通过 LacC 催化的 D-塔格糖 6-磷酸化磷酸化高效生物催化合成 D-塔格糖 1,6-二磷酸,Biocatalysis and Biotransformation

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通过 LacC 催化的 D-塔格糖 6-磷酸化磷酸化高效生物催化合成 D-塔格糖 1,6-二磷酸
Biocatalysis and Biotransformation ( IF 1.4 ) Pub Date : 2019-07-10 , DOI: 10.1080/10242422.2019.1634694
Bernhard Schoenenberger ₁ , Stefanie Kind ₂ , Roland Meier ₁ , Thorsten Eggert ₂ , Markus Obkircher ₁ , Roland Wohlgemuth _{1,

3}

Affiliation

摘要 D-塔格糖1,6-二磷酸是一种重要的代谢物，可以通过经典的塔格糖6-磷酸途径或最近发现的塔格糖1-磷酸途径形成。手性代谢物天然存在于各种生物的半乳糖代谢中，由磷酸二羟基丙酮和 D-甘油醛 3-磷酸可逆缩合形成，而 D-塔格糖 6-磷酸是化学合成的，没有化学合成迄今为止已经描述了进一步磷酸化为 D-塔格糖 1,6-二磷酸的方法。因此，选择了 D-塔格糖 6-磷酸中 1-位的位点特异性生物催化磷酸化。来自乳酸乳球菌亚种的 lacC 基因。乳酸被合成，克隆到合适的表达载体中以允许在大肠杆菌中生产 LacC，即 D-塔格糖 6-磷酸激酶。随后，通过使用重组 D-塔格糖 6-磷酸激酶和磷酸烯醇丙酮酸/丙酮酸激酶系统进行 ATP 再生，开发了一种有效的生物催化合成方法，用于在其 C1 位磷酸化 D-塔格糖 6-磷酸。D-塔格糖 1,6-二磷酸的这种直接且可扩展的一步生物催化合成已成功放大到克级。图形概要通过使用重组 D-塔格糖 6-磷酸激酶和磷酸烯醇丙酮酸/丙酮酸激酶系统进行 ATP 再生，已开发出一种有效的生物催化合成方法，用于在其 C1 位磷酸化 D-塔格糖 6-磷酸。D-塔格糖 1,6-二磷酸的这种直接且可扩展的一步生物催化合成已成功放大到克级。图形概要通过使用重组 D-塔格糖 6-磷酸激酶和磷酸烯醇丙酮酸/丙酮酸激酶系统进行 ATP 再生，已开发出一种有效的生物催化合成方法，用于在其 C1 位磷酸化 D-塔格糖 6-磷酸。D-塔格糖 1,6-二磷酸的这种直接且可扩展的一步生物催化合成已成功放大到克级。图形概要

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Efficient biocatalytic synthesis of D-tagatose 1,6-diphosphate by LacC-catalysed phosphorylation of D-tagatose 6-phosphate

Abstract D-tagatose 1,6-diphosphate is an important metabolite which can be formed via the classical tagatose 6-phosphate pathway or via the tagatose 1-phosphate pathway discovered more recently. The chiral metabolite naturally occurs as an intermediate in the galactose metabolism of various organisms, where it is formed by a reversible condensation of dihydroxyacetone phosphate and D-glyceraldehyde 3-phosphate, whereas D-tagatose 6-phosphate has been synthesized chemically, no chemical synthetic method for further phosphorylation to the D-tagatose 1,6-diphosphate has been described so far. Therefore, a site-specific biocatalytic phosphorylation of the 1-position in D-tagatose 6-phosphate has been chosen. The lacC gene from Lactococcus lactis subsp. lactis was synthesized, cloned into an appropriate expression vector to allow for the production of LacC, the D-tagatose 6-phosphate kinase in Escherichia coli. Subsequently, an efficient biocatalytic synthesis has been developed for the phosphorylation of D-tagatose 6-phosphate at its C1-position by using the recombinant D-tagatose 6-phosphate kinase and the phosphoenolpyruvate/pyruvate kinase-system for ATP regeneration. This straightforward and scalable one-step biocatalytic synthesis of D-tagatose 1,6-diphosphate was successfully scaled up to the gram scale. Graphical Abstract

更新日期：2019-07-10

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