site > International journal > (2005) Coexpression of folding accessory proteins for production of active cyclodextrin glycosyltransferase of Bacillus macerans in recombinant Escherichia coli.

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작성일 : 12-07-17 17:33

(2005) Coexpression of folding accessory proteins for production of active cyclodextrin glycosyltransferase of Bacillus macerans in recombinant Escherichia coli.

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http://www.sciencedirect.com/science/article/pii/S1046592805000288 [1086]

Coexpression of folding accessory proteins for production of active cyclodextrin glycosyltransferase of Bacillus macerans in recombinant Escherichia coli.

Sung-Gun Kim, Dae-Hyuk Kweon, Dae-Hee Lee, Yong-Cheol Park and Jin-Ho Seo*
Protein Expression and Purification 41(2): 426-432. 2005.06.01.
(SCI I/F: 1.587)

Coexpression of foldingaccessoryproteins, molecular chaperones, and human peptidyl-prolyl cis–trans isomerase (PPIase) increased production of activecyclodextringlycosyltransferase (CGTase) of Bacillusmacerans, which is otherwise mainly expressed as inclusion body in recombinantEscherichiacoli. The best partner for soluble expression of CGTase was found to be human PPIase followed by coexpression of DnaK-DnaJ-GrpE together with GroEL-GroES. Such a significant enhancement by human PPIase coexpression seemed to be due to dual functions of chaperone and peptidyl-prolyl cis–trans isomerization. Coexpression of GroEL-GroES or minichaperone alone did not influence the specific CGTase activity. For production of active CGTase in large amounts, a high cell density culture was achieved using a pH-stat fed-batch strategy. The optimized fed-batch fermentation resulted in dry cell weight of 103.4 g/L and CGTase activity of 1200 U/mL. Combination of human PPIase expression at a gene level and cell culture optimization at a process scale exerted a synergistic effect on the product yield of soluble CGTase expression in recombinantE. coli.

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