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Development of an efficient process intensification strategy for enhancing Pfu DNA polymerase production in recombinant Escherichia coli
Hu, Jian-Hua1,2; Wang, Feng1; Liu, Chun-Zhao1
2015-04-01
Source PublicationBIOPROCESS AND BIOSYSTEMS ENGINEERING
ISSN1615-7591
Volume38Issue:4Pages:651-659
Abstract

An efficient induction strategy that consisted of multiple additions of small doses of isopropyl-beta-D-thiogalactopyranoside (IPTG) in the early cell growth phase was developed for enhancing Pfu DNA polymerase production in Escherichia coli. In comparison to the most commonly used method of a single induction of 1 mM IPTG, the promising induction strategy resulted in an increase in the Pfu activity of 13.5 % in shake flasks, while simultaneously decreasing the dose of IPTG by nearly half. An analysis of the intracellular IPTG concentrations indicated that the cells need to maintain an optimum intracellular IPTG concentration after 6 h for efficient Pfu DNA polymerase production. A significant increase in the Pfu DNA polymerase activity of 31.5 % under the controlled dissolved oxygen concentration of 30 % in a 5 L fermentor was achieved using the multiple IPTG induction strategy in comparison with the single IPTG induction. The induction strategy using multiple inputs of IPTG also avoided over accumulation of IPTG and reduced the cost of Pfu DNA polymerase production.

KeywordCell Growth Escherichia Coli Pfu Dna Polymerase Iptg Induction
SubtypeArticle
Subject AreaBiotechnology & Applied Microbiology ; Engineering
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Technology
DOI10.1007/s00449-014-1304-4
Indexed BySCI
Language英语
WOS KeywordFed-batch Cultures ; Pyrococcus-furiosus ; E. Coli ; Expression ; Protein ; Oxygen ; Iptg ; Accumulation ; Purification ; Limitation
Funding ProjectNational Key Technology Research and Development Program of China [2012BAK25B01] ; Knowledge Innovation Program of the Chinese Academy of Sciences [KSZD-EW-Z-015] ; Chinese Academy of Sciences Fellowships for Young International Scientists [2011Y1GA01]
WOS Research AreaBiotechnology & Applied Microbiology ; Engineering
WOS SubjectBiotechnology & Applied Microbiology ; Engineering, Chemical
WOS IDWOS:000351513500005
Citation statistics
Cited Times:7[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/13794
Collection研究所(批量导入)
Corresponding AuthorLiu, Chun-Zhao
Affiliation1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Hu, Jian-Hua,Wang, Feng,Liu, Chun-Zhao. Development of an efficient process intensification strategy for enhancing Pfu DNA polymerase production in recombinant Escherichia coli[J]. BIOPROCESS AND BIOSYSTEMS ENGINEERING,2015,38(4):651-659.
APA Hu, Jian-Hua,Wang, Feng,&Liu, Chun-Zhao.(2015).Development of an efficient process intensification strategy for enhancing Pfu DNA polymerase production in recombinant Escherichia coli.BIOPROCESS AND BIOSYSTEMS ENGINEERING,38(4),651-659.
MLA Hu, Jian-Hua,et al."Development of an efficient process intensification strategy for enhancing Pfu DNA polymerase production in recombinant Escherichia coli".BIOPROCESS AND BIOSYSTEMS ENGINEERING 38.4(2015):651-659.
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