PMID: 9435077Jan 22, 1998Paper

Purification and characterization of a novel thermostable alpha-L-arabinofuranosidase from a color-variant strain of Aureobasidium pullulans

Applied and Environmental Microbiology
B C Saha, R J Bothast

Abstract

A color-variant strain of Aureobasidium pullulans (NRRL Y-12974) produced alpha-L-arabinofuranosidase (alpha-L-AFase) when grown in liquid culture on oat spelt xylan. An extracellular alpha-L-AFase was purified 215-fold to homogeneity from the culture supernatant by ammonium sulfate treatment, DEAE Bio-Gel A agarose column chromatography, gel filtration on a Bio-Gel A-0.5m column, arabinan-Sepharose 6B affinity chromatography, and SP-Sephadex C-50 column chromatography. The purified enzyme had a native molecular weight of 210,000 and was composed of two equal subunits. It had a half-life of 8 h at 75 degrees C, displayed optimal activity at 75 degrees C and pH 4.0 to 4.5, and had a specific activity of 21.48 mumol min-1. mg-1 of protein against p-nitrophenyl-alpha-L-arabinofuranoside (pNP alpha AF). The purified alpha-L-AFase readily hydrolyzed arabinan and debranched arabinan and released arabinose from arabinoxylans but was inactive against arabinogalactan. The K(m) values of the enzyme for the hydrolysis of pNP alpha AF, arabinan, and debranched arabinan at 75 degrees C and pH 4.5 were 0.26 mM, 2.14 mg/ml, and 3.25 mg/ml, respectively. The alpha-L-AFase activity was not inhibited at all by L-arabinose (1.2 M). The enzyme did...Continue Reading

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Citations

Sep 6, 2003·Applied and Environmental Microbiology·Abelardo Margolles, Clara G de los Reyes-Gavilán
Oct 26, 2005·Biotechnology Letters·H P Makkonen, J P Nakas
May 17, 2011·Applied Biochemistry and Biotechnology·Maíra Nicolau de AlmeidaSebastião Tavares de Rezende
Aug 3, 2000·Bioscience, Biotechnology, and Biochemistry·T Yanai, M Sato
Oct 24, 2007·Applied Microbiology and Biotechnology·Barend J M de WetBernard A Prior
Dec 31, 2005·Journal of Industrial Microbiology & Biotechnology·Mondher Th Numan, Narayan B Bhosle
Jul 29, 2011·Protein Science : a Publication of the Protein Society·Tatiana A C B SouzaMario T Murakami
Sep 6, 2003·Applied and Environmental Microbiology·Karl RumboldBernard A Prior
Sep 29, 2001·Canadian Journal of Microbiology·A K Shofiqur RahmanK Takamizawa

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