创伤弧菌FrsA蛋白的纯化及生化表征

为确定创伤弧菌FrsA蛋白(VvFrsA)在大肠杆菌中的表达条件及体外纯化步骤,并对其进行生化表征分析。通过分子生物学手段将合成的创伤弧菌的FrsA基因(VvFrsA)连接到pET28a载体上,构建pET28a-VvFrsA原核表达载体。将该载体转化至大肠杆菌BL21(DE3)感受态细胞后,经IPTG诱导实现了VvFrsA蛋白的可溶性表达。采用Ni-NTA亲和层析法纯化,得到重组VvFrsA蛋白。酶催化活性研究结果显示,VvFrsA具有酯酶功能,可以催化对硝基苯酚脂肪酸酯的水解,并且其最适底物为对硝基苯酚短链脂肪酸酯。综上,利用大肠杆菌BL21(DE3)菌株成功表达了可溶性VvFrsA蛋白,并且VvFrsA体外具有催化对硝基苯酚脂肪酸酯水解活性。

集胞藻PCC6803中N-乙酰鸟氨酸转氨酶的生化表征及结构分析

旨在对集胞藻PCC6803中slr1022基因编码的N-乙酰鸟氨酸转氨酶进行生化表征及结构分析,为进一步研究该酶的催化功能及机制奠定理论基础。以集胞藻PCC6803基因组为模板,通过PCR扩增获得slr1022基因,将其连接到表达载体pET-28a上,转化大肠杆菌BL21(DE3)感受态。经IPTG诱导,Ni-NTA亲和层析纯化后获得重组Slr1022蛋白,然后通过紫外分光光度法对Slr1022蛋白的催化功能进行表征,并运用生物信息学软件对该蛋白进行结构分析。成功构建pET28a-slr1022重组表达质粒,并诱导表达重组Slr1022蛋白,SDS-PAGE电泳鉴定该蛋白分子量约为50 kD,与理论大小相符。Slr1022蛋白与底物N-乙酰鸟氨酸的结合常数K_m和最大反应速度V_(max)分别是0.12 mmol/L和0.60 μmol/(L·s),并且Slr1022蛋白与另一底物α-酮戊二酸的K_(m)和V_(max)分别是0.039 mmol/L和0.65 μmol/(L·s)。Slr1022蛋白在pH 8.5时催化活性最强。Slr1022蛋白和其他来源的N-乙酰鸟氨酸转氨酶氨基酸序列有一定的同源性,而且活性位点的氨基酸残基高度保守。成功克隆并表达纯化出Slr1022蛋白,酶学性质和生物信息学研究表明Slr1022蛋白为N-乙酰鸟氨酸转氨酶。

Characterization of apparent sulfur oxidation activity of thermophilic archaea in bioleaching of chalcopyrite

The apparent sulfur oxidation activities of four pure thermophilic archaea, Acidianus brierleyi （JCM 8954）, Metallosphaera sedula （YN 23）, Acidianus manzaensis （YN 25） and Sulfolobus metallicus （YN 24） and their mixture in bioleaching chalcopyrite were compared, which were characterized indirectly by the evolution of the cells concentration, pH value and sulfate ions concentration in solution. The results show that the mixed culture contributed significantly to the raising of leaching rate, which suggests that the mixed culture had a higher sulfur oxidation activity than the pure culture. Meanwhile, the results also indicate that the changes of parameters characterizing the sulfur oxidation activity of thermophilic archaea are often influenced by many factors, so it is hard to reflect accurately the specific sulfur oxidation activities among the different sulfur-oxidizing microbes when bioleaching chalcopyrite at different conditions. Accordingly, an efficient method to characterize microbial sulfur oxidation activity appears to be desirable.

黄素单加氧酶及其在食品功能因子合成中的应用

黄素单加氧酶是一种重要的氧化还原酶,能够催化多种化合物的羟基化、拜尔-维利格氧化、硫氧化、环氧化和卤化反应等,被广泛应用于食品、医药和化工等领域。该文从分类、特性研究(包括表达条件优化和催化反应过程控制)、蛋白质工程及其在食品功能因子合成中的应用等方面对黄素单加氧酶进行综述,并对其发展前景进行展望。

Characterization of SiC nanowires prepared on C/C composite without catalyst by CVD

SiC nanowires were prepared on C/C composite surface without catalyst by chemical vapor deposition（CVD） using CH3 SiCl3 as precursor.SEM images of the CVD-product reveal that some long nanowires have grown to tens of micrometers with some gathered as a ball.Some short nanowires agglomerate like chestnut shell with many thorns accompanied by some deposited nano-particles.XRD,Raman-spectrum and FTIR patterns indicate that the product is a typical β-SiC.TEM images show that the nanowires have a wide diameter range from 10 to 100 nm,and some thin nanowires are bonded to the thick one by amorphous CVD-SiC.A SiC branch generates from an amorphous section of a thick one with an angle of 70° between them,which is consistent with the [111] axis stacking angle of the crystal.SAED and fast Fourier transform（FFT） patterns reveal that the nanowires can grow along with different axes,and the bamboo-nodes section is full of stacking faults and twin crystal.The twisted SiC lattice planes reveal that the screw dislocation growth is the main mechanism for the CVD-SiC nanowires.

Characterization of Catalytic Cracking Catalysts Regenerated by Gasifying Deposited Coke

Partially or fully regenerated catalytic cracking catalysts were prepared by gasifying the coke deposited on coked catalysts with a gaseous mixture of oxygen and steam in a fixed fluidized bed （FFB）. The resultant samples were characterized by different methods such as the nitrogen adsorption-desorption analysis, the X-ray diffractometry, the infrared spectroscopy, the ammonia temperature-programmed desorption （NH3-TPD） method, the X-ray fluorescence （XRF） analysis, the transmission electron microscopy and energy dispersive X-ray spectroscopy （TEM-EDX）, the thermal-gravimetric analysis （TGA） and the differential thermal analysis （DTA）. The results showed that exposure of catalyst to steam for about 10 minutes at temperature ≥ 800 ℃ could not cause too much destruction of the catalysts, and an amount of coke equating to about 0.27 m% was enough to block approximately all acid sites in micro-pores of the zeolite catalyst. Coke didn＇t show equal reactivity during coke burning-off that could be accelerated by the catalytic action of nearby metal atoms. However, when the carbon content on the catalyst reached about 2.44 m%, the catalytic action of metals on the catalyst was not evident. The severe thermal and hydrothermal environment during exposure of the catalyst to steam at a temperature in the range of about 860--880 ℃ for 30 minutes could lead to collapse of pore structure and transformation of crystal phase and consequently decrease of the surface area and acid amount on the catalyst.

Multiscale Characterization of Automotive Surface Coating Formation for Sustainable Manufacturing

Automotive surface coating manufacturing is one of the most sophisticated and expensive steps in automotive assembly. This step involves generating multiple thin layers of polymeric coatings on the vehicle surface through paint spray and curing in a multistage, dynamically changing environment. Traditionally, the quality control is solely post-process inspection based, and process operational adjustment is only experience based, thus the manufacturing may not be （highly） sustainable. In this article, a multiscale system modeling and analysis methodology is introduced for achieving a sustainable application of polymeric materials through paint spray and film curing in automotive surface coating manufacturing. By this methodology, the correlations among paint material, application processes and coating performance can be identified. The model-based analysis allows a comprehensive and deep study of the dynamic behaviors of the material, process, and product in a wide spectrum of length and time. Case studies illustrate the efficacy of the methodology for sustainable manufacturing.

Characterization and Genomic Analysis of a Plaque Purified Strain of Cyanophage PP

Cyanophages are ubiquitous and essential components of the aquatic environment and play an important role in the termination of algal blooms.As such,they have attracted widespread interest.PP was the first isolated cyanophage in China,which infects Plectonema boryanum and Phormidium foveolarum.In this study,this cyanophage was purified three times by a double-agar overlay plaque assay and characterized.Its genome was extracted,totally sequenced and analyzed.Electron microscopy revealed a particle with an icosahedral head connected to a short stubby tail.Bioassays showed that PP was quite virulent.The genome of PP is a 42,480 base pair(bp),linear,double-stranded DNA molecule with 222 bp terminal repeats.It has high similarity with the known Pf-WMP3 sequence.It contains 41 open reading frames(ORFs),17 of which were annotated.Intriguingly,the genome can be divided into two completely different parts,which differ both in orientation and function.

Characterization of tricalcium phosphate solubilization by Stenotrophomonas maltophilia YC isolated from phosphate mines

The phosphate solubilizing characteristics of a strain YC, which was isolated from phosphate mines (Hubei, China), were studied in National Botanical Research Institute’s phosphate (NBRIP) growth medium containing tricalcium phosphate (TCP) as sole phosphorus (P) source. The strain YC is identified as Stenotrophomonas maltophilia (S. maltophilia) based upon the results of morphologic, physiological and biochemical characteristics and 16S rRNA sequences analysis. The results show that the strain S. maltophilia YC can solubilize TCP and release soluble P in NBRIP growth medium. A positive correlation between concentration of soluble P and population of the isolate and a negative correlation between concentration of soluble P and pH in the culture medium are observed from statistical analysis results. Moreover, gluconic acid is detected in the culture medium by HPLC analysis. It indicates that the isolate can release gluconic acid during the solubilizing experiment, which causes acidification of the culture medium and then TCP solubilization. S. maltophilia YC has a maximal TCP solubilizing capability when using maltose as carbon source and ammonium nitrate as nitrogen source, respectively, in NBRIP growth medium.

Reading, Habitus, and Social Representations in a University Context

The lack of reading habits among Mexican population is an idea based on national and international indicators, which illustrate the danger of this situation; however, these indicators fail to go deeper into the problem. This project was aimed to obtain more information about the habitus and social representations of college students reading habits, using as theoretical elements the principles developed by Pierre Bourdieu and Serge Moscovici. The project was developed in a public university, located in Mexico＇s northeast region; 19 students were part of the sample group. Two research tools were used to obtain data： survey and focus group. The project analyzed academic and environmental aspects, beyond the ＂ought to be＂, that affected the development and provided characteristics on the daily sociocultural reality of the students, with a view of the real environment of the students without ＂make-up＂. The project deepened into the analysis of the reading practices and not only the usual numeric indicators, which traditionally condemn the reading practices in Mexico. The results of this study showed that students read a lot more than what they and their teachers think they do, this taking into consideration electronic documents, general interest publications, and commercial magazines, among others. The point of view of some teachers and researchers, when it comes to reading habits recognize non-academic, simple and interesting reading more important impact on their reading habits shows a tendency to be pedantic and canonic; it does not for young people, which in the long term, could have a

Partial Characterization of Chitinolytic Extract from Endophytic Streptomyces sp. and Its Effects on the Boll Weevil

The present study achieves the biochemistry partial characterization of the chitinolytic extract produced by an endophytic Streptomyces sp. strain （A8）. This extract was also tested against Anthonomus grandis, the cotton boll weevil aiming its control. The chitinase crude extract from the A8 strain was cultured for five days in a minimum liquid media supplemented with chitin. The extract was partially characterized by standard methods. The chitinolytic extract had an optimum temperature of 66 ＂C and an optimum pH between 4 to 9 （around 80% of relative activity）. We also characterized the temperature and pH stability and measured the effects of enzyme inhibitors. The filtered chitinolytic extract was added to an artificial boll weevil diet. Boll weevil development from the egg stage to the adult stage was prolonged, and the percentage of adults that emerged was approximately 66% less than on control diet. This study showed that the.larval development of A. grandis was inhibited by the presence of characterized chitinolytic extract in artificial diet. This work provides an experimental basis for using the chitinase from an endophytic bacterium Streptomyces sp. as a biocontrol alternativeto controlling the plant pest A. grandis.

Synthesis and characterization of Li Fe(PO_4)_(3-x)/3Br_x/C as a cathode material for lithium-ion battery

The olivine-typed cathode material of Li Fe PO4 was prepared via sol-gel method,and the bromine was doped into Li Fe PO4.The crystal structure,morphology,and electrochemical properties of the samples were investigated by X-ray diffraction,scanning electron microscopy and charge–discharge cycle measurements.The results showed that the electrochemical performance of Li Fe PO4 had been improved by bromine doping,and the best doping amount of bromine is 2%.The discharge capacity of this sample can reach 152 m Ah/g at 0.2 C.