Characterization of a thermostable cyclodextrin glucanotransferase from Pyrococcus furiosus DSM3638

A gene that encodes the enzyme Pyrococcus furiosus cyclodextrin glucanotransferase (PFCGT) was cloned in Escherichia coli. PFCGT was highly expressed in recombinant E. coli after compensation for codon usage bias using the pRARE plasmid. Purified PFCGT was extremely thermostable with an optimal temp...

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Published inExtremophiles : life under extreme conditions Vol. 11; no. 3; pp. 537 - 541
Main Authors Lee, Myoung-Hee, Yang, Sung-Jae, Kim, Jung-Woo, Lee, Hee-Seob, Kim, Jung-Wan, Park, Kwan-Hwa
Format Journal Article
LanguageEnglish
Published Heidelberg Tokyo : Springer-Verlag 01.05.2007
Tokyo Springer
Springer Nature B.V
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Summary:A gene that encodes the enzyme Pyrococcus furiosus cyclodextrin glucanotransferase (PFCGT) was cloned in Escherichia coli. PFCGT was highly expressed in recombinant E. coli after compensation for codon usage bias using the pRARE plasmid. Purified PFCGT was extremely thermostable with an optimal temperature and pH of 95°C and 5.0, respectively, retaining 97% of its activity at 100°C. Incubation at 60°C for 20 min during the purification process led to a 1.5-fold increase in enzymatic activity. A time course assay of the PFCGT reaction with starch indicated that cyclic α-1,4-glucans with DPs greater than 20 were produced at the beginning of the incubation followed by an increase in β-CD. The major final product of PFCGT cyclization was β-CD, and thus the enzyme is a β-CGTase.
Bibliography:http://dx.doi.org/10.1007/s00792-007-0061-6
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ISSN:1431-0651
1433-4909
DOI:10.1007/s00792-007-0061-6