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Purification and Characterization of Pectin Lyase Secreted by Penicillium citrinum

S. Yadav1*, P. K. Yadav2, D. Yadav2,3, and K. D. S. Yadav1

1Department of Chemistry, D. D. U. Gorakhpur University, Gorakhpur 273009, India; E-mail: sangeeta_rahul@rediffmail.com

2Department of Biotechnology, D. D. U. Gorakhpur University, Gorakhpur 273009, India

3Department of Molecular Biology and Genetic Engineering, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar (Uttarakhand) 263145, India

* To whom correspondence should be addressed.

Received August 5, 2008; Revision received October 20, 2008
The importance of various parameters such as sugarcane juice concentration, pH of the medium, and effects of different solid supports for maximum secretion of pectin lyase from Penicillium citrinum MTCC 8897 has been studied. The enzyme was purified to homogeneity by Sephadex G-100 and DEAE-cellulose chromatography. The molecular mass determined by SDS-PAGE was 31 kDa. The Km and kcat values were found to be 1 mg/ml and 76 sec–1, respectively. The optimum pH of the purified pectin lyase was 9.0, though it retains activity in the pH 9.0-12.0 range when exposed for 24 h. The optimum temperature was 50°C, and the pectin lyase was found to be completely stable up to 40°C when exposed for 1 h. The purified pectin lyase was found efficient in retting of Linum usitatissimum, Cannabis sativa, and Crotalaria juncea.
KEY WORDS: pectin lyase, retting, solid state fermentation, Penicillium citrinum, Cannabis sativa, Crotalaria juncea, Linum usitatissimum

DOI: 10.1134/S0006297909070141