食品与发酵工业2025，Vol.51Issue(10)：9-16,8.DOI:10.13995/j.cnki.11-1802/ts.039737
谷氨酰胺转氨酶耐热突变体在茂原链霉菌中的表达及酶学性质分析
Expression of a heat-resistant transglutaminase in Streptomyces mobaraensis and its enzymatic property analysis
摘要
Abstract
Transglutaminase(TGase)which can catalyze protein crosslinking has been widely used in food processing.To promote its industrial application,Streptomyces mobaraensis which can efficiently produce heat-resistant TGase was successfully constructed in this study.First,this study synthesized the expression cassette of a TGase thermostable mutant TGm3(1 745 bp),containing the natural promoter,pro-region,and terminator of S.mobaraensis TGase in addition to TGm3 coding sequence(optimized based on codon preference of streptomy-ces).Second,the expression cassette of TGm3 was integrated into the TGase gene locus of S.mobaraensis mutant smY2019-Δtg,yielding the TGm3-producing strain smY2019-TGm3.At last,the fermentation level of smY2019-TGm3 and the enzymatic properties of TGm3 were ana-lyzed.As indicated by the shake flask fermentation,the extracellular TGase activity of smY2019-tgm3 reached the maximum value(19.5 U/mL)after 60 h fermentation.After being purified by the cation exchange column,the specific activity of TGm3 reached 41.76 U/mg,2.7 times higher than that of wild-type TGase.The analysis of enzymatic properties showed that the 60℃ half-life(60.6 min)and the optimal temperature(60℃)of TGm3 were 33.6 times and 10℃ higher than those of wild-type TGase,respectively.These results show that smY2019-tgm3 can efficiently synthesize heat-resistant TGase and has a certain potential for industrial application.关键词
茂原链霉菌/谷氨酰胺转氨酶/高效表达/热稳定性Key words
Streptomyces mobaraensis/transglutaminase/efficient expression/thermostability引用本文复制引用
陈智恒,周灿,韦佐权,谭亚梦,叶佳才,崔世修,刘松..谷氨酰胺转氨酶耐热突变体在茂原链霉菌中的表达及酶学性质分析[J].食品与发酵工业,2025,51(10):9-16,8.基金项目
国家重点研发计划项目(2021YFC2101400) （2021YFC2101400）
国家自然科学基金项目(32071474) （32071474）
中央高校基本科研业务费专项资金资助项目(JUSRP221033) （JUSRP221033）
