Random mutagenesis of pullulanase from Klebsiella aerogenes for studies of the structure and function of the enzyme.

Random mutagenesis of pullulanase from Klebsiella aerogenes for studies of the structure and function of the enzyme. Research Abstract Details 

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Random mutagenesis of pullulanase from Klebsiella aerogenes for studies of the structure and function of the enzyme. Abstract Text:

M Yamashita,T Kinoshita,M Ihara,T Mikawa,Y Murooka,

To study the structure and function of pullulanase from Klebsiella aerogenes, a method involving random mutagenesis of the entire gene for pullulanase was used. Out of 50,000 clones screened at high temperature, seven genes for mutant proteins were identified by DNA sequencing. The amino acid substitutions in the seven mutant proteins were clustered on the NH2-terminal side of the four conserved regions found in alpha-amylases. These mutant pullulanases were classified into two types: those whose catalytic activity was altered and those whose thermal stability was increased. The results presented here and in previous reports suggest that pullulanase from K. aerogenes has similar active sites to those of alpha-amylases with the four conserved regions, as well as another substrate-binding site closer to the NH2-terminus. The plate assay method used for isolation of thermostable variants may be applicable to the generation of useful variants of other enzymes.

Random mutagenesis of pullulanase from Klebsiella aerogenes for studies of the structure and function of the enzyme. Publishing Authors By Initials

M Yamashita,T Kinoshita,M Ihara,T Mikawa,Y Murooka,

For similar investigative techniques: genetic techniques: gene transfer techniques: transfection: transformation, bacterial research abstracts see: investigative techniques: genetic techniques: gene transfer techniques: transfection: transformation, bacterial research

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Random mutagenesis of pullulanase from Klebsiella aerogenes for studies of the structure and function of the enzyme. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biochemistry

VOLUME: 116

Page Numbers: 1233-40

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Dec

YEAR: 1994

Random mutagenesis of pullulanase from Klebsiella aerogenes for studies of the structure and function of the enzyme. Information

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LANGUAGE: eng

NlmUniqueID: 376600

Random mutagenesis of pullulanase from Klebsiella aerogenes for studies of the structure and function of the enzyme. Keywords Mesh Terms:

KEYWORDS: Transformation, Bacterial

MESH TERMS: genetics

Chemical & Substance for Abstract: Random mutagenesis of pullulanase from Klebsiella aerogenes for studies of the structure and function of the enzyme. Information

Substance Name: pullulanase

Registry Number: EC 3.2.1.41

Grant and Affiliation Information for Random mutagenesis of pullulanase from Klebsiella aerogenes for studies of the structure and function of the enzyme.

AFFILIATION: Department of Fermentation Technology, Faculty of Engineering, Hiroshima University.

Country: JAPAN

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MEDLINETA: J Biochem

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