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Production of biocatalysts on the basis of recombinant heterologous xylanase producer strains in the Penicillium verruculosum fungus: Their application in the hydrolysis of timber and wood processing industry wastes
Authors:D O Osipov  A M Rozhkova  V Yu Matys  A V Koshelev  O N Okunev  E A Rubtsova  A G Pravil’nikov  I N Zorov  O A Sinitsyna  I N Oveshnikov  E R Davidov  A P Sinitsyn
Affiliation:1. Bach Institute of Biochemistry, Russian Academy of Sciences, Leninskii pr. 33/2, Moscow, 119071, Russia
2. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, pr. Nauki 5, Pushchino, Moscow oblast, 142290, Russia
3. Moscow State University, Moscow, 119992, Russia
4. State Research Institute for Biosynthesis of Proteins, ul. Aleksandra Solzhenitsyna 27, Moscow, 109004, Russia
Abstract:The aim of this work was to create biocatalysts with an increased heterologous expression of endo-β-1,4-xylanase of P. canescens using recombinant P. verruculosum strains, to analyze the properties of new enzyme preparations, and to study the saccharifying activity of these preparations in the hydrolysis of plant raw materials, such as hogged aspen and detarred pine wood wastes of the timber and wood processing industries. The xylanase activity of the existing enzymatic preparations is insufficiently high to hydrolyze a xylan-rich biomass. The creation of increasingly xylanolytically active P. verruculosum-based recombinant strains containing homologous or heterologous genes of xylanase and mannanase is therefore a problem of great interest. Using the methods of genetic engineering, we obtained enzymatic preparations that are biocatalysts for the hydrolysis of plant raw material wastes of the sawmilling and wood processing industries and, according to the data of chromatographic fractionation, have compositions of 45–60% cellulase and 20–50% xylanase (which is optimal for the saccharifying of bagasse, along with aspen and pine wood). The originality of our technique lies in the creation of biocatalysts with predetermined properties, thus reducing appreciably the cost of enzyme preparation by eliminating the need to mix components of the carbohydrase complex for the hydrolysis of plant raw materials, e.g., aspen and pine wood.
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