Advanced
Soluble Production of CMP-Neu5Ac Synthetase by Co-expression of Chaperone Proteins in Escherichia coli
Soluble Production of CMP-Neu5Ac Synthetase by Co-expression of Chaperone Proteins in Escherichia coli
Microbiology and Biotechnology Letters. 2014. Jun, 42(2): 190-193
Copyright © 2014, The Korean Society for Microbiology and Biotechnology
  • Received : March 08, 2014
  • Accepted : April 16, 2014
  • Published : June 28, 2014
Download
PDF
e-PUB
PubReader
PPT
Export by style
Share
Article
Author
Metrics
Cited by
TagCloud
About the Authors
화영 최
령 이
승기 조
원흥 이
진호 서
남수 한
namsoo@cbnu.ac.kr

Abstract
CMP-Neu5Ac synthetase is a key enzyme for the synthesis of CMP-Neu5Ac, which is an essential precursor of sialylated glycoconjugates. For the soluble expression of the CMP-Neu5Ac synthetase gene (neuA) from Escherichia coli K1, various heat shock proteins were co-expressed in E. coli BL21 (DE3) Star. In order to do this, a pG-KJE8 plasmid, encoding genes for GroEL-ES and DnaK-DnaJ-GrpE, was co-transformed with neuA and was expressed at 20°C by the addition of 0.01 mM IPTG and 0.005 mg/ml L-arabinose. The co-expression of a variety of heat shock proteins resulted in the remarkably improved production of soluble CMP-Neu5Ac synthetase in E. coli .
Keywords
PPT Slide
Lager Image
PPT Slide
Lager Image
PPT Slide
Lager Image
Acknowledgements
This study was supported by the research grant of Chungbuk National University in 2012.
View Fulltext  
Boehm G , Stahl B 2007 Oligosaccharides from milk J. Nutr. 137 847 - 849
Gilbert M , Cunningham AM , Watson DC , Martin A , Richards JC , Wakarchuk WW 1997 Characterization of a recombinant Neisseria meningitidis alpha-2,3-sialyltransferase and its acceptor specificity Eur. J. Biochem. 249 187 - 194    DOI : 10.1111/j.1432-1033.1997.t01-1-00187.x
Lee WH , Shin SY , Kim MD , Han NS , Seo JH 2012 Modulation of guanosine nucleotides biosynthetic pathways enhanced GDP-L-fucose production in recombinant Escherichia coli Appl. Microbiol. Biotechnol. 93 2327 - 2334    DOI : 10.1007/s00253-011-3776-3
Mogk A , Tomoyasu T , Goloubinoff P , Rüdiger S , Röder D , Langen H , et al. 1999 dentification of thermolabile Escherichia coli proteins: prevention and reversion of aggregation by DnaK and ClpB Embo. J. et al. 18 6934 - 6949    DOI : 10.1093/emboj/18.24.6934
Nakano T , Sugawara M , Kawakami H 2001 Sialic acid in human milk: composition and functions Acta Paediatr Taiwan 42 11 - 17
Preston A , Mandrell RE , Gibson BW , Apicella MA 1996 The lipooligosaccharides of pathogenic gram-negative bacteria Crit. Rev. Microbiol. 22 139 - 180    DOI : 10.3109/10408419609106458
Ringenberg M , Lichtensteiger C , Vimr E 2001 Redirection of sialic acid metabolism in genetically engineered Escherichia coli Glycobiology 11 533 - 539    DOI : 10.1093/glycob/11.7.533
Schein CH 1989 Production of Soluble Recombinant Proteins in Bacteria Nature 7 1141 - 1148
Sørensen HP , Mortensen KK 2005 Soluble expression of recombinant proteins in the cytoplasm of Escherichia coli Microb. Cell Fact. 4 1 -    DOI : 10.1186/1475-2859-4-1
Warrick JM , Chan HY , Gray-Board GL 1999 Suppression of polyglutamine-mediated neurodegeneration in Drosophila by the molecular chaperone HSP70 Nat. Genet. 23 425 - 428    DOI : 10.1038/70532
Weickert MJ , Doherty DH , Best EA , Olins PO 1996 Optimization of heterologous protein production in Escherichia coli Curr. Opin. Biotechnol. 7 494 - 499    DOI : 10.1016/S0958-1669(96)80051-6