Isolation and Characterization of Extracellular alpha-Galactosidases from Penicillium canescens

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O. A. Sinitsyna^1*, E. A. Fedorova¹, I. M. Vakar², E. G. Kondratieva³, A. M. Rozhkova³, L. M. Sokolova⁴, T. M. Bubnova⁴, O. N. Okunev⁴, A. M. Chulkin⁵, Y. P. Vinetsky⁵, and A. P. Sinitsyn^1,3

¹Faculty of Chemistry, Lomonosov Moscow State University, 119991 Moscow, Russia; fax: (495) 939-0997; E-mail: oasinitsyna@gmail.com

²Institute of Food Biotechnology, Russian Academy of Agricultural Sciences, ul. Samokatnaya 4B, 111033 Moscow, Russia; fax: (495) 362-3371

³Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33/2, 119071 Moscow, Russia; fax: (495) 954-2732

⁴Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, pr. Nauki 5, 142292 Pushchino, Moscow Region, Russia; fax: (495) 923-3602

⁵State Research Institute of Genetics and Selection of Industrial Microorganisms, 1-yi Dorozhnyi Proezd 1, 117545 Moscow, Russia; fax: (495) 315-0501

^* To whom correspondence should be addressed.

Received June 9, 2007; Revision received June 26, 2007

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Two alpha-galactosidases were purified to homogeneity from the enzymatic complex of the mycelial fungus Penicillium canescens using chromatography on different sorbents. Substrate specificity, pH- and temperature optima of activity, stability under different pH and temperature conditions, and the influence of effectors on the catalytic properties of both enzymes were investigated. Genes aglA and aglC encoding alpha-galactosidases from P. canescens were isolated, and amino acid sequences of the proteins were predicted. In vitro feed testing (with soybean meal and soybean byproducts enriched with galactooligosaccharides as substrates) demonstrated that both alpha-galactosidases from P. canescens could be successfully used as feed additives. alpha-Galactosidase A belonging to the 27th glycosyl hydrolase family hydrolyzed galactopolysaccharides (galactomannans) and alpha-galactosidase C belonging to the 36th glycosyl hydrolase family hydrolyzed galactooligosaccharides (stachyose, raffinose, etc.) of soybean with good efficiency, thus improving the digestibility of fodder.

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KEY WORDS: alpha-galactosidase, Penicillium canescens, HPLC, stachyose, raffinose, galactomannan, feed additives, soybean

DOI: 10.1134/S000629790801015X

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