Dataset Information

Prospecting for L-arabinose/D-xylose symporters from Pichia guilliermondii and Aureobasidium leucospermi.

ABSTRACT: With the strong trend toward sustainable technologies, such as the gradual substitution of fossil fuel consumption, improvement in the utilization of sugars from lignocellulosic biomass appears to be an alternative for bioenergy. However, from a number of C5 sugars, few are used in fermentative processes for ethanol production. One of the reasons is because wild-type Saccharomyces cerevisiae is unable to efficiently co-utilize hexoses and pentoses via specific transporters for each type of sugar. Thus, a system of pentose uptake that is not modulated by D-glucose is required. Here, we were able to identify the presence of sugar/H+ symporters for D-xylose and L-arabinose, especially for Pichia guilliermondii, where an uptake of D-glucose via symporter was not detected. The best D-xylose uptake route in P. guilliermondii exhibited a KM of 48 mM and VMAX of 0.48 mmol h-1 g-1 at the early stationary phase (24 h). For L-arabinose, the best route of uptake exhibited a KM of 109 mM and VMAX of 0.8 mmol h-1 g-1 on log phase (12 h). The highest kinetic uptake was observed when the final pH of the medium was below 7. In general, an alkaline medium limited the expression of symporters. The results obtained in this study will help in the further investigation of these symporters through their overexpression in engineered S. cerevisiae.

SUBMITTER: da Silva RR

PROVIDER: S-EPMC7058768 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Prospecting for L-arabinose/D-xylose symporters from Pichia guilliermondii and Aureobasidium leucospermi.

da Silva Ronivaldo Rodrigues RR Prista Catarina C Dias Maria Conceição Loureiro MCL Boscolo Mauricio M da Silva Roberto R Gomes Eleni E

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] 20190904 1

With the strong trend toward sustainable technologies, such as the gradual substitution of fossil fuel consumption, improvement in the utilization of sugars from lignocellulosic biomass appears to be an alternative for bioenergy. However, from a number of C<sub>5</sub> sugars, few are used in fermentative processes for ethanol production. One of the reasons is because wild-type Saccharomyces cerevisiae is unable to efficiently co-utilize hexoses and pentoses via specific transporters for each ty ...[more]

PMID: 31486050

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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:UnlabelledGlucose is known to inhibit the transport and metabolism of many sugars in Escherichia coli. This mechanism leads to its preferential consumption. Far less is known about the preferential utilization of nonglucose sugars in E. coli. Two exceptions are l-arabinose and d-xylose. Previous studies have shown that l-arabinose inhibits d-xylose metabolism in Escherichia coli. This repression results from l-arabinose-bound AraC binding to the promoter of the d-xylose metabolic genes and inhibiting their expression. This mechanism, however, has not been explored in single cells. Both the l-arabinose and d-xylose utilization systems are known to exhibit a bimodal induction response to their cognate sugar, where mixed populations of cells either expressing the metabolic genes or not are observed at intermediate sugar concentrations. This suggests that l-arabinose can only inhibit d-xylose metabolism in l-arabinose-induced cells. To understand how cross talk between these systems affects their response, we investigated E. coli during growth on mixtures of l-arabinose and d-xylose at single-cell resolution. Our results showed that mixed, multimodal populations of l-arabinose- and d-xylose-induced cells occurred at intermediate sugar concentrations. We also found that d-xylose inhibited the expression of the l-arabinose metabolic genes and that this repression was due to XylR. These results demonstrate that a strict hierarchy does not exist between l-arabinose and d-xylose as previously thought. The results may also aid in the design of E. coli strains capable of simultaneous sugar consumption.ImportanceGlucose, d-xylose, and l-arabinose are the most abundant sugars in plant biomass. Developing efficient fermentation processes that convert these sugars into chemicals and fuels will require strains capable of coutilizing these sugars. Glucose has long been known to repress the expression of the l-arabinose and d-xylose metabolic genes in Escherichia coli. Recent studies found that l-arabinose also represses the expression of the d-xylose metabolic genes. In the present study, we found that d-xylose also represses the expression of the l-arabinose metabolic genes, leading to mixed populations of cells capable of utilizing l-arabinose and d-xylose. These results further our understanding of mixed-sugar utilization and may aid in strain design.

Dataset Information

Prospecting for L-arabinose/D-xylose symporters from Pichia guilliermondii and Aureobasidium leucospermi.

Publications

Prospecting for L-arabinose/D-xylose symporters from Pichia guilliermondii and Aureobasidium leucospermi.

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