Purification and characterization of exo-α-d-mannosidase from a Cellulomonas sp.

Kaoru Takegawa, Satoshi Miki, Takayuki Jikibara, Shojiro Iwahara

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

A Gram-positive bacterium isolated from a soil sample was found to produce a high level of α-mannosidase in culture media. The organism was identified as Cellulomonas sp. from various bacteriological characteristics. The production of the enzyme was strongly induced by yeast extract. To purify the enzyme, the bacterium was first grown in a glucose medium, and then the cells were transferred to an inducing medium containing only yeast extract. The enzyme had two different molecular weight forms. The high-molecular-weight form was purified from the inducing medium by column chromatography. The enzyme was purified to homogeneity by ultracentrifugal analysis. The enzyme was strongly inactivated by Hg2+, Cu2+, Zn2+, EDTA and p-chloromercuribenzoic acid. The molecular weight of the enzyme was estimated to be about 450 000 by gel filtration. The purified enzyme could liberate only mannose from native yeast mannan and the optimum pH was 6.5-8.0. The enzyme rapidly cleaved α-1,2- and α-1,6-linked mannose chains, but the hydrolysis rate for α-1,3 linkage was very low. In addition, the purified enzyme showed only slight activity towards p-nitrophenyl-α-d-mannoside and did not hydrolyze O-α-d-mannopyranosyl(1 → 2)-d-mannitol.

Original languageEnglish
Pages (from-to)431-437
Number of pages7
JournalBBA - General Subjects
Volume991
Issue number3
DOIs
Publication statusPublished - Jun 27 1989

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Cellulomonas
Mannosidases
Purification
Enzymes
Yeast
Yeasts
Molecular Weight
Molecular weight
Mannose
Bacteria
p-Chloromercuribenzoic Acid
Mannosides
Mannans
Column chromatography
Gram-Positive Bacteria
Mannitol
Edetic Acid
Gel Chromatography
Culture Media
Chromatography

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Purification and characterization of exo-α-d-mannosidase from a Cellulomonas sp. / Takegawa, Kaoru; Miki, Satoshi; Jikibara, Takayuki; Iwahara, Shojiro.

In: BBA - General Subjects, Vol. 991, No. 3, 27.06.1989, p. 431-437.

Research output: Contribution to journalArticle

Takegawa, Kaoru ; Miki, Satoshi ; Jikibara, Takayuki ; Iwahara, Shojiro. / Purification and characterization of exo-α-d-mannosidase from a Cellulomonas sp. In: BBA - General Subjects. 1989 ; Vol. 991, No. 3. pp. 431-437.
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