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.
关键词
α-葡萄糖苷酶/克隆表达/低聚异麦芽糖/转糖苷/潘糖
Key words
α-glucosidase/cloning and expression/iso-malto-oligosaccharide/trans glycoside/panose
维普
万方数据