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Comparative one-factor-at-a-time, response surface (statistical) and bench-scale bioreactor level optimization of thermoalkaline protease production from a psychrotrophic Pseudomonas putida SKG-1 isolate

Microbial Cell Factories, ISSN: 1475-2859, Vol: 10, Issue: 1, Page: 114
2011
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Key words Central composite design (CCD), Raoultella ornithinolytica, Response surface methodology (RSM), Solid-state fermentation (SSF), Tannase INTRODUCTION Tannin is naturally occurring polyphenolic compound and 4th

Article Description

Background: Production of alkaline protease from various bacterial strains using statistical methods is customary now-a-days. The present work is first attempt for the production optimization of a solvent stable thermoalkaline protease by a psychrotrophic Pseudomonas putida isolate using conventional, response surface methods, and fermentor level optimization.Results: The pre-screening medium amended with optimized (w/v) 1.0% glucose, 2.0% gelatin and 0.5% yeast extract, produced 278 U protease ml at 72 h incubation. Enzyme production increased to 431 Uml when Mg (0.01%, w/v) was supplemented. Optimization of physical factors further enhanced protease to 514 Uml at pH 9.0, 25°C and 200 rpm within 60 h. The combined effect of conventionally optimized variables (glucose, yeast extract, MgSO and pH), thereafter predicted by response surface methodology yielded 617 U protease ml at glucose 1.25% (w/v), yeast extract 0.5% (w/v), MgSO 0.01% (w/v) and pH 8.8. Bench-scale bioreactor level optimization resulted in enhanced production of 882 U protease ml at 0.8 vvm aeration and 150 rpm agitation during only 48 h incubation.Conclusions: The optimization of fermentation variables using conventional, statistical approaches and aeration/agitation at fermentor level resulted in ~13.5 folds increase (882 Uml ) in protease production compared to un-optimized conditions (65 Uml ). This is the highest level of thermoalkaline protease reported so far by any psychrotrophic bacterium. © 2011 Singh et al; licensee BioMed Central Ltd.

Bibliographic Details

Santosh K. Singh; Sanjay K. Singh; Vinayak R. Tripathi; Satyendra K. Garg; Sunil K. Khare

Springer Science and Business Media LLC

Biochemistry, Genetics and Molecular Biology; Chemical Engineering; Immunology and Microbiology

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