Glycerolglycolipid is composed of sugar and glyceride linked by glycosidic bond. It has many biological activities, such as anti-tumor, anti-virus and anti-inflammatory, and has become the focus of current research. The traditional production method of glycerolglycolipid is extracted from plant, which has low extraction rate and purity. The chemical synthesis method requires high reaction conditions and separation equipment, the product is not easy to separate, and the waste liquid pollutes the environment. In this study, the recombinant mutant plasmids of glycosyltransferase Agt from marine bacterium Candidatus pelagibacter sp. HTCC7211 were constructed by genetic engineering method to realize the expression in Escherichia coli BL21-CodonPlus (DE3)-RIL. The enzyme activity, the optimum reaction conditions and extraction method of catalytic synthesis of monoglucosyl diacylglycerol were further researched. The results showed that the mutant T257S could efficiently convert UDP-glucose and diacylglycerol into monoglucosyl diacylglycerol, and catalytic efficiency was 1.56 times higher than that of wild-type enzyme. Monoglucosyl diacylglycerol was extracted by cyclohexane/ethyl acetate method, and the extraction rate was 84.7%. Under the optimum reaction conditions (enzyme 8 U/ml, UDP-glucose 500 mg/L, 35 ℃ and pH 8.5), the conversion rate and purity could reach 68.5% and 93.8%, receptively. Therefore, the preparation of monoglucosyl diacylglycerol by mutant T257S has the advantages of high conversion, high product purity and less synthetic steps, which has a good industrial application prospect.
ZHANG Zhiping
,
ZHAO Yuzhe
,
ZHAO Caimeng
,
YANG Xu
,
WEI Tao
. Preparation of mutants of glycosyltransferase Agt from Candidatus pelagibacter sp. HTCC7211 and its application in the catalytic synthesis of monoglucosyl diacylglycerol[J]. Food and Fermentation Industries, 2022
, 48(13)
: 9
-15
.
DOI: 10.13995/j.cnki.11-1802/ts.030745
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