化学
磷酸二羟丙酮
生物化学
磷酸化
磷酸烯醇丙酮酸羧激酶
磷酸盐
大肠杆菌
PEP群易位
酶
激酶
ATP合酶
基因
作者
Bernhard Schoenenberger,Stefanie Kind,R Meier,Thorsten Eggert,Markus Obkircher,Roland Wohlgemuth
标识
DOI:10.1080/10242422.2019.1634694
摘要
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.
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