Seung-Won Jang

Characterization of the starch degradation activity from newly isolated Rhizopus oryzae WCS-1 and mixed cultures with Saccharomyces cerevisiae for efficient ethanol production from starch

A starch degrading fungi was isolated from nuruk, a Korean traditional starter culture made with wheat flour. This newly isolated fungi was identified as Rhizopus oryzae WCS-1 via internal transcribed spacer region sequencing. Enzymatic assays for starch degradation activities indicated an optimum temperature and pH of 55°C and pH 4.0, respectively, using partially purified enzymes from R. oryzae WCS-1. Mixed cultures with S. cerevisiae CEN-PK 2-1D, L2612, BY4742, and D452-2 were performed. S. cerevisiae CEN-PK 2-1D has a maltose utilization pathway and showed the highest ethanol production rate with low maltose concentrations. Addition of 5 mM NaCl to mixed cultures of R. oryzae WCS-1 and S. cerevisiae CEN-PK 2-1D resulted in 21 and 33% improvements in the starch consumption rate and the ethanol production rate, respectively, because Na+ stimulated the starch degradation activity of partially purified enzymes from R. oryzae WCS-1.

Sequence analysis of KmXYL1 genes and verification of thermotolerant enzymatic activities of xylose reductase from four Kluyveromyces marxianus strains

Kluyveromyces marxianus has the capability of producing xylitol from xylose because of the endogenous xylose reductase (KmXYL1) gene. In this study, we cloned KmXYL1 genes and compared amino acid sequences of xylose reductase (XR) from four K. marxianus strains (KCTC 7001, KCTC 7155, KCTC 17212, and KCTC 17555). Four K. marxianus strains showed high homologies (99%) of amino acid sequences with those from other reported K. marxianus strains and around 60% homologies with that from Scheffersomyces stipitis. For XR enzymatic activities, four K. marxianus strains exhibited thermostable XR activities up to 45°C and K. marxianus KCTC 7001 showed the highest XR activity. When reaction temperatures were increased from 30 to 45°C, NADH-dependent XR activity from K. marxianus KCTC 7001 was highly increased (46%). When xylitol fermentations were performed at 30 or 45°C, four K. marxianus strains showed very poor xylitol production capabilities regardless fermentation temperatures. Xylitol productions from four K. marxianus strains might be limited because of low xylose uptake rate or cell growth although they have high thermostable XR activities.