碳水化合物酶- 50年的研究在罗蒙诺索夫莫斯科国立大学化学酶学系,历史和前景

Arkady P. Sinitsyn, Olga A. Sinitsyna, Ivan N. Zorov, Aleksandra M. Rozhkova
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摘要

这篇综述描述了从20世纪70年代中期到现在化学酶学系对碳水化合物研究的发展历史。多酶纤维素酶配合物作用下纤维素和可再生植物原料酶转化过程的机理和动力学研究结果;这些复合物的各个组成部分的作用-碱性(内切葡聚糖酶和纤维素生物水解酶)和辅助酶(多糖单加氧酶,β-葡萄糖苷酶,木聚糖酶)及其协同相互作用。介绍了不同设计的植物原料生物转化反应器的特点:周期性反应器、连续柱式反应器、恒电场水解反应器、磁场中铁磁颗粒强烈混合反应器。讨论了利用各种预处理方法提高植物原料反应性的可能性,以及纤维素的结构和理化性质对酶转化效率的影响。本文介绍了利用诱变方法产生纤维素酶和其他糖酶的微生物高活性菌株的数据——木霉(Hypocrea)、青霉(Talaromyces)、曲霉(Aspergillus)、黄孢(Myceliophtora)等,以及它们产生的酶复合物的组成和形成它们的酶的性质。它描述了基于P. canescens和P. verruculosum的表达系统的创建,以及在它们的帮助下重组生产菌株的生产,这使得获得确保植物原料生物转化过程高效率的酶制剂(EP)成为可能,并创造了广泛的糖酶生产者,用于工业和农业的各个领域。使用疣状芽孢杆菌表达系统获得的一些工业上重要的EP目前正在Agroferment LLC工厂生产。
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CARBOHYDRASES – 50 YEARS OF RESEARCH AT THE DEPARTMENT OF CHEMICAL ENZYMOLOGY OF LOMONOSOV MOSCOW STATE UNIVERSITY, HISTORY AND PROSPECTS
The review describes the history of the development of research on carbohydrasеs conducted at the Department of Chemical Enzymology from the mid-1970s to the present. The results concerning the study of the mechanism and kinetics of the processes of enzymatic conversion of cellulose and renewable plant raw materials under the action of a multi-enzyme cellulases complexes; the role of individual components of these complexes - basic (endoglucanases and cellobiohydrolases) and auxiliary enzymes (polysaccharide monooxygenase, β-glucosidase, xylanase) and their synergistic interaction. The features of using reactors of various designs for bioconversion of plant raw materials are described: periodic type, continuous column type, reactor for hydrolysis in a constant electric field, reactor with intensive mixing by ferromagnetic particles in magnetic field. The possibilities of increasing the reactivity of plant raw materials using various pretreatment methods, as well as the influence of the structural and physico-chemical properties of cellulose on the efficiency of its enzymatic conversion are discussed. Data on the creation of highly active strains of microscopic fungi-producers of cellulases and other carbohydrases using methods of induced mutagenesis - Trichoderma (Hypocrea), Penicillium (Talaromyces), Aspergillus, Chrysosporium (Myceliophtora) spp., as well as data on the composition of the enzyme complexes produced by them and the properties of the enzymes forming them are presented. It describes the creation of expression systems based on P. canescens and P. verruculosum and the production of recombinant producer strains with their help, which made it possible to obtain enzyme preparations (EP) that ensure high efficiency of bioconversion processes of plant raw materials, as well as to create producers of a wide range of carbohydrases for practical use in various fields of industry and agriculture. A number of industrially important EP obtained using the P. verruculosum expression system are currently being produced at the Agroferment LLC plant.
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