1.Metabolic engineering for improving ethanol fermentation of xylose by wild yeast.
Lingyan ZHANG ; Liang ZHANG ; Zhongyang DING ; Zhengxiang WANG ; Guiyang SHI
Chinese Journal of Biotechnology 2008;24(6):950-956
One yeast strain, which was isolated from 256 natural samples, was found to be able to utilize D-xylose effectively. On the basis of assimilation physiological and molecular biological tests, the yeast strain was identified as a strain of Candida tropicalis. Furthermore, metabolic engineering breeding strategy was applied to change the metabolic flux in order to increase ethanol productivity. In this study, the C. tropicalis was used as the host strain and the plasmid pYX212-XYL2, which was formerly constructed for over expression of XYL2 gene encoding xylitol dehydrogenase (XDH) from Pichia stipitis, was used as the backbone of the recombinant vector. A hygro gene was inserted into downstream position of XYL2 gene, meanwhile, the result plasmid pXY212-XYL2-Hygro transformed into C. tropicalis by electroporation. Thus, a recombinant yeast C. tropicalis XYL2-7 was obtained through hygromycin B resistance screening and its specific XDH activity was 0.5 u/mg protein, which was 3 times more than that of the parent strain. Additionally, the recombinant yeast was applied in the fermentation of xylose. Compared with the parent yeast, it was concluded that the xylitol yield in the broth decreased by 3 times, however, the ethanol yield increased by 5 times. The feasibility of ethanol production from xylose by C. tropicalis was firstly studied in this paper. These research results are helpful to advance the bioconversion of renewable resources (e. g. straw, wheat bran, and husk) to fuel ethanol.
Candida tropicalis
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genetics
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metabolism
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D-Xylulose Reductase
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genetics
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metabolism
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Electroporation
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Ethanol
;
metabolism
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Fermentation
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Pichia
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enzymology
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genetics
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Recombination, Genetic
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Xylose
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metabolism
2.Urine metabolomic analysis to detect metabolites associated with the development of contrast induced nephropathy.
Deborah B DIERCKS ; Kelly P OWEN ; Jeffrey A KLINE ; Mark E SUTTER
Clinical and Experimental Emergency Medicine 2016;3(4):204-212
OBJECTIVE: Contrast induced nephropathy (CIN) is a result of injury to the proximal tubules. The incidence of CIN is around 11% for imaging done in the acute care setting. We aim to analyze the metabolic patterns in the urine, before and after dosing with intravenous contrast for computed tomography (CT) imaging of the chest, to determine if metabolomic changes exist in patients who develop CIN. METHODS: A convenience sample of high risk patients undergoing a chest CT with intravenous contrast were eligible for enrollment. Urine samples were collected prior to imaging and 4 to 6 hours post imaging. Samples underwent gas chromatography/mass spectrometry profiling. Peak metabolite values were measured and data was log transformed. Significance analysis of microarrays and partial least squares was used to determine the most significant metabolites prior to CT imaging and within subject. Analysis of variance was used to rank metabolites associated with temporal change and CIN. CIN was defined as an increase in serum creatinine level of ≥ 0.5 mg/dL or ≥ 25% above baseline within 48 hours after contrast administration. RESULTS: We sampled paired urine samples from 63 subjects. The incidence of CIN was 6/63 (9.5%). Patients without CIN had elevated urinary citric acid and taurine concentrations in the pre-CT urine. Xylulose increased in the post CT sample in patients who developed CIN. CONCLUSION: Differences in metabolomics patterns in patients who do and do not develop CIN exist. Metabolites may be potential early identifiers of CIN and identify patients at high-risk for developing this condition prior to imaging.
Citric Acid
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Creatinine
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Humans
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Incidence
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Least-Squares Analysis
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Metabolomics*
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Spectrum Analysis
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Taurine
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Thorax
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Tomography, X-Ray Computed
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Xylulose