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1.
The e?ect of the bacterial strain CI4 on the germination of spores from the green alga Ulva pertusa was assayed and it was found that the bacterial bioˉlm and cell-free supernatant strongly inhibited spore germination. In attempts to deˉne the chemical nature of the antifouling substance in the supernatant of CI4, the culture supernatants were tested for activity after heat treatment, enzy- matic treatments, size fractionation, and separation into aqueous and organic fractions. Results suggest that this bacterium produces an extracellular component with speciˉc activity toward algal spores that was heat-sensitive and between 3 and 10 kDa in molecular size. The exposure of the organic phase fraction to spores showed inhibitive e?ect on spore germination. Pronase and car- boxypeptidase y did not signiˉcantly a?ect the activity of inhibitory component, suggesting that the component was not a protein or a peptide. The bacterium CI4 was identiˉed as Pseudoalteromonas haloplanktis based on the phenotypic characters and 16S rRNA gene analysis.  相似文献   

2.
Abstract. A new concept (‘enzyme sharing’) concerning the interaction of marine nematodes and microbes in the degradation of sedimentary detritus is presented. Elements of this concept are (1) the notorious tendency of many aquatic nematodes to agglutinate detrital particles by mucus secretions, (2) new observations on the stimulation of microbial growth by nematodes in agar plates, and (3) literature data on limited endogenous proteolytic capacities of aquatic nematodes.
Observations on nematode‐microbe associations in agar plates prompted the conceptual synthesis. In agar medium without the addition of any nutrients a spectacular growth of bacteria was visible on the sinusoidal crawling trails of nematodes only 2 – 3 days after introduction of the worms (species of Adoncholaimus, Anoplostoma and Sabatieria). Juveniles of Anoplostoma that hatched in the agar cultures left their minute trails in the medium and these were rapidly occupied by bacteria. The nematodes repeatedly visited their bacterial trails, which persisted as a peculiar biotic structure for more than one year and survived the nematodes.
In sterile agar preparations containing the fluorogenic methylumbelliferyl‐β‐glucoside in the presence of the nematode Adoncholaimus, an enhanced fluorescence of the medium was visible, indicating β‐glucosidase activity. We therefore assume that oncholaimid nematodes discharge enzymes that alone, or in concert with microbial activities, contribute to the hydrolytic cleavage of refractory polysaccharides containing β‐glucosidic bonds such as agar components and cellulose. The sugars thus produced may then be taken up by the nematodes and concomitantly support the conspicuous growth of microbes. Since we did not observe any feeding of the nematodes on the associated microbes in agar plates, we question the nutritive potential of intact microbial cells for a number of nematodes abounding in detrital habitats, and call attention to the significance of ambient dissolved or adsorbed organic monomeric nutrients.
Consequently, we perceive the puzzling perpetual accretion of detrital organic particles to sediment agglutinations by nematodes as an adaptation for operating an ‘enzymatic reactor’ for the production of dissolved nutrients. We hypothesise a relationship of mutual commensalism of nematodes and heterotrophic microbes in detrital habitats and propose the term ‘enzyme sharing’ for this relationship. Both parties invest in a common enzyme pool that decomposes organic detritus for their nutrition. We present here evidence that nematodes contribute β‐glucosidase, which is involved in the cellulase system. Data from the literature suggest that microbial enzymatic processing of detrital proteins yields amino acids available to nematodes, which apparently have no efficient proteolytic enzyme system in their intestines.  相似文献   

3.
本文首先通过MALDI-TOF/TOF-MS技术,测定金枪鱼(Thunnus sp.)胰腺酶解液中的多肽组分及其丰度,确定GQPVR、GLPPK和EHER是其中的优势肽;再利用Discovery Studio平台中的反向找靶和分子对接模块,发现这些优势肽均可以与Keap1蛋白结合,推测其可能具有抗氧化活性;最后,利用细胞实验对酶解液的抗氧化活性进行验证,发现其对H2O2氧化损伤的胰岛素瘤细胞(INS-1)具有剂量依赖型保护效果,高剂量和低剂量酶解液处理后细胞活力较模型组显著提高,凋亡率从79.17%分别下降到43.2%(P0.01)和28.9%(P0.01);胰岛素的分泌量从3.65mIU/L分别增加到5.75mIU/L(P0.01)和4.95mIU/L(P0.01)。DiscoveryStudio的抗氧化功能预测和细胞学实验相结合的研究方法,为酶解液的功能组分鉴定提供了新的思路。  相似文献   

4.
This work investigates the alkaline phosphatase activity in a littoral marine ecosystem (Toulon Bay and Le Niel Bay, France) in order to study its biochemical characteristics with respect to pH, sea water composition and phosphate sensitivity. We also characterise the active forms in sea water and determine the extent to which zooplankton generate phosphatase activity with respect to other plankton classes. In Toulon Bay, phosphatase was produced mostly by the microplankton fraction (>90 μm), accounting for more than 90% of total activity. In contrast, most of the phosphatase activity in Le Niel Bay was generated by the nanoplankton fraction (5–90 μm) and the picoplankton fraction (0.25–5 μm). The microplankton enzymes had non Michaelis-Menten kinetics suggesting the involvement of multiple enzyme processes with distinct kinetic constants. This activity is in major part secreted into the sea water and is stimulated by the ionic strength and the pH of the sea water. Cypris larvae of the genus Balanus played a special role in this release. For the nanoplankton and picoplankton, part of this activity was due to non-secreted enzymes, probably bound to membranes or occurring intracellularly. Moreover, nano and picoplankton phosphatase required higher pH than microplankton enzyme. For all plankton size classes, there was no activity at low pH, suggesting that acid phosphatases were not involved in reactions with substrates dissolved in water.  相似文献   

5.
In Toulon Bay (France), very high phosphatase activities have been found in the zooplankton fraction>90 microm. This work was intended to specify their origin. For that purpose, larvae, juvenile and adult Crustacea (Copepods: Calanoids, Cyclopoids, Branchiopods: Cladocera, and Cirripeds) were isolated. Their activities were measured using paranitrophenyl phosphate dissolved in sea water in order to calculate Km (the enzyme half saturation concentration) and Vmax (the reaction rate when the enzyme is saturated with substrate). Vmax were referred to protein contents of the isolated organisms to calculate specific activities. For all zooplankton groups high and low affinity phosphatase activities were found. The low affinity enzyme was responsible for at least 70% of the total phosphatase activity. Its specific activity was higher for larvae than for copepodites and adults. In Cirriped nauplii this activity was particularly high with values which were several hundred times higher than that in other Crustacea. These enzymes had optimum pH close to 8.4, magnesium requirement and were competitively inhibited by orthophosphate. Experiments with intact and lysed Cirriped nauplii confirmed that living organisms had only a weak external activity and showed that most of the activity of these larvae was primarily intracellular.  相似文献   

6.
氢和氧稳定同位素示踪湖泊蒸发的对比研究   总被引:1,自引:0,他引:1  
氢氧稳定同位素被广泛用于水文循环过程的研究。本文观测了2015年太湖湖水H~2HO和H_2~(18)O组分,分析了它们的时空变化规律及其控制因子,探讨亚热带大型浅水湖泊的同位素富集机制;基于稳定同位素质量守恒法计算太湖蒸发量;评价了动力分馏学系数的传统湖泊算法与海洋算法的适用性;重点分析了H~2HO和H_2~(18)O示踪湖泊蒸发的效果,对比二者之间的差异。研究结果表明,在空间上,太湖湖水和河水的氢氧同位素在南部特别是东南部较为富集但在北部区域较为贫化,这主要是受水流方向的控制,东南部湖水经历的蒸发时间较长,因此湖水中同位素累积较多;在季节上,冬季湖水同位素较贫化、春夏季较富集。对于2015年太湖的年蒸发量,用氢同位素示踪的结果与观测值较一致,为880mm;氧同位素的示踪结果略低,为690mm。使用传统湖泊研究中对动力学分馏系数的取值,会导致蒸发被显著低估,而氧稳定同位素的示踪结果对动力学分馏系数的取值更为敏感,同时氢稳定同位素在同位素分馏过程中主要是平衡分馏效应占主导,因此H~2HO在动力学分馏系数的参数化方案中影响较小,在实际应用中更为稳定。本文的研究结果表明了稳定同位素水文学研究中使用合适的动力学分馏系数的重要性。  相似文献   

7.
作者在已完成大气激光雷达系统夜间工作的基础上 ,对白天工作条件下的各种环境背景光的干扰进行了理论与实验分析 ,得出太阳直射光和天空光为白天工作的主要背景干扰。实验结果表明 ,经过激光发射与接收视场角严格匹配及使用干涉滤光器 (2 .6 nm和 0 .15nm)进行窄带滤波 ,背景干扰被明显地剔除 ,可进一步压低背景干扰约 2 0倍。证明采取视场匹配及干涉滤光器的措施可基本保证激光雷达系统在白天条件下工作。  相似文献   

8.
Although elasmobranchs are widely distributed species, commonly found on sandy, gravely or muddy bottoms, several biological aspects of their metabolism still remain poorly investigated. In this work the efficiency of antioxidant system and detoxification enzymes were investigated in the coastal spotted dogfish Scyliorhinus canicula and in the red mullet Mullus barbatus for comparison with a teleost species. Organisms were sampled during a bottom trawl survey and analyzed for the biotransformation activity (EROD), levels of metallothioneins, catalase, glutathione S-transferases and total oxyradical scavenging capacity (TOSC) toward peroxyl radicals and hydroxyl radicals. EROD activity in the elasmobranchs was more than one order of magnitude lower than in the red mullets, while similar levels of metallothioneins were measured in these species. S. canicula showed significantly lower antioxidant enzymes and a more reduced efficiency in neutralizing *OH; on the other hand the scavenging capability toward ROO* was comparable in S. canicula and M. barbatus.  相似文献   

9.
哈维氏弧菌对条纹斑竹鲨4种酶活性的影响   总被引:2,自引:0,他引:2  
以哈维氏弧菌(Vibrio harveyi)对条纹斑竹鲨(Chiloscyllium plagiosum)进行病原接种试验,分别在4、8、12、24、48、72、96h后测定肝脏、脾脏、鳃及血清的酸性磷酸酶(acid phosphatase,ACP)、碱性磷酸酶(alkaline phosphatase,AKP)、超氧化物歧化酶(superoxide dismutase,SOD)和血清中溶菌酶(lysozyme,LSZ)的活力.实验结果表明:对照组条纹斑竹鲨的这4种酶的活力呈现出明显的组织差异性,血清中SOD活力为124.32 U/cm3,显著高于其他3种酶的活力;其他组织中3种酶的活力由高到低依次为SOD、ACP、AKP.在实验组条纹斑竹鲨被感染4~72h期间,血清中SOD活力明显下降,ACP活力持续下降,感染96h后除LSZ外,其他3种酶的活力都呈回升趋势;其他组织中ACP与AKP活力变化均为被感染4h后下降,12h后明显回升,SOD则在被感染4h后活力上升,而后下降,96h后回升.这4种酶的活力的变化主要是应激作用与非特异性免疫防御作用共同作用导致的结果.  相似文献   

10.
Chemical and enzymatic reagents have been employed to determine available concentrations of Fe, Mn, Cu and Zn in contaminated estuarine sediment. Gastric and intestinal enzymes (pepsin, pH 2, and trypsin, pH 7·6, respectively) removed significantly more metal than was water-soluble or exchangeable (by seawater or ammonium acetate), while gastro-intestinal fluid of the demersal teleost, Pleuronectes platessa L. (plaice), employed to operationally define a bioavailable fraction of contaminants, generally solubilized more metal than the model enzymes. Manganese was considerably more available than Fe under these conditions and it is suggested that the principal mechanism of contaminant release is via surface complexation and reductive solubilization of Mn oxides, a process which is enhanced under conditions of low pH. Of the chemical reagents tested, acetic acid best represents the fraction of Mn (as well as Cu and Zn) which is available under gastro-intestinal conditions, suggesting that the reducing tendency of acetate is similar to that of the ligands encountered in the natural digestive environment. Although the precise enzymatic and non-enzymatic composition of plaice gastro-intestinal fluid may be different to that encountered in more representative, filter-feeding or burrowing organisms, a general implication of this study is that contaminants associated with Mn oxides are significantly more bioavailable than those associated with Fe oxides, and that contaminant bioavailability may be largely dictated by the oxidic composition of contaminated sediment.  相似文献   

11.
用组织学、组织化学和酶技术,对雌性莫桑比克罗非鱼在孵卵前后口腔粘膜的组织结构、抗菌酶的比活力变化进行初步研究。结果表明:在口腔粘膜的上皮中,杯状细胞是一个重要的结构,它随着雌鱼的生理状态的改变而变化。雄鱼、雌鱼和孵卵期的雌鱼口腔粘膜上皮中的杯状细胞数量上的差别显著。孵卵期的雌鱼口腔粘膜类似于哺乳动物的子宫粘膜也是受性激素影响的靶器官。该种鱼的口腔粘膜中存在着两种抗菌酶;蛋白质水解酶和壳多糖酶。孵卵前后,这两种酶的比活力分别提高1.4—2.03倍。说明孵卵时,口腔粘膜为使卵及仔鱼健康的发育,增强了抗菌能力。研究结果证明:抗菌力的增加与口腔粘膜中的杯状细胞的大量增生有着密切的关系。  相似文献   

12.
Extracellular peptide hydrolysis rates were measured in seawater and sediment from Flax Pond salt marsh using peptide analogs (LYA-peptides) as substrates. This technique allows the direct measurement of specific hydrolysis products and thus provides insights into enzymatic hydrolysis pathways. In sediments, hydrolysis rate constants of LYA-peptides varied seasonally and with depth. Highest activity was found in spring and summer, and most cores exhibited a subsurface maximum. Calculations using the concentrations of chemically-measured peptides suggested that extracellular hydrolysis of peptides is faster than the rate of free amino acids uptake. However, not all peptides may be available for enzymatic hydrolysis. In both seawater and sediment, extracellular hydrolysis of peptides of up to 8 amino acids yielded smaller peptides and free amino acids. Hydrolysis rates depended on size of the peptide substrate, although a clear relationship with number of amino acid constituents was not evident. Peptides containing >2 amino acids were hydrolyzed 10–400 times faster than dipeptides or the fluorogenic substrate Leucine-MCA. Thus, dipeptidases are either uncommon in nature, or hydrolysis is carried out by nonspecific hydrolases that with a low affinity for dipeptides. This is also suggested by the presence of a lag time before dipeptide hydrolysis begins, and the absence of dipeptide hydrolysis in 0.2-μm-filtered. One implication of this finding is that measurements of hydrolysis rates using substrates like Leu-MCA may not accurately predict the magnitude of hydrolysis rates of macromolecules in the marine environment. Even though dipeptide hydrolysis is slow compared to that of larger peptides, LYA-dipeptides are preferentially produced from the hydrolysis of larger substrates. LYA-dipeptides do not penetrate cell membranes of microorganisms because of their size, but natural dipeptides are smaller and can be transported across the cell membrane. Since dipeptides do not appear to accumulate in natural waters, they must be rapidly removed by microorganisms.  相似文献   

13.
Spermatozoal enzymes of fish (NAD+- and NADP-dependent dehydrogenases and creatine kinase (CK)) were previously determined to be sensitive to tributyltin (TBT) in laboratory experiments and were thus indicated for use as biomarkers for TBT exposure. However, the potential ability of spermatozoal enzymes as biomarkers of TBT exposure has never been recapitulated in a field study. For this purpose, the kinetic activities of spermatozoal enzymes of the natural turbot Scophthalmus maximus population from the Gulf of Gdańsk (GDA) and the Pomeranian Bay (POM) in the southern Baltic Sea were measured. Gas chromatography/high-resolution mass spectrometry was used to determine the concentrations of TBT and its breakdown products, dibutyltin (DBT) and monobutyltin (MBT), in the muscle, liver and testes of the male turbot. Males from GDA had significantly higher enzymatic activities and butyltin (BT) content in tissues than those from POM. A general linear model (GLM) showed that lactate dehydrogenase (LDH), malate dehydrogenase (MDH), glucose-6-phosphate dehydrogenase (G6PDH) and CK activities increased significantly with BT concentration in the testes and liver. We indicate the potential effects of TBT pollution on the spermatozoal enzymes of Baltic turbot.  相似文献   

14.
The present study clarifies the enzymatic properties of two avian cytochrome P4501A (CYP1A) paralogs, CYP1A4 and 1A5, using a yeast-based vector system. Recombinant CYP1A4 and 1A5 proteins from common cormorant (Phalacrocorax carbo) were expressed in yeast cells, and showed typical reduced CO-difference spectra with a peak at 446 nm. Kinetic analysis of O-dealkylase of methoxy-, ethoxy-, pentoxy- and benzyloxyresorufin catalyzed by the CYP1A enzymes revealed that Vmax value for ethoxyresorufin-O-deethylase (EROD) activity was much higher than that for the other three O-dealkylase activities for both isozymes. Interestingly, remarkable substrate specificity of the CYP1As was observed for O-dealkylation of benzyloxyresorufin and methoxyresorufin; CYP1A4 was highly specific for catalyzing benzyloxyresorufin-O-debenzylase activity, whereas CYP1A5 was more efficient in catalyzing methoxyresorufin-O-demethylase activity. The present study also measured CYP1A-dependent EROD activity in the presence of 2,3,7,8-tetrachlorodibenzofuran (TCDF) to evaluate the ability of this dioxin-like congener to inhibit the EROD activity. One hundred nanomolar TCDF noncompetitively inhibited CYP1A5-dependent EROD activity, although no inhibitory effect was detected for CYP1A4-dependent EROD activity. These results indicate that the avian CYP1A paralogs have different affinities for substrate and inhibitor, thus suggesting their distinct physiological and toxicological roles.  相似文献   

15.
An extensive subsurface oxygen minimum zone off northern and central-south Chile, associated with the Peru–Chile undercurrent, has important effects on the metabolism of the organisms inhabiting therein. Planktonic species deal with the hypoxic and anoxic environments by relying on biochemical as well as physiological processes related to their anaerobic metabolisms. Here we characterize, for the first time, the potential enzymatic activities involved in the aerobic and anaerobic energy production pathways of microplanktonic organisms (<100 μm), their relationship, and this relationship's association with the oxygen concentration and microplanktonic biomass in the oxygen minimum zone and adjacent areas of the Humboldt Current System water column. Our results demonstrate significant potential enzymatic activity of catabolic pathways in the oxygen minimum zone. Malate dehydrogenase had the highest oxidizing activity of nicotinamide adenine dinucleotide (reduced form) in the batch of catabolic enzymatic activities assayed, including potential pyruvate oxidoreductases activity, the electron transport system, and dissimilatory nitrate reductase. Malate dehydrogenase correlated significantly with almost all the enzymes analyzed within and above the oxygen minimum zone, and also with the oxygen concentration and microplankton biomass in the water column of the Humboldt Current System, especially in the oxygen minimum zone off Iquique. These results suggest a possible specific pattern for the catabolic activity of the microplanktonic realm associated with the oxygen minimum zone spread along the Humboldt Current System off Chile. We hypothesize that malate dehydrogenase activity could be an appropriate indicator of microplankton catabolism in the oxygen minimum zone and adjacent areas.  相似文献   

16.
为了解重金属Hg对锦州湾海域表层沉积物中异养细菌的主要代谢酶的影响,采用Hakanson潜在生态危害指数法对锦州湾表层沉积物中重金属Hg开展潜在生态风险评价,利用平板扩散法测定了9个站位沉积物中异养细菌四种水解酶活性。结果显示:锦州湾海域表层沉积物中重金属污染严重,Hg的含量和潜在生态危害河口最为严重,向外逐步递减,表明本海域表层沉积物中的Hg主要由陆源输入;表层沉积物中异养细菌水解酶活性与抗性异养菌比例均表现为西南部河口区高于东北部外海海域,与重金属Hg的含量分布及Hakanson潜在生态危害指数基本相似,这表明长期高浓度的重金属污染,改变了锦州湾海域表层沉积物中的异养菌的生理代谢过程,在生理或遗传物质上对重金属产生了一定的抗性;磷酸酶、蛋白酶、脂肪酶与沉积物中Hg的含量具有显著正相关性,说明其对重金属Hg较为敏感,在一定程度上可作为反映锦州湾海域沉积物中重金属Hg污染状况的生物学指标。  相似文献   

17.
褐藻寡糖具有多种生理活性,本研究通过酶解与分级条件的优化,获得了低分子质量褐藻寡糖的酶法制备工艺,并在体外模型上评价了不同分子质量组分的抗氧化活性。结果表明,优化后的褐藻寡糖制备条件为:底物浓度1.20%,酶底比4.35 U/mg,酶解时间4 h。进一步采用乙醇沉淀和超滤分离后,获得了重均分子质量分别为0.84、1.40、2.25和34.56 k Da的4种组分,其得率分别为50.82%、5.15%、11.48%和12.00%。各组分均具有一定的还原能力,可有效清除DPPH和羟自由基,其中低分子质量组分A的抗氧化活性最为显著,其清除羟自由基的能力与维生素C相近。  相似文献   

18.
Size‐fractionation by Nuclepore filters shows that about one‐third of the biomass of freshwater phytoplankton is 0.2–3 μm in size. Autoradiographs of natural water from North and South Island lakes of various trophic states, incubated with ‘'C‐bicarbonate, reveal the contribution of this ‘ultraplankton’ to primary production.

The lakes sampled in 1975–76 were Rotongaio, Rotorua, Tarawera, and Taupo (North Island); Brunner, Johnson, Kangaroo, Ohau, Pukaki, Rotoiti, and Wakatipu (South Island). In general, oligotrophic and mesotrophic lakes contain most ultraplankton, forming 11–35% of the biomass and 33–76% of the primary productivity. In both New Zealand and North America the importance of ultraplankton seems grossly underestimated, probably because it normally escapes detection in standard Utermohl and direct nitration examinations.  相似文献   

19.
The activities and substrate specificities of extracellular enzymes in natural systems are not well understood, despite their critical role in microbial remineralization of organic carbon. These enzymes initiate organic carbon degradation by selectively hydrolyzing high molecular weight substrates to lower molecular weight products which can be transported into cells. A set of single- and dual-labeled fluorescent polysaccharides was synthesized and characterized to explore a variety of approaches for measuring enzymatic hydrolysis of biopolymers via photophysical techniques, focusing particularly on rapid and robust optical techniques which are amenable to field measurements in remote locales. A shotgun-labeling approach yielded dual-labeled probes that exhibited substantial donor fluorophore quenching. The photophysical response of these probes to hydrolysis via purified enzymes was investigated in the lab, and fluorescence polarization proved to be a rapid and reliable technique for monitoring probe hydrolysis. Initial field results were also obtained from hydrolysis experiments in sediment porewaters. Because polarization measurements are rapid and simple, this approach could be used to follow the extracellular enzymatic hydrolysis of a wide range of biopolymers which fuel microbial metabolism.  相似文献   

20.
Extracellular enzymatic activity (EEA) of five enzymes (peroxidase, phenol oxidase, beta-glucosidase, beta-N-acetylglucosaminidase and acid phosphatase) was analysed in sediments colonised by Spartina maritima in two salt marshes (Rosário and Pancas) of the Tagus estuary (Portugal) with different characteristics such as sediment parameters and metal contaminant levels. Our aim was a better understanding of the influence of the halophyte on microbial activity in the rhizosphere under different site conditions, and its potential consequences for metal cycling and phytoremediation in salt marshes. Acid phosphatase and beta-N-acetylglucosaminidase presented significantly higher EEA in Rosário than in Pancas, whereas the opposite occurred for peroxidase. This was mainly attributed to differences in organic matter between the two sites. A positive correlation between root biomass and EEA of hydrolases (beta-glucosidase, beta-N-acetylglucosaminidase and acid phosphatase) was found, indicating a possible influence of the halophyte in sediment microbial function. This would potentially affect metal cycling in the rhizosphere through microbial reactions.  相似文献   

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