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中国精品科技期刊2020
于轶群,薛陆州,倪豪,等. 海洋α-葡萄糖苷酶的克隆表达及其制备高纯潘糖的研究[J]. 食品工业科技,2024,45(19):158−165. doi: 10.13386/j.issn1002-0306.2023110185.
引用本文: 于轶群,薛陆州,倪豪,等. 海洋α-葡萄糖苷酶的克隆表达及其制备高纯潘糖的研究[J]. 食品工业科技,2024,45(19):158−165. doi: 10.13386/j.issn1002-0306.2023110185.
YU Yiqun, XUE Luzhou, NI Hao, et al. Cloning and Expression of Marine α-Glucosidase and Its Preparation of High Purity Panose[J]. Science and Technology of Food Industry, 2024, 45(19): 158−165. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023110185.
Citation: YU Yiqun, XUE Luzhou, NI Hao, et al. Cloning and Expression of Marine α-Glucosidase and Its Preparation of High Purity Panose[J]. Science and Technology of Food Industry, 2024, 45(19): 158−165. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2023110185.

海洋α-葡萄糖苷酶的克隆表达及其制备高纯潘糖的研究

Cloning and Expression of Marine α-Glucosidase and Its Preparation of High Purity Panose

  • 摘要: 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个氨基酸,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10 h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: 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.

     

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