Purification and characterization of a CM-glycinin digesting protease from soybean seeds

Mohammad Akhtaruzzaman, Yoshinobu Kimura, Shigeaki Takagi

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A glycinin-digesting protease was purified from 4-h-imbibed soybean seeds to apparent homogeneity by a combination of OEAE-cellulose, gel filtration and Shodex IEC QA-824 chromatography. The purified soybean protease showed a single band on SOS–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 SOS–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 A5A4B3 complex; it corresponded to Ala(375)–Lys(381) of the B3 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 ESTA and the inactivated enzyme could be restored by the addition of MnCl2, suggesting that Mn2+ 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.

Original languageEnglish
Pages (from-to)1119-1124
Number of pages6
JournalBioscience, Biotechnology and Biochemistry
Volume57
Issue number7
DOIs
Publication statusPublished - 1993

Fingerprint

glycinin
Soybeans
Purification
Seed
Seeds
Peptide Hydrolases
proteinases
soybeans
seeds
Endopeptidases
Peptides
peptides
Molecular mass
Serine Proteases
Chromatography
Cellulose
Gel Chromatography
serine proteinases
Gels
chromatography

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Biochemistry
  • Organic Chemistry
  • Analytical Chemistry
  • Applied Microbiology and Biotechnology
  • Food Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry (miscellaneous)

Cite this

Purification and characterization of a CM-glycinin digesting protease from soybean seeds. / Akhtaruzzaman, Mohammad; Kimura, Yoshinobu; Takagi, Shigeaki.

In: Bioscience, Biotechnology and Biochemistry, Vol. 57, No. 7, 1993, p. 1119-1124.

Research output: Contribution to journalArticle

@article{041eeb40a6a6406c823e2763c6633b95,
title = "Purification and characterization of a CM-glycinin digesting protease from soybean seeds",
abstract = "A glycinin-digesting protease was purified from 4-h-imbibed soybean seeds to apparent homogeneity by a combination of OEAE-cellulose, gel filtration and Shodex IEC QA-824 chromatography. The purified soybean protease showed a single band on SOS–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 SOS–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 A5A4B3 complex; it corresponded to Ala(375)–Lys(381) of the B3 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 ESTA and the inactivated enzyme could be restored by the addition of MnCl2, suggesting that Mn2+ 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.",
author = "Mohammad Akhtaruzzaman and Yoshinobu Kimura and Shigeaki Takagi",
year = "1993",
doi = "10.1271/bbb.57.1119",
language = "English",
volume = "57",
pages = "1119--1124",
journal = "Bioscience, Biotechnology and Biochemistry",
issn = "0916-8451",
publisher = "Japan Society for Bioscience Biotechnology and Agrochemistry",
number = "7",

}

TY - JOUR

T1 - Purification and characterization of a CM-glycinin digesting protease from soybean seeds

AU - Akhtaruzzaman, Mohammad

AU - Kimura, Yoshinobu

AU - Takagi, Shigeaki

PY - 1993

Y1 - 1993

N2 - A glycinin-digesting protease was purified from 4-h-imbibed soybean seeds to apparent homogeneity by a combination of OEAE-cellulose, gel filtration and Shodex IEC QA-824 chromatography. The purified soybean protease showed a single band on SOS–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 SOS–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 A5A4B3 complex; it corresponded to Ala(375)–Lys(381) of the B3 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 ESTA and the inactivated enzyme could be restored by the addition of MnCl2, suggesting that Mn2+ 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.

AB - A glycinin-digesting protease was purified from 4-h-imbibed soybean seeds to apparent homogeneity by a combination of OEAE-cellulose, gel filtration and Shodex IEC QA-824 chromatography. The purified soybean protease showed a single band on SOS–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 SOS–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 A5A4B3 complex; it corresponded to Ala(375)–Lys(381) of the B3 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 ESTA and the inactivated enzyme could be restored by the addition of MnCl2, suggesting that Mn2+ 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.

UR - http://www.scopus.com/inward/record.url?scp=0027638537&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027638537&partnerID=8YFLogxK

U2 - 10.1271/bbb.57.1119

DO - 10.1271/bbb.57.1119

M3 - Article

C2 - 7763983

AN - SCOPUS:0027638537

VL - 57

SP - 1119

EP - 1124

JO - Bioscience, Biotechnology and Biochemistry

JF - Bioscience, Biotechnology and Biochemistry

SN - 0916-8451

IS - 7

ER -