Abstract
A gene encoding a putative UDP-glucose 4-epimerase (pGALE) in Pyrococcus horikoshii was cloned and expressed in Escherichia coli. The purified enzyme could reversibly catalyze both the synthesis of UDP-Gal and UDP-Glc but preferred the binding of UDP-Gal by approximately 10-fold. The optimum pH and temperature were 6.5 and 65°C. The enzyme acted effectively without the addition of nicotinamide adenine dinucleotide (NAD +), possibly due to the presence of tightly bound NAD +. In particular, pGALE could be coupled with trehalose synthase (TreT) from P. horikoshii to regenerate UDP-Gal from UDP. The possible byproduct of glycosyltransferase, UDP, was capable of being converted to UDP-Glc with trehalose by TreT, and UDP-Glc was simultaneously converted to UDP-Gal by pGALE. Conclusively, the results suggest that pGALE and TreT with trehalose is an effective one-pot two-enzyme system for the regeneration of UDP-Gal, a high-cost substrate of galactosyltransferase, to complete a sugar nucleotide cycle.
Original language | English |
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Pages (from-to) | 423-429 |
Number of pages | 7 |
Journal | Bioresource technology |
Volume | 110 |
DOIs | |
Publication status | Published - 2012 Apr |
Bibliographical note
Funding Information:This work was supported by the Marine and Extreme Genome Research Center Program, Ministry of Maritime Affairs and Fisheries, Republic of Korea.
All Science Journal Classification (ASJC) codes
- Bioengineering
- Environmental Engineering
- Renewable Energy, Sustainability and the Environment
- Waste Management and Disposal