Isolation and Properties of Extracellular β-Xylosidases from Fungi Aspergillus japonicus and Trichoderma reesei

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M. V. Semenova^1*, M. I. Drachevskaya², O. A. Sinitsyna², A. V. Gusakov², and A. P. Sinitsyn^1,2

¹Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, 119071 Moscow, Russia; E-mail: margs@mail.ru

²Chemical Faculty, Lomonosov Moscow State University, 119899 Moscow, Russia; fax: (495) 939-0997

^* To whom correspondence should be addressed.

Received April 2, 2009; Revision received April 14, 2009

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Homogeneous β-xylosidases with molecular mass values 120 and 80 kDa (as shown by SDS-PAGE), belonging to the third family of glycosyl hydrolases, were isolated by anion-exchange, hydrophobic, and gel-penetrating chromatography from enzyme preparations based on the fungi Aspergillus japonicus and Trichoderma reesei, respectively. The enzymes exhibit maximal activity in acidic media (pH 3.5-4.0), and temperature activity optimum was 70°C for the β-xylosidase of A. japonicus and 60°C for the β-xylosidase of T. reesei. Kinetic parameters of p-nitrophenyl β-xylopyranoside and xylooligosaccharide hydrolysis by the purified enzymes were determined, which showed that β-xylosidase of A. japonicus was more specific towards low molecular weight substrates, while β-xylosidase of T. reesei preferred high molecular weight substrates. The competitive type of inhibition by reaction product (xylose) was found for both enzymes. The interaction of the enzymes of different specificity upon hydrolysis of glucurono- and arabinoxylans was found. The β-xylosidases exhibit synergism with endoxylanase upon hydrolysis of glucuronoxylan as well as with α-L-arabinofuranosidase and endoxylanase upon hydrolysis of arabinoxylan. Addition of β-xylosidases increased efficiency of hydrolysis of plant raw materials with high hemicellulose content (maize cobs) by the enzymic preparation Celloviridine G20x depleted of its own β-xylosidase.

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KEY WORDS: xylanases, β-xylosidase, xylooligosaccharides, xylans, hydrolysis

DOI: 10.1134/S0006297909090089

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