Department of Food and Nutrition, Brain Korea 21 Project, Yonsei University, Seoul 120-749, Republic of Korea.
Enzyme Microb Technol. 2013 Oct 10;53(5):359-63. doi: 10.1016/j.enzmictec.2013.07.009. Epub 2013 Aug 13.
Useful nucleoside diphosphate (NDP)-sugars and α-galacto-oligosaccharides were synthesized by recombinant Escherichia coli whole cells and compared to those produced by enzyme-coupling. Production yields of NDP-glucoses (Glcs) by whole cells harboring trehalose synthase (TS) were 60% for ADP-Glc, 82% for GDP-Glc, and 27% for UDP-Glc, based on NDP used. Yield of UDP-galactose (Gal) by the whole-cell harboring a UDP-Gal 4-epimerase (pGALE) was 26% of the quantity of UDP-Glc. α-Galacto-oligosaccharides, α-Gal epitope (Galα-3Galβ-4Glu) and globotriose (Galα-4Galβ-4Glu), were produced by the combination of three recombinant whole cells harboring TS, pGALE, and α-galactosyltransferase, with production yields of 48% and 54%, based on UDP, respectively. Production yields of NDP-sugars and α-galacto-oligosaccharides by recombinant whole-cell reactions were approximately 1.5 times greater than those of enzyme-coupled reactions. These results suggest that a recombinant whole-cell system using cells harboring TS with trehalose as a substrate may be used as an alternative and practical method for the production of NDP-sugars and α-galacto-oligosaccharides.
利用重组大肠杆菌全细胞合成了有用的核苷二磷酸（NDP）-糖和α-半乳糖低聚糖，并将其与酶偶联法生产的产物进行了比较。携带海藻糖合成酶（TS）的全细胞生产 NDP-葡萄糖（Glc）的产率分别为 ADP-Glc 的 60%、GDP-Glc 的 82%和 UDP-Glc 的 27%，基于所用的 NDP。携带 UDP-Gal 4-差向异构酶（pGALE）的全细胞生产 UDP-半乳糖（Gal）的产率为 UDP-Glc 量的 26%。通过组合携带 TS、pGALE 和 α-半乳糖基转移酶的三个重组全细胞，生产了α-半乳糖低聚糖、α-半乳糖表位（Galα-3Galβ-4Glu）和Globotriose（Galα-4Galβ-4Glu），基于 UDP 的产率分别为 48%和 54%。重组全细胞反应生产 NDP-糖和α-半乳糖低聚糖的产率约为酶偶联反应的 1.5 倍。这些结果表明，使用携带海藻糖作为底物的 TS 的重组全细胞系统可能作为生产 NDP-糖和α-半乳糖低聚糖的替代和实用方法。
