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

Research

Sunil K Khare¹  Vinayak R Tripathi²  Santosh K Singh²  Sanjay K Singh²  Satyendra K Garg² 

[1] Department of Chemistry, Indian Institute of Technology, 110016, Hauz-Khas, New Delhi, India;Department of Microbiology, Center of Excellence,Dr. Ram Manohar Lohia Avadh University, 224001, Faizabad, UP, India;

关键词: Response Surface Methodology;    Enzyme Production;    Agitation Speed;    Alkaline Protease;    Aeration Rate;   

DOI  :  10.1186/1475-2859-10-114

 received in 2011-10-04, accepted in 2011-12-28,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundProduction 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.ResultsThe pre-screening medium amended with optimized (w/v) 1.0% glucose, 2.0% gelatin and 0.5% yeast extract, produced 278 U protease ml-1 at 72 h incubation. Enzyme production increased to 431 Uml-1 when Mg2+ (0.01%, w/v) was supplemented. Optimization of physical factors further enhanced protease to 514 Uml-1 at pH 9.0, 25°C and 200 rpm within 60 h. The combined effect of conventionally optimized variables (glucose, yeast extract, MgSO4 and pH), thereafter predicted by response surface methodology yielded 617 U protease ml-1 at glucose 1.25% (w/v), yeast extract 0.5% (w/v), MgSO4 0.01% (w/v) and pH 8.8. Bench-scale bioreactor level optimization resulted in enhanced production of 882 U protease ml-1 at 0.8 vvm aeration and 150 rpm agitation during only 48 h incubation.ConclusionsThe optimization of fermentation variables using conventional, statistical approaches and aeration/agitation at fermentor level resulted in ~13.5 folds increase (882 Uml-1) in protease production compared to un-optimized conditions (65 Uml-1). This is the highest level of thermoalkaline protease reported so far by any psychrotrophic bacterium.

【 授权许可】

Unknown   
© Singh et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.