The Mechanism of Binding of Glucoamylase I from Aspergillus awamori var. kawachi to Cyclodextrins and Raw Starch
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概要
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Glucoamylase I (GAI) bound to β-cyclodextrin (β-CD) with the binding constants of K_d = 17.7μM and n = 1.69. Binding resulted in formation of an inclusion complex, as indicated by the fact that organic guest compounds for β-CD inhibited the binding of GAI to β-CD. Titration of GAI (A^1-R^<615>) with β-CD or Amylose A showed critical spectral perturbations at 286 nm that reflected the aromatic side chains of Tyr and Trp, but no spectral changes were observed in GAI' (A^1-V^<469>) and Gp-I (A^<470>-V^<514>). Titration of GAI with derivatives of β-CD, namely, 6-hydroxypropyl-β-CD (H-CD), 2,6-O-dimethyl-β-CD, and 2,3,6-O-trimethyl-β-CD, yielded spectral changes only in the case of H-CD. Therefore, the Cp region (A^<515>-R^<615>) of GAI seems stereospecifically able to recognize the structure of the secondary OH-side but not the primary OH-side of β-CD. Chemical modifications specific for aromatic amino acids of the GAI-CD inclusion complexes were done with N-bromosuccinimide and tetranitromethane. In the presence of β-CD, one Trp residue was protected from oxidation but Tyr residues were not. The analysis of a Cp region/β-galactosidase fusion protein indicated that Trp^<562> contributed to formation of inclusion complexes.
- 社団法人日本農芸化学会の論文
- 1994-01-23
著者
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Goto M
Univ. Tokyo Tokyo Jpn
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Hayashida Shinsaku
Department Of Agricultural Chemistry Kyushu University:corresponding Author
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Hayashida Shinsaku
Laboratory Of Applied Microbiology Department Of Agricultural Chemistry Kyushu University
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Goto Masatoshi
Laboratory of Applied Microbiology, Department of Agricultural Chemistry, Kyushu University
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Kanlayakrit Werasit
Laboratory of Applied Microbiology, Department of Agricultural Chemistry, Kyushu University
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Tanigawa Kei
Laboratory of Applied Microbiology, Department of Agricultural Chemistry, Kyushu University
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Tanigawa Kei
Laboratory Of Applied Microbiology Department Of Agricultural Chemistry Kyushu University
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Kanlayakrit W
Kyushu Univ. Fukuoka Jpn
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Kanlayakrit Werasit
Laboratory Of Applied Microbiology Department Of Agricultural Chemistry Kyushu University
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Goto Masatoshi
Department Of Agricultural Chemistry Kyushu University
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Goto Masatoshi
Laboratory Of Applied Microbiology Department Of Agricultural Chemistry Kyushu University
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Hayashida S
Laboratory Of Applied Microbiology Department Of Agricultural Chemistry Kyushu University
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