Substrate specificity of endo-β-galactosidases from Flavobacterium keratolyticus and Escherichia freundii is different from that of Pseudomonas sp

Makoto Ito, Yoshio Hirabayashi, Tatsuya Yamagata

Research output: Contribution to journalArticle

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Abstract

The substrate specificity of endo-β-galactosidase of Pseudomonas sp. was found to differ from that of Flavobacterium keratolyticus or Escherichia freundii, based on the following experimental results. (a) The endo-β-galactosidases from these three bacteria released 6-O-sulfo-GlcNAcβ1-3Gal as one of the major products from keratan sulfates from different sources. In addition to the sulfated disaccharide, Flavobacterium and Escherichia enzymes produced GlcNAcβ1-3Gal, which is also an integral repeating unit of keratan sulfate, whereas the Pseudomonas enzyme did not release any non-sulfated disaccharide. (b) Tetrasaccharides were prepared from the teleost skin keratan sulfate by digestion with Pseudomonas enzyme followed by gel filtration on Sephadex G-50 chromatography. A part of the tetrasaccharide fraction was hydrolyzed by Flavobacterium enzyme to produce 6-O-sulfo-GlcNAcβ1- 3Gal and GlcNAcβ1-3Gal, whereas the fraction was completely resistant to retreatment with the Pseudomonas enzyme. (c) Endo-β-galactosidases from F. keratolyticus and E. freundii hydrolyzed the internal β-1,4-galactosyl linkage of various neolacto-type glycosphingolipids to produce glucosylceramides. However, these glycosphingolipids were completely resistant to the Pseudomonos enzyme. These findings clearly show that the sulfation on the N-acetylglucosamine adjacent to galactose in the lactosaminoglycans is essential for expression of the Pseudomonas enzyme, but not for that of the Flavobacterium or Escherichia enzyme.

Original languageEnglish
Pages (from-to)773-780
Number of pages8
JournalJournal of Biochemistry
Volume100
Issue number3
DOIs
Publication statusPublished - Jan 1 1986
Externally publishedYes

Fingerprint

Galactosidases
Flavobacterium
Escherichia
Substrate Specificity
Pseudomonas
Substrates
Enzymes
Keratan Sulfate
Glycosphingolipids
Disaccharides
Glucosylceramides
Retreatment
Acetylglucosamine
Chromatography
Galactose
Gel Chromatography
Digestion
Bacteria
Skin
Gels

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Substrate specificity of endo-β-galactosidases from Flavobacterium keratolyticus and Escherichia freundii is different from that of Pseudomonas sp. / Ito, Makoto; Hirabayashi, Yoshio; Yamagata, Tatsuya.

In: Journal of Biochemistry, Vol. 100, No. 3, 01.01.1986, p. 773-780.

Research output: Contribution to journalArticle

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