Novel Enzyme Preparations with High Pectinase and Hemicellulase Activity Based on Penicillium canescens Strains

摘要

Recombinant strains of Penicillium canescens producing homologous pectin lyase A and heterologous endo-1,5-alpha-arabinase A and endo-1,4-alpha-polygalacturonase, as well as enzymes of the host strain (alpha-L-arabinofuranosidases, xylanases, and others), were obtained by genetic engineering. The enzyme preparations (EPs) obtained from the cultural medium of recombinant P. canescens strains efficiently hydrolyzed raw plant material with a high content of pectin compounds. It was shown that the yield of reducing sugars and arabinose increased 16 and 22% in comparison with the control EP based on the host strain when one of the obtained EPs was used for beet pulp hydrolysis. It was established that the most active EP consisted of pectin lyase (10%), endo-1,5-arabinase (26%),alpha-L-arabinofuranosidase and arabinoxylan-arabinofuranohydrolase (12%), and xylanase (10%). The activities of pectin lyase, polygalacturonase, and arabinase of the EP in reactions with various substrates were determined. The specificity, pH and T-optima, and thermal stability of the homogenous recombinant endo-1,5-alpha-arabinase were investigated. The kinetic parameters (K-m, k(cat)) of the linear arabinan hydrolysis were determined.