Biochemical and crystallographic characterization of the starch branching enzyme I (BEI) from Oryza sativa L

Nhuan Thi Vu, Hiroaki Shimada, Yoshimitsu Kakuta, Takashi Nakashima, Hiroko Ida, Toshiro Omori, Aiko Nishi, Hikaru Satoh, Makoto Kimura

研究成果: ジャーナルへの寄稿記事

16 引用 (Scopus)

抄録

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.

元の言語英語
ページ(範囲)2858-2866
ページ数9
ジャーナルBioscience, Biotechnology and Biochemistry
72
発行部数11
DOI
出版物ステータス出版済み - 12 1 2008

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1,4-alpha-Glucan Branching Enzyme
Amylopectin
Amylases
Enzymes
Amylose
Crystals
Synchrotrons
Glucans
Escherichia coli
Catalyst activity
X-Rays
X rays
Mutation
Substrates
Oryza
N-acetylglucosaminyltransferase IGnT

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Applied Microbiology and Biotechnology
  • Analytical Chemistry
  • Organic Chemistry

これを引用

Biochemical and crystallographic characterization of the starch branching enzyme I (BEI) from Oryza sativa L. / Vu, Nhuan Thi; Shimada, Hiroaki; Kakuta, Yoshimitsu; Nakashima, Takashi; Ida, Hiroko; Omori, Toshiro; Nishi, Aiko; Satoh, Hikaru; Kimura, Makoto.

:: Bioscience, Biotechnology and Biochemistry, 巻 72, 番号 11, 01.12.2008, p. 2858-2866.

研究成果: ジャーナルへの寄稿記事

Vu, Nhuan Thi ; Shimada, Hiroaki ; Kakuta, Yoshimitsu ; Nakashima, Takashi ; Ida, Hiroko ; Omori, Toshiro ; Nishi, Aiko ; Satoh, Hikaru ; Kimura, Makoto. / Biochemical and crystallographic characterization of the starch branching enzyme I (BEI) from Oryza sativa L. :: Bioscience, Biotechnology and Biochemistry. 2008 ; 巻 72, 番号 11. pp. 2858-2866.
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abstract = "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 {\AA} on a synchrotron X-ray source.",
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AU - Vu, Nhuan Thi

AU - Shimada, Hiroaki

AU - Kakuta, Yoshimitsu

AU - Nakashima, Takashi

AU - Ida, Hiroko

AU - Omori, Toshiro

AU - Nishi, Aiko

AU - Satoh, Hikaru

AU - Kimura, Makoto

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N2 - 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.

AB - 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.

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