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首页> 外国专利> A method for the biotechnological production of citric acid with a genetically modified yeast yarrowia lipolytica

A method for the biotechnological production of citric acid with a genetically modified yeast yarrowia lipolytica

机译：用转基因酵母解脂耶氏酵母生物技术生产柠檬酸的方法

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摘要

A method for the biotechnological production of citric acid in a product ratio citric acid / isocitric acid of at least 94: 6 in yeast cells of yarrowia lipolytica, with the steps of:a) transformation of yarrowia lipolytica - cells with a for the multiple genomic integration suitable vector of an expression cassette with a for the enzyme isocitrate lyase and a coding sequence, which acts as a selection marker gene,b) selecting yarrowia lipolytica cells, the at least 10 additional copies of the for the enzyme isocitrate lyase and coding sequence of the selective agent stable in their genome have integrated, in a suitable nutrient medium,c) cultivation of yarrowia lipolytica - cells, the at least 10 additional copies of the for the enzyme isocitrate lyase stable in their genome coding sequence have integrated, under nitrogen -, phosphorus -, sulfur -, magnesium - or in the case of the use of auxotrophic strains under amino acid deficiency and simultaneously excess of a usable carbon source,d) purification of the citric acid.
机译：一种在解脂耶氏酵母酵母细胞中柠檬酸/异柠檬酸产物比为至少94：6的柠檬酸的生物技术生产方法，该方法包括以下步骤：a）转化解脂耶氏酵母-具有用于多基因组的α整合表达盒的合适载体，其具有用于异柠檬酸裂合酶的酶和编码序列，其用作选择标记基因，b）选择解脂耶氏酵母细胞，用于异柠檬酸裂合酶的至少10个另外的拷贝和编码序列在适当的营养培养基中整合了在其基因组中稳定的选择剂的一部分，c）在解脂耶氏酵母-细胞中，在氮下整合了在其基因组编码序列中稳定的异柠檬酸裂解酶的至少10个额外拷贝-，磷-，硫-，镁-或在氨基酸缺乏且同时过量使用可用碳酸的情况下使用营养缺陷型菌株的情况ce，d）柠檬酸的纯化。

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