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- Title
Screening of a thiamine-auxotrophic yeast for α-ketoglutaric acid overproduction.
- Authors
Jingwen Zhou; Haiyan Zhou; Guocheng Du; Liming Liu; Jian Chen
- Abstract
Aims: To obtain a thiamine-auxotrophic yeast strain that overproduces α-ketoglutaric acid (α-KG) from glycerol and to investigate nutrient effects on α-KG production. Methods and Results: Yeast strain WSH-Z06, a thiamine auxotroph that gave high yields of α-KG from glycerol, was obtained by screening for ampicillin/kanamycin resistance and thiamine auxotrophy. The strain was identified as Yarrowia lipolytica based on physiological, chemical, and phylogenetic analysis. The ability of the strain to convert glycerol to α-KG was analysed by investigating the effects of nutritional factors, including thiamine, riboflavin, nitrogen sources, and calcium ion. Conclusions: Thiamine and calcium ion concentration had the greatest effect on α-KG accumulation. Under optimal conditions, a yield of 39·2 g l−1α-KG was obtained from 100 g l−1 glycerol, with 16·84 g l−1 pyruvate as a by-product. Significance and Impact of the Study: The current work provides a method for screening for an α-KG overproducer. Nutrients have a significant impact on α-KG production in the yeast strain presented here. The α-KG-overproducing yeast strain Y. lipolytica WSH-Z06 is a promising parent strain for further metabolic engineering to lower by-product accumulation and accelerate glycerol utilization.
- Subjects
THIAMINE-binding proteins; GLYCERIN; LEAVENING agents; CALCIUM ions; BACTERIAL metabolism; PROTEIN kinases
- Publication
Letters in Applied Microbiology, 2010, Vol 51, Issue 3, p264
- ISSN
0266-8254
- Publication type
Article
- DOI
10.1111/j.1472-765X.2010.02889.x