海洋α-葡萄糖苷酶的克隆表达及其制备高纯潘糖的研究

TS201.3; 潘糖是含α-1,6糖苷键的低聚异麦芽糖,α-葡萄糖苷酶是制备潘糖的关键酶制剂.本研究对深海热液口超嗜热古菌(Thermococcus siculi HJ21)中的α-葡萄糖苷酶序列进行合成,并克隆入载体pET29a,在大肠杆菌BL21中表达,通过His Trap HP层析柱纯化获得纯酶,以SDS-PAGE电泳测定α-葡萄糖苷酶分子量,并研究了酶的性质和其转糖苷作用.结果表明:该基因为729 bp,编码242个氦基酸,分子量约为27.2 kDa.该酶最适催化温度和pH分别为40℃和6.0.在以果糖为受体,与麦芽糖的比例为1∶9,反应10h时,其潘糖的生成量最高可以达到79.1％...

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Published in食品工业科技 Vol. 45; no. 19; pp. 158 - 165
Main Authors 于轶群, 薛陆州, 倪豪, 蒋蕾, 亢新鑫, 盘赛昆, 王淑军
Format Journal Article
LanguageChinese
Published 江苏省海洋生物资源与环境重点实验室,江苏海洋大学,江苏连云港 222005 01.10.2024
江苏省海洋生物技术重点实验室,江苏海洋大学,江苏连云港 222005
Subjects
低聚异麦芽糖
α-glucosidase
潘糖
iso-malto-oligosaccharide
转糖苷
cloning and expression
克隆表达
panose
α-葡萄糖苷酶
trans glycoside
Online AccessGet full text
ISSN1002-0306
DOI10.13386/j.issn1002-0306.2023110185

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Abstract TS201.3; 潘糖是含α-1,6糖苷键的低聚异麦芽糖,α-葡萄糖苷酶是制备潘糖的关键酶制剂.本研究对深海热液口超嗜热古菌(Thermococcus siculi HJ21)中的α-葡萄糖苷酶序列进行合成,并克隆入载体pET29a,在大肠杆菌BL21中表达,通过His Trap HP层析柱纯化获得纯酶,以SDS-PAGE电泳测定α-葡萄糖苷酶分子量,并研究了酶的性质和其转糖苷作用.结果表明:该基因为729 bp,编码242个氦基酸,分子量约为27.2 kDa.该酶最适催化温度和pH分别为40℃和6.0.在以果糖为受体,与麦芽糖的比例为1∶9,反应10h时,其潘糖的生成量最高可以达到79.1％.研究结果为古细菌α-葡萄糖苷酶制备高纯度潘糖提供了依据.
AbstractList TS201.3; 潘糖是含α-1,6糖苷键的低聚异麦芽糖,α-葡萄糖苷酶是制备潘糖的关键酶制剂.本研究对深海热液口超嗜热古菌(Thermococcus siculi HJ21)中的α-葡萄糖苷酶序列进行合成,并克隆入载体pET29a,在大肠杆菌BL21中表达,通过His Trap HP层析柱纯化获得纯酶,以SDS-PAGE电泳测定α-葡萄糖苷酶分子量,并研究了酶的性质和其转糖苷作用.结果表明:该基因为729 bp,编码242个氦基酸,分子量约为27.2 kDa.该酶最适催化温度和pH分别为40℃和6.0.在以果糖为受体,与麦芽糖的比例为1∶9,反应10h时,其潘糖的生成量最高可以达到79.1％.研究结果为古细菌α-葡萄糖苷酶制备高纯度潘糖提供了依据.
Abstract_FL Panose is iso-malto-oligosaccharide contained α-1,6 glycosidic bonds,and α-glucosidase is the key enzyme to produce panose.In this study,the α-glucosidase gene in deep-sea hydrothermal vents thermophilic archaea Thermococcus siculi HJ21 was synthesized,and cloned into the vector pET29a.Then expressed in Escherichia coli BL21 and purified by His Trap HP column.The molecular weight of α-glucosidase was determined by SDS-PAGE.The enzymatic properties and transglycosylation were also investigated.The results showed that the gene of α-glucosidase was 729 bp and encoded 242 amino acids.The molecular weight of α-glucosidase was about 27.2 kDa.Its optimal temperature and pH were 40 ℃ and 6.0 respectively.When the receptor was fructose,and the ratio of fructose and maltose was 1∶9,the highest yield of panose could reached 79.1％after 10 h reaction.The results provide a basis for producing high-purity panose by α-glucosidase from archaea.
Author 蒋蕾
于轶群
亢新鑫
倪豪
王淑军
薛陆州
盘赛昆
AuthorAffiliation 江苏省海洋生物资源与环境重点实验室,江苏海洋大学,江苏连云港 222005;江苏省海洋生物技术重点实验室,江苏海洋大学,江苏连云港 222005
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Author_FL KANG Xinxin
WANG Shujun
YU Yiqun
PAN Saikun
JIANG Lei
XUE Luzhou
NI Hao
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Copyright Copyright © Wanfang Data Co. Ltd. All Rights Reserved.
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DocumentTitle_FL Cloning and Expression of Marine α-Glucosidase and Its Preparation of High Purity Panose
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Issue 19
Keywords 低聚异麦芽糖
α-glucosidase
潘糖
iso-malto-oligosaccharide
转糖苷
cloning and expression
克隆表达
panose
α-葡萄糖苷酶
trans glycoside
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PublicationTitle 食品工业科技
PublicationTitle_FL Science and Technology of Food Industry
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Publisher 江苏省海洋生物资源与环境重点实验室,江苏海洋大学,江苏连云港 222005
江苏省海洋生物技术重点实验室,江苏海洋大学,江苏连云港 222005
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Snippet TS201.3; 潘糖是含α-1,6糖苷键的低聚异麦芽糖,α-葡萄糖苷酶是制备潘糖的关键酶制剂.本研究对深海热液口超嗜热古菌(Thermococcus siculi HJ21)中的α-葡萄糖苷酶序列进行合成,并克隆入载体pET29a,在大肠杆菌BL21中表达,通过His Trap...
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StartPage 158
Title 海洋α-葡萄糖苷酶的克隆表达及其制备高纯潘糖的研究
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