The electronic structure of protein chains L and M in photosynthetic reaction center (PRC) of Rhodobacter sphaeroides (Van Niel) Imhoff, Truper et Pfennig) was studied by using the Overlapping Dimer Approximation meth...The electronic structure of protein chains L and M in photosynthetic reaction center (PRC) of Rhodobacter sphaeroides (Van Niel) Imhoff, Truper et Pfennig) was studied by using the Overlapping Dimer Approximation method and the Extended Negative Factor Counter method at ab initio level. The result indicated that: (1) Amino acid residues, the molecular orbitals of which composed the main components of frontier orbitals of protein chain L (M), are located at the random coil areas of chain L (alpha helix areas of chain M). Since the random coil is flexible and more easy to change its conformation in the electron transfer process and to reduce the energy of the system, and the structure of the alpha helix is reletively stable, this difference might be one of the causes for the electron transfer in photosynthetic reaction center (PRC) only takes place along the L branch. (2) The His residues which axially coordinated to the 'special pair' P and accessory chlorophyll molecules (ABChls) are essentially important for the E-LUMO levels of P and ABChl. But, the corresponding molecular orbitals of these His residues do not appear in the composition of frontier orbitals of protein chains. It means that the interaction between pigment molecules and protein chains do not influence the contribution to the frontier orbitals of protein chains explicitly, but influences the corresponding E-LUMO levels significantly.展开更多
[Objective]The experiment aimed to explore a new way for observing surface structure of Nostoc sphaeroides Kutzing. [Method] The scanning electron microscope was used to observe the epidermal ultrastructure of wild an...[Objective]The experiment aimed to explore a new way for observing surface structure of Nostoc sphaeroides Kutzing. [Method] The scanning electron microscope was used to observe the epidermal ultrastructure of wild and cultured Nostoc sphaeroides Kutzing. [ Result] The epidermis of wild and cultured Nostoc sphaeroides Kutzing showed mixture structure of fibril colloid which was reticular arranged. The difference between wild and cultured Nostoc sphaeroides Kutzing was that the outer epidermis of cultured Nostoc sphaeroides Kutzing had trichome distribution but the wild Nostoc sphaeroides Kutzing did not has such distribution. The obsevation results of under smaller than 10 μm by scanning electron microscope was touched thick and showed many folds and distortions. [ Conclusion] The scanning electron microscope was an effective way to study development of Nostoc sphaeroides Kutzing colony and it was worth popularizing.展开更多
Bacillus thuringiensis/cereus L2 was added as a biostimulant to enhance the biomass accumulation and carotenoid yield of Rhodobacter sphaeroides using wastewater as the culturing medium. Results showed that biostimula...Bacillus thuringiensis/cereus L2 was added as a biostimulant to enhance the biomass accumulation and carotenoid yield of Rhodobacter sphaeroides using wastewater as the culturing medium. Results showed that biostimulation could significantly enhance the R. sphaeroides biomass production and carotenoid yield. The optimal biostimulant proportion was 40 μL（about 6.4 × 10^5CFU）. Through the use of biostimulation, chemical oxygen demand removal, R. sphaeroides biomass production, carotenoid concentration, and carotenoid yield were improved by 178%, 67%, 214%, and 70%, respectively. Theoretical analysis revealed that there were two possible reasons for such increases. One was that biostimulation enhanced the R. sphaeroides wastewater treatment efficiency. The other was that biostimulation significantly decreased the peroxidase activity in R. sphaeroides. The results showed that the highest peroxidase activity dropped by 87% and the induction ratio of the RSP_3419 gene was 3.1 with the addition of biostimulant. The enhanced carotenoid yield in R. sphaeroides could thus be explained by a decrease in peroxidase activity.展开更多
Coenzyme Q10 is widely used in food,cosmetics and pharmaceuticals,possessing a broad market.Rhodobacter sphaeroides is enriched in natural coenzyme Q10 and is becoming an important microorganism for producing natural ...Coenzyme Q10 is widely used in food,cosmetics and pharmaceuticals,possessing a broad market.Rhodobacter sphaeroides is enriched in natural coenzyme Q10 and is becoming an important microorganism for producing natural coenzyme Q10.The paper reviewed the biosynthesis pathways of coenzyme Q10 in R.sphaeroides and the advances in enhancement of coenzyme Q10 production in R.sphaeroides based on metabolic engineering.展开更多
This study aimed to increase bacterial growth and 5-aminolevulinic acid（ALA） biosynthesis of Rhodobacter sphaeroides in wastewater treatment through adding ferrous ion（ Fe2＋ ）. Results demonstrated that Fe2＋ eff...This study aimed to increase bacterial growth and 5-aminolevulinic acid（ALA） biosynthesis of Rhodobacter sphaeroides in wastewater treatment through adding ferrous ion（ Fe2＋ ）. Results demonstrated that Fe2＋ effectively enhanced the biomass production and ALA yield of R. sphaeroides. Moreover, the optimal Fe2＋ dosage was found to be 400 μmol/L, which was associated with the highest biomass of 4015.3 mg/L and maximum ALA yield of 15.9 mg/g-dry cell weight（mg/g-DCW）. Mechanism analysis revealed that Fe2＋ vastly improved Adenosine Triphosphate（ATP） production by up-regulating the nif gene expression, and increasing ATP enhanced the biomass and ALA yield by supplying energy for bacterial growth and ALA biosynthesis, respectively. Correlation analysis showed that the ALA and ATP yields had positive relation with nifA and nifU gene expression. In addition, the nifA and nifU gene expression displayed high consistency of co-transcription at the optimal Fe2＋ dosage.展开更多
In the present study,single factors including inoculation amount,fermentation temperature,fermentation duration and ratio of fermentation medium volume to total flask volume were optimized for enhancing the production...In the present study,single factors including inoculation amount,fermentation temperature,fermentation duration and ratio of fermentation medium volume to total flask volume were optimized for enhancing the production of coenzyme Q10 from genetic engineered Rhodobacter sphaeroides overexpressing UbiE.The experimental results suggested that optimal single factors were: inoculation amount 2%,fermentation temperature 30 ℃,fermentation duration 48 h,and fermentation medium volume to total flask volume 80%.The present study will promote the large scale production of CoQ10 from microorganisms.展开更多
Rhodobacter sphaeroides is a purple non-sulfur bacterium that belongs to the α-3 subdivision of Proteobacteria. R. sphaeroides is a model bacterial species because of its complex genome structure and expanded metabol...Rhodobacter sphaeroides is a purple non-sulfur bacterium that belongs to the α-3 subdivision of Proteobacteria. R. sphaeroides is a model bacterial species because of its complex genome structure and expanded metabolic capabilities. The genome of R. sphaeroides consists of two circular chromosomes and five endogenous plasmids. It has the ability to grow under a wide variety of environmental conditions. It grows aerobically (~20% O2), semi-aerobically (~2% O2), and photosynthetically (under anaerobic condition plus light). It has been previously shown that many bacterial species utilize a number of alternate carbon sources for their optimal growth under a variety of growth conditions. We hypothesize that different or an additional carbon source in the minimal medium differentially affects the bacterial growth under dark-aerobic conditions. The bacterial growth kinetics and the number of cells in the bacterial culture were analyzed by measuring the optical density (OD at 600 nm) and the colony forming units (CFUs) at regular intervals of bacterial cultures. Results reveal that sodium succinate is the preferred sole carbon source for the optimal growth of R. sphaeroides. The results of growth kinetics and CFUs together concluded that from the tested carbon sources, sodium succinate is the best single carbon source in the minimal media for the optimal growth of R. sphaeroides. Interestingly, cell culture grown in SIS supplemented with sodium acetate exhibits a prolonged lag phase with the lowest ODs and CFUs that later switches to the growth-burst phase support previously discovered similar phenomenon of the growth-rate switch in the presence of acetate metabolism. Future work will utilize the aerobically grown R. sphaeroides’ cells as a biocatalyst to deplete the oxygen levels from natural gas streams and industrial gas pipelines.展开更多
Carotenoids act as precursors of vitamin A,antioxidants,enhancers of immunity,and are thus widely used in food and pharmaceutical industry.Microbial fermentation is one of the most important solutions for production o...Carotenoids act as precursors of vitamin A,antioxidants,enhancers of immunity,and are thus widely used in food and pharmaceutical industry.Microbial fermentation is one of the most important solutions for production of natural carotenoids.Rhodobacter sphaeroides is one of most promising bacteria employed for large scale production of carotenoids.In the present study,crtA located in the carotenoids biosynthesis pathway in R.sphaeroides was amplified by PCR.The overexpression vector pRKcrtA was constructed and subsequently transferred into R.sphaeroides,producing the genetically engineered strain R.sphaeroides 2.4.1/pRKcrtA overexpressing crtA.The carotenoid production from the genetically engineered strain was significantly increased.Fermentation procedure was optimized for further enhanced carotenoids production.展开更多
The photosynthetic activities between two main developmental stages, colony and hormogonium, of the edible cyanobacterium Nostoc sphaeroides Kutzing, were compared. Hormogonia have a higher content of chlorophyll than...The photosynthetic activities between two main developmental stages, colony and hormogonium, of the edible cyanobacterium Nostoc sphaeroides Kutzing, were compared. Hormogonia have a higher content of chlorophyll than that of colonies. It showed that the ratios of phycocyain （PC）, allophycocyain （APC） and phycoerythrocyanin （PEC） in hormogonia and colonies were different. The room temperature chlorophyll fluorescence, 77 K chlorophyll fluorescence, measurements of PSⅠand PS Ⅱ activities all showed that colony has higher photosynthetic competence than hormogonia. Hormogonia had a higher respiration rate than colony, while their maximum photosynthetic oxygen evolution rates were very close. The responses of hormogonia and colonies to high light illuminations also were different. Both of their oxygen evolution rates decreased quickly with the prolonged high light illumination, but hormogonia can keep relatively higher PSⅡ activity （Fv/Fm） than that of colonies. The results suggested that colony was photosynthetically more competent than hormogonia, while the ability of hormogonia to tolerate high light illumination was higher than that of colony.展开更多
In order to enhance the degrading protein capability of purple non-sulfur bacteria(PNSB),an effective strain,L2,was used to co-culture with Rhodobacter sphaeroides ATCC17023.The effects of added strain on protein remo...In order to enhance the degrading protein capability of purple non-sulfur bacteria(PNSB),an effective strain,L2,was used to co-culture with Rhodobacter sphaeroides ATCC17023.The effects of added strain on protein removal of R.sphaeroides were investigated.Results showed that strain L2,being identified as Bacillus thuringiensis/cereus,had a high potential for producing protease with a production of 295 U/m L.The optimal B.thuringiensis/cereus(40 μL) could significantly increase protein degradation of R.sphaeroides.Protein removal and biomass production were improved by 483% and 67%,respectively.R.sphaeroides/total biomass production was more than 95%.Theoretical analysis revealed that R.sphaeroides syntrophically interacted with B.thuringiensis/cereus.Protein degradation of B.thuringiensis/cereus provided small molecule substrates(VFAs) for R.sphaeroides growth and cells materials synthesis.展开更多
文摘The electronic structure of protein chains L and M in photosynthetic reaction center (PRC) of Rhodobacter sphaeroides (Van Niel) Imhoff, Truper et Pfennig) was studied by using the Overlapping Dimer Approximation method and the Extended Negative Factor Counter method at ab initio level. The result indicated that: (1) Amino acid residues, the molecular orbitals of which composed the main components of frontier orbitals of protein chain L (M), are located at the random coil areas of chain L (alpha helix areas of chain M). Since the random coil is flexible and more easy to change its conformation in the electron transfer process and to reduce the energy of the system, and the structure of the alpha helix is reletively stable, this difference might be one of the causes for the electron transfer in photosynthetic reaction center (PRC) only takes place along the L branch. (2) The His residues which axially coordinated to the 'special pair' P and accessory chlorophyll molecules (ABChls) are essentially important for the E-LUMO levels of P and ABChl. But, the corresponding molecular orbitals of these His residues do not appear in the composition of frontier orbitals of protein chains. It means that the interaction between pigment molecules and protein chains do not influence the contribution to the frontier orbitals of protein chains explicitly, but influences the corresponding E-LUMO levels significantly.
文摘[Objective]The experiment aimed to explore a new way for observing surface structure of Nostoc sphaeroides Kutzing. [Method] The scanning electron microscope was used to observe the epidermal ultrastructure of wild and cultured Nostoc sphaeroides Kutzing. [ Result] The epidermis of wild and cultured Nostoc sphaeroides Kutzing showed mixture structure of fibril colloid which was reticular arranged. The difference between wild and cultured Nostoc sphaeroides Kutzing was that the outer epidermis of cultured Nostoc sphaeroides Kutzing had trichome distribution but the wild Nostoc sphaeroides Kutzing did not has such distribution. The obsevation results of under smaller than 10 μm by scanning electron microscope was touched thick and showed many folds and distortions. [ Conclusion] The scanning electron microscope was an effective way to study development of Nostoc sphaeroides Kutzing colony and it was worth popularizing.
基金supported by the National Natural Science Foundation of China (No. 51278489)
文摘Bacillus thuringiensis/cereus L2 was added as a biostimulant to enhance the biomass accumulation and carotenoid yield of Rhodobacter sphaeroides using wastewater as the culturing medium. Results showed that biostimulation could significantly enhance the R. sphaeroides biomass production and carotenoid yield. The optimal biostimulant proportion was 40 μL（about 6.4 × 10^5CFU）. Through the use of biostimulation, chemical oxygen demand removal, R. sphaeroides biomass production, carotenoid concentration, and carotenoid yield were improved by 178%, 67%, 214%, and 70%, respectively. Theoretical analysis revealed that there were two possible reasons for such increases. One was that biostimulation enhanced the R. sphaeroides wastewater treatment efficiency. The other was that biostimulation significantly decreased the peroxidase activity in R. sphaeroides. The results showed that the highest peroxidase activity dropped by 87% and the induction ratio of the RSP_3419 gene was 3.1 with the addition of biostimulant. The enhanced carotenoid yield in R. sphaeroides could thus be explained by a decrease in peroxidase activity.
基金Supported by Talent Project of Sichuan University of Science&Engineering(2015RC27)Meat Processing Key Laboratory of Sichuan Province(16R-27)
文摘Coenzyme Q10 is widely used in food,cosmetics and pharmaceuticals,possessing a broad market.Rhodobacter sphaeroides is enriched in natural coenzyme Q10 and is becoming an important microorganism for producing natural coenzyme Q10.The paper reviewed the biosynthesis pathways of coenzyme Q10 in R.sphaeroides and the advances in enhancement of coenzyme Q10 production in R.sphaeroides based on metabolic engineering.
基金supported by the National Natural Science Foundation of China(No.51708214)the High-level Personnel Research Startup Project of North China University of Water Resources and Electric Power(No.40550)the Treatment Technology Integration and Demonstration for Domestic Sewage of Typical Villages and Towns in Henan Province(No.161100310700)
文摘This study aimed to increase bacterial growth and 5-aminolevulinic acid（ALA） biosynthesis of Rhodobacter sphaeroides in wastewater treatment through adding ferrous ion（ Fe2＋ ）. Results demonstrated that Fe2＋ effectively enhanced the biomass production and ALA yield of R. sphaeroides. Moreover, the optimal Fe2＋ dosage was found to be 400 μmol/L, which was associated with the highest biomass of 4015.3 mg/L and maximum ALA yield of 15.9 mg/g-dry cell weight（mg/g-DCW）. Mechanism analysis revealed that Fe2＋ vastly improved Adenosine Triphosphate（ATP） production by up-regulating the nif gene expression, and increasing ATP enhanced the biomass and ALA yield by supplying energy for bacterial growth and ALA biosynthesis, respectively. Correlation analysis showed that the ALA and ATP yields had positive relation with nifA and nifU gene expression. In addition, the nifA and nifU gene expression displayed high consistency of co-transcription at the optimal Fe2＋ dosage.
基金Supported by the Research Project of Sichuan University of Science&Engineering(2015RC27)Scientific Research Foundation of the Education Department of Sichuan Province(15ZA0222)Research Project of Meat Processing Key Laboratory of Sichuan Province(16R-27)
文摘In the present study,single factors including inoculation amount,fermentation temperature,fermentation duration and ratio of fermentation medium volume to total flask volume were optimized for enhancing the production of coenzyme Q10 from genetic engineered Rhodobacter sphaeroides overexpressing UbiE.The experimental results suggested that optimal single factors were: inoculation amount 2%,fermentation temperature 30 ℃,fermentation duration 48 h,and fermentation medium volume to total flask volume 80%.The present study will promote the large scale production of CoQ10 from microorganisms.
文摘Rhodobacter sphaeroides is a purple non-sulfur bacterium that belongs to the α-3 subdivision of Proteobacteria. R. sphaeroides is a model bacterial species because of its complex genome structure and expanded metabolic capabilities. The genome of R. sphaeroides consists of two circular chromosomes and five endogenous plasmids. It has the ability to grow under a wide variety of environmental conditions. It grows aerobically (~20% O2), semi-aerobically (~2% O2), and photosynthetically (under anaerobic condition plus light). It has been previously shown that many bacterial species utilize a number of alternate carbon sources for their optimal growth under a variety of growth conditions. We hypothesize that different or an additional carbon source in the minimal medium differentially affects the bacterial growth under dark-aerobic conditions. The bacterial growth kinetics and the number of cells in the bacterial culture were analyzed by measuring the optical density (OD at 600 nm) and the colony forming units (CFUs) at regular intervals of bacterial cultures. Results reveal that sodium succinate is the preferred sole carbon source for the optimal growth of R. sphaeroides. The results of growth kinetics and CFUs together concluded that from the tested carbon sources, sodium succinate is the best single carbon source in the minimal media for the optimal growth of R. sphaeroides. Interestingly, cell culture grown in SIS supplemented with sodium acetate exhibits a prolonged lag phase with the lowest ODs and CFUs that later switches to the growth-burst phase support previously discovered similar phenomenon of the growth-rate switch in the presence of acetate metabolism. Future work will utilize the aerobically grown R. sphaeroides’ cells as a biocatalyst to deplete the oxygen levels from natural gas streams and industrial gas pipelines.
基金Supported by the Project of Sichuan Science and Technology Department(2019YJ0673)National Modern Agriculture Industry System/Sichuan Live Pig Innovation Team(SCSZTD-3-007)。
文摘Carotenoids act as precursors of vitamin A,antioxidants,enhancers of immunity,and are thus widely used in food and pharmaceutical industry.Microbial fermentation is one of the most important solutions for production of natural carotenoids.Rhodobacter sphaeroides is one of most promising bacteria employed for large scale production of carotenoids.In the present study,crtA located in the carotenoids biosynthesis pathway in R.sphaeroides was amplified by PCR.The overexpression vector pRKcrtA was constructed and subsequently transferred into R.sphaeroides,producing the genetically engineered strain R.sphaeroides 2.4.1/pRKcrtA overexpressing crtA.The carotenoid production from the genetically engineered strain was significantly increased.Fermentation procedure was optimized for further enhanced carotenoids production.
文摘The photosynthetic activities between two main developmental stages, colony and hormogonium, of the edible cyanobacterium Nostoc sphaeroides Kutzing, were compared. Hormogonia have a higher content of chlorophyll than that of colonies. It showed that the ratios of phycocyain （PC）, allophycocyain （APC） and phycoerythrocyanin （PEC） in hormogonia and colonies were different. The room temperature chlorophyll fluorescence, 77 K chlorophyll fluorescence, measurements of PSⅠand PS Ⅱ activities all showed that colony has higher photosynthetic competence than hormogonia. Hormogonia had a higher respiration rate than colony, while their maximum photosynthetic oxygen evolution rates were very close. The responses of hormogonia and colonies to high light illuminations also were different. Both of their oxygen evolution rates decreased quickly with the prolonged high light illumination, but hormogonia can keep relatively higher PSⅡ activity （Fv/Fm） than that of colonies. The results suggested that colony was photosynthetically more competent than hormogonia, while the ability of hormogonia to tolerate high light illumination was higher than that of colony.
基金Sponsored by the National Natural Science Foundation of China(Grant No.51278489)
文摘In order to enhance the degrading protein capability of purple non-sulfur bacteria(PNSB),an effective strain,L2,was used to co-culture with Rhodobacter sphaeroides ATCC17023.The effects of added strain on protein removal of R.sphaeroides were investigated.Results showed that strain L2,being identified as Bacillus thuringiensis/cereus,had a high potential for producing protease with a production of 295 U/m L.The optimal B.thuringiensis/cereus(40 μL) could significantly increase protein degradation of R.sphaeroides.Protein removal and biomass production were improved by 483% and 67%,respectively.R.sphaeroides/total biomass production was more than 95%.Theoretical analysis revealed that R.sphaeroides syntrophically interacted with B.thuringiensis/cereus.Protein degradation of B.thuringiensis/cereus provided small molecule substrates(VFAs) for R.sphaeroides growth and cells materials synthesis.
