L-天冬酰胺酶的补料分批发酵

Fed-Batch Fermentation of L-Asparaginase

DOI：10.3969/j.issn.1673-1689.2020.03.001

中文关键词: 枯草芽孢杆菌 L-天冬酰胺酶 发酵优化 分批发酵 补料分批发酵

英文关键词: / Bacillus subtilis,L-asparaginase,fermentation optimization,batch fermentation,fed-batch fermentation

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中文摘要:

L-天冬酰胺酶（L-Asparaginase，EC 3.5.1.1，L-ASNase）广泛应用于食品和医药领域。为实现重组L-ASNase的高产，在3 L罐水平研究了搅拌转速与补料分批发酵条件对Bacillus subtilis/ASNΔ25/B2菌体生长与产酶的影响。通过优化确定补料分批发酵条件如下：发酵0 ~ 8 h搅拌转速：700 r/min；发酵8 h后搅拌转速：900 r/min；发酵16~28 h：恒速流加（18.75 mL/h）蔗糖（800 g/L），恒速流加（32 mL/h）酵母蛋白胨（200 g/L）和玉米浆（80 g/L）混合氮源。基于以上发酵条件，L-ASNase酶活在48 h可达到1 413.6 U/mL，较分批发酵提高了66.2%，生产强度提高了24.6%。研究结果为重组L-ASNase工业化生产提供了基础数据。

英文摘要:

L-asparaginase（EC 3.5.1.1，L-ASNase）has been widely used in the fields of food and medicine. In order to achieve the efficient production of L-ASNase，the effects of stirring speed and fed-batch fermentation conditions on the cell growth and enzyme production of/Bacillus subtilis//ASNΔ25/B2 were investigated in 3 L fermenter. After the optimization，conditions used for fed-batch fermentation were determined as follows：the stirring speed was set at 700 r/min for the first 8 h and shifted to 900 r/min till the end of the fermentation；for fermentation at 16~28 h，sucrose solution（800 g/L）and nitrogen sources（yeast peptone：200 g/L；corn syrup：80 g/L）were fed at 18.75 mL/h and 32 mL/h，respectively. Based on this fermentation condition，the L-ASNase yield reached 1 413.6 U/mL at 48 h，which was 66.2% higher than that of batch fermentation. The final production intensity obtained here was 24.6% higher than that of the latter. The results provide the basic data for the industrial production of the recombinant L-ASNase.

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