摘要
对从深海沉积物宏基因组文库中获得的产低温脂肪酶基因工程菌LIP001进行了发酵条件优化。通过单因素试验对LIP001产脂肪酶的主要影响条件进行了探讨,确定了培养条件为30℃、pH7.0、接种量5%、装液量50ml。在单因素的基础上通过正交试验优化了影响重组菌LIP001产酶主要因素:橄榄油、酵母粉、磷酸盐、MgSO4,确定了培养基为橄榄油1%、酵母粉0.5%、蛋白胨1%、硫酸铵0.5%、磷酸盐0.5%、MgSO4为0.2%、氯霉素12.5μg/ml,优化后的脂肪酶活为1980U/ml,比优化前提高了54.7%,为大规模发酵奠定了基础。采用5升发酵罐方法试验,酶活达到2420U/ml。
One clone（LIP001） producing cold-active lipase was selected from metagenomic library of deep sea sediment for the optimization study of fermentation conditions.The effects of main factors on the growth and lipase-producing of LIP001 were determined by single-factor test.The results showed that the optimal condition were 30℃,pH7.0,and inoculum size 5%,liquid volume 50ml in 250ml flask,respectively.The fermentation conditions for the lipase-producing of LIP001 were further optimized by orthogonal experiment on the basis of single factor test.The major factors and their concentrations were as follows：olive oil（1%）,yeast extract（0.5%）,peptone（1%）,（NH4）2SO4（0.5%）,phosphate（0.5%）,MgSO4（0.2%）,and chloromycetin（12.5μg/ml）,respectively.Under the optimized condition,the lipase activity was 1980U/ml which was increased by 54.7%.For 5L fermentor research,the lipase activity was 2420U/ml.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2011年第4期65-70,共6页
China Biotechnology
基金
国家"863"计划(2007AA091407)
中国大洋矿产资源研究开发协会(DYXM-115-02-2-04)
福建省教育厅科技项目(JA09280)资助项目
关键词
深海
宏基因组文库
低温脂肪酶
发酵优化
Deep-sea Metagenomic library Cold-active lipase Fermentation optimization
