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Construction and characterization of a Saccharomyces cerevisiae strain able to grow on glucosamine as sole carbon and nitrogen source

机译:酿酒酵母菌株的构建和鉴定该菌株能够以氨基葡萄糖为唯一碳源和氮源生长

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摘要

Saccharomyces cerevisiae can transport and phosphorylate glucosamine, but cannot grow on this amino sugar. While an enzyme catalyzing the reaction from glucosamine-6-phosphate to fructose-6-phosphate, necessary for glucosamine catabolism, is present in yeasts using N-acetylglucosamine as carbon source, a sequence homology search suggested that such an enzyme is absent from Saccharomyces cerevisiae. The gene YlNAG1 encoding glucosamine-6-phosphate deaminase from Yarrowia lipolytica was introduced into S. cerevisiae and growth in glucosamine tested. The constructed strain grew in glucosamine as only carbon and nitrogen source. Growth on the amino sugar required respiration and caused an important ammonium excretion. Strains overexpressing YlNAG1 and one of the S. cerevisiae glucose transporters HXT1, 2, 3, 4, 6 or 7 grew in glucosamine. The amino sugar caused catabolite repression of different enzymes to a lower extent than that produced by glucose. The availability of a strain of S. cerevisiae able to grow on glucosamine opens new possibilities to investigate or manipulate pathways related with glucosamine metabolism in a well-studied organism.
机译:酿酒酵母可以转运和磷酸化葡萄糖胺,但不能在这种氨基糖上生长。虽然使用N-乙酰氨基葡糖作为碳源的酵母中存在一种酶,该酶催化从6磷酸氨基葡萄糖到6磷酸果糖的反应,这是葡萄糖胺分解代谢所必需的,但序列同源性研究表明,酿酒酵母中不存在这种酶。 。将来自解脂耶氏酵母的编码氨基葡萄糖6-磷酸脱氨酶的基因Y1NAG1引入酿酒酵母中,并测试其在氨基葡萄糖中的生长。所构建的菌株在葡糖胺中生长,仅作为碳和氮源。氨基糖上的生长需要呼吸并引起重要的铵排泄。过表达Y1NAG1和啤酒糖酵母转运蛋白HXT1、2、3、4、6或7之一的菌株在葡糖胺中生长。氨基糖引起的不同酶的分解代谢阻遏作用的程度低于葡萄糖产生的作用。能够在葡糖胺上生长的酿酒酵母菌株的可用性为研究或操纵与研究透彻的生物体中的葡糖胺代谢相关的途径提供了新的可能性。

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