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| Optimization Fermentation Conditions of the Nitrile Hydratase Fusant Strain Tolerance to the Substrate |
| ZHANG Li-ling1,2, WANG Xiao-lan 1,2 |
1. School of life Science ,Jiangxi Normal University, Nanchang 330022,China;
2. Key Laboratory of Functional Small Organic Molecule, Ministry of Education, Nanchang 330022,China |
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Abstract Orthogonal design was used to optimized the fermentation conditions of the nitrile hydratase(NHase) fusant strain tolerance to the substrate, Fermented liquid starting pH, fermentation period, inoculation quantity, pack liquid coefficient as the investigation factors,Finally determined the best fermentation conditions for: start pH8.0, fermentation period 54 h, Inoculation quantity 12%, pack fluid coefficient 12%. Under the optimized conditions, activity of nitriles hydratase of the fusant strain increase to 11 millionU/ml, is up 83.3% before optimization. Response surface methodology was used to optimize the fermentation medium. A Plackett-Burman design was used to evaluate the influence of eight factors firstly. Results showed that the concentration of glucose, urea, K2HPO4 and KH2PO4 played important roles in influencing the activity of nitrile hydratase. Then, the path of steepest ascent and the Central composite design were adopted for further optimization, and the optimum concentration levels and the relationships among these factors was found out by quadratic regression model equation with Design-Expert statistic methods, the optimal concentration of the variables were determined as: glucose 22.62 g/L, urea 9.76g/L, K2HPO4 1.22g/L,KH2PO4 1.268g/L. Under such conditions, the activity of nitrile hydratase was increased to 12.80 millionU/ml, which was 16.4% higher than old medium composition.
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Received: 19 March 2012
Published: 25 August 2012
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