1.Bacteriocin from Purple Nonsulfur Phototrophic Bacteria, Rhodobacter capsulatus.
Sang Seob LEE ; Tae Jung OH ; Jaisoo KIM ; Jong Bae KIM ; Hyun Soon LEE
Journal of Bacteriology and Virology 2009;39(4):269-276
To find whether productivity of bacteriocin is controlled between different species under unusual cultural conditions, we used Rhodobacter capsulatus ATCC 17016 as a producer and Rhodopseudomonas palustris ATCC 17003 as an indicator. Rhodobacter capsulatus was cultured under aerobic conditions in the dark in Lascelles medium containing 0.3% Triton X-100. As a result, bacteriocin productivity increased enormously. The optimal pH range of bacteriocin production was 6~7.8. Through partial purification of bacteriocin, the molecular weight was roughly estimated at 14 kDa. Plasmid had no influence on bacteriocin production by Rhodobacter capsulatus. Our findings indicate that culture conditions affect bacteriocin productivity between more distantly related species, and bacteriocin of Rhodobacter capsulatus is not encoded by a plasmid.
Bacteria
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Efficiency
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Hydrogen-Ion Concentration
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Molecular Weight
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Octoxynol
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Plasmids
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Rhodobacter
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Rhodobacter capsulatus
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Rhodopseudomonas
2.Engineering the C4 pathway of Corynebacterium glutamicum for efficient production of 5-aminolevulinic acid.
Lijun WANG ; Sihan YAN ; Taowei YANG ; Meijuan XU ; Xian ZHANG ; Minglong SHAO ; Huazhong LI ; Zhiming RAO
Chinese Journal of Biotechnology 2021;37(12):4314-4328
5-aminolevulinic acid (5-ALA) plays an important role in the fields of medicine and agriculture. 5-ALA can be produced by engineered Escherichia coli and Corynebacterium glutamicum. We systematically engineered the C4 metabolic pathway of C. glutamicum to further improve its ability to produce 5-ALA. Firstly, the hemA gene encoding 5-ALA synthase (ALAS) from Rhodobacter capsulatus and Rhodopseudomonas palustris were heterologously expressed in C. glutamicum, respectively. The RphemA gene of R. palustris which showed relatively high enzyme activity was selected. Screening of the optimal ribosome binding site sequence RBS5 significantly increased the activity of RphemA. The ALAS activity of the recombinant strain reached (221.87±3.10) U/mg and 5-ALA production increased by 14.3%. Subsequently, knocking out genes encoding α-ketoglutarate dehydrogenase inhibitor protein (odhI) and succinate dehydrogenase (sdhA) increased the flux of succinyl CoA towards the production of 5-ALA. Moreover, inhibiting the expression of hemB by means of sRNA reduced the degradation of 5-ALA, while overexpressing the cysteine/O-acetylserine transporter eamA increased the output efficiency of intracellular 5-ALA. Shake flask fermentation using the engineered strain C. glutamicum 13032/∆odhI/∆sdhA-sRNAhemB- RBS5RphemA-eamA resulted in a yield of 11.90 g/L, which was 57% higher than that of the original strain. Fed-batch fermentation using the engineered strain in a 5 L fermenter produced 25.05 g/L of 5-ALA within 48 h, which is the highest reported-to-date yield of 5-ALA from glucose.
Aminolevulinic Acid/metabolism*
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Corynebacterium glutamicum/metabolism*
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Fermentation
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Metabolic Engineering
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Rhodobacter capsulatus/enzymology*
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Rhodopseudomonas/enzymology*
3.In vitro assessment of gastrointestinal viability of two photosynthetic bacteria, Rhodopseudomonas palustris and Rhodobacter sphaeroides.
Xu-xia ZHOU ; Yuan-jiang PAN ; Yan-bo WANG ; Wei-fen LI
Journal of Zhejiang University. Science. B 2007;8(9):686-692
The objectives of this study were to assess the potential of two photosynthetic bacteria (PSB), Rhodopseudomonas palustris HZ0301 and Rhodobacter sphaeroides HZ0302, as probiotics in aquaculture. The viability of HZ0301 and HZ0302 in simulated gastric transit conditions (pH 2.0, pH 3.0 and pH 4.0 gastric juices) and in simulated small intestinal transit conditions (pH 8.0, with or without 0.3% bile salts) was tested. The effects of HZ0301 and HZ0302 on the viability and permeability of intestinal epithelial cell in primary culture of tilapias, Oreochromis nilotica, were also detected. All the treatments were determined with three replicates. The simulated gastric transit tolerance of HZ0301 and HZ0302 strains was pH-dependent and correspondingly showed lower viability at pH 2.0 after 180 min compared with pH 3.0 and pH 4.0. Both HZ0301 and HZ0302 were tolerant to simulated small intestine transit with or without bile salts in our research. Moreover, there was no significant difference (P>0.05) among three treatments including the control and the groups treated with HZ0301 or HZ0302 both in intestinal epithelial cell viability and membrane permeability, showing no cell damage. In summary, this study demonstrated that HZ0301 and HZ0302 had high capacity of upper gastrointestinal transit tolerance and were relatively safe for intestinal epithelial cells of tilapias.
Animals
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Gastrointestinal Tract
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microbiology
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Microbial Viability
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Phototrophic Processes
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Rhodobacter sphaeroides
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isolation & purification
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physiology
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Rhodopseudomonas
;
isolation & purification
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physiology
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Species Specificity
;
Tilapia
;
microbiology
4.Hydrogen photoproduction from acetate by Rhodopseudomonas palustris.
Su-Ping YANG ; Chun-Gui ZHAO ; Rui-Tian LIU ; Yin-Bo QU ; Xin-Min QIAN
Chinese Journal of Biotechnology 2002;18(4):486-491
Based on the characteristics of metabolism of photosynthetic bacteria and the major kinds of organic compounds produced in wastewater degradation, eleven kinds of organic compounds were chosen for hydrogen photoproduction using Rhodopseudomonas palustris Z strain. The maximal volumetric H2 productivity was obtained using acetate as the sole carbon source and electron donor. The kinetics of cell growth and H2 liberation, and the influences of several major limiting factors on photoevolution of H2 were examined using acetate as carbon source. It was shown that hydrogen production was partially correlated with cell growth. The medium composition of the preculture, the preculture time, and inoculation volume were confirmed to have big effects on hydrogen photoevolution. The time delay of H2 production was evidently shortened using the inoculum of late exponential growth phase or stationary phase using ammonium sulfate as nitrogen source or with the inoculum of middle exponential growth phase using glutamate as the nitrogen source. The identity of temperature and light intensity for H2 evolution and cell growth has significant potential application in the technology of splitting organic acid into H2 by photosynthetic bacteria. The concentrations of acetate and glutamate in the medium affected hydrogen photoevolution and cell growth significantly. The productivity of H2 increased with substrate concentrations when substrate concentrations of sodium acetate and sodium glutamate were lower than 70 mmol/L and 15 mmol/L, respectively. Hydrogen production was inhibited but the cell growth was faster when the concentration of sodium glutamate over 15 mmol/L due to forming free NH4+. The highest rate of hydrogen production was 19.4 mL.L-1.h-1 using 30 mmol/L of sodium acetate as hydrogen donor under the standard conditions, respectively. The optimal conditions for hydrogen production were 35-37 degrees C, 6000-8000 lx and pH 7.3-8.3. The effects of oxygen and inoculation volume on photoproduction of hydrogen were also discussed.
Acetates
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metabolism
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pharmacology
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Cell Division
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drug effects
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radiation effects
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Dose-Response Relationship, Drug
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Glutamic Acid
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metabolism
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pharmacology
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Hydrogen
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metabolism
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Hydrogen-Ion Concentration
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Light
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Oxygen
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pharmacology
;
Rhodopseudomonas
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drug effects
;
metabolism
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radiation effects
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Temperature
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Time Factors