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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. Apr, 42(2): 190-193
Copyright © 2014, The Korean Society for Microbiology and Biotechnology
  • Received : March 08, 2014
  • Accepted : April 16, 2014
  • Published : April 28, 0214
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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 .
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Acknowledgements
This study was supported by the research grant of Chungbuk National University in 2012.
References
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