Hydrolysis of a-d-galactosyl oligosaccharides in soymilk by a-d-galactosidase of bifidobacterium breve 203

Kenji Sakai, Takashi Tachiki, Hidehiko Kumagai, Tatsurokuro Tochkura

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Bifidobacterium breve 203 grew well on soymilk, reaching a cell number of 2x 109 cells/ml on 24 hr cultivation, at which time the medium had solidified and its pH had decreased to 4.0. Stachyose and raffinose in the medium were assimilated in preference to sucrose. A crude extract of B. breve 203 hydrolyzed the oligosaccharides in soymilk almost completely into their constituent monosaccharides. The a-D-galactosidase responsible for the first reaction in the degradation of the a-D-galactosyl oligosaccharides was isolated from a crude extract of the organism (500-fold purification, 2% yield) by means of ammonium sulfate-fractionation and column chromatographies on DEAE-cellulose, DEAE-cellulofine, Sepharose 6B, hydroxylapatite, Sephacryl S-300 and Phenyl-Sepharose 6B. The enzyme was a homo-octamer with a molecular weight of about 330,000, and its isoelectric point was 3.7. It reacted optimally at pH 5.5 and hydrolyzed stachyose (Km 8.6 ium), raffinose (Km 4.4mM) and melibiose (Km 3.0 mM), with 50 -60% of its reactivity towardp-nitrophenyl a-D-galactoside (Km 0.16 him). The soymilk and guar gum was also investigated. reaction of the purified a-D-galactosidase.

Original languageEnglish
Pages (from-to)315-322
Number of pages8
JournalAgricultural and Biological Chemistry
Volume51
Issue number2
DOIs
Publication statusPublished - Feb 1987
Externally publishedYes

Fingerprint

Galactosidases
Bifidobacterium breve
Raffinose
galactosidases
guar gum
soymilk
Complex Mixtures
Oligosaccharides
oligosaccharides
Hydrolysis
stachyose
hydrolysis
raffinose
Melibiose
Galactosides
DEAE-Cellulose Chromatography
DEAE-Cellulose
agarose
Column chromatography
Monosaccharides

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Hydrolysis of a-d-galactosyl oligosaccharides in soymilk by a-d-galactosidase of bifidobacterium breve 203. / Sakai, Kenji; Tachiki, Takashi; Kumagai, Hidehiko; Tochkura, Tatsurokuro.

In: Agricultural and Biological Chemistry, Vol. 51, No. 2, 02.1987, p. 315-322.

Research output: Contribution to journalArticle

Sakai, Kenji ; Tachiki, Takashi ; Kumagai, Hidehiko ; Tochkura, Tatsurokuro. / Hydrolysis of a-d-galactosyl oligosaccharides in soymilk by a-d-galactosidase of bifidobacterium breve 203. In: Agricultural and Biological Chemistry. 1987 ; Vol. 51, No. 2. pp. 315-322.
@article{964f4c7e74b946b18188798181a76c9a,
title = "Hydrolysis of a-d-galactosyl oligosaccharides in soymilk by a-d-galactosidase of bifidobacterium breve 203",
abstract = "Bifidobacterium breve 203 grew well on soymilk, reaching a cell number of 2x 109 cells/ml on 24 hr cultivation, at which time the medium had solidified and its pH had decreased to 4.0. Stachyose and raffinose in the medium were assimilated in preference to sucrose. A crude extract of B. breve 203 hydrolyzed the oligosaccharides in soymilk almost completely into their constituent monosaccharides. The a-D-galactosidase responsible for the first reaction in the degradation of the a-D-galactosyl oligosaccharides was isolated from a crude extract of the organism (500-fold purification, 2{\%} yield) by means of ammonium sulfate-fractionation and column chromatographies on DEAE-cellulose, DEAE-cellulofine, Sepharose 6B, hydroxylapatite, Sephacryl S-300 and Phenyl-Sepharose 6B. The enzyme was a homo-octamer with a molecular weight of about 330,000, and its isoelectric point was 3.7. It reacted optimally at pH 5.5 and hydrolyzed stachyose (Km 8.6 ium), raffinose (Km 4.4mM) and melibiose (Km 3.0 mM), with 50 -60{\%} of its reactivity towardp-nitrophenyl a-D-galactoside (Km 0.16 him). The soymilk and guar gum was also investigated. reaction of the purified a-D-galactosidase.",
author = "Kenji Sakai and Takashi Tachiki and Hidehiko Kumagai and Tatsurokuro Tochkura",
year = "1987",
month = "2",
doi = "10.1080/00021369.1987.10868063",
language = "English",
volume = "51",
pages = "315--322",
journal = "Bioscience, Biotechnology and Biochemistry",
issn = "0916-8451",
publisher = "Japan Society for Bioscience Biotechnology and Agrochemistry",
number = "2",

}

TY - JOUR

T1 - Hydrolysis of a-d-galactosyl oligosaccharides in soymilk by a-d-galactosidase of bifidobacterium breve 203

AU - Sakai, Kenji

AU - Tachiki, Takashi

AU - Kumagai, Hidehiko

AU - Tochkura, Tatsurokuro

PY - 1987/2

Y1 - 1987/2

N2 - Bifidobacterium breve 203 grew well on soymilk, reaching a cell number of 2x 109 cells/ml on 24 hr cultivation, at which time the medium had solidified and its pH had decreased to 4.0. Stachyose and raffinose in the medium were assimilated in preference to sucrose. A crude extract of B. breve 203 hydrolyzed the oligosaccharides in soymilk almost completely into their constituent monosaccharides. The a-D-galactosidase responsible for the first reaction in the degradation of the a-D-galactosyl oligosaccharides was isolated from a crude extract of the organism (500-fold purification, 2% yield) by means of ammonium sulfate-fractionation and column chromatographies on DEAE-cellulose, DEAE-cellulofine, Sepharose 6B, hydroxylapatite, Sephacryl S-300 and Phenyl-Sepharose 6B. The enzyme was a homo-octamer with a molecular weight of about 330,000, and its isoelectric point was 3.7. It reacted optimally at pH 5.5 and hydrolyzed stachyose (Km 8.6 ium), raffinose (Km 4.4mM) and melibiose (Km 3.0 mM), with 50 -60% of its reactivity towardp-nitrophenyl a-D-galactoside (Km 0.16 him). The soymilk and guar gum was also investigated. reaction of the purified a-D-galactosidase.

AB - Bifidobacterium breve 203 grew well on soymilk, reaching a cell number of 2x 109 cells/ml on 24 hr cultivation, at which time the medium had solidified and its pH had decreased to 4.0. Stachyose and raffinose in the medium were assimilated in preference to sucrose. A crude extract of B. breve 203 hydrolyzed the oligosaccharides in soymilk almost completely into their constituent monosaccharides. The a-D-galactosidase responsible for the first reaction in the degradation of the a-D-galactosyl oligosaccharides was isolated from a crude extract of the organism (500-fold purification, 2% yield) by means of ammonium sulfate-fractionation and column chromatographies on DEAE-cellulose, DEAE-cellulofine, Sepharose 6B, hydroxylapatite, Sephacryl S-300 and Phenyl-Sepharose 6B. The enzyme was a homo-octamer with a molecular weight of about 330,000, and its isoelectric point was 3.7. It reacted optimally at pH 5.5 and hydrolyzed stachyose (Km 8.6 ium), raffinose (Km 4.4mM) and melibiose (Km 3.0 mM), with 50 -60% of its reactivity towardp-nitrophenyl a-D-galactoside (Km 0.16 him). The soymilk and guar gum was also investigated. reaction of the purified a-D-galactosidase.

UR - http://www.scopus.com/inward/record.url?scp=0001604114&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0001604114&partnerID=8YFLogxK

U2 - 10.1080/00021369.1987.10868063

DO - 10.1080/00021369.1987.10868063

M3 - Article

AN - SCOPUS:0001604114

VL - 51

SP - 315

EP - 322

JO - Bioscience, Biotechnology and Biochemistry

JF - Bioscience, Biotechnology and Biochemistry

SN - 0916-8451

IS - 2

ER -