[1]王文研,张光亚.ELPs-PDOR融合基因表达条件的优化[J].华侨大学学报(自然科学版),2012,33(3):296-299.[doi:10.11830/ISSN.1000-5013.2012.03.0296]
　WANG Wen-yan,ZHANG Guang-ya.Optimization of Cultivation Conditions of ELPs-PDOR Fusion Gene[J].Journal of Huaqiao University(Natural Science),2012,33(3):296-299.[doi:10.11830/ISSN.1000-5013.2012.03.0296]
点击复制

ELPs-PDOR融合基因表达条件的优化()
分享到：

《华侨大学学报（自然科学版）》[ISSN:1000-5013/CN:35-1079/N]

卷:
第33卷
期数:
2012年第3期
页码:
296-299
栏目:
出版日期:
2012-05-20

文章信息/Info

Title:
Optimization of Cultivation Conditions of ELPs-PDOR Fusion Gene
文章编号:
1000-5013(2012)03-0296-04
作者:
王文研; 张光亚
华侨大学化工学院
Author(s):
WANG Wen-yan;  ZHANG Guang-ya
College of Chemical Engineering, Huaqiao University, Xiamen 361021, China
关键词:
类弹性蛋白多肽;  1; 3-丙二醇氧化还原酶;  大肠杆菌;  重组蛋白;  均匀设计
Keywords:
eastin-like polypeptides;  1; 3-propanediol oxidoreductase;  Escherichia coli;  recombinant protein;  uniform-design
分类号:
Q78
DOI:
10.11830/ISSN.1000-5013.2012.03.0296
文献标志码:
A
摘要:
利用均匀设计法和二次多项式逐步回归,对重组大肠杆菌生产类弹性蛋白多肽-1,3-丙二醇氧化还原酶(ELPs-PDOR)重组蛋白的培养条件进行优化.结果表明:在装液量为30%,诱导剂浓度为6.3mmol·L-1,诱导温度为30℃,诱导时间为2h的优化条件下进行培养,重组菌融合蛋白表达量是原始表达量的3.3倍,酶活力提高到10.84mkat·L-1,提高2.1倍.
Abstract:
The optimal parameters of cultivation condition of eastin-like polypeptides-1,3-propanediol oxidoreductase(ELPs-PDOR) by recombinant Escherichia coli were obtained through uniform-design and two polynomial stepwise regression.The optimal conditions of experiment were as follows: the medium volume was 30%,the induction concentration was 6.3 mmol·L-1,the cultural temperature was 30 ℃ and the induction temperature was 2 h.On the basis of this,the yield of ELPs-PDOR is 3.3 times higher than the primitive expression,and the enzymatic activity of PDOR is 10.84 mkat·L-1 which is 2.1 times of the enzymatic activity of primitive expression.

参考文献/References:

[1] URRY D W. Physical chemistry of biological free energy transduction as demonstrated by elastic protein-based polymers [J]. Journal of Physical Chemistry, 1997, (51):11007-11028.
[2] URRY D W. Free energy transduction in polypeptides and proteins based on inverse temperature transitions [J]. Progress in Biophysics and Molecular Biology, 1992(1):23-57.doi:10.1016/0079-6107(92)90003-O.
[3] LIM D W, KIMBERLY T C, MACKAY J A. Improved non-chromatographic purification of a recombinant protein by cationic elastin-like polypeptides [J]. Biomacromolecules, 2007(5):1417-1424.
[4] 黄凯宗, 李晶晶, 李巍. 类弹性蛋白多肽的从头设计、非色谱纯化及盐效应 [J]. 生物工程学报, 2011(4):553-658.
[5] LI Wei, FANG Bai-shan, ZHANG Guang-ya. Codon optimization of 1, 3-propanediol oxidoreductase expression in Escherichia coli and enzymatic properties [J]. Electronic Journal of Biotechnology,.
[6] AHRENS K, MENZEL K, ZENG A. Kinetic, dynamic, and pathway studies of glycerol metabolism by Klebsiella pneumoniae in anaerobic continuous culture (Ⅲ):Enzymes and fluxes of glycerol dissimilation and 1, 3-propanediol formation [J]. Biotechnology and Bioengineering, 1998(5):544-552.doi:10.1002/(SICI)1097-0290(19980905)59:5< 544::AID-BIT3> 3.0.CO; 2-A.
[7] 陈宏文, 聂金峰, 方柏山. 克雷伯杆菌1, 3-丙二醇氧化还原酶的分离纯化及其酶学性质 [J]. 华侨大学学报(自然科学版), 2009(1):62-66.
[8] MIROUX B, WALKER J E. Overproduction of proteins in Escherichia coli:Mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels [J]. Journal of Molecular Biology, 1996(3):289-298.doi:10.1006/jmbi.1996.0399.
[9] 李志西. 试验优化设计与统计分析 [M]. 北京:科学出版社, 1994.
[10] KWEON D H, HAN N S, PARK K M. Overproduction of phytolacca insularis protein in batch and fed-batch culture of recombinant Escherichia coli [J]. Process Biochemistry, 2001(6):537-542.doi:10.1016/S0032-9592(00)00237-5.

相似文献/References:

[1]兰琳,林锡煌,彭益强,等.耐高浓度甘油的1,3-丙二醇生产菌的诱变选育[J].华侨大学学报(自然科学版),2007,28(4):399.[doi:10.3969/j.issn.1000-5013.2007.04.017]
　LAN Lin,LIN Xi-huang,PENG Yi-qiang,et al.Breeding of Strain Producing 1,3-Propanediol in High Concentration of Glycerol[J].Journal of Huaqiao University(Natural Science),2007,28(3):399.[doi:10.3969/j.issn.1000-5013.2007.04.017]
[2]陈国,陈宏文,彭益强,等.NaCS/PDMDAAC微囊化肺炎克雷伯氏菌发酵产1,3-丙二醇[J].华侨大学学报(自然科学版),2008,29(4):554.[doi:10.11830/ISSN.1000-5013.2008.04.0554]
　CHEN Guo,CHEN Hong-wen,PENG Yi-qiang,et al.Primary Study on Culture of Encapsulated Klebsiella pneumoniae for Production of 1,3-Propanediol in Shaking Flask[J].Journal of Huaqiao University(Natural Science),2008,29(3):554.[doi:10.11830/ISSN.1000-5013.2008.04.0554]
[3]葛慧华,张光亚,王士斌.氨基酸对类弹性蛋白多肽产量的影响[J].华侨大学学报(自然科学版),2013,34(5):542.[doi:10.11830/ISSN.1000-5013.2013.05.0542]
　GE Hui-hua,ZHANG Guang-ya,WANG Shi-bin.Influence of Amino Acids on the Production of Elastin-Like Polypeptides ［KV8F-40］[J].Journal of Huaqiao University(Natural Science),2013,34(3):542.[doi:10.11830/ISSN.1000-5013.2013.05.0542]

备注/Memo

备注/Memo:
国家自然科学基金资助项目(20806031); 福建省自然科学基金资助项目(2009J01030); 福建省高校新世纪优秀人才支持项目(07176C02)
更新日期/Last Update: 2014-03-23