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1.
An alkaline protease from Acetes chinensis was purified and characterized in this study. The steps of purification include ammonium sulfate precipitation, ion-exchange
chromatography with Q-sepharose Fast Flow, gel filtration chromatography with S300 and the second ion-exchange chromatography
with Q-sepharose Fast Flow. The protease was isolated and purified, which was present and active on protein substrates (azocasein
and casein). The specific protease activity was 17.15 folds and the recovery was 4.67. The molecular weight of the protease
was estimated at 23.2 kD by SDS-PAGE. With azocasein as the susbstrate, the optimal temperature was 55°C and the optimal pH
value was 5.5. Ion Ca2+ could enhance the proteolytic activity of the protease, while Cu2+, EDTA and PMSF could inhibit its activity. 相似文献
2.
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
3.
We isolated a bacterial strain (HC4) that is able to degrade κ-carrageenan from a live specimen of the red alga Hyalosiphonia caespitosa. With 16S rRNA gene sequencing, we identified the strain as Tamlana sp., and then purified an extracellular κ-carrageenase from a culture of Tamlana sp. HC4 by ammonium sulfate precipitation, Sephadex G-200 gel filtration chromatography, and DE-cellulose 52 anion-exchange
chromatography. The purified enzyme yields a single band on SDS-PAGE with a molecular mass of 66.4 kDa. The optimal pH and
temperature for κ-carrageenase activity are at 8.0 and 30°C, respectively. The enzyme is stable over the range of pH 7.2–8.6
below 45°C. The enzyme activity is strongly inhibited by Zn2+ and Cu2+ at 1 mmol/L. The enzyme-catalyzed reaction follows Michaelis-Menten kinetics with the Michaelis constant (K
m
) at 7.63 mg/ml. Analysis of the degradation products of the κ-carrageenase by ESI-MS and 13C-NMR spectroscopy indicates that the enzyme degrades κ-carrageenan down to the level of κ-neocarrabiose sulfate. 相似文献
4.
Vibrio harveyi cells (dose—<103 cells mL−1) and extracellular products (ECP; >25 μmg mL−1 of total protein concentration) destroyed haematopoietic cultures of Nephrops norvegicus within 24 h of exposure. Cytopathic effects (CPE) started after 4h of exposure to the bacterial cells, with some granularity
in the cytoplasm, mostly in cells in the outer periphery of the explant growth. At the end of the infection, a considerable
number of nuclei remained attached to the substrate, apparently unaffected. Following exposure to ECP, initial deterioration
was observed at 2 h with the presence of granularity in the cytoplasm of<20% cells, and few cells displayed small vacuoles
around the nuclei. Parallel results were obtained using whole animal experiments, with V. harveyi cells being lethal to nephrops within 24 h. 相似文献
5.
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
6.
A new kinetic spectrophotometric method has been developed for the determination of vanadium (V). The method is based on the
catalytic effect of vanadium (V) on the oxidation of weak acid brilliant blue dye (RAWL) by KBrO3 using the citric acid as activation reagent. The obtained optimum conditions are: c (RAWL) = 1×10−4 molL−1, c (KBrO3) = 3×10−2 molL−1, c (citric acid) = 9×10−3 molL−1, pH = 2.50, the reaction time being 7.0 min and the temperature being 25.0°C. Under the optimum conditions, the proposed
method allows the determination of vanadium (V) in the range of 0–70.0 ng mL−1 and the detection limit is down to 0.407 ng mL−1. For standard vanadium (V) solution determination, the recovery efficiency is in the range of 98.5%–102% and the RSD ranges
from 0.76%–1.25%. Moreover, it is demonstrated that most cations and anions do not interfere with the determination of vanadium
(V) under the analytical condition. The new method was successfully applied in the determination of vanadium (V) in fresh
water and seawater samples with satisfactory results. Vanadium (V) in the seawater samples from Qingdao offshore was determined
using the method and the distribution of vanadium (V) was mapped. Compared with other instrumental analytical methods, the
proposed method shows good selectivity, sensitivity, simplicity, lower cost and rapidity. It can be employed on shipboard
easily. 相似文献
7.
Ustadi K. Y. Kim S. M. Kim 《中国海洋大学学报(英文版)》2005,4(3):198-204
The protease inhibitor was purified from five different fish eggs. The molecular weights of Pacific herring, chum salmon, pond smelt, glassfish, and Alaska pollock egg protease inhibitors were 120, 89, 84.5, 17, and 16.8kDa, respectively. The specific inhibitory activity of glassfish egg protease inhibitor was the highest followed by those of Pacific herring and Alaska pollock in order. The specific inhibitory activity and purity of glassfish egg protease inhibitor were 19.70 U mg^- 1 protein and 164.70 folds of purification, respectively. Glassfish egg protease inhibitor was reasonably stable at 50 - 65℃ and pH 8, which was more stable at high temperature and pH than protease inhibitors from the other fish species. Glassfish egg protease inhibitor was noncompetitive with inhibitor constant （Ki） of 4.44 nmol L^-1 相似文献
8.
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10.
A new extracellular κ-carrageenase, namely CgkP, 34.0 kDa in molecular weight, was purified from Pseudoalteromonas sp. QY203. CgkP showed relatively high activity at acidities ranging from pH6.0 to pH9.0 and temperatures ranging from 30℃ to 50℃ with the highest activity at 45℃ and pH7.2. Sodium chloride increased its activity markedly, and KCl increased its activity slightly. The divalent and trivalent metal ions including Cu2+ , Ni2+ , Zn2+ , Mn2+ , Al3+ and Fe3+ significantly inhibited its activity, while Mg2+ did not. CgkP remained 70% of original activity after being incubated at 40℃ for 48 h, and remained 80% of the activity after being incubated at 45℃ for 1 h. It exhibited endo-κ-carrageenase activity, mainly depolymerizing the κ-carrageenan into disaccharide and tetrasaccharide. CgkP was more thermostable than most of previously reported κ-carrageenases with a potential of being used in industry. 相似文献
11.
ＩＮＴＲＯＤＵＣＴＩＯＮＴｈｅＪａｐａｎｅｓｅｓｅａｂａｓｓ (ＬａｔｅｏｌａｂｒａｘｊａｐｏｎｉｃｕｓＣ＆Ｖ)ｄｉｓｔｒｉｂｕｔｅｓｗｉｄｅｌｙｉｎｔｈｅｃｏａｓｔａｌｗａｔｅｒｓａｌｏｎｇｔｈｅＢｏｈａｉＳｅａ ,ＹｅｌｌｏｗＳｅａ ,ＥａｓｔＣｈｉｎａＳｅａ,ａｎｄＳｏｕｔｈＣｈｉｎａＳｅａ .Ａｓａｃａｒｎｉｖｏｒｏｕｓｆｉｓｈ ,ｉｔｏｃｃｕｐｉｅｓｔｈｅｈｉｇｈｔｒｏｐｈｉｃｌｅｖｅｌｉｎｔｈｅｍａｒｉｎｅｅｃｏｓｙｓｔｅｍａｎｄｈａｓａｄａｐｔｅｄｔｏｖａｒｉｏｕｓｅｎｖｉｒｏｎｍｅｎｔａｌｒ… 相似文献
12.
1 INTRODUCTION Chironomid larvae are main groups in most aquatic ecosystems, playing a crucial ecological role in decomposition of detritus and material ex- changes between water column and the sediment (Liang et al., 1995a, b; Chen, et al., 1982). They a… 相似文献
13.
ZHOU Wenjun LI Yun DAI Jixun 《中国海洋大学学报(英文版)》2007,6(3):299-302
Cryopreservation of Porphyra yezoensis conchocelis was conducted with cryoprotectants and a proposed pretreatment procedure and thawing methods explored. Six cryoprotectants combined by DMSO with ethylene glycol （EG）, propylene glycol （PEG）, sorbitol and sucrose were developed. The effect of prefreezing at -40℃ or -20℃ for different time durations was compared and the thawing methods were screened. It was shown that the cryoprotectant including 10% DMSO with 0.5 molL^-1 sorbitol exhibited the optimal effect. The ideal pretreatment was that conchocelis segments were stayed for 20min at -40℃ before stored in liquid nitrogen, and 40℃ water bath was proper for quick thawing. The highest recovery rate of cryopreserved P. yezoensis conchocelis reached 89.41%. 相似文献
14.
Effects of salinity, light and temperature on growth rates of two species of Gracilaria （Rhodophyta） 总被引：2，自引：0，他引：2
Effects of temperature, salinity and light intensity on growth rates of Gracilaria lichenoides and G. tenuistipitata var. liui Zhang et Xia were tested. Eight to ten levels of each factor were first tested separately. The best growth rate was obtained
under the conditions of 32°C, 30 and 240 μmol/(m2·s) for G. lichenoides, and 24°C, 20 and 200 μmol/(m2·s) for G. tenuistipitata, respectively. Then a uniform design was used to evaluate the optimal combinations of the three factors. The best conditions
for the highest daily specific growth rates (% increase in wet weight) are determined to be 31.30°C, 32.10, and 287.23 μmol/(m2·s) for G. lichenoides (16.26%/d), and 25.38°C, 21.10, and 229.07 μmol/(m2·s) for G. tenuistipitata (14.83%/d), respectively.
Supported by the 908 Special Program (908-02-04-07), the National Basic Research Program of China (973 Program, No. 2006CB400608),
and K. C. Wong Magna Fund in Ningbo University 相似文献
15.
ＩＮＴＲＯＤＵＣＴＩＯＮＴｈｅｍｅｔａｂｏｌｉｓｍｉｎｆｉｓｈｉｎｃｌｕｄｅｓｓｔａｎｄａｒｄｍｅｔａｂｏｌｉｓｍ(ＲＳ)ｒｏｕｔｉｎｅｍｅｔａｂｏｌｉｓｍ(ＲＲ),ｓｐｅｃｉｆｉｃｄｙｎａｍｉｃａｃｔｉｏｎ(ＳＤＡ)ａｎｄａｃｔｉｖｅｍｅｔａｂｏｌｉｓｍ(ＲＡ),ｒｅｌａｔｅｄａｓ: ＲＴ=ＲＳ ＲＲ ＳＤＡ ＲＡｗｈｅｒｅＲＳｉｓｔｈｅｍｅｔａｂｏｌｉｓｍｏｆｔｈｅｆｉｓｈａｔｒｅｓｔ;ＲＲｔｈｅｍｅｔａｂｏｌｉｓｍｏｆｔｈｅｒｏｕｔｉｎｅｌｙａｃｔｉｖｅｆｉｓｈ;ＳＤＡｔｈｅｍｅｔａｂｏｌｉｓｍｏｆｔｈ… 相似文献
16.
Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10℃ and under continuous irradiance of 30 μmol m-2 s-1 for 3 weeks were encapsulated in calcium alginate beads. The beads were dehydrated in 0.4 molL-1 sucrose prepared with seawater for 6 h, desiccated in an incubator set at 10℃ and 70% relative humidity for 4 h, pre-frozen at either -40℃ or -60℃ for 30 min, and stored in liquid nitrogen for ＞24 h. As high as 43% of survival rate was observed when gametophytes were thawed by placing the beads in 40℃seawater and re-hydrated in 0.05 molL-1 citrate sodium prepared using 30‰ NaC1 7 d later. More cells of male gametophytes survived the whole procedure in comparison with female gametophytes. The cells of gametophytes surviving the preservation were able to grow asexually and produce morphologically normal sporophytes. 相似文献
17.
Extracellular products (ECP) produced byVibrio anguillarum strain M3 originally isolated from diseased flounder (Paralichthys olivaceus) were prepared. ECP of M3 showed gelatinase, casinase, amylase and haemolytic activity on agarose plates. High protease activity
against azocasin was detected. Bacterium M2 showed highest growth and protease activity at 25°C. The protease present in ECP
showed maximal activity at pH 8 and 55°C; was completely inactivated by application of 80°C heat for 30 min; was completely
inhibited by EDTA and HgCl2, and was partially inhibited by PMSF, SDS, MnCl2 and iodoacetic acid; but not inhibited by CaCl2 and MgCl2. The ECP was toxic to flounder fish at LD50 values of 3.1 μg protein/g body weight. The addition of HgCl2 and application of heat at 50°C decreased the lethal toxicity of ECP. When heated at 100°C, ECP lethality to flounder was
completely inhibited. After intramuscular injection of ECP into flounder, it showed evident histopathological changes including
necrosis of muscle, extensive deposition of haemosiderin in the spleen, dilated blood vessels congested with numerious lymphocytes
in the liver. These results showed that ECP protease was a lethal factor produced by the bacteriumV. anguillarum M3.
Contribution No. 4213 from the Institute of Oceanology, Chinese Academy of Sciences.
Funded by projects under the Major State Basic Research Development Program, Grant G1999012003. 相似文献
18.
STUDIES ON CULTURE CONDITIONS OF BENTHIC DIATOMS FOR FEEDING ABALONE I.EFFECTS OF TEMPERATURE AND LIGHT INTENSITY ON GROWTH RATE 总被引：2，自引：0，他引：2
INTRODUCTIONBenthicdiatomscomPrisethemaininitialfoodforjuvenileabalone.ItisimPortanttocIJlturehighqualitybenthicdiatornsduringtheeariystageofjuvenileabaloneculture.AtpresentthespeCesofbenthicdiatomsusedinabalonebedingfannsprirnarilycomefromnaturalpopulations.SomespedesinseveralgenerasuchasNavicula,Ntzchia,Caroneis,Achnanthes,AmPhoraareusuallythedothenantspch(Chen,etal.,l977,Ukiandaxuchi,l979,Nonnan-Boudnauetal.,l986,Austinetal.l990).Inordertoselodsornespedeswithhighadaptability,fas… 相似文献
19.
Chondrus is a type of commercially produced red seaweed that widely used for food and carrageen extraction. Although the natural life
history of the alga had been well understood, the factors influencing development of the tetraspore and carpospore remain
poorly understood. In the perspective of seedling resources, the regulation of early development is crucial for the seedling
nursing; therefore, it is necessary to understand the physiological influences during its early development. In this study,
we studied the effects of temperature and irradiance on the early development of Chondrus ocellatus Holm under laboratory conditions. The released tetraspores and carpospores were cultivated at different temperatures (10–28°C)
and irradiances (10, 60 μmol photons m−2s−1) with a photoperiod of 12L:12D. The results indicate that both tetraspores and carpospores are tolerant to temperatures of
10–25°C, and have the highest relative growth rate at 20°C. Irradiance variances influenced the growth of the discoid crusts,
and the influence was more significant with increasing temperature; 60 μmol photons m−2s−1 was more suitable than 10 μmol photons m−2s−1. The optimum temperature and irradiance for the development of seedlings was 20°C and 60 μmol photons m−2s−1, respectively. 相似文献
20.
A cell line,SHK,was derived from the kidney of spotted halibut Verasper variegates.The cell line was subcultured more than 40 passages in minimum essential medium(MEM)supplemented with fetal bovine serum(FBS)and 10 ng ml-1 basic fibroblast growth factor(bFGF).Cell morphology from primary culture and subculture was observed continuously by microscopy.The SHK cell line consisted predominantly of fibroblast-like cells.The cell line was able to grow between 20℃ and 30℃ with the optimum growth at 24℃ and with a reduced growth between 12℃ and 20℃.The growth rate of the cells increased as the proportion of FBS increased from 10% to 20% at 28℃ with optimum growth at the concentration of 20%.The doubling time of the cells was determined to be 44.8 h.Chromosome analysis revealed that 52% of the SHK cells maintained a normal diploid chromosome number (2n=46).The cells were successfully transfected with green fluorescent protein(GFP)reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies.The cells were infected by lymphosystis disease virus(LCDV)and found to be susceptible to the virus in cytopathic effect(CPE)observation.The infection was confirmed by PCR and electron microscopy experiments,which proved the existence of the viral particles in the cytoplasm of the virus-infected cells. 相似文献