A one-pot approach to bio-synthesize globotriose and its derivatives from simpler substrates

• Multi-enzyme one-pot system is designed to achieve globotriose on a large scale.
• We exploit a cheaper substrate, galactose, for donor UDP-galactose production.
• Globotriose derivatives are achieved by this one-pot system.
• Various donors can be obtained for LgtC by SpGalK and SpGalU.

Globotriose is involved in numerous pathogenic processes and drug development strategies. Recent studies have demonstrated that globotriosylceramide could be used in colon cancer therapy and as a crucial indicator for susceptibility to HIV-1 infection. Therefore, the cost-effective and facile approaches for large-scale production of globotiose as potential drugs are highly required. Here, a multi-enzyme one-pot system containing a galactokinase (SpGalK, E.C.2.7.1.6), a UDP-glucose pyrophosphorylase (SpGalU, E.C.2.7.7.9), a α-1,4-galactosyltransferase (LgtC, E.C. 2.4.1.44) and a commercial inorganic pyrophosphatase (PPase, EC 3.6.1.1) was designed to achieve globotriose on preparative scales. This method exploits a cheaper initial substrate, galactose, for donor UDP-galactose production. More importantly, the substrate specificity of SpGalK and SpGalU is highly promiscuous and various UDP-galactose derivatives obtained could be used as the donor substrates for LgtC. This pointcut of rapid preparation of globotriose derivatives is proposed for the first time. Finally, three globotriose analogs were achieved by this one-pot multi-enzyme system in our study.

Enzymatic synthesis of globotriose and its derivatives.Figure optionsDownload as PowerPoint slide