食品科学

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短乳杆菌发酵生产尿苷磷酸化酶条件的优化

王伟洁1,李红梅1,*,邓龙华1,高露姣2,黄艳青2   

  1. 1.上海理工大学医疗器械与食品学院,上海 200093;2.中国水产科学研究院东海水产研究所,上海 200090
  • 出版日期:2013-09-15 发布日期:2013-09-27
  • 通讯作者: 李红梅
  • 基金资助:

    上海市研究生创新基金项目(54-11-115-004);上海市大学生创新基金项目(52-12-308-201)

Optimization of Fermentation Medium for Producing Uridine Phosphorylase by Lactobacillus brevis

WANG Wei-jie1,LI Hong-mei1,*,DENG Long-hua1,GAO Lu-jiao2,HUANG Yan-qing2   

  1. 1. School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China;
    2. East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, China
  • Online:2013-09-15 Published:2013-09-27
  • Contact: LI Hong-mei

摘要:

采用响应面法对短乳杆菌产尿苷磷酸化酶发酵培养基及发酵条件进行优化。以葡萄糖酵母膏培养基为基础培养基,通过单因素和Plackett-Burman设计法确定促进尿苷磷酸化酶合成的4个较为重要的有效因子:葡萄糖(P=0.002)、酵母粉(P=0.054)、磷酸根(P=0.059)和尿苷(P=0.001);在此基础上采用最陡爬坡试验逼近最大响应区域,并利用Box-Behnken设计对影响显著因素进行优化,得到最适培养基组成和培养条件为:NaCl 5g/L、葡萄糖18.30g/L、酵母粉15.71g/L、蛋白胨15g/L、磷酸根3.99g/L、肌苷20mmol/L、尿苷21.38mmol/L、初始pH8.0、摇床转速110r/min、发酵温度32℃、培养时间14h、接种量1.0%。在此优化条件下,短乳杆菌产尿苷磷酸化酶能力达到1.325U/mg,较优化前(0.883U/mg)提高了50.0%。

关键词: 短乳杆菌, 尿苷磷酸化酶, Box-Behnken设计, 响应面分析

Abstract:

The culture medium and fermentation conditions for producing uridine phosphorylase by Lactobacillus brevis was
optimized by response surface methodology. On the basis of glucose-yeast extract culture medium, four factors such as glucose
concentration (P = 0.002), yeast extract (P = 0.054), phosphate group (P = 0.059) and uridine (P = 0.001) were identified as
important factors that affect the production of uridine phosphorylase by single factor and Placket-Burman designs. These factors
were further examined through steepest ascent path method to find the maximum response region and optimized by response
surface Box-Behnken design. The optimal fermentation medium comprised NaCl 5 g/L, glucose 18.30 g/L, yeast extract 15.71 g/L,
peptone 15 g/L, phosphate group 3.99 g/L, carnine 20 mmol/L and uridine concentration 21.38 mmol/L at initial pH of 8.0.
The optimal culture conditions were shaking speed of 110 r/min, culture temperature of 32 ℃, fermentation time of 14 h and
inoculum amount of 1.0%. Under the optimized conditions, the production of uridine phosphorylase by Lactobacillus brevis
was 1.325 U/mg wet bacteria, which exhibited an increase by 50.0% compared with non-optimization.

Key words: Lactobacillus brevis, uridine phosphorylase, Box-Behnken design, response surface methodology

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