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Screening and evolution of a novel protist xylose isomerase from the termite Reticulitermes speratus for efficient xylose fermentation in Saccharomyces cerevisiae

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    • 바이오플라스틱
      1. 고무
      2. 플라스틱
      3. 기타
    • 바이오정밀화학
      1. 용매
      2. 화학제품
      3. 연료
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    • 화장품용 기능성소재
      1. 기능성
      2. 계면활성제⁄증점제
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논문

Screening and evolution of a novel protist xylose isomerase from the termite Reticulitermes speratus for efficient xylose fermentation in Saccharomyces cerevisiae

학술지

Biotechnology for biofuels

저자명

Katahira, Satoshi; Muramoto, Nobuhiko; Moriya, Shigeharu; Nagura, Risa; Tada, Nobuki; Yasutani, Noriko; Ohkuma, Moriya; Onishi, Toru; Tokuhiro, Kenro

초록

<P><B>Background</B></P><P>The yeast <I>Saccharomyces cerevisiae</I>, a promising host for lignocellulosic bioethanol production, is unable to metabolize xylose. In attempts to confer xylose utilization ability in <I>S. cerevisiae</I>, a number of xylose isomerase (XI) genes have been expressed heterologously in this yeast. Although several of these XI encoding genes were functionally expressed in <I>S. cerevisiae</I>, the need still exists for a <I>S. cerevisiae</I> strain with improved xylose utilization ability for use in the commercial production of bioethanol. Although currently much effort has been devoted to achieve the objective, one of the solutions is to search for a new XI gene that would confer superior xylose utilization in <I>S. cerevisiae</I>. Here, we searched for novel XI genes from the protists residing in the hindgut of the termite <I>Reticulitermes speratus</I>.</P><P><B>Results</B></P><P>Eight novel XI genes were obtained from a cDNA library, prepared from the protists of the <I>R. speratus</I> hindgut, by PCR amplification using degenerated primers based on highly conserved regions of amino acid sequences of different XIs. Phylogenetic analysis classified these cloned XIs into two groups, one showed relatively high similarities to <I>Bacteroidetes</I> and the other was comparatively similar to <I>Firmicutes</I>. The growth rate and the xylose consumption rate of the <I>S. cerevisiae</I> strain expressing the novel XI, which exhibited highest XI activity among the eight XIs, were superior to those exhibited by the strain expressing the XI gene from <I>Piromyces</I> sp. E2. Substitution of the asparagine residue at position 337 of the novel XI with a cysteine further improved the xylose utilization ability of the yeast strain. Interestingly, introducing point mutations in the corresponding asparagine residues in XIs originated from other organisms, such as <I>Piromyces</I> sp. E2 or <I>Clostridium phytofermentans</I>, similarly improved xylose utilization in <I>S. cerevisiae</I>.</P><P><B>Conclusions</B></P><P>A novel XI gene conferring superior xylose utilization in <I>S. cerevisiae</I> was successfully isolated from the protists in the termite hindgut. Isolation of this XI gene and identification of the point mutation described in this study might contribute to improving the productivity of industrial bioethanol.</P><P><B>Electronic supplementary material</B></P><P>The online version of this article (doi:10.1186/s13068-017-0890-1) contains supplementary material, which is available to authorized users.</P>

발행연도

2017

발행기관

BioMed Central

라이선스

cc-by

ISSN

1754-6834

10

페이지

pp.203

주제어

Xylose isomerase; Termite; Cloning and functional expression of XI gene; Saccharomyces cerevisiae; Xylose fermentation; Ethanol production; Mutagenesis

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1 2023-12-11
2 2023-12-11
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논문; 2017-08-23

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