The original strain HY-7 was isolated from the bauxite mine drainage(BMD) taken from a reservoir in Sanmenxia Mine,Henan Province,China.The optimum temperature and pH for the growth of strain HY-7 were 30 ℃ and 7.0...The original strain HY-7 was isolated from the bauxite mine drainage(BMD) taken from a reservoir in Sanmenxia Mine,Henan Province,China.The optimum temperature and pH for the growth of strain HY-7 were 30 ℃ and 7.0,respectively.The optimum UV radiating time was 20 s and the positive mutation rate was 23.0%.The growth curves show that strain HY-7 needs144 h to reach the stationary phase after its mutagenesis,which is 24 h earlier than that of the original strain.Sequence homology analysis indicated that this community consisted of mainly two branches:one sharing high homology with Paenibacillus stellifer and the other sharing high homology with Sporolactobacillus laevolacticus.The experimental results showed that the TiO2 grade of mtile concentrate increased from 78.21%to 91.80%and the recovery of TiO2 reached 95.24%after 7 d of bioleaching.The bio-desilication process can not only effectively improve the TiO2 grade of rutile concentrate but also meet the requirements of environmental protection.展开更多
In this study, 8 kinds of desiccation models and 3 kinds of collection models were studied for banana stems. The results showed that the crushing-com- pression-air drying processing model showed the best desiccation e...In this study, 8 kinds of desiccation models and 3 kinds of collection models were studied for banana stems. The results showed that the crushing-com- pression-air drying processing model showed the best desiccation effect for banana stems. Under this desiccation model, the moisture content was reduced from 90.5% to 19.93% with weight reduced by 75.2%; after 3 to 6 months of storage, the mois- ture content was still remained within 17%-20%, and the volume was decreased to 30%-40% of the original volume of fresh banana stems. This desiccation model made banana stems basically meet the requirements by drying storage. The plant- cutting down-crushing-compression-transportation-desiccation-storage production model was initially developed for desiccation and collection of banana stems, which would lay a certain foundation for the utilization of banana stem resource by enterprises.展开更多
The strain Lv(1- z) isolated from the Henan bauxite was characterized by morphological observation, biochemical and physiological identification, and 16S rDNA sequence analysis. The influences of temperature, initia...The strain Lv(1- z) isolated from the Henan bauxite was characterized by morphological observation, biochemical and physiological identification, and 16S rDNA sequence analysis. The influences of temperature, initial pH value, the volume of medium, shaking speed and illite concentration on the desilicating ability of the strain Lv(1- z) were investigated. The results show that the bacterium is a Gram-negative rod-shaped bacterium with oval endspores and thick capsule, but without flagellum. The biochemical and physiological tests indicate that the strain Lv(1- z) is similar to Bacillus rnucilaginosus. In GenBank the 16S rDNA sequence similarity of the strain Lv(1- z) and the B. rnucilaginosus YNUCC0001(AY571332) is more than 99%. Based on the above results, the strain Lv(1- z) is identified as B. rnucilaginosus. The optimum conditions for the strain L(1- z) to remove silicon from illite are as follows., temperature is 30℃ ;initial pH value is 7.5; medium volume in 200 mL bottle is 60 mL; shaking speed of rotary shaker is 220 r/m ; illite concentration is 1%.展开更多
A Lactobacillus brevis CGMCC 1306 isolated from fresh milk without pasteurization was found to have higher glutamate decarboxylase (GAD) activity. An effective isolation and purification procedure of GAD from a cell...A Lactobacillus brevis CGMCC 1306 isolated from fresh milk without pasteurization was found to have higher glutamate decarboxylase (GAD) activity. An effective isolation and purification procedure of GAD from a cell-free extract of Lactobacillus brevis was developed, and the procedure included four steps: 30%-90% saturation (NH4)2SO4 fractional precipitation, Q sepharose FF anion-exchange chromatography, sephacryl S-200 gel filtration, and resource Q anion-exchange chromatography. Using this protocol, the purified GAD was demonstrated to possess electrophoretic homogeneity via SDS-PAGE. The purification fold and activity recovery of GAD were 43.78 and 16.95%, respectively. The molecular weight of the purified GAD was estimated to be approximately 62 kDa via SDS-PAGE. The optimum pH and temperature of the purified GAD were 4.4 and 37℃, respectively. The purified GAD had a half-life of 50rain at 45℃ and the Km value of the enzyme from Lineweaver-Burk plot was found to be 8.22.5'-pyridoxal phosphate (PLP) had little effect on the regulation of its activity.展开更多
Using the unsteady incompressible Navier-Stokes equation as the governing equation, the large eddy simulation (LES) model is implemented to investigate the shedding of vortices, the flow pattern of turbulence, the uns...Using the unsteady incompressible Navier-Stokes equation as the governing equation, the large eddy simulation (LES) model is implemented to investigate the shedding of vortices, the flow pattern of turbulence, the unsteady pressure fluctuation and the time history of the lift coefficient and drag coefficient of hoistable masts with various mast shapes and various arrangements in this paper. Combining the FFT, combined time-frequency transform and wavelet power spectrum analysis, the characteristics of unsteady pressure can be obtained in both time and frequency domain. It shows that the main frequency of pressure fluctuation is near the frequency of vortex shedding in time domain using the FFT method. It can be inferred from the combined time-frequency transform that the unsteady pressure fluctuation has obviously the peak value and the second peak value in time domain. It could indicate that the fluctuation power varies from the fluctuation frequency through the power spectrum analysis. By the data analysis, it shows that the vortex shedding is the dominant cause of the periodically pressure fluctuation. And the interaction pattern of wake and interplay between wake and the walls of masts under different arrangements are also discussed in this paper.展开更多
The activity of whole-cell biocatalysts is strongly compromised by the cell envelope, which is a permeability harrier against the diffusion of substrates and products. Although common chemical or physical permeahiliza...The activity of whole-cell biocatalysts is strongly compromised by the cell envelope, which is a permeability harrier against the diffusion of substrates and products. Although common chemical or physical permeahilization methods used in cultured cells enhance cell permeability, these methods inevitably add several extra processing steps after cell cultivation, as well as impede large scale processing. To increase membrane permeability and cell- bound glutamate decarboxylase (GAD) activity of recombinant Escherichia coil (BL21 (DE3)-pET28a-gadB) cells without the need for an additional permeabilization step, we investigated the permeabilizing effects of adding cell wall synthesis inhibitors or suffactants to the culture media. Ampidllin was the most effective at improving cell-bound GAD activity of the BL21 (DE3)-pET28a-gadB, although it decreased the cell biomass yield. The best permeabilization effect was observed using an ampicillin concentration of 5 pg. ml-1. Using this concentration, the cell hiomass did decrease by 40.58%, but the cell-bound GAD activity of BL21 (DE3)-pET28a-gadB and total cell-bound GAD activity per milliliter of culture was enhanced by 6.24- and 3.64-fold, respectively. Treatment ofBL21 (DE3)-pET28a-gadB cells with 5 tag.ml 1 ampicillin resulted in structural changes to the cell envelope, but did not substantially affect GAD expression. By entrapping the ampicillin-treated cells in an open pore gelation matrix, which is a polymer derived from polyvinyl alcohol (PVA), alginate, and boric acid, the transfor- mation rate of γ-aminobutyric acid (GABA) at the 10th cycle produced by immobilized and permeabilized cells remained 46% of the first cycle. GAD activity of the immobilized, permeabilized cells remained over 90% after 30 days of storage at 4 ℃.展开更多
The catalytic activity of two common bacterial enzymes, lactate dehydrogenase (LDH) and cytochrome c oxidase (COX) from Escherichia coli was examined following bacterial exposure to microwave (MW) radiation unde...The catalytic activity of two common bacterial enzymes, lactate dehydrogenase (LDH) and cytochrome c oxidase (COX) from Escherichia coli was examined following bacterial exposure to microwave (MW) radiation under well-defined experimental conditions. The experiments were conducted in a specialized microwave processing apparatus, with an exposure frequency of 18 GHz, and a temperature profile that was restricted to below 40℃ to avoid thermal degradation of the bacteria. The absorbed power was calculated to be 1,500 kW/m3 and the electric field was determined to be 300 Wm. Both values were theoretically confirmed using Computer Simulation Technology (CST) Microwave Studio 3D Electromagnetic Stimulation Software. Results showed that the activity of both enzymes was increased following MW radiation compared to negative controls and thermally treated samples subjected to similar temperature profiles. It is suggested that the increase in COX and LDH enzyme activity could not be explained by conventional heating alone, but was rather a result of micro-thermal effects that incorporated 'undetectable' thermal mechanisms.展开更多
Riemerella anatipestifer is the causative agent of septicemia anserum exsudativa in ducks. Its pathogenesis and virulence factors are still unclear. The glycelytic enzyme, glyceraldehyde-3-phosphate dehydrogenase (GA...Riemerella anatipestifer is the causative agent of septicemia anserum exsudativa in ducks. Its pathogenesis and virulence factors are still unclear. The glycelytic enzyme, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an anchorless and multifunctional protein on the surface of several pathogenic microorganisms, is involved in virulence and adhesion. Whether homologs of GAPDH exist, and display similar characteristics in R. anatipestifer (RaGAPDH) has not been determined. In our research, the RaGAPDH activity from various R. anatipestifer isolates was confirmed. Twenty-two gapdh genes from genornic DNA of R. anatipestifer isolates were cloned and sequenced for phylogenetic analysis. The distribution of RaGAPDH in R. anatipestifer CZ2 strain was confirmed by antisera to recombinant RaGAPDH. The ability of purified RaGAPDH to bind host proteins was analyzed by solid-phase ligandbinding assay. Results revealed that all R. anatipestifer isolates showed different levels of GAPDH activity except four strains, which contained a gapdh-like gene. The gapdh of R. anatipestifer, which is located phylogenetically in the same branch as enterohemorrhagic Escherichia coil (EHEC), belonged to class I GAPDH, and encoded a 36.7-kDa protein. All RaGAPDH-encoding gene sequences from field isolates of R. anatipestiferdisplayed 100% homology. The RaGAPDH localized on the extracellular membrane of several R. anatipestifer strains. Further, it was released into the culture medium, and exhibited GAPDH enzyme activity. We also confirmed the binding of RaGAPDH to plasminogen and fibrinogen. These results demonstrated that GAPDH was present in R. anatipestifer, although not in all strains, and that RaGAPDH might contribute to the microorganism's virulence.展开更多
The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many com...The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles(MNPs) was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease(DNase) and Ribonucleases(RNase) free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction(PCR) and reverse transcription PCR(RT-PCR) to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria.展开更多
基金Project(2011-622-40)supported by the Mineral Exploration Foundation of Henan Province,ChinaProject(51104189)supported by the National Natural Science Foundation of ChinaProject(2013M531814)supported by the 53rd China Postdoctoral Science Foundation
文摘The original strain HY-7 was isolated from the bauxite mine drainage(BMD) taken from a reservoir in Sanmenxia Mine,Henan Province,China.The optimum temperature and pH for the growth of strain HY-7 were 30 ℃ and 7.0,respectively.The optimum UV radiating time was 20 s and the positive mutation rate was 23.0%.The growth curves show that strain HY-7 needs144 h to reach the stationary phase after its mutagenesis,which is 24 h earlier than that of the original strain.Sequence homology analysis indicated that this community consisted of mainly two branches:one sharing high homology with Paenibacillus stellifer and the other sharing high homology with Sporolactobacillus laevolacticus.The experimental results showed that the TiO2 grade of mtile concentrate increased from 78.21%to 91.80%and the recovery of TiO2 reached 95.24%after 7 d of bioleaching.The bio-desilication process can not only effectively improve the TiO2 grade of rutile concentrate but also meet the requirements of environmental protection.
基金Supported by Key Project of the National Twelfth-Five Year Research Program of China(2012BAC18B03)Natural Science Foundation of Hainan Province(414195,313106)~~
文摘In this study, 8 kinds of desiccation models and 3 kinds of collection models were studied for banana stems. The results showed that the crushing-com- pression-air drying processing model showed the best desiccation effect for banana stems. Under this desiccation model, the moisture content was reduced from 90.5% to 19.93% with weight reduced by 75.2%; after 3 to 6 months of storage, the mois- ture content was still remained within 17%-20%, and the volume was decreased to 30%-40% of the original volume of fresh banana stems. This desiccation model made banana stems basically meet the requirements by drying storage. The plant- cutting down-crushing-compression-transportation-desiccation-storage production model was initially developed for desiccation and collection of banana stems, which would lay a certain foundation for the utilization of banana stem resource by enterprises.
基金Project (50321402) supported by the National Natural Science Foundation of China project (2004CB619204) supportedby the National Basic Research Program
文摘The strain Lv(1- z) isolated from the Henan bauxite was characterized by morphological observation, biochemical and physiological identification, and 16S rDNA sequence analysis. The influences of temperature, initial pH value, the volume of medium, shaking speed and illite concentration on the desilicating ability of the strain Lv(1- z) were investigated. The results show that the bacterium is a Gram-negative rod-shaped bacterium with oval endspores and thick capsule, but without flagellum. The biochemical and physiological tests indicate that the strain Lv(1- z) is similar to Bacillus rnucilaginosus. In GenBank the 16S rDNA sequence similarity of the strain Lv(1- z) and the B. rnucilaginosus YNUCC0001(AY571332) is more than 99%. Based on the above results, the strain Lv(1- z) is identified as B. rnucilaginosus. The optimum conditions for the strain L(1- z) to remove silicon from illite are as follows., temperature is 30℃ ;initial pH value is 7.5; medium volume in 200 mL bottle is 60 mL; shaking speed of rotary shaker is 220 r/m ; illite concentration is 1%.
基金Supported by the National Natural Science Foundation of China (No.30570411)the Research Plan of Zhejiang Province, China.
文摘A Lactobacillus brevis CGMCC 1306 isolated from fresh milk without pasteurization was found to have higher glutamate decarboxylase (GAD) activity. An effective isolation and purification procedure of GAD from a cell-free extract of Lactobacillus brevis was developed, and the procedure included four steps: 30%-90% saturation (NH4)2SO4 fractional precipitation, Q sepharose FF anion-exchange chromatography, sephacryl S-200 gel filtration, and resource Q anion-exchange chromatography. Using this protocol, the purified GAD was demonstrated to possess electrophoretic homogeneity via SDS-PAGE. The purification fold and activity recovery of GAD were 43.78 and 16.95%, respectively. The molecular weight of the purified GAD was estimated to be approximately 62 kDa via SDS-PAGE. The optimum pH and temperature of the purified GAD were 4.4 and 37℃, respectively. The purified GAD had a half-life of 50rain at 45℃ and the Km value of the enzyme from Lineweaver-Burk plot was found to be 8.22.5'-pyridoxal phosphate (PLP) had little effect on the regulation of its activity.
文摘Using the unsteady incompressible Navier-Stokes equation as the governing equation, the large eddy simulation (LES) model is implemented to investigate the shedding of vortices, the flow pattern of turbulence, the unsteady pressure fluctuation and the time history of the lift coefficient and drag coefficient of hoistable masts with various mast shapes and various arrangements in this paper. Combining the FFT, combined time-frequency transform and wavelet power spectrum analysis, the characteristics of unsteady pressure can be obtained in both time and frequency domain. It shows that the main frequency of pressure fluctuation is near the frequency of vortex shedding in time domain using the FFT method. It can be inferred from the combined time-frequency transform that the unsteady pressure fluctuation has obviously the peak value and the second peak value in time domain. It could indicate that the fluctuation power varies from the fluctuation frequency through the power spectrum analysis. By the data analysis, it shows that the vortex shedding is the dominant cause of the periodically pressure fluctuation. And the interaction pattern of wake and interplay between wake and the walls of masts under different arrangements are also discussed in this paper.
基金Supported by the grants from the National Natural Science Foundation of China(21176220,20876143,31470793)the Natural Science Foundation of Zhejiang Province(Z13B060008)the Key Technology Research and Development Project of Ningbo(2011C11023)
文摘The activity of whole-cell biocatalysts is strongly compromised by the cell envelope, which is a permeability harrier against the diffusion of substrates and products. Although common chemical or physical permeahilization methods used in cultured cells enhance cell permeability, these methods inevitably add several extra processing steps after cell cultivation, as well as impede large scale processing. To increase membrane permeability and cell- bound glutamate decarboxylase (GAD) activity of recombinant Escherichia coil (BL21 (DE3)-pET28a-gadB) cells without the need for an additional permeabilization step, we investigated the permeabilizing effects of adding cell wall synthesis inhibitors or suffactants to the culture media. Ampidllin was the most effective at improving cell-bound GAD activity of the BL21 (DE3)-pET28a-gadB, although it decreased the cell biomass yield. The best permeabilization effect was observed using an ampicillin concentration of 5 pg. ml-1. Using this concentration, the cell hiomass did decrease by 40.58%, but the cell-bound GAD activity of BL21 (DE3)-pET28a-gadB and total cell-bound GAD activity per milliliter of culture was enhanced by 6.24- and 3.64-fold, respectively. Treatment ofBL21 (DE3)-pET28a-gadB cells with 5 tag.ml 1 ampicillin resulted in structural changes to the cell envelope, but did not substantially affect GAD expression. By entrapping the ampicillin-treated cells in an open pore gelation matrix, which is a polymer derived from polyvinyl alcohol (PVA), alginate, and boric acid, the transfor- mation rate of γ-aminobutyric acid (GABA) at the 10th cycle produced by immobilized and permeabilized cells remained 46% of the first cycle. GAD activity of the immobilized, permeabilized cells remained over 90% after 30 days of storage at 4 ℃.
文摘The catalytic activity of two common bacterial enzymes, lactate dehydrogenase (LDH) and cytochrome c oxidase (COX) from Escherichia coli was examined following bacterial exposure to microwave (MW) radiation under well-defined experimental conditions. The experiments were conducted in a specialized microwave processing apparatus, with an exposure frequency of 18 GHz, and a temperature profile that was restricted to below 40℃ to avoid thermal degradation of the bacteria. The absorbed power was calculated to be 1,500 kW/m3 and the electric field was determined to be 300 Wm. Both values were theoretically confirmed using Computer Simulation Technology (CST) Microwave Studio 3D Electromagnetic Stimulation Software. Results showed that the activity of both enzymes was increased following MW radiation compared to negative controls and thermally treated samples subjected to similar temperature profiles. It is suggested that the increase in COX and LDH enzyme activity could not be explained by conventional heating alone, but was rather a result of micro-thermal effects that incorporated 'undetectable' thermal mechanisms.
基金Project supported by the Fundamental Research Funds of the Central Universities of China(No.XDJK2013C009)
文摘Riemerella anatipestifer is the causative agent of septicemia anserum exsudativa in ducks. Its pathogenesis and virulence factors are still unclear. The glycelytic enzyme, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an anchorless and multifunctional protein on the surface of several pathogenic microorganisms, is involved in virulence and adhesion. Whether homologs of GAPDH exist, and display similar characteristics in R. anatipestifer (RaGAPDH) has not been determined. In our research, the RaGAPDH activity from various R. anatipestifer isolates was confirmed. Twenty-two gapdh genes from genornic DNA of R. anatipestifer isolates were cloned and sequenced for phylogenetic analysis. The distribution of RaGAPDH in R. anatipestifer CZ2 strain was confirmed by antisera to recombinant RaGAPDH. The ability of purified RaGAPDH to bind host proteins was analyzed by solid-phase ligandbinding assay. Results revealed that all R. anatipestifer isolates showed different levels of GAPDH activity except four strains, which contained a gapdh-like gene. The gapdh of R. anatipestifer, which is located phylogenetically in the same branch as enterohemorrhagic Escherichia coil (EHEC), belonged to class I GAPDH, and encoded a 36.7-kDa protein. All RaGAPDH-encoding gene sequences from field isolates of R. anatipestiferdisplayed 100% homology. The RaGAPDH localized on the extracellular membrane of several R. anatipestifer strains. Further, it was released into the culture medium, and exhibited GAPDH enzyme activity. We also confirmed the binding of RaGAPDH to plasminogen and fibrinogen. These results demonstrated that GAPDH was present in R. anatipestifer, although not in all strains, and that RaGAPDH might contribute to the microorganism's virulence.
基金supported by the National Basic Research Program of China(2014CB744501)the National High Technology Research and Development Program of China(2012AA022703)+8 种基金the National Key Special Science Program(2013ZX10004103-002)the National Natural Science Foundation of China(61201033,21205013,61271056,61527806)Projects of Development of Science and Medical Technology(201208038)Projects of Health Ministry of Nanjing(ZKX12038)the Clinical Science and Technology Special Projects in Jiangsu Province(BL2012067,BL2014094)the Talents Planning of Six Summit Fields of Jiangsu Province(2013-WSN-056)China Postdoctoral Science Foundation Funded Project(2014M551491,2015T80487)Jiangsu Planned Projects for Postdoctoral Research Funds(1302007A)the Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province
文摘The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles(MNPs) was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease(DNase) and Ribonucleases(RNase) free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction(PCR) and reverse transcription PCR(RT-PCR) to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria.