广西科学院学报2026，Vol.42Issue(2)：228-239,12.DOI:10.13657/j.cnki.gxkxyxb.20260604.001
重组蔗糖异构酶一步法催化合成甘油葡萄糖苷的研究
Study on the One-step Catalytic Synthesis of Glucosylglycerol by Recombinant Sucrose Isomerase
摘要
Abstract
To establish an efficient and environmentally friendly enzymatic synthesis route for glucosylglycerol(GG),a site-saturation mutagenesis strategy was applied to modify two phenylalanine residues(F297 and F321)of Erwinia rhapontici Sucrose Isomerase(SI,EC 5.4.99.11).Mutants exhibiting reduced intramolecular transglycosylation activity but enhanced hydrolytic activity were selected.Following mutant screening,ex-pression conditions were systematically optimized.The selected mutants were then employed as biocatalysts for GG synthesis,and the resulting products were quantitatively analyzed by High-Performance Liquid Chro-matography(HPLC).The results showed that,using ZYM-5052/Amp auto-induction medium,recombinant expression was enhanced,yielding an overall enzymatic activity approximately 16-fold higher than that in-duced by isopropyl β-D-thiogalactopyranoside(IPTG).Among the mutants,F321M demonstrated a 7-fold in-crease in GG yield compared to the wild-type enzyme.Under optimized reaction conditions—sucrose concen-tration at 1.0 mol/L,glycerol concentration at 2.0-2.5 mol/L,and a reaction time of 20 h the molar conver-sion rate of GG reached 24.54％.Furthermore,when the initial glycerol concentration was raised to 5.0 mol/L,the GG yield increased to 133.6 g/L.This study successfully demonstrates a one-step enzymatic synthesis of GG using immobilized engineered cells,with sucrose as the glycosyl donor and glycerol as the acceptor,providing both a simplified production route and refined reaction conditions.关键词
甘油葡萄糖苷/蔗糖异构酶/点饱和突变/酶催化/固定化细胞Key words
glucosylglycerol/sucrose isomerase/site-saturation mutagenesis/enzyme catalysis/matrix-en-trapped cells分类
化学化工引用本文复制引用
侯芳芳,成春燕,杨祥开,韦星明,周兴,杨克迪..重组蔗糖异构酶一步法催化合成甘油葡萄糖苷的研究[J].广西科学院学报,2026,42(2):228-239,12.基金项目
南宁高新区大健康产业—南宁国家高新区大健康产业联合创新服务平台建设与能力提升项目(桂科 AD25069031)和广西非粮生物质能源技术创新中心项目(桂科ZY23055011)资助. （桂科 AD25069031）
