产酯酶B1海洋枯草芽孢杆菌C5发酵条件优化  

Optimization of Fermentation Conditions for Marine Bacillus subtilis C5 Producing Esterase B1

作　　者：包兴艳[1,2] 郝建华[1] 陈世建[1,2] 王芳[1] 王伟[1] 孙谧[1] 

机构地区：[1]中国水产科学研究院黄海水产研究所海洋产物资源与酶工程实验室,青岛266071 [2]上海海洋大学食品学院,上海201306

出　　处：《应用与环境生物学报》2012年第6期999-1003,共5页Chinese Journal of Applied and Environmental Biology

基　　金：国家"十二五""863"项目(No.2011AA090703);国际科技合作与交流专项(No.2011DFB30250);青岛市科技计划(No.11-2-4-20-hz);基本科研业务费(No.20603022011019)资助~~

摘　　要：为提高芽孢杆菌C5产酯酶B1的能力,采用响应面法对其发酵条件进行优化.首先通过单因素实验筛选氮源、碳源、接种量、起始发酵pH、装液量、发酵温度和转速这7个因素,当酶活达到最高值时,各单因素的值分别为氮源玉米浆30 mL/L,碳源麦芽糖35 g/L,接种量4%(φ),起始pH 7.0,装液量15 mL,培养温度36℃,转速在实验室现有条件下最高选择220 r/min;再通过Plackett-Burman(PB)设计法,评价了这7个因素对酯酶B1产量影响的大小,确定氮源玉米浆的浓度、发酵起始pH和发酵温度为酯酶产生的3个主要影响因素;利用中心组合设计(CCD)及SAS软件分析获得了主要因素的最优条件,即玉米浆浓度28.70 mL/L,起始发酵pH为7.10,发酵温度为35.8℃.预测最高酶活力为139.18U/mL,实验最终酶活达到138.40 U/mL,与原发酵条件相比提高了228.15%.其实验值与预测值基本相符,说明预测模型可应用于酯酶发酵条件的优化.The fermentation conditions of marine Bacillus subtilis C5 were optimized in order to improve the yield of the esterase B1. Firstly, single factor experiment was applied to screen the following seven factors： the nitrogen source, carbon source, inoculation volume, initial pH, broth content, fermentation temperature and rotating speed. The results indicated that when the production of esterase reached the highest yield, the value of each single factor was as follows： corn syrup 30 mL/ L, maltose 35 g/L, 4% （~0）, pH 7.0, 15 mL, 36 ~C, 220 r/min, respectively. Secondly, those variables were valuated through the Plackett-Burman （PB） design method. The results showed that nitrogen source, pH and fermentation temperature were the main affecting variables. Then, the central composite design （CCD） and SAS software were adopted to determine the optimal level of the three main factors. The most suitable variables were identified as follows： corn syrup 28.70 mL/L, pH 7.10, fermentation temperature 35.8 ~C, respectively. The esterase activity was predicted to be 139.18 U/mL. The result of verification experiment under the optimum conditions showed that the maximum esterase activity was 138.40 U/mL, and the yield of esterase B1 was raised by 228.15% through optimizing the fermentation conditions. These results suggested that the predicted model was reliable and available for the optimization of esterase fermentation conditions.

关 键 词：枯草芽孢杆菌 酯酶 发酵 优化 PLACKETT-BURMAN 响应面法 甲基对硫磷 

分 类 号：TQ925.9[轻工技术与工程—发酵工程]