Purification and Characterization of a CM-Glycinin Digesting Protease from Soybean Seeds
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概要
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A glycinin-digesting protease was purified from 4-h-imbibed soybean seeds to apparent homogeneity by a combination of DEAE-cellulose, gel filtration and Shodex IEC QA-824 chromatography. The purified soybean protease showed a single band on SDS-PAGE and degraded the carboxymethylated glycinin molecule at neutral or alkaline pH. The glycinin-digesting protease has an apparent molecular mass of 98 kilodaltons as estimated by SDS-PAGE under both reduced and non-reduced conditions. A peptidic substrate for the soybean protease was detected and purified from a tryptic digest of glycinin A_5A_4B_3 complex ; it corresponded to Ala(375)-Lys(381) of the B_3 subunit. It was found that the protease could hydrolyze the peptide bond between Tyr(378) and Asn(389) of that peptide. The soybean protease was significantly inhibited by PMSF, indicating the endopeptidase is a serine protease. Moreover, the glycinin-digesting activity is also susceptible to EDTA and the inactivated enzyme could be restored by the addition of MnCl_2, suggesting that Mn^<2+> is an important factor for the endopeptidase activity. To our knowledge, this is the first report of purification and characterization of a glycinin-digesting protease from soybean seeds.
- 社団法人日本農芸化学会の論文
- 1993-07-23
著者
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KIMURA Yoshinobu
Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University
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Kimura Yoshinobu
Department Of Agricultural Science Faculty Of Agriculture Okayama University
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TAKAGI Shigeaki
Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University
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Takagi S
Okayama Univ. Okayama Jpn
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Takagi Shigeaki
Department Of Bioresources Chemistry Faculty Of Agriculture Okayama University
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Akhtaruzzaman Mohammad
Department of Agricultural Science, Faculty of Agriculture, Okayama University
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Akhtaruzzaman Mohammad
Department Of Agricultural Science Faculty Of Agriculture Okayama University
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Takagi Shigeaki
Department of Agricultural Chemistry, Faculty of Agriculture Okayama University
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