Genomic and transcriptomic analysis of Candida intermedia reveals the genetic determinants for its xylose-converting capacity

Table 2 Kinetics of C. intermedia xylose reductases in crude cell extracts

	Specific activity (U (mg protein)⁻¹)
Construct	NADH	NADPH	Ratio NADH/NADPH
p426Ø	0.010 ± 0.079	0.067 ± 0.131	0.15
p426-SsXYL1_K270R	0.481 ± 0.046	0.721 ± 0.018	0.67
p426-CiXYL1	0.008 ± 0.008	0.351 ± 0.070	0.02
p426-CiXYL1_2	0.449 ± 0.016	0.456 ± 0.062	0.99
p426-CiXYL1_3	0.004 ± 0.128	0.312 ± 0.103	0.01

All of the enzymes were expressed in a S. cerevisiae strain deleted of the native aldose reductase GRE3. Thus, the basal cofactor reduction detected for the strain transformed with empty plasmid (0.010 and 0.067 U (mg protein⁻¹) for NADH and NADPH, respectively) is due to spontaneous NAD(P)H oxidation over time, and/or to background enzymatic activities in the crude protein extract. Mean values ± SD from two independent experiments are represented, including cell cultivation, protein extraction and activity quantification

ISSN: 2731-3654