1.Effect of short-chain thioesterase deficiency on P(3HB-co-LA) biosynthesis in Escherichia coli.
Xiangju WEI ; Ju WU ; Pengye GUO ; Shengmin ZHOU ; Hui WU
Chinese Journal of Biotechnology 2021;37(1):196-206
Polyhydroxyalkanoates (PHAs) have obtained much attention in biomaterial fields due to their similar physicochemical properties to those of the petroleum-derived plastics. Poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)] is one member of the PHAs family, and has better toughness and transparency compared to existing polylactic acid (PLA) and poly[(R)-3-hydroxybutyrate] [P(3HB)]. First, we confirmed the one-step biosynthesis of P(LA-co-3HB) with the lactate fraction of 23.8 mol% by introducing P(3HB-co-LA) production module into Escherichia coli MG1655. Then, the lactate fraction was increased to 37.2 mol% in the dld deficient strain WXJ01-03. The genes encoding the thioesterases, ydiI and yciA, were further knocked out, and the lactate fraction in the P(3HB-co-LA) was improved to 42.3 mol% and 41.1 mol% respectively. Strain WXJ03-03 with dld, ydiI and yciA deficient was used for the production of the LA-enriched polymer, and the lactate fraction was improved to 46.1 mol%. Notably, the lactate fraction in P(3HB-co-LA) from xylose was remarkably higher than from glucose, indicating xylose as a potent carbon source for P(3HB-co-LA) production. Therefore, the deficiency of thioesterase may be considered as an effective strategy to improve the lactate fraction in P(3HB-co-LA) in xylose fermentation.
Escherichia coli/genetics*
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Hydroxybutyrates
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Lactic Acid
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Polyesters
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Polyhydroxyalkanoates
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Xylose
2.Progress in studies on production of chemicals from xylose by Saccharomyces cerevisiae.
Ming WANG ; Tao LUAN ; Jianzhi ZHAO ; Hongxing LI ; Xiaoming BAO
Chinese Journal of Biotechnology 2021;37(3):1042-1057
Effective utilization of xylose is a basis for economic production of biofuels or chemicals from lignocellulose biomass. Over the past 30 years, through metabolic engineering, evolutionary engineering and other strategies, the metabolic capacity of xylose of the traditional ethanol-producing microorganism Saccharomyces cerevisiae has been significantly improved. In recent years, the reported results showed that the transcriptome and metabolome profiles between xylose and glucose metabolism existed significant difference in recombinant yeast strains. Compared with glucose, the overall process of xylose metabolism exhibits Crabtree-negative characteristics, including the limited glycolytic pathway activity, which reduces the metabolic flux of pyruvate to ethanol, and the enhanced cytosolic acetyl-CoA synthesis and respiratory energy metabolism. These traits are helpful to achieve efficient synthesis of downstream products using pyruvate or acetyl-CoA as precursors. This review provides a detailed overview on the modification and optimization of xylose metabolic pathways in S. cerevisiae, the characteristics of xylose metabolism, and the construction of cell factories for production of chemicals using xylose as a carbon source. Meanwhile, the existed difficulties and challenges, and future studies on biosynthesis of bulk chemicals using xylose as an important carbon source are proposed.
Biofuels
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Ethanol
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Fermentation
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Metabolic Engineering
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Saccharomyces cerevisiae/genetics*
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Xylose
3.Evaluation of the Self-Testing Blood Glucose Monitoring System GlucoDr.S According to ISO 15197:2013 Guidelines.
Namhee KIM ; Bo Gyung KIM ; Sun Hee JUN ; Kyunghoon LEE ; Tae Jung OH ; Sung Hee CHOI ; Soo LIM ; Sang Hoon SONG ; Woon Heung SONG ; Junghan SONG ; Hak Chul JANG
Laboratory Medicine Online 2018;8(3):77-86
BACKGROUND: The performance of the self-monitoring of blood glucose in patients with diabetes should be properly evaluated to ensure strict glycemic control. This study evaluated the self-testing Blood Glucose Monitoring System GlucoDr.S™ (All Medicus Co., Ltd., Korea). METHODS: This study recruited 120 patients. Use of the glucometer was evaluated according to ISO 15197:2013 guidelines. The YSI 2300 STAT PLUS Glucose Analyzer (YSI Life Sciences, USA) was used as the reference device. RESULTS: The standard deviation and coefficients of variation ranges for measurement repeatability and intermediate measurement precision conducted with 10 meters and 3 reagent lots on the same day were 2.7–3.2 mg/dL (<100 mg/dL) and 3.4–3.7% (≥100 mg/dL), respectively, and 3.7 mg/dL (<100 mg/dL) and 2.1–2.6% (≥100 mg/dL), respectively. Each coefficient of determination (R2) for linearity of the 3 reagent lots was >0.99. The influence effect of hematocrit and the 24 interference agents was not significant, except for xylose. A system accuracy test was conducted with 100 subjects taking duplicate measurements from each of the 3 reagent lots. When glucose levels were <100 mg/dL and ≥100 mg/dL, >95% of the samples were within ±15 mg/dL and within ±15% of the average measured values of the reference measurement, respectively. In Consensus Error grid analysis, all results were distributed in zone A and B. The results of the user performance evaluation using 115 lay persons were also included in the acceptance range. CONCLUSION: The GlucoDr.S™ showed acceptable performance according to the ISO 15197:2013 guidelines and could be a clinically useful self-testing glucometer.
Biological Science Disciplines
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Blood Glucose*
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Consensus
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Glucose
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Hematocrit
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Humans
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Xylose
4.Optimization of xylose fermentation for ethanol production by Candida shehatae HDYXHT-01.
Jingping GE ; Guoming LIU ; Xiaofeng YANG ; Hongbing SUN ; Hongzhi LING ; Wenxiang PING
Chinese Journal of Biotechnology 2011;27(3):404-411
Plackett-Burman (PB) design and central composite design (CCD) were applied to optimize of xylose fermentation for ethanol production by Candida shehatae HDYXHT-01. The PB results showed that (NH4)2SO4, KH2PO4, yeast extract and inoculum volume were the main affecting factors. With ethanol productivity as the target response, the optimal fermentation was determined by CCD and response surface analysis (RSM). The optimal fermentation conditions were (NH4)2SO4 1.73 g/L, KH2PO4 3.56 g/L, yeast extract 2.62 g/L and inoculum volume 5.66%. Other fermentation conditions were xylose 80 g/L, MgSO47H20 0.1 g/L, pH 5.0 and 250 mL flask containing 100 mL medium and cultivated at 30 degrees C for 48 h and the agitation speed was 140 r/min. Under this fermentation conditions, ethanol productivity was 26.18 g/L, which was 1.15 times of the initial.
Candida
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metabolism
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Ethanol
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metabolism
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Fermentation
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Industrial Microbiology
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Xylose
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metabolism
5.Biocatalysis of formaldehyde to L-xylose.
Zhailin CHU ; Xiaoyun LU ; Yuwan LIU ; Bo CUI ; Meidong JING ; Huifeng JIANG
Chinese Journal of Biotechnology 2020;36(5):942-948
It is of great significance to use biosynthesis to transform the inorganic substance formaldehyde into organic sugars. Most important in this process was to find a suitable catalyst combination to achieve the dimerization of formaldehyde. In a recent report, an engineered glycolaldehyde synthase was reported to catalyze this reaction. It could be combined with engineered D-fructose-6-phosphate aldolase, a "one-pot enzyme" method, to synthesize L-xylose using formaldehyde and the conversion rate could reach up to 64%. This process also provides a reference for the synthesis of other sugars. With the increasing consumption of non-renewable resources, it was of great significance to convert formaldehyde into sugar by biosynthesis.
Biocatalysis
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Formaldehyde
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chemistry
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Fructose-Bisphosphate Aldolase
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metabolism
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Xylose
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chemical synthesis
6.Coconut-derived D-xylose affects postprandial glucose and insulin responses in healthy individuals.
Yun Jung BAE ; Youn Kyung BAK ; Bumsik KIM ; Min Sun KIM ; Jin Hee LEE ; Mi Kyung SUNG
Nutrition Research and Practice 2011;5(6):533-539
Metabolic alterations including postprandial hyperglycemia have been implicated in the development of obesity-related diseases. Xylose is a sucrase inhibitor suggested to suppress the postprandial glucose surge. The objectives of this study were to assess the inhibitory effects of two different concentrations of xylose on postprandial glucose and insulin responses and to evaluate its efficacy in the presence of other macronutrients. Randomized double-blind cross-over studies were conducted to examine the effect of D-xylose on postprandial glucose and insulin response following the oral glucose tolerance test (OGTT). In study 1, the overnight-fasted study subjects (n = 49) consumed a test sucrose solution (50 g sucrose in 130 ml water) containing 0, 5, or 7.5 g D-xylose powder. In study 2, the overnight-fasted study subjects (n = 50) consumed a test meal (50 g sucrose in a 60 g muffin and 200 ml sucrose-containing solution). The control meal provided 64.5 g of carbohydrates, 4.5 g of fat, and 10 g of protein. The xylose meal was identical to the control meal except 5 g of xylose was added to the muffin mix. In study 1, the 5 g xylose-containing solutions exhibited significantly lower area under the glucose curve (AUCg) and area under the insulin curve (AUCi) values for 0-15 min (P < 0.0001, P < 0.0001), 0-30 min (P < 0.0001, P < 0.0001), 0-45 min (P < 0.0001, P < 0.0001), 0-60 min (P < 0.0001, P < 0.0001), 0-90 min (P < 0.0001, P < 0.0001) and 0-120 min (P = 0.0071, P = 0.0016). In study 2, the test meal exhibited significantly lower AUCg and AUCi values for 0-15 min (P < 0.0001, P < 0.0001), 0-30 min (P < 0.0001, P < 0.0001), 0-45 min (P < 0.0001, P = 0.0005), 0-60 min (P = 0.0002, P = 0.0025), and 0-90 min (P = 0.0396, P = 0.0246). In conclusion, xylose showed an acute suppressive effect on the postprandial glucose and insulin surges.
Carbohydrates
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Cross-Over Studies
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Glucose
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Glucose Tolerance Test
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Hyperglycemia
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Insulin
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Meals
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Sucrase
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Sucrose
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Xylose
7.Inhibitory Effects of D-mannose on Streptococcus mutans in the Presence of Sucrose.
Journal of Bacteriology and Virology 2016;46(2):57-62
This study aimed to examine the inhibitory effect of rare sugars on Streptococcus mutans (S. mutans) in the presence of sucrose. Xylitol and three rare sugars (D-xylose, D-lyxose and D-mannose) were used in this study. S. mutans KCTC 3065 was cultured in Brain Heart Infusion (BHI) medium containing xylitol, D-xylose, D-lyxose, or D-mannose in the presence of sucrose, and the effect on S. mutans growth was assessed by measuring solution turbidity at different time points after inoculation. To assess effects on pH, sucrose was added at different concentrations, and solution pH was measured at different time points after inoculation. All sugars significantly inhibited the growth of S. mutans in the presence of sucrose. Especially, D-lyxose and D-mannose exhibited significantly greater inhibition than that of xylitol. Furthermore, unlike D-lyxose, D-mannose significantly inhibited the decrement of pH, and its effect was greater than that of xylitol. Taken together, D-mannose has strong inhibitory effect on S. mutans in the presence of sucrose.
Brain
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Carbohydrates
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Dental Caries
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Heart
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Hydrogen-Ion Concentration
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Mannose*
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Streptococcus mutans*
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Streptococcus*
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Sucrose*
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Xylitol
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Xylose
8.Microbial production of poly (glycolate-co-lactate-co-3-hydroxybutyrate) from glucose and xylose by Escherichia coli.
Yangyang DA ; Wei LI ; Lilong SHI ; Zhengjun LI
Chinese Journal of Biotechnology 2019;35(2):254-262
Escherichia coli was metabolically engineered to produce poly(glycolate-co-lactate-co-3-hydroxybutyrate) using glucose and xylose as carbon sources. The combinatorial biosynthetic route was constructed by the overexpression of a series of enzymes including D-tagatose 3-epimerase, L-fuculokinase, L-fuculose-phosphate aldolase, aldehyde dehydrogenase, propionyl-CoA transferase, β-ketothiolase, acetoacetyl-CoA reductase, and polyhydroxyalkanoate synthase. Overexpression of polyhydroxyalkanoate granule associated protein significantly improved biopolymer synthesis, and the recombinant strain reached 3.73 g/L cell dry weight with 38.72% (W/W) biopolymer content. A co-culture engineering strategy was developed to produce biopolymer from a mixture of glucose and xylose, achieving 4.01 g/L cell dry weight containing 21.54% (W/W) biopolymer. The results of this work offer an approach for simultaneously utilizing glucose and xylose and indicate the potential for future biopolymer production from lignocellulosic biomass.
3-Hydroxybutyric Acid
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Escherichia coli
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Glucose
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Glycolates
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Lactates
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Metabolic Engineering
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Polyesters
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Xylose
9.Composition and Partial Structure Characterization of Tremella Polysaccharides.
Mycobiology 2009;37(4):286-294
Heteropolysaccharides isolated from liquid cultures of nine Tremella species contained 0.3 to 1.2% protein, 2.7 to 5% ash, 0.9 to 3.4% acetyl groups, 76.5 to 84.2% carbohydrates and trace amounts of starch. The polysaccharides in aqueous solution were slightly acidic (pH 5.1 to 5.6). They consisted of the following monomeric sugars: fucose, ribose, xylose, arabinose, mannose, galactose, glucose and glucuronic acid. The backbones of the polysaccharide structures consisted of alpha-(1-->3)-links while the side chains were beta-linked.
Arabinose
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Carbohydrates
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Fucose
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Galactose
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Glucose
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Glucuronic Acid
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Mannose
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Polysaccharides
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Ribose
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Starch
;
Xylose
10.The Optimal Culture Conditions for the Mycelial Growth of Oudemansiella radicata.
Sang Beom KIM ; Seong Hwan KIM ; Kyung Rim LEE ; Jae Ouk SHIM ; Min Woong LEE ; Mi Ja SHIM ; U Youn LEE ; Tae Soo LEE
Mycobiology 2005;33(4):230-234
Oudemansiella radicata, one of edible mushrooms belonging to Tricholomataceae of Basidiomycota, has been known to exhibit outstanding therapeutic effects on the hypertension caused by high blood pressure and inhibitory effects on the sarcoma 180 and Erhrlich carcinoma of mice. As one of preliminary experiments for producing fruiting-body of O. radicata, this study was carried out to obtain the basic information for culture conditions of mycelial growth of the fungus. The optimal temperature and pH for the mycelial growth were 25degrees C and pH 6, respectively. The medium for favorable mycelial growth of O. radicata was shown in the Lilly medium, whereas compact mycelial density was found in Hamada medium. The carbon and nitrogen sources promoting for mycelial growth of O. radicata were xylose and alanine, respectively. The optimum C/N ratio was about 20 : 1 in case that 3% glucose was supplimented to the basal medium as a carbon source.
Agaricales
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Alanine
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Animals
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Basidiomycota
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Carbon
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Fungi
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Glucose
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Hydrogen-Ion Concentration
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Hypertension
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Mice
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Nitrogen
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Sarcoma 180
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Xylose