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	β-1,3-1,4-葡聚糖酶在毕赤酵母中的克隆与表达; Clone and expression of β-1,3-1,4-glucanase in Pichia pastoris
	陈龙军 ; 陈济琛 ; 陈亚兰 ; 林戎斌 ; 林新坚
	2013-05-15
关键词	毕赤酵母X-33 β-1 3-1 4-葡聚糖酶 诱导 Pichia pastoris X-33 β-1 3-1 4-glucanase induction
英文摘要	β-1,3-1,4-d-葡聚糖酶是一类专一性降解β-1,3-1,4-葡糖苷键中的β-1,4-糖苷键,产生小分子还原糖的水解酶,广泛应用于啤酒工业和饲料工业中。本研究根据毕赤酵母密码子偏好性优化β-1,3-1,4-葡聚糖酶基因序列,采用PCr法将其插入毕赤酵母表达载体PPICzαA,经SACI线性化后电击整合入毕赤酵母X-33基因组,构建重组酵母;经菌落PCr验证和摇瓶筛选,获得一株X-33/PPICzαA-bgl,甲醇诱导96H后,酶活力达308.5u/Ml,经SdS-PAgE电泳,实际蛋白分子量约为33ku。β-1,3-1,4-d-葡聚糖酶最适反应PH为5.0,最适反应温度为50℃。; β-1,3-1,4-glucanase,which can hydrolyzeβ-1,3-1,4-glucan by cutting off theβ-1,4-glycosidic linkages and release shorter reducing sugar,has been applied in the brewing and animal feed additive industry.It can effectively improve digestibility of barley-based diets and reduce enteritis.In this report,the gene ofβ-1,3-1,4-glucanase,which was optimized according to Pichia pastoris condon usage,was amplified by polymerase chain reaction and ligated into the expression vector pPICZαA.The recombinant vector was lineared by Sac I and transformed into Pichia pastoris X-33 by electroporation.Then the optimal positive transformant,named X-33/pPICZαA-bgl,was screened by colony PCR and shake flask fermentation.The enzyme activity of recombinantβ-1,3-1,4-glucanase couldreach 308.5U/mL after 96h methanol induction.Results of SDS-PAGE showed that about 33ku protein of theβ-1,3-1,4-glucanase was expressed.Acidity and temperature optimal for this recombinant enzyme was pH5.0 and 50℃,respectively.; 福建省农业科学院青年基金(2012DQC-11); 公益性行业(农业)科研专项(201303094-05)
语种	zh_CN
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/106755]
专题	化学化工－已发表论文
推荐引用方式 GB/T 7714	陈龙军,陈济琛,陈亚兰,等. β-1,3-1,4-葡聚糖酶在毕赤酵母中的克隆与表达, Clone and expression of β-1,3-1,4-glucanase in Pichia pastoris[J],2013.
APA	陈龙军,陈济琛,陈亚兰,林戎斌,&林新坚.(2013).β-1,3-1,4-葡聚糖酶在毕赤酵母中的克隆与表达..
MLA	陈龙军,et al."β-1,3-1,4-葡聚糖酶在毕赤酵母中的克隆与表达".(2013).

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