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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 (?CAT8) strain, while the fermentative pathway genes were significantly upregulated. The respiratory activities in the ?CAT8 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 ?CAT8 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 ?CAT8 strain. A further transcriptional comparison between ?CAT8 and ?HAP4 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.

SUBMITTER: Qi K

PROVIDER: S-EPMC4054228 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature

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Triggering respirofermentative metabolism in the crabtree-negative yeast Pichia guilliermondii by disrupting the CAT8 gene.

Qi Kai K Zhong Jian-Jiang JJ Xia Xiao-Xia XX

Applied and environmental microbiology 20140418 13

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 ...[more]

PMID: 24747899

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Project description:BackgroundChromium is one of the most used toxic heavy metals. A large amount of chromium waste is discharged into the environment every year, causing serious environmental pollution, especially the pollution of soil and water by hexavalent chromium. Eliminating hexavalent chromium is the primary challenge to achieve a pollution-free environment.ObjectivesThis study aims to understand the mechanism of Pichia guilliermondii's reduction of hexavalent chromium through enzymatic characteristic, oxidative stress response, and reduction product.Material and methodsThe strain Pichia guilliermondii ZJH-1 was isolated and stored in our laboratory. The hexavalent chromium uses 1,5-diphenyl carbazide method (DPC) to measure. The UV spectrophotometer was used to measure the intracellular antioxidant enzyme activity, and the kit was used to measure the activity of catalase and glutathione reductase. The reduction products were analyzed by ultraviolet full-wavelength scanning and FTIR.ResultsThe reduction of hexavalent chromium by ZJH-1 is accompanied by an increase in active oxygen and antioxidant levels. Chromate reductase mainly exists in the extracellular fluid, and the carboxyl, amide, hydroxide and other groups of the cell wall are involved in the bioremediation of Cr(VI) by complexing with Cr(VI) and Cr(III). After ZJH-1 was treated with different concentrations of Cr(VI), the expression of proteins with molecular weights of 15 kDa, 18 kDa, 35 kDa, 62 kDa, and 115 kDa increased significantly. This strain is the most suitable for chromate reductase (CChR). The optimum temperature is 40℃ and the optimum pH is 7.0. Cu2+ can enhance the activity of chromate reductase. At the optimum temperature and pH, the chromate reductase Km of this strain is 0.40 μmol and Vmax is 14.47 μmoL.L-1·min-1.ConclusionsThe bioremediation of Cr(VI) by Pichia guilliermondii ZJH-1 is attributable to the reduction product (Cr(III)) that can be removed in the precipitate and can be fixed on the cell surface and accumulated in the cell.

Project description:A study was conducted using 3 groups of gestating gilts and sows (n = 98) to determine the effects of Pichia guilliermondii (Pg), a whole cell-inactivated yeast product (CitriStim; ADM Alliance Nutrition), on performance and immune parameters of dams and litters. Within 24 h of breeding, gilts and sows were allotted to 1 of 3 treatments consisting of a control (SC) diet or SC diet supplemented with 0.1 (S1) or 0.2% (S2) Pg. Dietary treatments were maintained through lactation. Colostrum and milk (day 14) samples were collected for IgA, IgG, and IgM analysis. Blood samples were collected from sows on day 110 of gestation (group 3 only), while at weaning for all 3 groups, and from piglets at 14 d of age for peripheral white blood cell counts and serum IgA, IgG, and IgM analysis. Inclusion of Pg resulted in an increase in number born alive as the level of Pg increased (12.49, 13.33, and 13.43 born alive per litter for SC, S1, and S2, respectively; linear effect [LS], P = 0.003). Additionally, the percentage of piglets weighing less than 0.9 kg at birth was reduced in sows provided Pg at 0.1% or 0.2% compared with control (LS, P = 0.006). Sows receiving Pg during gestation and lactation also weaned a greater number of piglets (10.31, 10.55, and 10.60 weaned per litter in control, 0.1% and 0.2% Pg, respectively; LS, P = 0.02). However, percent preweaning mortality was 17.58%, 19.38%, and 19.61% for control, 0.1%, and 0.2% Pg, respectively (LS, P = 0.02). There were no differences in gestation BW gain, farrowing (days 110 to 48 h postfarrowing) or lactation (day 110 to weaning) BW loss, number of mummies or stillborn, or piglets' individual birth or weaning weight. On day 110 of gestation, the neutrophil concentration (quadratic effect [QS], P = 0.03) and neutrophil:lymphocyte ratio (QS, P = 0.04) in peripheral blood were greater in S1 than SC, with S2 being intermediate. At weaning there was a linear increase in neutrophil concentration (P = 0.03), neutrophil:lymphocyte ratio (P = 0.01), and percentage of neutrophils in the leukocyte population (P = 0.01) as level of Pg increased in sow diets. In conclusion, Pg inclusion in sow diets linearly increased total number born alive and weaned, with no change in average birth or weaning weight, and decreased the number of lightweight pigs at birth. However, inclusion of Pg had no effect on immune parameters measured in milk, colostrum, or day 14 piglet serum, but increased the peripheral blood neutrophil concentration of gilts and sows.

Dataset Information

Triggering respirofermentative metabolism in the crabtree-negative yeast Pichia guilliermondii by disrupting the CAT8 gene.

Publications

Triggering respirofermentative metabolism in the crabtree-negative yeast Pichia guilliermondii by disrupting the CAT8 gene.

Similar Datasets

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