One-pot enzymatic synthesis of 2-deoxy-scyllo-inosose from d-glucose and polyphosphate

Abstract

2-Deoxy-scyllo-inosose (2DOI, [2S,3R,4S,5R]-2,3,4,5-tetrahydroxycyclohexan-1-one) is a biosynthetic intermediate of 2-deoxystreptamine-containing aminoglycoside antibiotics, including butirosin, kanamycin, and neomycin. In producer microorganisms, 2DOI is constructed from d-glucose 6-phosphate (G6P) by 2-deoxy-scyllo-inosose synthase (DOIS) with the oxidized form of nicotinamide adenine dinucleotide (NAD+). 2DOI is also known as a sustainable biomaterial for production of aromatic compounds and a chiral cyclohexane synthon. In this study, a one-pot enzymatic synthesis of 2DOI from d-glucose and polyphosphate was investigated. First, 3 polyphosphate glucokinases (PPGKs) were examined to produce G6P from d-glucose and polyphosphate. A PPGK derived from Corynebacterium glutamicum (cgPPGK) was found to be suitable for G6P production under ordinary enzymatic conditions. Next, 7 DOISs were examined for the one-pot enzymatic reaction. As a result, cgPPGK and BtrC, the latter of which is a DOIS derived from the butirosin producer Bacillus circulans, achieved nearly full conversion of d-glucose to 2DOI in the presence of polyphosphate.

Improved Synthesis of Furfurylamine from a High Titer of Biomass-Derived Furfural by a Thermostable Triple Mutant ω-Transaminase in a Three-Component Deep Eutectic Solvent ChCl/Lactic Acid/Malonic Acid System

“…Very recently, one-pot chemoenzymatic synthesis reactions have attracted extensive attention. This combined process can not only improve the reaction efficiency but also use less equipment . In this work, FLA was obtained from biomass-derived d -xylose in a combination conversion by sequential DES (ChCl/MA/LA) chemocatalysis and HNILQE cell biocatalysis.…”

“…This combined process can not only improve the reaction efficiency but also use less equipment. 47 In this work, FLA was obtained from biomass-derived D-xylose in a combination conversion by sequential DES (ChCl/MA/LA) chemocatalysis and HNILQE cell biocatalysis. In 50 mL of DES ChCl/MA/LA (15 wt %) at 190 °C, 2.252 g of D-xylose formed 211.83 mM FAL after 45 min (Figure 9).…”

Constructing an Intensified UDP Recycling System for the Glycosylation of Natural Products by Phosphorylation of Byproduct Fructose

“…CgPPGK can be stably expressed in E. coli with high activity toward glucose, so it was selected to construct the enzyme cascade reaction system. All of the four enzymes were successfully expressed in E.…”

Improving the production of carbamoyltobramycin by an industrial Streptoalloteichus tenebrarius through metabolic engineering

“…Streptoalloteichus tenebrarius Tb used in this study is an industrialized bacterium, mainly producing apramycin, carbamoyltobramycin, and a small amount of carbamoylkanamycin B. Based on the research regarding the aminoglycoside secondary metabolites with 2-deoxystreptamine structure, apramycin, tobramycin, and kanamycin B were all synthesized from the initial substrate D-glucose (Kudo et al 2021 ). The key intermediate paromamine was obtained through a series of reactions from D-glucose.…”