Highly active xylose reductase from
View Patent ↗A new xylose reductase encoding gene from Neurspora crassa was heterologously expressed in E. coli as a His-tag fusion protein and subsequently purified in high yield. This xylose reductase was shown to have a high turnover rate and catalytic efficiency, high stability at room temperature, broad pH profile, and a preference of NADPH over NADH. This enzyme is utilized in production of xylitol and other sugar alcohols such as sorbitol and also in the metabolic enhancement of organisms used for fermentation of plant biomass into ethanol.
1. A purified recombinant xylose reductase comprising the amino acid sequence of SEQ ID NO: 2.
2. The purified xylose reductase of claim 1 , wherein the xylose reductase is at least 95% pure.
3. The xylose reductase of claim 1 further characterized as:
(a) having a molecular weight of about 36 kDa;
(b) active with NADH and NADPH as cofactors with a preference for NADPH;
(c) having D-ribose, L-arabinase, D-galactase and D-glucose as substrates;
(d) having a pH optima in the range of about pH 4.5-pH 6.0;
(e) having a K m of 34 mM for xylose and 1.8 μM for NADPH; and
(f) stable at room temperature.
4. The xylose reductase of claim 1 , wherein the xylose reductase comprises a fusion protein.
5. The xylose reductase of claim 1 , wherein the xylose reductase is purified from a heterologous host.
6. The xylose reductase of claim 5 , wherein the heterologous host is selected from the group consisting of bacteria, yeast, and plants.
7. The xylose reductase of claim 5 , wherein the heterologous host is E. coli.