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Identification and Biochemical Characterization of Major β-Mannanase in Talaromyces cellulolyticus Mannanolytic System.
Applied Biochemistry and Biotechnology ( IF 3.1 ) Pub Date : 2020-06-05 , DOI: 10.1007/s12010-020-03350-6
Keiko Uechi 1 , Masahiro Watanabe 2 , Tatsuya Fujii 2 , Saori Kamachi 2 , Hiroyuki Inoue 2
Affiliation  

Talaromyces cellulolyticus is a promising fungus for providing a cellulase preparation suitable for the hydrolysis of lignocellulosic material, although its mannan-degrading activities are insufficient. In the present study, three core mannanolytic enzymes, including glycosyl hydrolase family 5–7 (GH5–7) β-mannanase (Man5A), GH27 α-galactosidase, and GH2 β-mannosidase, were purified from a culture supernatant of T. cellulolyticus grown with glucomannan, and the corresponding genes were identified based on their genomic sequences. Transcriptional analysis revealed that these genes were specifically induced by glucomannan. Two types of Man5A products, Man5A1 and Man5A2, were found as major proteins in the mannanolytic system. Man5A1 was devoid of a family 1 carbohydrate-binding module (CBM1) at the N-terminus, whereas Man5A2 was devoid of both CBM1 and Ser/Thr-rich linker region. The physicochemical and catalytic properties of both Man5A1 and Man5A2 were identical to those of recombinant Man5A (rMan5A) possessing CBM1, except for the cellulose-binding ability. Man5A CBM1 had little effect on mannan hydrolysis of pretreated Hinoki cypress. The results suggest that an improvement in Man5A CBM1 along with the augmentation of identified mannanolytic enzyme components would aid in efficient hydrolysis of softwood using T. cellulolyticus cellulase preparation.



中文翻译:

纤维素酶解纤菌甘露聚糖系统中主要β-甘露聚糖酶的鉴定及生化特性。

踝节菌属纤维热是用于提供适合于木质纤维素材料水解的纤维素酶制剂有希望的真菌,尽管它的甘露聚糖降解活性是不够的。在本研究中,从纤溶解球菌的培养上清液中纯化了三种核心甘露聚糖酶,包括糖基水解酶5-7(GH5-7)β-甘露聚糖酶(Man5A),GH27α-半乳糖苷酶和GH2β-甘露糖苷酶用葡甘露聚糖生长,并根据其基因组序列鉴定相应的基因。转录分析表明,这些基因是由葡甘露聚糖特异性诱导的。发现两种类型的Man5A产物Man5A1和Man5A2是甘露聚糖分解系统中的主要蛋白质。Man5A1在N末端不含家庭1碳水化合物结合模块(CBM1),而Man5A2不含CBM1和富含Ser / Thr的接头区域。Man5A1和Man5A2的物理化学和催化特性与具有CBM1的重组Man5A(rMan5A)相同,除了具有纤维素结合能力。Man5A CBM1对预处理的扁柏的甘露聚糖水解影响很小。溶纤梭菌纤维素酶制剂。

更新日期:2020-06-05
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