Reinvestigation of N-Bromosuccinimide Oxidation of Glucoamylases from Aspergillus saitoi
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概要
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The reaction of Gluc M_2,the minor glucoamylase [EC 3.2.1.3,α-D-(1→4)glucan glucohydrolase] from Aspergillus saitoi, with N-bromosuccinimide (NBS) was reinvestigated in order to elucidate the structure-activity relationship. Some data were also obtained for Gluc M_1. The tryptophan residues in Gluc M_2 were oxidized at various NBS/Gluc M_2 rations. The enzymatic activity decreased to about 75% of that of native Gluc M_2 with the oxidation of the first 6 tryptophan residues at an NBS/Gluc M_2 ratio of 10. In the presence of 10% soluble starch, about 4.5 tryptophan residues in Gluc M_2 were oxidized at an NBS/Gluc M_2 ratio of 10. The remaining activity of Gluc M_2 at this stage of oxidation was about 7.5%. On further oxidation after removal of soluble starch, the enzymatic activity decreased to zero with the concomitant oxidation of 3 tryptophan residues. The results indicated that the essential tryptophan residue(s) is amongst these 3 tryptophan residues.The ultraviolet difference spectrum induced by addition of maltose and maltitol to Gluc M_2 showed four troughs at 280,288,295 and 310-313nm, The latter 3 troughs were probably due to tryptophan residues of Gluc M_2,and the magnitude of the troughs decreased with the progress of oxidation, but to a far lesser extent than teh decrease in activity, These results indicate that there are two types of tryptophan residues is in Gluc M_2. One of them is located near subsites 1 and 2 and is related to catalysis, while the other is located at subsites 2 and 3,and contributes to the maltose binding.
- 社団法人日本薬学会の論文
- 1986-07-25
著者
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入江 昌親
Department of Microbiology, Hoshi College of Pharmacy
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井口 法男
Department of Microbiology, Hoshi College of Pharmacy
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小山 隆
Department of Pharmacy, College of Science and Technology, Nihon University
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島田 弘子
Department of Pharmacy, College of Science and Technology, Nihon University
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入江 昌親
星薬大・微生物
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入江 昌親
Department Of Chemistry Institute For Infectious Diseases University Of Tokyo
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島田 弘子
Department Of Microbiology College Of Pharamacy
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井口 法男
日大・薬・微生物
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小山 隆
日大・薬・微生物
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Koyama T
Shinshu Univ. Nagano Jpn
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