Enhanced secretion of recombinant α-cyclodextrin glucosyltransferase from E. coli by medium additives
Enhanced secretion of recombinant α-cyclodextrin glucosyltransferase from E. coli by medium additives
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DOI：
10.1016/j.procbio.2010.02.009
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发表时间：
2010-06-01
影响因子：
4.4
通讯作者：
Chen, Jian
中科院分区：
文献类型：
--
作者：
Ding, Runrong;Li, Zhaofeng;Chen, Jian
研究培养基添加剂对大肠杆菌分泌重组α-环糊精葡萄糖基转移酶（α-CGTase）的影响。结果表明，补充E. SDS、甘氨酸、Ca ~（2+）、Na ~+对α-CGT酶的分泌均有促进作用。正交试验结果表明，在添加0.03%SDS、400 mM Na+、0.3%甘氨酸和10 mM Ca 2+的条件下，α-CGTase的分泌量最大。在此条件下，胞外酶活力达到12.89U/ml，比无添加物培养提高了15倍。进一步分析表明，E.在优化条件下，大肠杆菌细胞膜的渗透率比对照条件下显著增加。这些可能是增加α-CGTase从周质区室分泌到培养基中的潜在机制。(C)2010爱思唯尔有限公司版权所有。
The purpose of this study was to investigate the effect of medium additives on the secretion of recombinant a-cyclodextrin glucosyltransferase (alpha-CGTase) into the culture media of Escherichia coli. It is found that supplementation of the E. coli culture with SDS, glycine, Ca2+ or Na+, individually, facilitated the secretion of alpha-CGTase. Orthogonal experiment showed that the optimal condition to achieve maximal secretion of alpha-CGTase was the supplementation with 0.03% SDS, 400 mM Na+, 0.3% glycine and 10 mM Ca2+ together. Under this condition, extracellular enzyme activity reached 12.89 U/ml, which is 15 times higher than that of the culture without any additives. Further analysis showed that the permeability, fluidity and phosphatidylglycerol content of the E. coli cell membrane under the optimized condition were significantly increased in comparison to those under the control condition. These might be the potential mechanisms for the increased secretion of alpha-CGTase from the periplasmic compartment into the culture medium. (C) 2010 Elsevier Ltd. All rights reserved.