Production of Highly Effective Enzyme Complexes of Cellulases and Hemicellulases on the basis of Penicillium verruculosum Strain for Hydrolysis of Plant Raw Materials

Автор: A.P. Sinitsyn, D.O. Osipov, A.M Rozhkova, E.B. Bushina, G.S. Dotsenko, O.A. Sinitsyna, E.G. Kondratieva, I.N. Zorov, O.N. Okunev, V.A. Nemashkalov, V.Yu. Matys, and A.V. Koshelev

Страница: 40-53

Production of Highly Effective Enzyme Complexes of Cellulases and Hemicellulases on the basis of Penicillium verruculosum Strain for Hydrolysis of Plant Raw Materials

Russian Journal of Biotechnology, 2013, N 5, P. 40-53

UDC 577.152, 577.113, 579.252

Section:  “Producers, Biology, Selection, and Gene Engineering”

A.P. Sinitsyn ^{1, 2},  D.O. Osipov ¹,  A.M Rozhkova ^{1, 2},  E.B. Bushina ¹,  G.S. Dotsenko ^{1, *},  O.A. Sinitsyna ^{1, 2},  E.G. Kondratieva ¹,  I.N. Zorov ^{1, 2},  O.N. Okunev ³,  V.A. Nemashkalov ³,  V.Yu. Matys ³,  and  A.V. Koshelev ³

¹  The Bach Institute of Biochemistry, Russ. Acad. Sci.,  119071, Moscow Russia

²  The Department of Chemistry, Lomonosov Moscow State University,  119991, Moscow Russia

³  The Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russ. Acad. Sci.,  142292, Pushchino, Moskovskaya oblast Russia

e-mail:  gsdotsenko@gmail.com

Methods for production and analysis of cellulose- and hemicellulase-containing enzyme preparations of various component structures based on the Penicillium verruculosum carbohydrase complex and intended for the effective hydrolysis of different types of cellulosic raw materials have been developed. New recombinant strains of P. verruculosum producing multienzyme carbohydrase complexes with increased activities of cellulases (due to the expression of endo-β-1,4-glucanases I and IV and cellobiohydrolase II of Trichoderma reesei), and also hemicellulases (due to the expression of P. canescens and T. reesei endo-β-1,4-xylanases and T. reesei endo-β-1,4-mannanase) were constructed. The hydrolytic efficiency of the enzyme preparations produced by the new recombinant strains during continuous hydrolysis of three types of various cellulosic raw materials (milled aspen, depitched pine wood and milled bagasse) was studied. It was shown that the new enzyme preparations that included both recombinant proteins and their own basic cellulase complex are characterized by the highest hydrolytic ability exceeding that of the enzyme preparations based on the original P. verruculosum strain. The recombinant enzyme preparations were characterized by high stability; optimal рН and temperature values for the cellulase, xylanase and mannanase activities were in the range 3,5—5,5 and 50—80°C, respectively.

Key words:  cellulases,  cellulosic raw materials,  enzymatic hydrolysis,  hemicellulases,  Penicillium verruculosum.

01.04.2015, 1596 просмотров.