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1.
2-haloacid dehalogenases constitute a group of dehalogenases which are capable of dehalogenating the halogenated organic compounds. So far, the 2-haloacid dehalogenases have been found in many bacteria, but not in Paracoccus genus. In the present study, one enzyme 2-haloacid dehalogenase(designated as Deh99), induced by DL-2-chloropropionate(DL-2-CPA), was purified from the marine bacterium Paracoccus sp. DEH99, isolated from marine sponge Hymeniacidon perlevis. The enzyme of Deh99 was purified to homogeneity by ammonium sulfate precipitation, ion exchange chromatography(Q-Sepharose HP), and Superdex 200 gel filtration chromatography. The molecular weight of Deh99 was estimated to be 25.0 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE), and 50.0 kDa natively by gel filtration chromatography. The enzyme of Deh99 stereospecifically dehalogenated L-2-CPA to produce D-lactate, with an apparent Michaelis-Menten constant(Km) value of 0.21 mmol L-1 for L-2-CPA. The optimal pH and temperature for Deh99 activity were 10.0 and 40℃, respectively. The enzyme of Deh99 acted on short-carbon-chain 2-haloacids, with the highest activity towards monochloroacetate. The activity of Deh99 was slightly affected by DTT and EDTA, but strongly inhibited by Cu2+ and Zn2+. The enzyme of Deh99 shows unique substrate specificity and inhibitor sensitivities compared to previously characterized 2-haloacid dehalogenases and is the reported one about purified 2-haloacid dehalogenase isolated from the bacteria of Paracoccus genus. 相似文献
2.
A new chemically mutagenic mutant ofSynechococcus PCC7942 named high-CO2 requiring mutant, which could grow at 4% CO2 but could not grow at air levels of CO2, was isolated. Comparative studies on some physiological aspects of the mutant and high-CO2 growing cells (growing at 4% CO2) were conducted. The result showed that the mutant had lower growing rate, about 1/40th photosynthetic affinity to inorganic carbon, 25% lower carbonic anhydrase (CA) activity, lower quenching rate of chlorophyll fluorescence, and about 1/2 alkalinization rate of the medium. The CA activity responses of the two types of cells to different concentration of CO2 were determined. Upon the addition, of inorganic carbon (Ci), the rate of active Ci uptake described by the rate of chlorophyll fluorescence quenching of the mutant was obviously lower compared with that of the high-CO2 growing cells; the size of the internal inorganic carbon pool size detemined by the extent of fluorescence quenching of the mutant was also smaller.
相似文献3.
A superoxide dismutase was purified from Enteromorpha linza using a simple and safe procedure, which comprised phosphate buffer extraction, ammonium sulphate precipitation, ion exchange chromatography on Q-sepharose column, and gel filtration chromatography on Superdex 200 10/300GL. The E. linza superoxide dismutase （E/SOD） was purified 103.6-fold, and a yield of 19.1% and a specific activity of 1 750 U/rag protein were obtained. The SDS-PAGE exhibited E/SOD a single band near 23 kDa and the gel filtration study showed E/SOD＇s molecular weight is near 46 kDa in nondenatured condition, indicating it＇s a homodimeric protein. E/SOD is an iron-cofactored superoxide dismutase （Fe-SOD） because it was inhibited by hydrogen peroxide, insensitive to potassium cyanide. The optimal temperature for its maximal enzyme activity was 35℃, and it still had 29.8% relative activity at 0℃, then E/SOD can be classified as a cold-adapted enzyme. E/SOD was stable when temperature was below 40℃ or the pH was within the range of 5 10. The first 11 N-terminal amino acids orE/SOD were ALELKAPPYEL, comparison of its N-terminal sequence with other Fe-SOD N-terminal sequences at the same position suggests it is possibly a chloroplastic Fe-SOD. 相似文献
4.
High-precision isotope composition determinations using multicollector, magnetic-sector inductively coupled plasma mass spectrometry (MC-ICPMS) have recently revealed that some transition metal isotopes such as those of Mo, Fe, Cu, Zn etc. can be used as biogeochemical tracers in global change research.The Mo isotope system may be useful in paleoredox investigations indicating that δ^97 ^95Mo in seawater may co-vary with changes in the relative proportions of anoxic and oxic sedimentation in the ocean, and that this variation may be recorded in δ^97 ^95Mo of anoxic sediments. The Mo continental flux into the oceans and the global Mo isotope budget can be estimated from δ^97 ^95M0 values. The Fe isotope composition in seawater is an important issue because Fe plays a controlling role in biological productivity in the oceans and its abundance in seawater may have substantial effect on climate changes. Iron isotope fractionations could result from bio- and abio-processes and have about 0.1% variation ( δ^56 ^54Fe), so Fe isotopes considered alone cannot be used to distinguish the products of abiotic and biotic Fe processing in geological records. Cu and Zn isotopes are also used as biogeochemical tracers, but the researches are relatively less. This review mainly focuses on the methods for preparation, purification and determination of new isotope tracer samples, and on isotope applications in marine environmental changes. 相似文献
5.
Cao Ji-xiang 《中国海洋湖沼学报》1996,14(4):378-386
This study on the effect of different concentrations of germanium dioxide (GeO2) on the specific growth rate (SGR), pigment contents, protein content and amino acid composition ofSpirulina platensis showed that Ge was not the essential element of this alga; that GeO2 could speed up growth and raise protein content ofS. platensis, and could possibly influence the photosynthesis system. The concentration range of GeO2 beneficial to growth ofS. platensis is from 5–100mg/l. GeO2 is proposed to be utilized to remove contamination byChlorella spp. usually occurring in the cultivation ofSpirulina. 相似文献
6.
Allelic variation in a total of 7 microsatellites was examined between elvers of freshwater eels （Anguillajaponica and Anguilla anguilla）. The number of alleles at these loci ranged from 8 to 26. A single test of each locus revealed significant deficits of heterozygotes （P〈0.01）. Significant departure from expectations of Hardy-Weinberg equilibrium （HWE） was found for all loci within four subpopulations of A. japonica, which opposes the panmixia hypothesis of Schmidt. Also exact tests of population differentiation based on allelic frequency distribution disagree the hypothesis of random distribution of individuals among populations. Population structure among four populations of A. japonica was revealed with FST value of 0.009 8 （P=-0.000 48; 10 000 iteration）. Pairwise matrixes of FST and RST showed a significant difference between two distantly related species-A, japonica and A. anguilla. Divergent time of the two species calculated by Goldstein method is over 2 million years. The results may challenge the Schmidt＇s theory about the distribution of freshwater eels. 相似文献
7.
INTRODUCTIONManystudiesonvascularplantSduringthenontwperiodhavedrireP0redinChinaandabroad.TherearefeWreportSonwhethertherearesomevasctilarplantS,andhowmanyspedessurvive,underice,andontheirbiomassesandphotosyntheticproduCtivitiesduringwinter,espedal1yduringbeAfpinchLillyamepe0ylenctal.,l976,Liuetal.,l994).WeimtlycarriedoutaseadriontheunderforvascularplantsdurinwinderinsomewaterbodieswherevascularplantSgrowluxdrianhydurinInildareonsinnorthemareas,soastoimproveourknowedgeofvascularplaflt… 相似文献
8.
The effects of a diet containing Hanseniaspora opuntiae C21 on growth and digestive enzyme activity were estimated in juvenile Apostichopus japonicus. Groups of sea cucumbers were fed diets containing H. opuntiae C21 at 0(control), 10 4, 10 5, and 10 6 CFU(colony-forming units)/g feed. Results showed that after 45 d the specific growth rate(SGR) of sea cucumbers fed a C21-supplemented diet at 10 4 CFU/g feed was significantly higher than that of the control( P 0.05). Intestinal trypsin and lipase activities were significantly enhanced by C21 administration at 10 4 and 10 5 CFU/g feed compared with the control( P 0.05). After feeding for 23–42 d, C21 was demonstrated by denaturing gradient gel electrophoresis to be present in the intestine of sea cucumbers. In addition, after feeding the C21-supplemented diets for 15 d, the sea cucumbers were switched to an unsupplemented diet and C21 was confirmed to be capable of colonizing the intestine for at least 31 d after cessation of feeding. In conclusion, C21 was shown to successfully colonize the intestine of juvenile A. japonicus via dietary supplementation, and improve growth and digestive enzyme activity. 相似文献
9.
LI Hao MIAO Jinlai CUI Fengxia LI Guangyou 《中国海洋大学学报(英文版)》2007,6(4):365-368
Two-dimensional electrophoresis(2-DE) of protein extracted and purified from Alexandrium sp. LC3 was conducted. In the SDS-PAGE study, the relative molecular weights of the proteins were mainly in the range of 14 kDa?31 kDa and 43 kDa?66 kDa, and more proteins were detected between 14 kDa and 31 kDa. With the improved protein preparation, the two-dimensional electrophoresis patterns indicated that the relative molecular weights of the proteins were between 14 kDa and 100 kDa, and most of them ranged from 14 kDa to 31 kDa. This was consistent with the result of the SDS-PAGE analysis. The isoelectric points were found to lie between 3.0 and 8.0, and most of them were in the range of 3.0?6.0. Better separation effect was acquired with pre-prepared immobilized gradient (IPG) strip (pH 3?5.6), and about 320 protein spots could be visualized on the 2-DE map by staining. Within pH 3?10 and pH 3?5.6 strips, the protein samples of Alexandrium sp. LC3 could be separated well. 相似文献
10.
A bacterium hydrolyzing carboxymethylcellulose, isolated from Antarctic sea ice, was identified as Pseudoalteromonas sp. based on 16S rDNA gene sequences and named as Pseudoalteromonas sp. AN545. The extracellular endo-1,4-β-glucanase AN-1 was purified successively by ammonium sulfate precipitation, DEAE-Sepharose ion exchange chromatography and Sephadex G-75 gel filtration chromatography. The molecular mass of AN-1 was estimated to be 47.5 kDa utilizing SDS-PAGE and gel chromatography analysis. AN-1 could hydrolyze caboxymethylcellulose, avicel and β-glucan, but not cellobiose, xylan and p-Nitrophenyl-β-D-glucopyranoside. The optimal temperature and pH for the β-glucanase activity of AN-1 were determined to be at 30°C and pH 6.0, respectively. AN-1 was stable at acidic solutions of pH 5.0-6.5 and temperatures below 30°C for 1 h. Moreover, the specific activity was enhanced by Ca2+ and Mg2+, and inhibited by Cu2+. The kinetic parameters Michaelis constant (Km) and maximum velocity (Vmax) of AN-1 were 3.96 mg/mL and 6.06×10-2 mg/(min·mL), respectively. 相似文献
11.
Cultures of a blue-green algaSpirulina maxima (Setch. et Gard.) Geitler with various concentrations of Se in Zarrouk's medium showed that not higher than 40 mg/L Se could
promote its growth. The present experiments showed thatS. maxima grown under normal conditions, has an oxidant stress defence system for hydrogen peroxide (H2O2) removal, which is the Halliwell-Asada pathway. When 4 to 20 mg/L Se was added to the algal medium, this pathway was replaced
by a so-called Sestressed pathway containing GSH peroxidase (GSH-POD). As a result of the occurrence of both higher activity
of GSH-POD and lower levels of hydroxyl radical (OH·), the Se-stressed pathway scavenged H2O2 so effectively that the growth ofS. maxima was promoted by 4 to 20 mg/L Se. While GSH-POD activity of the alga disappeared at 40 mg/L Se, the recovery of ascorbate
peroxidase was observed. The lower levels of ascorbic acid and GSH made the Halliwell-Asada pathway for scavenging H2O2 less effective, while the highest activity of catalase might be responsible in part for the H2O2 removal, causing the level of OH· inS. maxima grown at 40 mg/L Se to be much higher than the OH· level in this alga grown at 4 to 20 mg/L Se, but lower than that in the
control. The OH· level changes caused the growth ofS. maxima cultured at 40 mg/L Se to increase slightly to close to that of the control. 相似文献
12.
A study on the nonspecific immunity of Litopenaeus vannamei ever inhabiting freshwater and seawater was carried out at different molt stages by comparing their total hemocyte count(THC) and respiratory burst(RB) and activity of phenol oxidase(PO), nitric oxide synthase(NOS) and lysozyme(LY). Two-way ANOVA showed that salinity and molt stage independently affected THC and RB and the activity of PO, NOS and LY of juvenile L. vannamei significantly(P 0.05). The THC and RB and the activity of NOS gradually increased from the post-molt stages(A and B) to the pre-molt stages(D0–D3), which were common in shrimps inhabiting freshwater and seawater. The activity of PO peaked at the inter-molt stage(C), and touched the lowest at the post-molt stage in freshwater and pre-molt stage in seawater. The activity of LY was stable over the molt cycle. The RB and the activity of PO, NOS and LY of juvenile L. vannamei were significantly lower in freshwater than in seawater; whereas THC was significantly higher in freshwater than in seawater(P 0.05). It was concluded that the post-molt stage(especially stage A) was critical to shrimp culture, which should be intensively attended when L. vannamei was cultured in freshwater. 相似文献
13.
Serum immunoglobulin of the mandarin fish, Siniperca chuatsi with development of polyclonal antibody
Serum immunoglobulin from the mandarin fish, or the so-called Chinese perch,Siniperca chuatsi (Basilewsky), was successfully purified using affinity chromatography. Heavy and light chains were detected on electrophoresis
gel, with molecular weights being estimated at 72 and 29 kDa, respectively. The tetrameric IgM ofS. chuatsi was calculated to be 808 kDa. The rabbit polyclonal antisera against the purifed immunoglobulin were developed and tested
by Western blot analysis. The antisera reacted strongly with the heavy chains ofS. chuatsi immunoglobulin. Humoral immune responses of the mandarin fish can then be examined using the developed polyclonal antibody.
Project No. 30025035 supported by NSFC, State Key Laboratory of Freshwater Ecology and Biotechnology (2000FB02), and the Chinese
Academy of Sciences (KZ9511-B1-111). 相似文献
14.
Trace and RE element geochemistry and genesis have been studied with respect to ferromanganese nodules from the sediments of the Pacific, Atlantic and Indian oceans. 相似文献
15.
Membrane preconcentration of iron in seawater samples and on-site determination in spectrophotometry
A novel method for on-site determination of trace iron was developed using membrane preconcentration and spectrophotometric detection. Fe(Ⅱ)-ferrozine complex was reacted with cetyltrimethylammonium bromide (CTAB) to form a Fe(Ⅱ)-ferrozine-CTAB paired compound, which was collected on a membrane by filtration under vacuum. The membrane was immersed in 2 mL of ethanol-nitric acid and the absorbance of the solution measured for quantitative analysis. Various factors affecting the iron collection and determination were investigated. With different sample preconcentration volumes, the range of determination was broadened to 0.5-120 g/L. The detection limit of this method reached 0.19 g/L and the recoveries were between 97.2 and 109% when the concentration enrichment was about 45. The relative standard deviation (n = 7) was 1.9% for samples containing 10 g/L Fe. Twelve seawater samples were analyzed on-site using the proposed method, and two were also analyzed in inductively coupled plasma mass spectrometry. No significant difference was shown between the two methods by the Student’s t-test. The method has also been used on-site for iron enrichment experiments with phytoplankton and concluded to be simple, accurate and inexpensive. 相似文献
16.
A component (s-factor) with obvious promoting effect on hydrogen evolution of hydrogenase has been isolated and extracted
from a cell-free preparation ofSpirulina platensis. The effect of the s-factor in the reaction system is similar to that of Na2S2O4, but is coupled with light. The s-factor has the maximum absorption peak at 620 nm in the oxidized state, at 590 nm in the
reduced state. The partially purified s-factor showed two bands by SDS-PAGE and is distinctly different from phycocyanin,
which has no change of oxidized state and reduced state absorption spectra, and also has no promoting effect on hydrogenase
ofSpirulina platensis under the light.
The research supported by the NSFC. 相似文献
17.
Zhang Cheng Han Xiaotian Zou Jingzhong Yu Zhiming Song Xiuxian Christian Schuett 《中国海洋湖沼学报》2005,23(3):317-322
A fragment of a large sub-unit ribosomal DNA (LrDNA) of 12 strains ofProrocentrum species was amplified by polymerase chain reaction (PCR). The PCR products were digested by 3 restriction endonucleases (Cfo
I, Hae III, and RSA I) and then resolved in agarose gels. Results show that different species had different RFLP patterns,
except forP. arcuatum (ME 131), which had the same pattern toP. micans (ME160 and 04). The same fragment of 19 strains of the genus was also amplified and subjected to denaturing gradient gel
electrophoresis (DGGE). 11 different patterns were resolved. Different cultures of a same species had the same pattern. The
results of RFLP and DGGE analyses showed that eight newly isolated epibenthicProrocentrum species were different from each other, and also from other cultured ones examined in this study.P arcuatum (ME132) could not be differentiated fromP. micans (ME160 and 04), it was probably mis-identified, since they are quite different morphologically.P. redfieldii (ME138) could also not be distinguished formP. triestinium (ME132), it should be regarded as a synonym ofP. triestinium. Unexpectedly, a restriction site was found inP. micans, compared with previous sequence data.
Project supported by National Basic Research Priorities Program (2001CB409701, 2001CB409710) and supported by NSFC (40376040,
40025614) 相似文献
18.
Modern sedimentation rates and sedimentation feature in the Huanghe River estuary based on210Pb technique 总被引：1，自引：0，他引：1
Li Fengye 《中国海洋湖沼学报》1993,11(4):333-342
Based on210Pb technique, sedimentation rates of 11 cores collected from the Huanghe River estuary were determined. CaCO3 distribution of 4 cores chosen from among these was measured. Profiles of210Pb in the cores showed that the distribution of210Pb activity decayed with depth, appeared in stages and in more than one segment. The sedimentation rates,210Pb and CaCO3 data have similar distribution trend in the 4 cores. The profiles of210Pb and CaCO3 were used to study sedimentation intensity, transportation trend of material and sedimentation features in the corring area.
Contribution No. 2230 from the Institute of Oceanology, Academia Sinica.
This research was supported by the National Natural Science Foundation of China. 相似文献
19.
Flavobacterium columnare causes columnaris disease in freshwater fish. In the present study, the antigenic regions of five outer membrane proteins (OMPs), including zinc metalloprotease, prolyl oligopeptidase, thermolysin, collagenase and chondroitin AC lyase, were bioinformatically analyzed, fused together, and then expressed as a recombinant fusion protein in Escherichia coli. The expressed protein of 95.6 kDa, as estimated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was consistent with the molecular weight deduced from the amino acid sequence. The purified recombinant protein was used to vaccinate the grass carp, Ctenopharyngodon idella. Following vaccination of the fish their IgM antibody levels were examined, as was the expression of IgM, IgD and IgZ immunoglobulin genes and other genes such as MHC Iα and MHC IIβ, which are also involved in adaptive immunity. Interleukin genes (IL), including IL-1β, IL-8 and IL-10, and type I and type II interferon (IFN) genes were also examined. At 3 and 4 weeks post-vaccination (wpv), significant increases in IgM antibody levels were observed in the fish vaccinated with the recombinant fusion protein, and an increase in the expression levels of IgM, IgD and IgZ genes was also detected following the vaccinations, thus indicating that an adaptive immune response was induced by the vaccinations. Early increases in the expression levels of IL and IFN genes were also observed in the vaccinated fish. At four wpv, the fish were challenged with F. columnare, and the vaccinated fish showed a good level of protection against this pathogen, with 39% relative percent survival (RPS) compared with the control group. It can be concluded, therefore, that the five OMPs, in the form of a recombinant fusion protein vaccine, induced an immune response in fish and protection against F. columnare. 相似文献