Several CoA transferases from Clostridium beijerinckii, C. perfringens and Klebsiella pneumoniae were examined for biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) in recombinant Escherichia coli XL1-Blue strain. The CB3819 gene and the CB4543 gene from C. beijerinckii, the pct gene from C. perfringens and the pct gene from K. pneumoniae, which encodes putative CoA transferase gene, respectively, was co-expressed with the Pseudomonas sp. MBEL 6-19 phaC1437 gene encoding engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 (PhaC1 Ps6-19 ) to examine its activity for the construction of key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli XL1-Blue expressing the phaC1437 gene and CB3819 gene synthesized poly(3-hydroxybutyrate) [P(3HB)] homopolymer to the P(3HB) content of 60.5 wt% when it was cultured in a chemically defined medium containing 20 g/L of glucose and 2 g/L of sodium 3-hydroxybutyrate. Expression of the phaC1437 gene and CB4543 gene in recombinant E. coli XL1-Blue also produced P(3HB) homopolymer to the P(3HB) content of 51.2 wt% in the same culture condition. Expression of the phaC1437 gene and the K. pneumoniae pct gene in recombinant E. coli XL1-Blue could not result in the production of PHAs in the same culture condition. However, the recombinant E. coli XL1-Blue expressing the phaC1437 gene and the C. perfringens gene could produce poly(3-hydroxybutyrate-co-lactate [P(86.4mol%3HB-co-13.7 mol%LA) up to the PHA content of 10.6 wt% in the same culture condition. Newly examined CoA transfereases in this study may be useful for the construction of engineered E. coli strains to produce PHA containing novel monomer such lactate.
: Bacillus amyloliquefaciens M27 was isolated from the cotton-waste compost for cultivation of oyster mushroom (Pleurotus ostreatus). B. amyloliquefaciens M27 is a biocontrol agent with antagonistic activities against a wide range of fungal pathogens. The aim of this work was to evaluate the possibility of exploiting antagonistic bacteria, B. amyloliquefaciens M27, in the biological control of the cucumber powdery mildew fungus, Podosphaera fusca. In greenhouse tests, the isolate was found to be very effective to control powdery mildew on cucumber leaves showing 4.0% diseased area, whereas diseased area in the control was 80.5%. The filtrate of the isolate cultured on MH and LB media were more effective for control of the disease than those cultured on TSB, NB, and KB media. When two, five, ten, 20, 50 and 100-fold diluted culture broth of isolate on LB media were treated, disease areas were 0%, 0%, 0%, 1.3%, 3.1% and 5.0%, respectively, whereas diseased area in the control was 60.0%. The filtrate of the isolate cultured on LB media was treated to cucumber plants on July, October and December just before the outbreak of the powdery mildew occurred. When 10-fold diluted filtrate of the isolate was treated, control efficacy was 88.9~98.9% in the treated seasons. The results showed that the culture filtrate of B. amyloliquefaciens M27 was very effective to control powdery mildew of cucumber.
Biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) was examined in recombinant Escherichia coli W strain from sucrose. The Pseudomonas sp. MBEL 6-19 phaC1437 gene and the Clostridium propionicum pct540 gene, which encode engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 (PhaC1 ) and engineered C. propionicum propionyl-CoA transferase (Pct Cp ), respectively, were expressed in E. coli W to construct key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)].The recombinant E. coli W expressing the phaC1437 gene and the pct540 gene could synthesize P(3HB-co-13mol%LA) up to the polymer content of 31.3 wt% when it was cultured in chemically defined MR medium containing 20 g/L of sucrose and 2 g/L of sodium 3-hydroxybutyrate. When Ralstonia eutropha phaAB genes were additionally expressed to provide 3-hydroxybutyrate-CoA (3HB-CoA) from sucrose, P(3HB-co-16mol%LA) could be synthesized from sucrose as a sole carbon source without supplement of sodium 3-hydroxybutyrate in culture medium, but the PHA content was decreased to 12.2 wt%. The molecular weight of P(3HB-co-16 mol%LA) synthesized in E. coli W using sucrose as carbon source were 1.53×10 4 (M n ) and 2.78×10 4 (M w ), respectively, which are not different from those that have previously been reported by other recombinant E. coli strains. Engineered E. coli strains developed in this study should be useful for the production of lactate-containing PHAs from sucrose, one of the most abundant and least expensive carbon sources.
Skipping breakfast is a risk factor closely related to metabolic syndrome and obesity. We analyzed the relationship between breakfast size, metabolic syndrome and obesity. The study included 5,548 adults who visited a health promotion center at Pusan National University from January to November of 2006. Subjects were divided into four groups according to breakfast size -skipper group (no breakfast), small intake group, medium intake group and large intake group. 959 (17.3%) of the 5548 subjects were included in the Skipper group. Intake of daily calories, proteins, carbohydrates and fats was the lowest in the Skipper group. Breakfast size increased linearly with an increased intake of daily calories, proteins, carbohydrates and fats. Body mass index (23.4 kg/m 2 ) and waist circumference (79.6 cm) were the lowest in the Small intake group. In the Small intake group, triglycerides, fasting plasma glucose, systolic blood pressure and diastolic blood pressure were the lowest, and high density lipoprotein cholesterol levels were the highest. The number of metabolic risk factors was the lowest in Small intake group. Odds ratio of metabolic syndrome (Odds ratio=0.612) was the lowest in Small intake group. Along with increasing breakfast size, the odds ratio also increased. In this study, breakfast size was found to influence metabolic risk factors. Skipping breakfast worsened metabolic risk factors, while a small breakfast size had a favorable effect on metabolic risk factors.
In order to select antagonists for biological control of downy mildew of cucumber, 126 bacteria were isolated from cucumber plants collected from several locations in Korea. Among them, Five isolates were selected as potential biocontrol agents of cucumber downy mildew using a leaf disc bioassay method. In preventive and curative effect tests, the isolate CC110 was found to be effective to control downy mildew on cucumber showing diseased area by 0% whereas that of control was 15.0~18.0%. A bacterium isolate CC110 was identified as Bacillus amyloliquefaciens subsp. plantarum based on phylogenetic analysis using gyrB gene sequence. The culture liquid of isolate CC110 in TSB media were more effective for the control of the disease than those cultured in LB, NB, and KB media in leaf disk bioassay. when undiluted liquid, two-fold, five-fold diluted culture broth, and undiluted liquid, two-fold, five-fold diluted filtrate of isolate CC110 in TSB media were treated, diseased area of cucumber powdery mildew were 0%, 3.0%, 8.0%, 0%, 4.0% and 7.0%, respectively, whereas diseased area in the control was 21.0%. In the cucumber seedling tests, when the culture broth of isolate CC110 in TSB media was treated, diseased area were 35.0%, whereas that of control was 82.0%. When B. amyloliquefaciens CC110 was treated four times at five-day interval in the vinylhouse test, the control effect of cucumber downy mildew was higher than that treated three at seven-day interval.
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