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1.
The gene(741 bp) encoding carboxylesterase from the thermophilic bacterium Geobacillus sp.ZHl was cloned and overexpressed in Escherichia coli.The purified recombinant protein presented a molecular mass of about 40 kDa by SDS-PAGE analysis.Enzyme assays using p-nitrophenyl esters with different acyl chain lengths as the substrates confirmed its esterase activity,yielding highest specific activity with p-nitrophenyl acetate.Among the p-nitrophenyl esters tested,the carboxylesterase presented preference for p-nitrophenyl caprylate,but hydrolyzed p-nitrophenyl butyrate more efficiently.When p-nitrophenyl butyrate was used as a substrate,the recombinant carboxylesterase exhibited highest activity at pH 8.0 and 60℃.Almost no decrease in esterase activity was observed at 60℃for 3 h,and over 40% of activity was still maintained after incubation at 90℃for 3 h.These results indicate that Geobacillus sp.ZH1 recombinant esterase was thermostable.The enzymatic activity was inhibited by the addition of phenylmethylsulfonyl fluoride,indicating that it contains serine residue,which plays a key role in the catalytic mechanism.Except SDS and xylene,this esterase showed stability toward other tested detergents and organic solvents.Cloning,expression,and biochemical characterization of Geobacillus sp.ZH1 carboxylesterase lay a good foundation for its structural characterization and industrial application.  相似文献   

2.
A thermostable superoxide dismutase (SOD) from the inshore thermophile Thermus sp. JM1 was purified to homogeneity by steps of fractional ammonium sulfate precipitation, DEAE-Sepharose chromatography and Phenyl-Sepharose chromatography. The specific activity of the purified native enzyme was 1 656 U/mg. A sod gene from this strain was cloned and overexpressed in Escherichia coli (E. coli). The prepared apo-enzyme of the purified recombinant SOD (rSOD) was reconstituted with either Fe or Mn by means of incubation with appropriate metal salts. As a result, only Mn 2+ - reconstituted rSOD (Mn-rSOD) exhibited the specific activity of 1 598 U/mg. SOD from Thermus sp. JM1 was Mn-SOD, judging by the specific activities analysis of Fe or Mn reconstituted rSODs and the insensitivity of the native SOD to both cyanide and H 2 O 2 . Both the native SOD and Mn- rSOD were determined to be homotetramers with monomeric molecular mass of 26 kDa and 27.5 kDa, respectively. They had high thermostability at 50 ° C and 60 ° C, and showed striking stability across a wide pH span from 4.0 to 11.0.  相似文献   

3.
The study investigated the effects of salinity on growth,antimicrobial activities and secondary metabolites of 47 marine filamentous fungi isolated from the East China Sea near the western shore of the Taiwan Straits.The results indicate that NaCl promoted the growth up to 91.5% of test strains.However,only 14.9% of them showed a significant increase of antimicrobial activity against Candida albicans.When incubated in different concentrations of NaCl,the colony growth, antimicrobial activities and composition of secondary metabolites of the strain Ty01b-8 of Penicillium sp.varied.Treatment with KCl also showed a similar effect.An alkaloid isolated from the fermentation broth of Ty01b-8 was identified as chrysogine,inhibition activity of which against Hela cells was 15.05% at 20 μ g/ml,and yield was 4.4 and 4.9 times higher in 3 percent and 6 percent NaCl treatments,respectively, compared with the non-salt culture condition. These findings prove that salinity is an important factor influencing growth and secondary metabolites of some marine fungi,which can be used to screen for new metabolites from marine fungi,and to enhance their metabolites production in industrial fermentation.  相似文献   

4.
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5.
Bacillus sp. JM7, a strain isolated from the deep-sea of the South China Sea, was found to efficiently degrade 79.4% native chicken feather within 30 h. Scanning electron microscopy analysis showed that JM7 strain could gradually degrade feather by modifying the microstructure of feather keratin. A total of 25 protease genes were predicted from the draft genome of JM7 strain, among which a predicted subtilisin-like serine protease(designated as Ker02562) was further characterized for its keratinolytic activity. The recombinant Ker02562 functioned at a wide range of temperatures from 30℃ to 60℃, with an optimum at 40–50℃. Ker02562 was highly active at various pHs ranging from 5.0 to 13.0, with a maximum activity observed at pH 7.0–9.0. Remarkably, recombinant Ker02562 was stable in extreme alkaline environments(pH 10–13), which was much better than most other reported keratinases. Collectively, these favorable properties could make Bacillus sp. JM7 and Ker02562 attractive to be applied in the detergent formulation and feather bioconversion.  相似文献   

6.
Enzymatic properties of UFE, a novel marine fibrinolytic enzyme   总被引：2，自引：0，他引：2  
A novel potent protease, Urechis unicinctus fibrinolytic enzyme (UFE), was firstly discovered. The enzymatic properties of UFE were further investigated. As a low molecular mass protein, UFE appeared to be very stable to heat and pH. When temperature was below 50 ℃, the remnant enzyme activity remained almost unchanged, but when temperature was raised to 60 ℃, the remnant enzyme activity began to decrease rapidly. UFE was quite stable in the range of pH value from 3 to 12, especially in slightly alkaline pH value. Mn2 , Cu2 and Fe2 ions were activators of UFE, while Fe3 and Ag ions were inhibitors of UFE. Fe2 ion along with Fe3 ion might regulate UFE activity in vivo. The optimum pH and temperature of UFE were about 8 and 50 ℃, respectively. Other characteristics of this enzyme were also studied. Systematic research results are significant when UFE is applied for medical and industrial purposes.  相似文献   

7.
mRNA was isolated from the hepatopancrease of shrimp Penaeus monodon with a PolyAT-tract System 1000 Kit. By using mRNA as template, double - strand cDNA with EcoR I/Xho I ends was synthesized by using a ZAP Express cDNA Synthesis Kit. The cDNA was inserted into the lambda ZAP Express vector predigested with EcoR I/Xho I, and the recombinant DNA was in vitro packaged into lambda phage with GigapackIII Gold packaging extracts. These recombinant phages were then used to transfect E. coli XL1 - Blue MRF', and finally a cDNA expression library was constructed. The library is 7.2 × 10~5 pfu in capacity and its recombination ratio is higher than 99 % . The size of the inserted cDNAs was determined by EcoR I/Xho I digestion of 9 phagemids prepared by in vivo excision of plaques selected randomly from amplified cDNA library . The longest inserted cDNA is about 1.6 kb in length. The complete sequence (about 1.2 kb) of actin cDNA was amplified from the library by PCR reveals that this library contains full-length  相似文献   

8.
A total of 117 agar-decomposing cultures were isolated from coastal seawater around Qingdao,China.The phenotypic and agarolytic features of an agarolytic isolate,QM38,were investigated.The strain was gram negative,strictly aerobic,curved rod and polar flagellum.On the basis of several phenotypic characters,biochemical and morphological characters and phylogenetic analysis of the gene coding for the 16S rRNA,the strain was identified as Agarivorans albus strain QM38.This strain can liquefy the agar on the solid agar plate.An excellular agarase activity was determined in liquid culture.The enzyme exhibited maximal activity at 40℃,pH 7.6.Its activity was greatly affected by different concentrations of agarose.The highest activity 32 U/ml was achieved in the culture supernatant.The hydrolytic product was analyzed by fluorophore-assisted carbohydrate electrophoresis (FACE).After complete hydrolysis of agarose,a series of agaro-oligosaccharides were produced.The main products of the enzymes were oligosaccharides in the degree of polymerization (DP) of 2,4,6 and 8.Three genes agaD01,agaD02 and agaD03,encoding β-agarases,had been cloned from genomic DNA of Agarivorans albus strain QM38.The open reading frame of agaD01,consisted of 2 988 bp,and shared 95.5%-98.9% identity to the β-agarase genes of some strains of Vibrio and Agarivorans.Gene agaD02 comprised 2 868 bp and encoded a 955-amino-acid protein.It showed 97.4% and 98.7% identity to the β-agarase genes of strain Vibrio sp.PO-303 and strain Vibrio sp.JT0107,respectively.Only partial sequence of agaD03 gene has been cloned.It showed 96.5% identity to β-agarase gene (agaB) of Pseudoalteromonas sp.CY24,and shared 96.8% identity to β-agarase-c gene of Vibrio sp.PO-303.  相似文献   

9.
A new member of antimicrobial protein genes of the Crustin family was cloned from haemocytes of the Chinese shrimp Fennero- penaeus chinensis by 3 ′and 5′ RACE. The full-length cDNA of Crustin-like gene contains a 390 bp open reading frame, encoding 130 amino acids. The deduced peptide contains a putative signal peptide of 17 amino acids and mature peptide of 113 amino acids. The molecular mass of the deduced mature peptide is 12. 3 ku. It is highly cationic with a theoretical isoelectric point of 8.5. The deduced amino acids sequence of this Crustin showed high homology with those of Penaeus （ Litopenaeus ） setferus. Northern blotting showed that the cloned Crustin gene was mainly expressed in haemocytes, gill, intestine, and RNA in situ hybridization indicated that the Crustin gene was constitutively expressed exclusively in haemocytes of these tissues. Capillary elec- trophoresis RT-PCR analysis showed that Crustin was up-regulated dramatically from 12 to 48 h after a brief decrease of mRNA during first 6 h in response to microbe infection. The level of Crustin mRNA began to restore at 72 h post-challenge. This indicated that Crustin gene might play an important role when shrimps are infected by bacterial pathogen.  相似文献   

10.
2018年北极太平洋区域夏季海冰物理及光学性质的研究   总被引：2，自引：1，他引：1  
The reduction in Arctic sea ice in summer has been reported to have a significant impact on the global climate. In this study, Arctic sea ice/snow at the end of the melting season in 2018 was investigated during CHINARE-2018, in terms of its temperature, salinity, density and textural structure, the snow density, water content and albedo, as well as morphology and albedo of the refreezing melt pond. The interior melting of sea ice caused a strong stratification of temperature, salinity and density. The temperature of sea ice ranged from –0.8℃ to 0℃, and exhibited linear cooling with depth. The average salinity and density of sea ice were approximately 1.3 psu and 825 kg/m~3, respectively, and increased slightly with depth. The first-year sea ice was dominated by columnar grained ice. Snow cover over all the investigated floes was in the melt phase, and the average water content and density were 0.74% and 241 kg/m~3, respectively. The thickness of the thin ice lid ranged from 2.2 cm to 7.0 cm, and the depth of the pond ranged from 1.8 cm to 26.8 cm. The integrated albedo of the refreezing melt pond was in the range of 0.28–0.57. Because of the thin ice lid, the albedo of the melt pond improved to twice as high as that of the mature melt pond. These results provide a reference for the current state of Arctic sea ice and the mechanism of its reduction.  相似文献   

11.
It was found that the marine yeast strain YF07b could secrete a large amount of killer toxin against a pathogenic yeast strain WCY which could cause milky disease in Portunus trituberculatus.The marine yeast strain YF07b was identified to be Pichia anomala according to the results of routine yeast identification and 18S rDNA and ITS sequences.The gene encoding killer toxin in the marine yeast strain YF07b was amplified by PCR technology.After sequencing,the results show that an open reading frame,consisting of 1 281 bp,encoded a presumed protein of 427 amino acids.The sequence of the cloned gene was found to have 99% match with that of the gene encoding killer toxin in Pichia anomalas strain K.A signal peptide including 17 amino acids appeared in the N-terminal domain of the killer toxin.Therefore,the mature protein consisted of 410 amino acids,its molecular mass was estimated to be 47.4 ku and its isoelctronic point was 4.5.  相似文献   

12.
Abstract-The growth rate of the hyalid amphipod Hyale perieri was studied on the bases of Ikeda'sgrowth model which is based on the inter moult period (IP) and moult increament (ΔBL). To applythis approach, laboratory experiments were carried out at three temperatures regimes (15℃, 20℃,25℃) to gain accurate data of IP and BL. The total number of specimens used in this study was 86 at15℃, 24 at 20℃ and 70 at 25℃. The number of flagellar segments of both antennae of the Hyaleperieri could not be used as an index of growth (instar criterion). The obtained results indicated that,the predicted IP of the specimens was inversely related to temperature and in good agreement with theobserved values at the experimental temperatures. IP data obtained from laboratory-reared specimes arecombined with ΔBL data to establish a growth model for Hyale perieri from its release from the mar-supium (1.64 mm BL) to the maximum size (12.67 mm BL) as a function of temperature. The maxi-mum numbers of consecutive moults  相似文献   

13.
One strain of unicellular greenish algae embedded by mucilage was successfully isolated from equatorial area in the Indian Ocean. Microscopic observation, ultrastructure features and genetic identification confirmed that the strain was closely related to Cyanothece sp., which was a cyanobacteria species with great ecological significance.Cells were solitary with oval or bacilliform shapes. Diameters of this strain were relatively small, ranging from 2.5 to 6.5 μm on average. Ultrastructure of cells was simple. Thylakoids were arranged parietal and keritomized content were observed in the thylakoid region. Various electron-transparent granules with low electron-dense region as well as cyanophycin or glycogen granules-like organelle and carbonxysomes were also observed. For pigment composition, the dominant pigments were chlorophyll a, β-Carotene, Zeaxanthin and an unknown pigment, contributing 23.8%, 26.1%, 14.7% and 15.7% to total pigments respectively. The phylogenetic analysis of16 S rRNA gene and nif H gene confirmed that Strain EIO409 was closely related with Cyanothece sp..  相似文献   

14.
A sediment core MD05-2902 was collected from the deep-sea basin of the Xisha Trough. The vertical distribution and diversity of bacteria in the core was investigated through ten sub-sampling with an interval of 1 m using bacterial 16 S rRNA gene as a phylogenetic bio-marker. Eighteen phylogenetic groups were identified from 16 S rRNA gene clone libraries. The dominant bacterial groups were JS1, Planctomycetes and Chloroflexi, which accounted for 30.6%, 16.6%, and 15.6% of bacterial clones in the libraries, respectively. In order to reveal the relationship between biotic and abiotic data, a nonmetric multidimensional scaling analysis was performed. The result revealed that the δ15N, δ13C, total organic carbon and total organic nitrogen possibly influenced the bacterial community structure. This study expanded our knowledge of the biogeochemical cycling in the Xisha Trough sediment.  相似文献   

15.
The neutralizing activities of eight monoclonal antibodies （MAbs） against white spot syndrome virus （WSSV） （2D2, 2B2,1D2, 1D5, 1C2, 4A1, 6A4 and 6B4） were analyzed by in vivo experiments. Gills from WSSV-infected shrimp were homogenized and ten-fold serially diluted by PBS, and then incubated with MAbs （hybridoma culture supernatant）, respectively. The mixture of WSSV and MAbs were injected into crayfish （Procambarus clarkii）. After challenge, the death rates of crayfish were counted to determine the neutralizing activities of MAbs. At the same time, the mixture of myeloma culture supernatant and WSSV or PBS was served as positive or negative control, respectively. The results showed that, at each virus dilution, the mean time to death of the crayfish injected with MAb-treated virus was significantly longer than that in the positive control, though they all showed 100% mortality within 25 d, and meanwhile, few crayfish died in the negative control. Among the eight MAbs, 2D2, 2B2, 1D2 and 1D5, especially the former two, delayed the mortality significantly, and 1 C2, 4A1 and 6A4 delayed the mortality as well but not so efficiently, while MAb 6IM was efficient only when the virus concentration increased. The results indicated that the anti-WSSV MAbs can neutralize WSSV in different virus dilutions.  相似文献   

16.
Genetic diversity of two wild Penaeus monodon populations sampled from the coastal waters of Qinglan （ Hainan Province of China, HN） and Malaysia （KD）, and the F1 generation of a Thailand broodstock population （CP） were examined by vertical polyacrylamide gel electrophoresis. Of 21 loci encoded by ten enzymes, 11 were polymorphic. The mean proportions of polymorphic loci of HN, KD and CP were 36. 36%, 45.45% and 50.00% , with the average heterozygosities of 0.135, 0.181 and 0.191, and the effective numbers of alleles per loci were 1. 300, 1. 330 and 1. 329, respectively. The divergent indexes of HN, KD and CP were0.023, 0.124, and 0.117, respectively. The genetic distance between the two wild populations was 0.005, and the gene differentiation coefficient was also very low （0.014）. The results indicated that the F1 generation population had a higher genetic diversity than the two wild populations, and that the HN population had the lowest one. There was no significant differentiation between HN and KD populations and all populations existed with a slight heterozygote excess.  相似文献   

17.
The diversity of modular polyketide synthase (PKS) genes in sediments of Ardley Island in Antarctica, was studied by restriction fragment length polymorphism (RFLP) analysis. Phylogenetic analysis of 14 amino acid (AA) sequences indicates that the identified ketosynthase (KS) domains were clustered with those from diverse bacterial groups, including Cyanobacteria, γ-Proteobacteria, Actinobacteria, Firmicutes, and some unidentified microorganisms from marine sponge, bryozoan and other environmental samples. The obtained KS domains showed 43%-81% similarity at the AA level to reference sequences in GenBank. Six identified KS domains showed diverse sequences of the motif (VQTACSTS) that was used to identify the hybrid PKS/nonribosomal peptide synthetase (NRPS) enzyme complex, and formed a new branch. These results reveal a high diversity and novelty of PKS genes in antarctic sediments.  相似文献   

18.
A deep-sea bacterium from the Pacific Ocean identified as Tenacibaculum sp. HMG1 was found to have strong malachite green(MG) degradation activity. The MG tolerance and decolorizing activities of strain HMG1 were confirmed by bacterial growth and high-performance liquid chromatography(HPLC) analyses. Strain HMG1 was capable of removing 98.8% of the MG in cultures within 12 h and was able to grow vigorously at 20 mg/L MG. A peroxidase gene detected in the genome of strain HMG1 was found to be involved in the MG biodegradation process. The corresponding recombinant peroxidase(r POD) demonstrated high degradative activity at 1 000 mg/L MG. Based on the common candidate intermediates, strain HMG1 was inferred to have one primary MG degradation pathway containing r POD. In addition, five other candidate intermediates of the r POD-MG degradative process were detected. The optimal conditions for MG degradation were determined and showed that strain HMG1 and the r POD enzyme could maintain high bioactivity at a low temperature(20°C), variable p H values(6.0–9.0), higher salinities(100 mmol/L) and other factors, such as multiple metal ions, H_2O_2 and EDTA.MG-tolerant strain Tenacibaculum sp. HMG1 and its peroxidase have prospective applications as environmental amendments for MG degradation during coastal remediation.  相似文献   

19.
The Taiwan Strait is a transition zone between the East China Sea and the South China Sea with unique hydrology and a geographical environment that creates special marine community features.To analyze the spatial structure and seasonal changes of the nekton assemblages in the Taiwan Strait,seasonal experimental trawl surveys were conducted during 2006–2007.The results showed that there were two assemblages in the area with different sets of species,and the average similarity within each group varies between 39.38% and 74.20%.By using multivariate statistical analysis and analyzing the distribution of dominant species,we found that the structures of the assemblages had obvious seasonal variation.The middle region from the Putian transect to the Xiamen transect could be considered a mixing area for the two assemblages.The analysis of the relationship between species assemblages and environmental factors indicated that temperature was the most important factor affecting the community structure in cold seasons,and 22.5°C and 17°C could be considered dividing lines for spring and winter,respectively.In warm seasons,the most important factor was water depth,but the relationship with depth was not as significant,with a correlation between 0.264 and 0.399.The seasonal changes of nekton assemblages basically reflected the dynamic currents in the Taiwan Strait.The south coastal assemblage extended to nearly the entire area of the Taiwan Strait along with a strong and intense warm current that shrinks in spring and winter when the southward intrusion of the cold Zhejiang-Fujian coastal current becomes stronger.The impact of shortterm and long-term environmental changes,such as extreme weather,global warming and human activity on nekton assemblages,had been recognized but need further research.Our study on nekton assemblages could be used as a baseline for measuring future changes.  相似文献   

20.
Subtropical sediment cores(QA09-1 and QA12-9) from the coastal zone of Qi'ao Island in the Zhujiang River Estuary were used to determine the rates of sulfate reduction and their response to experimental temperature changes. The depth distribution of the sulfate reduction rates was measured from whole-core incubations with radioactive tracer 35SO42-, and peaks of 181.19 nmol/(cm3·d) and 107.49 nmol/(cm3·d) were exhibited at stations QA09-1 and QA12-9, respectively. The profiles of the pore water methane and sulfate concentrations demonstrated that anaerobic oxidation of methane occurred in the study area, which resulted in an increase in the sulfate reduction rate at the base of the sulfate-reducing zone. Meanwhile, the sulfate concentration was not a major limiting factor for controlling the rates of sulfate reduction. In addition, the incubation of the sediment slurries in a block with a temperature gradient showed that the optimum temperature for the sulfate reduction reaction was 36℃. The Arrhenius plot was linear from the lowest temperature to the optimum temperature, and the activation energy was at the lower end of the range of previously reported values. The results suggested that the ambient temperature regime of marine environments probably selected for the microbial population with the best-suited physiology for the respective environment.  相似文献   

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