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Xylitol production is increased by expression of codon-optimized Neurospora crassa xylose reductase gene in Candida tropicalis

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논문

Xylitol production is increased by expression of codon-optimized Neurospora crassa xylose reductase gene in Candida tropicalis

학술지

Bioprocess and biosystems engineering

저자명

Jeon, Woo Young; Yoon, Byoung Hoon; Ko, Byoung Sam; Shim, Woo Yong; Kim, Jung Hoe

초록

<P>Xylose reductase (XR) is the first enzyme in <SMALL>D</SMALL>-xylose metabolism, catalyzing the reduction of <SMALL>D</SMALL>-xylose to xylitol. Formation of XR in the yeast <I>Candida tropicalis</I> is significantly repressed in cells grown on medium that contains glucose as carbon and energy source, because of the repressive effect of glucose. This is one reason why glucose is not a suitable co-substrate for cell growth in industrial xylitol production. XR from the ascomycete <I>Neurospora crassa</I> (NcXR) has high catalytic efficiency; however, NcXR is not expressed in <I>C</I>. <I>tropicalis</I> because of difference in codon usage between the two species. In this study, NcXR codons were changed to those preferred in <I>C</I>. <I>tropicalis</I>. This codon-optimized NcXR gene (termed NXRG) was placed under control of a constitutive glyceraldehyde-3-phosphate dehydrogenase (GAPDH) promoter derived from <I>C</I>. <I>tropicalis</I>, and integrated into the genome of xylitol dehydrogenase gene (<I>XYL2</I>)-disrupted <I>C</I>. <I>tropicalis</I>. High expression level of NXRG was confirmed by determining XR activity in cells grown on glucose medium. The resulting recombinant strain, LNG2, showed high XR activity (2.86 U (mg of protein)<SUP>&#x2212;1</SUP>), whereas parent strain BSXDH-3 showed no activity. In xylitol fermentation using glucose as a co-substrate with xylose, LNG2 showed xylitol production rate 1.44&nbsp;g&nbsp;L<SUP>&#x2212;1</SUP>&nbsp;h<SUP>&#x2212;1</SUP> and xylitol yield of 96% at 44&nbsp;h, which were 73 and 62%, respectively, higher than corresponding values for BSXDH-3 (rate 0.83&nbsp;g&nbsp;L<SUP>&#x2212;1</SUP>&nbsp;h<SUP>&#x2212;1</SUP>; yield 59%).</P>

발행연도

2012

발행기관

Springer-Verlag

라이선스

cc-by-nc

ISSN

1615-7591

ISSN

1615-7605

35

1

페이지

pp.191-198

주제어

Xylitol; Candida tropicalis; Xylose reductase; Glucose repression; Codon optimization

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1 2023-12-11

논문; 2011-09-16

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