大连工业大学学报2026，Vol.45Issue(1)：27-34,8.DOI:10.19670/j.cnki.dlgydxxb.2026.0103
毕赤酵母中糖基化修饰对持续性内切葡聚糖酶SmCel5A的影响
Impact of glycosylation on processive endoglucanase SmCel5A in Pichia pastoris
摘要
Abstract
The endoglucanase SmCel5A from Sporocytophaga sp.CX11 was heterologously expressed in Pichia pastoris X-33,and the N-glycosylation modification was identified at its Asn258 site by mass spectrometry.To investigate the effect of glycosylation modification in Sm Cel5A-pPICZaA-X33,the mutant Sm Cel5A-N258D-pPICZaA-X33 was constructed and expressed in P.pastoris.Characterization of enzymatic properties revealed the activity of deglycosylated Sm Cel5A-N258D-pPICZaA-X33 against sodium carboxymethyl cellulose 95％decreasd compared with Sm Cel5 A-pPICZaA-X33,and its activities against microcrystalline cellulose and filter paper decreased by 75％and 50％,respectively.The optimal pH shifted from 7.0 to 6.0,with reduced stability under alkaline conditions.The optimal temperature decreased from 40 ℃ to 35 ℃,accompanied by poorer thermal stability.The results indicate that the glycosylation modification at the Asn258 site plays a crucial role in maintaining the catalytic activity,thermal stability,and alkali tolerance of SmCel5A.关键词
内切葡聚糖酶/生孢噬纤维菌/N-糖基化修饰/毕赤酵母Key words
endoglucanase/Sporocytophaga sp./N-glycosylation modification/Pichia pastoris分类
生物科学引用本文复制引用
王曦泽,王梓懿,陈晓艺..毕赤酵母中糖基化修饰对持续性内切葡聚糖酶SmCel5A的影响[J].大连工业大学学报,2026,45(1):27-34,8.基金项目
辽宁省教育厅重点攻关项目(LJKZZ20220061). （LJKZZ20220061）
