重组枯草芽孢杆菌高产氨肽酶策略与提取工艺优化

Strategy to Obtain High Yield of Aminopeptidase from Recombinant Bacillus subtilis and Its Extraction Process Optimization

DOI：10.3969/j.issn.1673-1689.2015.08.012

中文关键词: 氨肽酶 发酵优化 提取工艺

英文关键词: aminopeptidase, optimization of fermentation, extraction process

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中文摘要:

研究不同的产酶促进剂（渗透压调节剂、表面活性剂、有机酸盐）对重组枯草芽孢杆菌产氨肽酶的影响，并对氨肽酶的提取工艺进行优化。获得产酶促进剂优化组合为：氯化钾0.1 mol/L、吐温802.4 μL/mL、L-谷氨酸钠0.6%、酒石酸钾0.7%；优化培养条件为：装液量60 mL/250 mL、初始pH 7.5、接种体积分数5%、转速220 r/min、培养时间36 h。在上述优化条件下，该重组菌株产氨肽酶酶活达232.5 U/mL，是优化前的3.32倍。该重组氨肽酶发酵液经低速离心、双水相萃取、超滤浓缩提取后，回收率达67.23%，纯化倍数为8.29。

英文摘要:

This study aimed to investigate the effects of different enzyme accelerants on the production of aminopeptidase by recombinant Bacillus subtilis and develop an efficient extract process of aminopeptidase. The results indicated that the enzyme accelerants（KCl at 0.1 mol/L，Tween-80 at 2.4 μL/mL，L-glutamate at 0.6%，potassium tartrate at 0.7%） significantly increased the production of aminopeptidase. The cultural conditions were of 60 mL/250 mL of liquid，5% inoculation size，at 37 ℃，initial pH 7.5，shake flask rotation 220 r/min，and cultured time 36 h. Under these conditions，the aminopeptidase activity of recombinant Bacillus subtilis could reach 232.5 U/mL. Using the efficient extract process of aminopeptidase，the recovery and the purification fold of the L-leucine aminopeptidase could reach 67.23% and 8.29，respectively.

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