title: Triggering respirofermentative metabolism in the crabtree-negative yeast Pichia guilliermondii by disrupting the CAT8 gene.
abstract: Pichia guilliermondii is a Crabtree-negative yeast that does not normally exhibit respirofermentative metabolism under aerobic conditions, and methods to trigger this metabolism may have applications for physiological study and industrial applications. In the present study, CAT8, which encodes a putative global transcriptional activator, was disrupted in P. guilliermondii. This yeast's ethanol titer increased by >20-fold compared to the wild type (WT) during aerobic fermentation using glucose. A comparative transcriptional analysis indicated that the expression of genes in the tricarboxylic acid cycle and respiratory chain was repressed in the CAT8-disrupted (ΔCAT8) strain, while the fermentative pathway genes were significantly upregulated. The respiratory activities in the ΔCAT8 strain, indicated by the specific oxygen uptake rate and respiratory state value, decreased to one-half and one-third of the WT values, respectively. In addition, the expression of HAP4, a transcriptional respiratory activator, was significantly repressed in the ΔCAT8 strain. Through disruption of HAP4, the ethanol production of P. guilliermondii was also increased, but the yield and titer were lower than that in the ΔCAT8 strain. A further transcriptional comparison between ΔCAT8 and ΔHAP4 strains suggested a more comprehensive reprogramming function of Cat8 in the central metabolic pathways. These results indicated the important role of CAT8 in regulating the glucose metabolism of P. guilliermondii and that the regulation was partially mediated by repressing HAP4. The strategy proposed here might be applicable to improve the aerobic fermentation capacity of other Crabtree-negative yeasts.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Qi
authors.affiliation: State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai, China State Key Laboratory of Microbial Metabolism, and Laboratory of Molecular Biochemical Engineering, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China jjzhong@sjtu.edu.cn xiaoxiaxia@sjtu.edu.cn.
authors.forename: Jian-Jiang
authors.initials: JJ
authors.lastname: Zhong
authors.affiliation: State Key Laboratory of Microbial Metabolism, and Laboratory of Molecular Biochemical Engineering, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China jjzhong@sjtu.edu.cn xiaoxiaxia@sjtu.edu.cn.
authors.forename: Xiao-Xia
authors.initials: XX
authors.lastname: Xia
mesh_terms: D000431:Ethanol / Q000378:metabolism; D005285:Fermentation; D020869:Gene Expression Profiling; D055786:Gene Knockout Techniques; D005947:Glucose / Q000378:metabolism; D060847:Metabolic Engineering; D064688:Metabolic Flux Analysis; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*
chemical_list: D014157:Transcription Factors; D000431:Ethanol; D005947:Glucose; D010100:Oxygen
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