|本期目录/Table of Contents|

[1]李倩,王梦,刘珞,等.L-乳酸脱氢酶在大肠杆菌BL-21(DE3)中的表达[J].生物加工过程,2011,9(06):21-25.[doi:doi:10.3969/j.issn.1672-3678.2011.06.005]
 LI Qian,WANG Meng,LIU Luo,et al.Expression of L-lactate dehydrogenase in Escherichia coli BL-21(DE3)[J].Chinese Journal of Bioprocess Engineering,2011,9(06):21-25.[doi:doi:10.3969/j.issn.1672-3678.2011.06.005]
点击复制

L-乳酸脱氢酶在大肠杆菌BL-21(DE3)中的表达()
分享到:

《生物加工过程》[ISSN:1672-3678/CN:32-1706/Q]

卷:
9
期数:
2011年06期
页码:
21-25
栏目:
出版日期:
2011-11-30

文章信息/Info

Title:
Expression of L-lactate dehydrogenase in Escherichia coli BL-21(DE3)
文章编号:
1672-3678(2011)06-0021-05
作者:
李倩王梦刘珞谭天伟
北京化工大学 生命科学与技术学院 北京市生物加工过程重点实验室,北京100029
Author(s):
LI QianWANG MengLIU LuoTAN Tianwei
Beijing Key Laboratory of Bioprocess, College of Life Science and Technology,Beijing University of Chemical Technology, Beijing 100029, China
关键词:
大肠杆菌L-乳酸脱氢酶基因鼠李糖乳杆菌
分类号:
TQ028.8
DOI:
doi:10.3969/j.issn.1672-3678.2011.06.005
文献标志码:
A
摘要:
为了在大肠杆菌中构建利用葡萄糖生产L-乳酸的途径,以鼠李糖乳杆菌(Lactobacillus rhamnosus)LA-04-01基因组为模板,设计引物扩增L乳酸脱氢酶基因L-ldh。将该基因连接到表达载体pET-28a(+)上,并转化大肠杆菌Top10。通过卡那霉素抗性平板筛选,提取重组质粒pET28a-L-ldh并测序,结果正确。将pET28a-L-ldh转化大肠杆菌BL-21(DE3),通过卡那霉素抗性平板筛选,得到产乳酸的大肠杆菌基因工程菌。经IPTG诱导后,SDS-PAGE电泳,检测到目的蛋白条带,L-乳酸脱氢酶比酶活力达到9.44 U/mL。该基因工程菌通过摇瓶发酵,L-乳酸产量达到3 g/L,成功构建出一条在大肠杆菌中生产L-乳酸的新途径。

参考文献/References:

[1]Hábová V,Melzoch K,Rychtera M.Electrodialysis as a useful technique for lactic acid separation from a model solution and a fermentation broth[J].Desalination,2004,162:361-372.
[2]于雷,雷霆,裴晓林,等.应用基因组改组选育耐糖L乳酸高产菌株[J].食品科学,2009,28(9):369-373.
[3]Hofvendahl K,Hahn-Hagerdal B. Factors affecting the fermentative lactic acid production from renewable resources[J]. Enzym Microb Technol,2000,26:87-107.
[4]Skory D C.Isolation and expression of lactate dehydrogenase genes from Rhizopus oryzae[J]. Appl Environ Microbiol,2000,66(6):2343-2348.
[5]Adachi E,Torigoe M,Sugiyama M,et al. Modification of metabolic pathways of Saccharomyces cerevisiae by the expression of lactate dehydrogenase and deletion of pyruvate decarboxylase genes for the lactic acid fermentation at low pH value[J]. Ferment Bioeng,1998,86(3):284-289.
[6]Dien S,Nichols N,Bothast J. Recombinant Escherichia coli engineered for the production of L-lactic acid from hexose and pentose sugars[J]. Ind Microbiol Biotechnol,2001,27(4):259-264.
[7]吴新宇,董英.乳酸脱氢酶高产菌株的分离筛选与初步鉴定[J].食品与机械,2010,26(1):35-37.
[8]李建武,萧能,余瑞元,等.生物化学实验原理和方法[M].北京:北京大学出版社,1998.
[9]Liu Luo,Aigner A,Schmid R D. Identification,cloning,heterologous expression and characterization of a NADPH-dependent 7β-hydroxysteroid dehydrogenase from Collinsella aerofaciens[J].Appl Microbiol Biotechnol,2011,90(1):127-135.
[10]张克旭,陈宁,张蓓.代谢控制发酵[M].北京:中国轻工业出版社,1998.

相似文献/References:

[1]成成,李兆丰,李彬,等.利用重组大肠杆菌生产α-环糊精葡萄糖基转移酶[J].生物加工过程,2009,7(03):56.
 CHENG Cheng,LI Zhao-feng,LI Bin,et al.Production of α-cyclodextrin glycosyltransferase in recombinant Escherichia coli[J].Chinese Journal of Bioprocess Engineering,2009,7(06):56.
[2]尹强,康振,钟盛华,等.产琥珀酸工程菌株的发酵工艺条件优化[J].生物加工过程,2010,8(04):1.[doi:10.3969/j.issn.1672-3678.2010.04.001]
 YIN Qiang,KANG Zhen,ZHONG Shenghua,et al.Optimization of fermentation production of succinateengineered by? Escherichia coli[J].Chinese Journal of Bioprocess Engineering,2010,8(06):1.[doi:10.3969/j.issn.1672-3678.2010.04.001]
[3]赵清风,陈介南,张林,等.AgNO3对大肠杆菌和金黄色葡萄球菌的抗菌作用及机制[J].生物加工过程,2011,9(03):52.[doi:10.3969/j.issn.1672-3678.2011.03.011]
 ZHAO Qingfeng,CHEN Jienan,ZHANG Lin,et al.Antimicrobial activity and mechanism of silver nitrate on Escherichia coli and Staphylococcus aureus[J].Chinese Journal of Bioprocess Engineering,2011,9(06):52.[doi:10.3969/j.issn.1672-3678.2011.03.011]
[4]魏淼,刘欢,李艳,等.共表达甘油脱氢酶和二羟丙酮激酶对大肠杆菌生长及甘油代谢的影响[J].生物加工过程,2011,9(05):59.[doi:doi:10.3969/j.issn.1672-3678.2011.05.012]
 WEI Miao,LIU Huan,LI Yan,et al.Effects on growth and glycerol metabolism in E.coli by coexpression protein GldA and DhaKLM[J].Chinese Journal of Bioprocess Engineering,2011,9(06):59.[doi:doi:10.3969/j.issn.1672-3678.2011.05.012]
[5]刘嵘明,梁丽亚,王光明,等.基于辅因子调控对大肠杆菌两阶段发酵产丁二酸的影响[J].生物加工过程,2012,10(03):1.[doi:10.3969/j.issn.1672-3678.2012.03.001]
 LIU Rongming,LIANG Liya,WANG Guangming,et al.Effect of cofactor regulation on succinate production in Escherichia coli during dual-phase fermentation[J].Chinese Journal of Bioprocess Engineering,2012,10(06):1.[doi:10.3969/j.issn.1672-3678.2012.03.001]
[6]王珊珊,高璐,严明,等.过表达氨基葡萄糖脱氨酶对大肠杆菌氨基葡萄糖合成及中心碳代谢的影响[J].生物加工过程,2013,11(04):42.[doi:10.3969/j.issn.1672-3678.2013.04.008]
 WANG Shanshan,GAO Lu,YAN Ming,et al.Effects of overexpression glucosamine deaminase on glucosamine synthesis and central carbon metabolism in Escherichia coli[J].Chinese Journal of Bioprocess Engineering,2013,11(06):42.[doi:10.3969/j.issn.1672-3678.2013.04.008]
[7]郑志永,关怡新,林东强,等.重组基因表达对大肠杆菌生理的影响[J].生物加工过程,2004,2(02):13.[doi:10.3969/j.issn.1672-3678.2004.02.003]
[8]杨雪莲,严明,许琳,等.AspC基因导入前后E.coli BL21蛋白质组的解析[J].生物加工过程,2005,3(04):49.[doi:10.3969/j.issn.1672-3678.2005.04.011]
 YANG Xue-lian,YAN Ming,XU Lin,et al.Comparative proteome analysis of AspC gene overexpression in E.coli BL21[J].Chinese Journal of Bioprocess Engineering,2005,3(06):49.[doi:10.3969/j.issn.1672-3678.2005.04.011]
[9]叶逢春,陈银,邢新会.重组大肠杆菌生产可溶性MBP融合肝素酶的培养条件优化[J].生物加工过程,2006,4(03):28.[doi:10.3969/j.issn.1672-3678.2006.03.005]
 YE Feng-chun,CHEN Yin,XING Xin-hui.Optimization of cultivation conditions of recombinant E. coli for production of soluble MBP-fused heparinase Ⅰ[J].Chinese Journal of Bioprocess Engineering,2006,4(06):28.[doi:10.3969/j.issn.1672-3678.2006.03.005]
[10]胡永红,汤天羽,周华,等.改性胶原蛋白膜用于大肠杆菌EP8-10组合固定化[J].生物加工过程,2007,5(02):73.[doi:10.3969/j.issn.1672-3678.2007.02.014]
 HU Yong-hong,TANG Tian-yu,ZHOU Hua,et al.Integrated immobilization of E. coli EP8-10 by modified collagen membrane[J].Chinese Journal of Bioprocess Engineering,2007,5(06):73.[doi:10.3969/j.issn.1672-3678.2007.02.014]

备注/Memo

备注/Memo:
收稿日期:2011-02-18
基金项目:国家自然科学基金资助项目(20876011);北京市自然科学基金资助项目(2071002);国家重点基础研究发展计划(973计划)资助项目(2007CB707804);国家高技术研究发展计划(863计划)资助项目(2006AA020103,2006AA020102,2006AA020201,2007AA100404,2007CB714304)
作者简介:李倩(1986—),女,湖北咸宁人,硕士研究生,研究方向:基因工程;谭天伟(联系人),教授,E-mail:twtan@mail.buct.edu.cn
更新日期/Last Update: