生物转化法制备L-天冬酰胺

发布时间：2018-04-14 11:18

本文选题：生物转化 + L-天冬氨酸　；参考：《中国生物工程杂志》2016年01期

【摘要】：探索生物转化法制备L-天冬酰胺的技术与工艺。通过分子生物学方法,克隆来源于大肠杆菌(Escherichia coli,E.coli)JM109的天冬酰胺合成酶A基因asn A,并于E.coli BL21(DE3)中表达,利用构建的E.coli基因工程菌E.coli BL21(DE3)/p ET28a(+)-asn A全细胞高密度催化L-天冬氨酸生产L-天冬酰胺,以PITC柱前衍生-高效液相检测底物和产物。表达的蛋白质分子质量约为37k Da,与预期大小相符,比酶活力为1786.6U/g。L-天冬氨酸转化率为95.8%,L-天冬酰胺产量可达126.5g/L,生产速率为15.81g/(L·h)。结果表明,已成功构建高效表达天冬酰胺合成酶A基因工程菌株,并用于催化L-天冬氨酸转化生产L-天冬酰胺,解决了L-天冬酰胺生物转化生产工艺中ATP成本过高的难题,为L-天冬酰胺制备提供新的绿色途径。
[Abstract]:To explore the technology and technology of preparing L-asparagine by biotransformation.By molecular biological method, the asparagine synthase A gene asn A was cloned from Escherichia coli E. colisiae JM109 and expressed in E.coli BL21DE3.Lasparagine was produced by using the constructed E.coli gene engineering strain E.coli BL21(DE3)/p ET28a (Ca-asnA) as a whole cell high density catalyst. The substrates and products were detected by PITC pre-column derivatization and high performance liquid chromatography.The molecular weight of the expressed protein is about 37kDa. the specific enzyme activity is 1786.6Ug.L- aspartic acid conversion rate is 95.8L- asparagine yield can reach 126.5 g / L, the production rate is 15.81g/(L HX.The results showed that the genetically engineered strain expressing asparagine synthase A was successfully constructed and used to catalyze the conversion of Laspartic acid to Lasparagine, which solved the problem of high cost of ATP in the production process of L- asparagine biotransformation.It provides a new green way for the preparation of L-asparagine.
【作者单位】：中国食品发酵工业研究院中国工业微生物菌种保藏管理中心;山东省富马酸生物转化工程技术研究中心;
【基金】：烟台市科技发展计划资助项目(2014SF151)
【分类号】：Q78;TQ929

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