Related Experiment Video
Updated: Jun 10, 2026

High-throughput Yeast Plasmid Overexpression Screen
Published on: July 27, 2011
Screening of a thiamine-auxotrophic yeast for alpha-ketoglutaric acid overproduction
Jingwen Zhou1, Haiyan Zhou, Guocheng Du
1State Key Laboratory of Food Science and Technology, Jiangnan University, Lihu Road, Wuxi, Jiangsu, China.
Aims:
To obtain a thiamine-auxotrophic yeast strain that overproduces alpha-ketoglutaric acid (alpha-KG) from glycerol and to investigate nutrient effects on alpha-KG production.
Methods And Results:
Yeast strain WSH-Z06, a thiamine auxotroph that gave high yields of alpha-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 alpha-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 alpha-KG accumulation. Under optimal conditions, a yield of 39.2 g l(-1)alpha-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 alpha-KG overproducer. Nutrients have a significant impact on alpha-KG production in the yeast strain presented here. The alpha-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.
More Related Videos
12:12Transformation of Probiotic Yeast and Their Recovery from Gastrointestinal Immune Tissues Following Oral Gavage in Mice
Published on: February 8, 2016
11:57Saccharomyces cerevisiae Models of Alzheimer's Disease to Screen Genes, Mutations, and Chemicals Affecting Amyloid Beta Production by γ-Secretase
Published on: June 24, 2025
Related Concept Videos
Bioreactor Controls-III
Production of Alcohol