Isolation and Characterization of Extracellular Pectin Lyase from Penicillium canescens

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O. A. Sinitsyna^1*, E. A. Fedorova¹, M. V. Semenova¹, A. V. Gusakov¹, L. M. Sokolova², T. M. Bubnova², O. N. Okunev², A. M. Chulkin³, E. A. Vavilova³, Y. P. Vinetsky³, and A. P. Sinitsyn^1,4

¹Chemical Faculty, Lomonosov Moscow State University, 119992 Moscow, Russia; fax: (495) 939-0997; E-mail: oasinitsyna@hotmail.com

²Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, pr. Nauki 5, 142290 Pushchino, Moscow Region, Russia; fax: (495) 923-3602

³State Research Institute of Genetics and Selection of Industrial Microorganisms, 1-yi Dorozhnyi Proezd 1, 117545 Moscow, Russia; fax: (495) 315-0501

⁴Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, 119071 Moscow, Russia; fax: (495) 954-2732

^* To whom correspondence should be addressed.

Received December 22, 2006; Revision received January 22, 2007

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Pectin lyase A (molecular weight 38 kD by SDS-PAGE, pI 6.7) was purified to homogeneity from culture broth of the mycelial fungus Penicillium canescens using chromatographic techniques. During genomic library screening, the gene encoding pectin lyase A from P. canescens (pelA) was isolated and sequenced, and the amino acid sequence was generated by applying the multiple alignment procedure (360 residues). A theoretical model for the three dimensional structure of the protein molecule was also proposed. Different properties of pectin lyase A were investigated: substrate specificity, pH- and temperature optimum of activity, stability under different pH and temperature conditions, and the effect of Ca²⁺ on enzyme activity. In the course of the laboratory trials, it was demonstrated that pectin lyase A from P. canescens could be successfully applied to production and clarification of juice.

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KEY WORDS: pectin lyase, Penicillium canescens, pectin, FPLC, liquefaction process, juice clarification

DOI: 10.1134/S0006297907050148

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