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| Optimization of Lipase Production Conditions by Bacillus subtilis Using Surface Methodology |
| SONG Ping,QI Xiao-ling,HU Yi,XIE Ning-chang |
| College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China |
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Abstract The fermentation conditions of lipase production by Bacillus subtilis CICC 20034 were optimized. Initially, the most suitable intruder tributyrin, nitrogen source urea, carbon glucose, and MgSO4 were selected according to single factorial experiments respectively. Based on the result, screening methodology Plackett-Burman design was used to evaluate the effects of twelve factors related to lipase production and four statistically significant factors tributyrin, urea, KH2PO4 and pH were selected. The path of steepest ascent was used to approach the optimal region of lipase production subsequently. Then, the optimal combined concentration for maximum enzyme activity were further optimized by response surface methodology and determined as follows: tributyrin 2. 62%, urea 8.57g/L, KH2PO4 2.59g/L and pH 9.47. The optimization of culture conditions of B.subtilis CICC 20034 led to a 6.7-fold increase in lipase production relative to initial result 0.072 /ml, which indicate that single factor in combination with response surface methodology, is an effective method for optimization of lipase production conditions by B.subtilis CICC 20034.
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Received: 21 January 2010
Published: 25 August 2010
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