Alpha-L-arabinofuranosidase from Ruminococcus Albus 8: Purification and Possible Role in Hydrolysis of Alfalfa Cell Wall

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 1984 May 1

PMID 6742828

Citations 34

Authors

L C Greve

J M Labavitch

R E HUNGATE

Affiliations

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Abstract

An alpha-L- arabinofuranosidase has been purified from the extracellular broth of cultures of Ruminococcus albus 8. The purification procedure utilized gel filtration, (NH4)2SO4 precipitation, and isoelectric focusing. The purified enzyme appeared to be homogeneous when chromatographed on disc and analytical isoelectric focusing gels. The estimated molecular weight of the native protein was 305,000 to 310,000. Sodium dodecyl sulfate-gel electrophoresis analysis suggested that the native protein is a tetramer composed of 75,000-molecular-weight subunits. The enzyme appeared to have no metal cofactor requirement but was sensitive to several sulfhydryl reagents. The pH optimum with p-nitrophenyl-alpha-L-arabinofuranoside as the substrate was 6.9 and the Km was 1.3 mM. Several lines of evidence indicated that the enzyme is a glycoprotein. When assayed against alfalfa cell wall material, the enzyme hydrolyzed only small amounts of arabinose from the substrate. When assayed together with hemicellulolytic or pectinolytic enzymes against the same substrate, the arabinosidase significantly enhanced the hydrolytic action of the glycanases .

Citing Articles

Molecular modeling and MM-PBSA free energy analysis of endo-1,4-β-xylanase from Ruminococcus albus 8.

Zhan D, Yu L, Jin H, Guan S, Han W Int J Mol Sci. 2014; 15(10):17284-303.

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Exocellular β-mannanases from hemicellulolytic fungi.

Johnson K World J Microbiol Biotechnol. 2014; 6(2):209-17.

PMID: 24429995 DOI: 10.1007/BF01200943.

Xylan-hydrolysing enzymes from thermophilic and mesophilic fungi.

Smith D, Bhat K, Wood T World J Microbiol Biotechnol. 2014; 7(4):475-84.

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Biochemical analyses of multiple endoxylanases from the rumen bacterium Ruminococcus albus 8 and their synergistic activities with accessory hemicellulose-degrading enzymes.

Moon Y, Iakiviak M, Bauer S, Mackie R, Cann I Appl Environ Microbiol. 2011; 77(15):5157-69.

PMID: 21666020 PMC: 3147433. DOI: 10.1128/AEM.00353-11.

Cell surface enzyme attachment is mediated by family 37 carbohydrate-binding modules, unique to Ruminococcus albus.

Ezer A, Matalon E, Jindou S, Borovok I, Atamna N, Yu Z J Bacteriol. 2008; 190(24):8220-2.

PMID: 18931104 PMC: 2593223. DOI: 10.1128/JB.00609-08.

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