Biochemical and Crystallographic Characterization of the Starch Branching Enzyme I (BEI) from Oryza sativa L

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概要

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• Starch branching enzyme (SBE) catalyzes the cleavage of α-1.4-linkages and the subsequent transfer of α-1.4 glucan to form an α-1.6 branch point in amylopectin. We overproduced rice branching enzyme I (BEI) in Escherichia coli cells, and the resulting enzyme (rBEI) was characterized with respect to biochemical and crystallographic properties. Specific activities were calculated to be 20.8 units/mg and 2.5 units/mg respectively when amylose and amylopectin were used as substrates. Site-directed mutations of Tyr235, Asp270, His275, Arg342, Asp344, Glu399, and His467 conserved in the α-amylase family enzymes drastically reduced catalytic activity of rBEI. This result suggests that the structures of BEI and the other α-amylase family enzymes are similar and that they share common catalytic mechanisms. Crystals of rBEI were grown under appropriate conditions and the crystals diffracted to a resolution of 3.0 Å on a synchrotron X-ray source.

• 社団法人 日本農芸化学会の論文

著者

• Omori Toshiro

  Frontier Res. Center Sanwa Shurui Co. Ltd.

• SATOH Hikaru

  Institute of Genetic Resources, Faculty of Agriculture, Kyushu University

• KIMURA Makoto

  Laboratory of Structural Biology, Graduate School of Systems Life Sciences, Kyushu University

• Nakashima Takashi

  Laboratory of Structural Biology, Graduate School of Systems Life Sciences, Kyushu University

• SHIMADA Hiroaki

  Laboratory of Structural Biology, Graduate School of Systems Life Sciences, Kyushu University

• KAKUTA Yoshimitsu

  Laboratory of Structural Biology, Graduate School of Systems Life Sciences, Kyushu University

• Vu Nhuan

  Laboratory of Biochemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Grad

• NISHI Aiko

  Institute of Genetic Resources, Faculty of Agriculture, Kyushu University

• IDA Hiroko

  Frontier Research Center, Sanwa Shurui Co., Ltd.

• Shimada Hiroaki

  Laboratory Of Structural Biology Graduate School Of Systems Life Sciences Kyushu University

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