ACETYL-COA PRODUCING ENZYMES IN YEAST
    1.
    发明申请
    ACETYL-COA PRODUCING ENZYMES IN YEAST 审中-公开
    在乙烯生产酶的乙酰辅酶A

    公开(公告)号:US20140030730A1

    公开(公告)日:2014-01-30

    申请号:US14045683

    申请日:2013-10-03

    CPC classification number: C12Q1/32 C07K14/33 C12N9/001 C12P7/16 Y02E50/10

    Abstract: The present invention relates to a method of identifying a heterologous polypeptide having enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA in (the cytosol of) a yeast cell comprising: a) providing a mutated yeast cell comprising a deletion of at least one gene of the (PDH) by-pass, selected from the genes encoding the enzymes pyruvate decarboxylase (PDC), acetaldehyde dehydrogenase (ALD), and acetyl-CoA synthetase (ACS); b) transforming said mutated yeast cell with an expression vector comprising a heterologous nucleotide sequence encoding a candidate polypeptide having potential enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA; c) testing said recombinant mutated yeast cell for its ability to grow on minimal medium containing glucose as sole carbon source, and d) identifying said candidate polypeptide as a heterologous polypeptide having enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA in (the cytosol of) said yeast cell when growth of said cell is observed. The invention further relates to a method of producing a fermentation production such as butanol.

    Abstract translation: 本发明涉及一种鉴定具有用于将丙酮酸,乙醛或乙酸酯转化成酵母细胞的(细胞溶质)中的乙酰辅酶A的酶活性的异源多肽的方法,包括:a)提供包含至少一个缺失的突变的酵母细胞 选自编码丙酮酸脱羧酶(PDC),乙醛脱氢酶(ALD)和乙酰辅酶A合成酶(ACS)的基因的(PDH)旁路的一个基因; b)用包含异源核苷酸序列的表达载体转化所述突变的酵母细胞,所述异源核苷酸序列编码具有将丙酮酸,乙醛或乙酸转化为乙酰辅酶A的潜在酶活性的候选多肽; c)测试所述重组突变酵母细胞在含有葡萄糖作为唯一碳源的基本培养基上生长的能力,以及d)将所述候选多肽鉴定为异源多肽,其具有将丙酮酸,乙醛或乙酸酯转化成乙酰辅酶A 当所述细胞的生长被观察时,所述酵母细胞的胞质溶胶。 本发明还涉及生产发酵生产如丁醇的方法。

    METABOLIC ENGINEERING OF XYLOSE-FERMENTING EUKARYOTIC CELLS
    2.
    发明申请
    METABOLIC ENGINEERING OF XYLOSE-FERMENTING EUKARYOTIC CELLS 审中-公开
    XYLOSE发酵的真核细胞的代谢工程

    公开(公告)号:US20150010943A1

    公开(公告)日:2015-01-08

    申请号:US14313025

    申请日:2014-06-24

    CPC classification number: C12P7/06 C12N1/18 C12N9/90 C12P7/00 C12P7/08 Y02E50/17

    Abstract: The present invention relates to genetic modifications in eukaryotic host cells that have been transformed to express a xylose isomerase that confers on the host cell the ability to isomerize xylose to xylulose. These genetic modifications are aimed at improving the efficiency of xylose metabolism and include, e.g., reduction of nonspecific aldose reductase activity, increased xylulose kinase activity and increased flux of the pentose phosphate pathway. The modified host cells of the invention are suitable for the production of a wide variety of fermentation products, including ethanol, in fermentation processes in which a source of xylose or a source of xylose and glucose are used as carbon source.

    Abstract translation: 本发明涉及真核宿主细胞中的遗传修饰,其已被转化以表达木糖异构酶,赋予宿主细胞将木糖异构化成木酮糖的能力。 这些遗传修饰旨在提高木糖代谢的效率,并且包括例如降低非特异性醛糖还原酶活性,增加的木酮糖激酶活性和增加戊糖磷酸途径的通量。 本发明的修饰的宿主细胞适用于在发酵过程中生产各种各样的发酵产物,包括乙醇,其中木糖来源或木糖和葡萄糖来源用作碳源。

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