本文選題：生物轉(zhuǎn)化 + L-天冬氨酸��； 參考：《中國(guó)生物工程雜志》2016年01期

【摘要】：探索生物轉(zhuǎn)化法制備L-天冬酰胺的技術(shù)與工藝。通過(guò)分子生物學(xué)方法,克隆來(lái)源于大腸桿菌(Escherichia coli,E.coli)JM109的天冬酰胺合成酶A基因asn A,并于E.coli BL21(DE3)中表達(dá),利用構(gòu)建的E.coli基因工程菌E.coli BL21(DE3)/p ET28a(+)-asn A全細(xì)胞高密度催化L-天冬氨酸生產(chǎn)L-天冬酰胺,以PITC柱前衍生-高效液相檢測(cè)底物和產(chǎn)物。表達(dá)的蛋白質(zhì)分子質(zhì)量約為37k Da,與預(yù)期大小相符,比酶活力為1786.6U/g。L-天冬氨酸轉(zhuǎn)化率為95.8%,L-天冬酰胺產(chǎn)量可達(dá)126.5g/L,生產(chǎn)速率為15.81g/(L·h)。結(jié)果表明,已成功構(gòu)建高效表達(dá)天冬酰胺合成酶A基因工程菌株,并用于催化L-天冬氨酸轉(zhuǎn)化生產(chǎn)L-天冬酰胺,解決了L-天冬酰胺生物轉(zhuǎn)化生產(chǎn)工藝中ATP成本過(guò)高的難題,為L(zhǎng)-天冬酰胺制備提供新的綠色途徑。
[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.
【作者單位】： 中國(guó)食品發(fā)酵工業(yè)研究院中國(guó)工業(yè)微生物菌種保藏管理中心;山東省富馬酸生物轉(zhuǎn)化工程技術(shù)研究中心;
【基金】：煙臺(tái)市科技發(fā)展計(jì)劃資助項(xiàng)目(2014SF151)
【分類號(hào)】：Q78;TQ929

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本文編號(hào)：1749077