Heterologous expression of cyclodextrin glycosyltransferase from Paenibacillus macerans in Escherichia coli and its application in 2-O-α-D-glucopyranosyl-L-ascorbic acid production

BMC Biotechnology
Yujia JiangMin Jiang

Abstract

Cyclodextrin glucanotransferase (CGTase) can transform L-ascorbic acid (L-AA, vitamin C) to 2-O-α-D-glucopyranosyl-L-ascorbic acid (AA-2G), which shows diverse applications in food, cosmetic and pharmaceutical industries. In this study, the cgt gene encoding α-CGTase from Paenibacillus macerans was codon-optimized (opt-cgt) and cloned into pET-28a (+) for intracellular expression in E. coli BL21 (DE3). The Opt-CGT was purified by Ni2+-NTA resin with a 55% recovery, and specific activity was increased significantly from 1.17 to 190.75 U·mg- 1. In addition, the enzyme was adopted to transform L-AA into 9.1 g/L of AA-2G. Finally, more economic substrates, including β-cyclodextrin, soluble starch, corn starch and cassava starch could also be used as glycosyl donors, and 4.9, 3.5, 1.3 and 1.5 g/L of AA-2G were obtained, respectively. N-terminal amino acid is critical to the activity of CGTase suggested by its truncation study. Furthermore, when the Opt-CGT was flanked by His6-tags on the C- and N-terminal, the recovery of purification by Ni2+-NTA resin is appreciably enhanced. α-cyclodextrin was the ideal glycosyl donor for AA-2G production. In addition, the selection of low cost glycosyl donors would make the process of AA-2G produ...Continue Reading

References

Oct 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·R JanknechtH G Stunnenberg
Jul 24, 2002·Nutrition in Clinical Care : an Official Publication of Tufts University·Robert A Jacob, Gity Sotoudeh
Apr 22, 2008·Nucleic Acids Research·Andreas R GruberIvo L Hofacker
Aug 30, 2008·Journal of Industrial Microbiology & Biotechnology·Rui Min OngRosli Md Illias
Feb 6, 2010·Proceedings of the National Academy of Sciences of the United States of America·Tamir TullerEytan Ruppin
May 29, 2012·Applied Microbiology and Biotechnology·Ruizhi HanJian Chen
Aug 18, 2012·Journal of Industrial Microbiology & Biotechnology·Hua LiuJian Chen
Feb 7, 2015·Bioprocess and Biosystems Engineering·Vanderson Carvalho FenelonGraciette Matioli

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Methods Mentioned

BETA
transglycosylation
PCR
electrophoresis
Assay

Software Mentioned

CGT
ViennaRNA
Opt

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