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丙谷二肽的生物合成及关键酶研究进展
程坤1,2,卫禾耕3,杨伟强4,盛清1,杨仲毅2
1.浙江理工大学 生命科学与医药学院,浙江 杭州 310018;2.台州学院 药学院,浙江 台州 318000;3.浙江永太科技股份有限公司,浙江 台州 317016;4.浙江海洲制药股份有限公司,浙江 台州 317016
摘要:
丙谷二肽(L-alanyl-L-glutamine,Ala-Gln)是目前发现的一种在现代医疗健康领域极其重要的二肽,其进入人体后能够迅速水解成L-谷氨酰胺,促进蛋白质合成。生物合成丙谷二肽具有绿色、高效、安全的优点,主要的生物合成途径有微生物发酵法和生物催化法,其关键酶分别是L-氨基酸连接酶和α-氨基酸酯酰基转移酶。通过敲除dpppep等基因来抑制二肽的降解,发酵法中Ala-Gln的最高水平为24.70 g/L,发酵时间为47 h。α-氨基酸酯酰基转移酶以L-丙氨酸甲酯和L-谷氨酰胺为原料合成Ala-Gln,相关研究主要集中在酶的表达体系、定向进化、固定化等领域。α-氨基酸酯酰基转移酶可以在25 min转化生成106.61 g/L的Ala-Gln,相较于微生物发酵法,该方法的反应时间更短,产物浓度更高,且无须ATP供能,是Ala-Gln工业化生产的首选工艺技术。
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DOI:10.14188/j.ajsh.20240902001
分类号:TQ464
基金项目:浙江理工大学绍兴生物医药研究院开放基金(SXAB202007)
Progress in biosynthesis and key enzymes of Ala-Gln
CHENG Kun1,2, WEI Hegeng3, YANG Weiqiang4, SHENG Qing1, YANG Zhongyi2
1.College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, Zhejiang, China;2.School of Pharmaceutical Science, Taizhou University, Taizhou 318000, Zhejiang, China;3.Zhejiang Yongtai Technology Co., Ltd., Taizhou 317016, Zhejiang, China;4.Zhejiang Haizhou Pharmaceutical Co., Ltd., Taizhou 317016, Zhejiang, China
Abstract:
L-alanyl-L-glutamine (Ala-Gln) is an important dipeptide in modern medical and health care. After entering the human body, it can be rapidly hydrolyzed into L-glutamine, which promotes protein synthesis. The synthesis of Ala-Gln dipeptide by biological method has the advantages of green, high efficiency and safety. The main biosynthetic pathways are microbial fermentation and biocatalysis, whose key enzymes are L-amino acid ligase and α-amino acid ester acyltransferase. The degradation of dipeptide can be inhibited by knocking out genes such as dpp and pep, and the highest level of Ala-Gln in the fermentation method can reach 24.70 g/L after a fermentation time of 47 h. α-amino acid ester acyltransferase synthesizes Ala-Gln from L-alanine methyl ester and L-glutamine, and the related research mainly focuses on the enzyme expression system, directional evolution, and immobilisation. α-amino acid ester acyltransferase can convert and produce 106.61 g/L of Ala-Gln in 25 min. Compared with microbial fermentation, this method has a shorter reaction time, higher product concentration, and does not need ATP for energy supply, making it the preferred process technology for industrial production of Ala-Gln.
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