新型里氏木霉β-葡萄糖苷酶BGLII（cel1A）的酶学性质及细胞内定位
Enzymatic properties and intracellular localization of the novel Trichoderma reesei beta-glucosidase BGLII (cel1A).
作者信息
Saloheimo Markku, Kuja-Panula Juha, Ylösmäki Erkko, Ward Michael, Penttilä Merja
机构信息
VTT Biotechnology, 02044 VTT, Finland.
出版信息
Appl Environ Microbiol. 2002 Sep;68(9):4546-53. doi: 10.1128/AEM.68.9.4546-4553.2002.
Abstract
This paper describes the characterization of an intracellular beta-glucosidase enzyme BGLII (Cel1a) and its gene (bgl2) from the cellulolytic fungus Trichoderma reesei (Hypocrea jecorina). The expression pattern of bgl2 is similar to that of other cellulase genes known from this fungus, and the gene would appear to be under the control of carbon catabolite repression mediated by the cre1 gene. The BGLII protein was produced in Escherichia coli, and its enzymatic properties were analyzed. It was shown to be a specific beta-glucosidase, having no beta-galactosidase side activity. It hydrolyzed both cellotriose and cellotetraose. BGLII exhibited transglycosylation activity, producing mainly cellotriose from cellobiose and sophorose and cellobiose from glucose. Antibodies raised against BGLII showed the presence of the enzyme in T. reesei cell lysates but not in the culture supernatant. Activity measurements and Western blot analysis of T. reesei strains expressing bgl2 from a constitutive promoter further confirmed the intracellular localization of this beta-glucosidase.
摘要
本文描述了来自纤维素分解真菌里氏木霉（枝状腐质霉）的一种细胞内β-葡萄糖苷酶BGLII（Cel1a）及其基因（bgl2）的特性。bgl2的表达模式与该真菌中已知的其他纤维素酶基因相似，并且该基因似乎受cre1基因介导的碳分解代谢物阻遏的控制。BGLII蛋白在大肠杆菌中产生，并对其酶学性质进行了分析。结果表明它是一种特异性β-葡萄糖苷酶，没有β-半乳糖苷酶的副活性。它能水解纤维三糖和纤维四糖。BGLII表现出转糖基化活性，主要从纤维二糖和槐糖产生纤维三糖，从葡萄糖产生纤维二糖。针对BGLII产生的抗体表明该酶存在于里氏木霉细胞裂解物中，但不存在于培养上清液中。对从组成型启动子表达bgl2的里氏木霉菌株进行的活性测定和蛋白质免疫印迹分析进一步证实了这种β-葡萄糖苷酶的细胞内定位。
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