT value. The expression and structure prediction of the enzyme offers potential value for commercial application in agriculture and industry. |
| Databáze: | OpenAIRE |
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