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首页> 外文期刊>Bioprocess and Biosystems Engineering >Trans-glycosylation capacity of a highly glycosylated multi-specific beta-glucosidase from Fusarium solani
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Trans-glycosylation capacity of a highly glycosylated multi-specific beta-glucosidase from Fusarium solani

机译：茄镰刀菌高度糖基化的多特异性β-葡萄糖苷酶的反式糖基化能力

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摘要

An extracellular beta-glucosidase from Fusaruim solani cultivated on wheat bran was purified by only two chromatographic steps. The purified enzyme exhibited optimal temperature and pH at 60 A degrees C and pH 5, respectively. The purified beta-glucosidase behaves as a very large protein due to its high degree of glycosylation. More interestingly, the endoglycosidase H (Endo H) treatment led to 97.55% loss of its initial activity after 24 h of treatment. Besides, the addition of Tunicamycin (nucleoside antibiotic blocking the N-glycosylation first step) during the culture of the fungus affected seriously the glycosylation of the enzyme. Both treatments (endo H and Tunicamycin) strengthened the idea that the hyperglycosylation is involved in the beta-glucosidase activity and thermostability. This enzyme was also shown to belong to class III of beta-glucosidases (multi-specific) since it was able to act on either cellobiose, gentiobiose or sophorose which are disaccharide composed of two units of d-glucose connected by beta 1-4, beta 1-6 and beta 1-2 linkage, respectively. The beta-glucosidase activity was strongly enhanced by ferrous ion (Fe2+) and high ionic strength (1 M KCl). The purified enzyme exhibited an efficient transglycosylation capacity allowing the synthesis of cellotriose and cellotetraose using cellobiose as donor.
机译：仅通过两个色谱步骤纯化了在小麦麸皮上培养的来自Fusaruim solani的细胞外β-葡萄糖苷酶。纯化的酶分别在60 A℃和5的pH下显示最佳温度和pH。纯化的β-葡萄糖苷酶由于其高度的糖基化作用，表现为非常大的蛋白质。更有趣的是，内切糖苷酶H（Endo H）处理在处理24小时后导致其初始活性降低了97.55％。此外，在真菌的培养过程中加入衣霉素（核苷抗生素阻断了N-糖基化的第一步）严重影响了酶的糖基化。两种处理方法（endo H和Tunicamycin）都增强了这种想法，即高糖基化与β-葡萄糖苷酶的活性和热稳定性有关。该酶还被证明属于β-葡萄糖苷酶的III类（多特异性），因为它能够作用于纤维二糖，龙胆二糖或槐糖糖，它们是由两个通过β1-4连接的d-葡萄糖单元组成的二糖， beta 1-6和beta 1-2链接。亚铁离子（Fe2 +）和高离子强度（1 M KCl）大大增强了β-葡萄糖苷酶的活性。纯化的酶表现出有效的转糖基化能力，从而允许使用纤维二糖作为供体来合成纤维三糖和纤维四糖。

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