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1.
《中国海洋大学学报(自然科学版)》2015,(8)
以壳聚糖/蒙脱土插层复合物为载体,采用吸附法和戊二醛交联法固定多酚氧化酶(PPO),考察了固定化酶的最佳制备条件及其酶学性质。结果表明,吸附法固定化多酚氧化酶(A-PPO)的最佳制备条件为pH=5.0,酶与载体质量比20mg/g,固定化时间6h,固定化酶的载酶量为12.12mg/g,单位载体酶活为12.76×103 U/g,酶活回收率为81.7%。交联法固定化酶(C-PPO)的最佳制备条件为pH=4.0,酶与载体质量比75 mg/g,固定化时间6h,固定化酶的载酶量为18.33mg/g,单位载体酶活为2.78×103 U/g,酶活回收率为12.9%。A-PPO和C-PPO的最适酶促反应条件为pH=8.0,温度分别为40和30℃。与游离多酚氧化酶相比,固定化酶的操作稳定性、贮存稳定性均有所提高。固定化酶重复使用5次后,可保留37.6%以上的相对酶活。PPO,A-PPO和C-PPO的米氏常数Km值分别为0.48,0.56和3.06mmol/L,较游离酶的Km值均有所增加;最大反应速率Vmax分别为2.70,2.08和0.18U/g,较游离酶的Vmax值均有所减小。 相似文献
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不结球白菜叶片壳聚糖酶的理化特性研究 总被引：3，自引：0，他引：3
通过不结球白菜叶片壳聚糖酶对壳聚糖的降解,研究了温度、pH、时间及金属离子等对此壳聚糖酶的影响。结果表明,壳聚糖酶的最适温度为45℃,最适pH为5.5,酶促反应速度在6 h内达到最大值;Mn2 和Cu2 对壳聚糖酶促反应分别起到促进和抑制的作用。经薄板层析分析证实,壳聚糖酶降解壳聚糖为低聚物。 相似文献
3.
烟曲霉产壳聚糖酶液体发酵条件研究 总被引：6，自引：0，他引：6
以烟曲霉(Aspergillus fumigatus)为出发菌,通过选择适合的培养条件,筛选出产壳聚糖酶的菌株。实验结果表明,烟曲霉是良好的产壳聚糖酶出发菌。烟曲霉液体发酵产壳聚糖酶最适培养基组成为:壳聚糖1%,KH2PO40.06%,尿素0.2%,(NH4)2SO40.1%,MgSO40.012%,20%土豆汁3%,NaCl 0.05%。在pH 5.0、温度28℃、摇床转速120 r/min培养条件下振荡培养3 d,烟曲霉产壳聚糖酶的能力最强,达25.1 U/mL。 相似文献
4.
壳聚糖酶基因在解脂耶氏酵母中的表达及重组酶性质研究 总被引：1，自引：0，他引：1
为构建高效表达壳聚糖酶工程菌株,以Microbacteriumsp.基因组为模板扩增壳聚糖酶基因,目的基因与表达载体pINA1317连接构建重组质粒,重组质粒NotⅠ酶切线性化后转化解脂耶氏酵母Y.liPolytica Po1h。所得结果为PCR扩增得到约1000bp的特异性片段,序列分析表明含有壳聚糖酶全长基因,开放阅读框801bp,编码266个氨基酸残基。转化子酶活力为10.4U/mL,是原菌株酶活力的3.15倍。重组蛋白经DEAE-Sepharose FF分离纯化,达到电泳纯,SDS-PAGE显示单一条带,分子量约41kDa。重组酶反应最适温度为45℃,最适pH为5.6,Km和Vmax分别是0.926mg/mL和6.15U/mL。质谱分析酶解产物主要为三糖和五糖。实现了壳聚糖酶在解脂耶氏酵母中的分泌表达,为壳聚糖酶的工业化生产奠定了理论基础。 相似文献
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产壳聚糖酶菌株选育及培养条件优化 总被引：6，自引：0，他引：6
以假单胞菌Y8．0为出发菌株,分别通过亚硝基胍(NTG),Co^60,UV(紫外)诱变,采用透明圈法筛选,获得了产壳聚糖酶较好的突变菌株假单胞菌Y8,其所产酶活力达到3．0U／mL,酶活力提高近6倍,并具有较好的遗传稳定性。3种诱变方法中,UV诱变效果最好。对假单胞菌Y8产壳聚糖酶培养条件进行研究,得到1个比较优化的培养条件为：pH值6．5,温度32℃,壳聚糖3g／L,酵母膏0．3％,培养时间3d。不同金属离子及不同诱导物对酶合成的影响表明：Fe^2 ,Mg^2 激活作用显著。而Zn^2 ,Mn^2 则具有一定的抑制作用,壳聚糖和氨基葡萄糖对壳聚糖酶的产生都有较好的诱导作用。 相似文献
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双酶分步水解金枪鱼(Eleotridae)碎肉制备高F值酶解液的工艺研究 总被引：2，自引：2，他引：0
以金枪鱼碎肉为原料, 采用双酶分步水解法制备高F值酶解液, 通过Box-Behnken试验设计, 分别确定两步酶解的最佳条件, 酶解液经活性炭静态吸附去除游离芳香族氨基酸, 对脱芳后的酶解液进行氨基酸组成分析并测定F值。结果表明, 胃蛋白酶为第一步水解用酶, 酶解的最佳工艺条件为酶用量650U/g, 料水比1∶7(g/mL), 温度35.9℃; 风味蛋白酶为第二步水解用酶, 酶解的最佳工艺条件为酶用量50700U/g, pH 6.51, 温度51℃, 最终水解度达到36.87%±0.54%; 酶解液在pH 3.0, 温度35℃条件下, 经5%(质量体积分数)的活性炭吸附时间3h后, 脱芳率达到63.18%, F值为30.33, 符合高F值肽的要求。 相似文献
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以金枪鱼碎肉为原料, 采用双酶分步水解法制备高F值酶解液, 通过Box-Behnken试验设计, 分别确定两步酶解的最佳条件, 酶解液经活性炭静态吸附去除游离芳香族氨基酸, 对脱芳后的酶解液进行氨基酸组成分析并测定F值。结果表明, 胃蛋白酶为第一步水解用酶, 酶解的最佳工艺条件为酶用量650U/g, 料水比1∶7(g/mL), 温度35.9℃; 风味蛋白酶为第二步水解用酶, 酶解的最佳工艺条件为酶用量50700U/g, pH 6.51, 温度51℃, 最终水解度达到36.87%±0.54%; 酶解液在pH 3.0, 温度35℃条件下, 经5%(质量体积分数)的活性炭吸附时间3h后, 脱芳率达到63.18%, F值为30.33, 符合高F值肽的要求。 相似文献
8.
用壳聚糖酶降解法对壳聚糖进行降解制备壳低聚糖，研究了温度、pH、底物浓度、脱乙酰度、反应时间对酶促反应的影响。对酶解产物进行HPLC分析，用Bio—Gel P-4柱对酶解产物进行初步分离。结果表明，壳聚糖酶降解壳聚糖的最适温度为45℃，pH为6．0，最适底物质量分数为6％，脱乙酰度越高酶解反应所得产物的数均相对分子质量越小，24h后酶解反应达到平衡，酶解产物壳寡糖水溶性极好。P-4柱分离得到聚合度20以上，10～20，10以下3种壳寡糖。 相似文献
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壳聚糖酶的分离纯化及性质研究 总被引：4，自引：0，他引：4
以本实验室分离的产壳聚糖酶YJ02菌株出发,制备壳聚糖酶发酵液,经离心、(NH4)2SO4分级盐析、Q-SepharoseFast Flow阴离子交换、Sephacry S-100凝胶过滤分离纯化步骤,SDS-PAGE显示为一条带,壳聚糖酶纯化了28.9倍,收率为47.6%。该酶的性质研究表明,该酶的分子量为66.2KD,最适反应温度为50℃,最适反应pH为6.0;Mn2 对酶活力有明显的促进作用,Cu2 ,Fe3 ,Hg2 对酶活力有抑制作用;小分子有机物EDTA和SDS对酶活力有抑制作用,而IAA和β-巯基乙醇对酶活力稍有促进作用。该壳聚糖酶对高脱乙酰度壳聚糖底物水解作用强,酶解位点可能为NGlc-NGlc,酶最大反应速度Vmax=4.1μmol/min,常数Km为64mmol/L。此壳聚糖酶水解壳聚糖制备的壳寡糖数均分子量为2 400Da。 相似文献
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Chandralata Raghukumar G. Sheelu P. A. Loka Bharathi Shanta Nair C. Mohandass 《Marine Georesources & Geotechnology》2001,19(1):1-16
In order to assess the impact of deep-sea mining on the in situ benthic life, we measured the microbial standing stock and concentration of organic nutrients in the deep-sea sediments of the Central Indian Ocean Basin in the Indian pioneer area. Sediments were collected using box core and grab samples during September 1996. The total bacterial numbers ranged from 10 10 -10 11 cells per g -1 dry weight sediment. There was a marginal decrease in the number of bacteria from surface to 30 cm depth, though the subsurface section registered a higher number than did the surface. The highest numbers were encountered at depths of 4-8 cm. The retrievable number of bacteria were two orders less in comparison with the direct total counts of bacteria. An almost homogeneous distribution of bacteria, total organic carbon, living biomass, and lipids throughout the depth of cores indicates active microbial and benthic processes in the deep sea sediments. On the other hand, a uniform distribution of total counts of bacteria, carbohydrates, and total organic carbon in all the cores indicates their stable nature and suggests that they can serve as useful parameters for long-term monitoring of the area after the benthic disturbance. Further studies on temporal variability in this region would not only verify the observed norms of distribution of these variables but would also help to understand restabilization processes after the simulated benthic disturbance. 相似文献
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Derek V. Ellis 《Marine Georesources & Geotechnology》2001,19(1):51-63
This article reviews information recently available from existing marine and coastal mining for responses to environmental issues affecting marine mining at different depths. It is particularly but not exclusively concerned with those issues affecting seabed biodiversity impact and recovery. Much information has been gathered in the past 10 years from shallow mining operations for construction aggregate, diamonds, and gold, from coastal mines discharging tailings to shallow and deep water, and from experimental deep mining tests. The responses to issues identified are summarized in a series of eight tables intended to facilitate site-specific consideration. Since impacts can spread widely in the surface mixing layer SML, and can affect the biologically productive euphotic zone, the main issues considered arise from the depth of mining relative to the SML of the sea. Where mining is below the SML, the issue is whether it is environmentally better to bring the extraction products to the surface vessel for processing (and waste discharge), or to process the extraction products as much as possible on the seabed. Responses to the issues need to be sitespecific, and dependent on adequate preoperational environmental impact and recovery prediction. For deep tailings disposal from a surface vessel, there are four important environmental unknowns: (1) the possible growth of "marine snow" (bacterial flocs) utilizing the enormous quantities of fine tailings particles (hundreds or thousands of metric tons per day) as nuclei for growth, (2) the possibility that local keystone plankton and nekton species may migrate diurnally down to and beyond the depth of deep discharge and hence be subjected to tailings impact at depth, (3) the burrow-up capability of deep benthos and their ability to survive high rates of tailings deposition, and (4) the pattern and rate of dispersion of a tailings density current through the deep water column from discharge point to seabed. Actions to obtain relevant information in general and site-specifically are suggested. 相似文献
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G. Parthiban 《Marine Georesources & Geotechnology》2000,18(3):223-235
Particle fluxes were measured 7 m above the sea bottom during the predisturbance, disturbance, and postdisturbance periods by using time series sediment traps attached to seven deep-sea moorings deployed in the INDEX experiment site in the Central Indian Basin. The predisturbance particle fluxes varied between 22.3 to 55.1 mg m -2 day -1 . Increased and variable particle fluxes were recorded by the sediment traps during the disturbance period. The increase observed was 0.5 to 4 times more than the background predisturbance fluxes. The increases in particle fluxes (~4 times) recorded by the sediment trap located in the southwestern direction (DMS-1) were the greatest, which could be the result of preferential movement of resuspended particles generated during the deep-sea benthic disturbance along the general current direction prevailing in this area during the experimental period. Also, the traps located closer to the disturbance area recorded greater fluxes than did the traps far away, across the Deep Sea Sediment Resuspension System path. This variability in recorded particle fluxes by the traps around the disturbance area clearly indicates that physical characteristics such as grain size and density of the resuspended particles produced during the disturbance had an important effect on particle movement. The postdisturbance measurements during ~5 days showed a reduction in particle fluxes of ~50%, indicating rapid particle settlement. 相似文献
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Arne Schnack-Friedrichsen Angela M. Davis Jim D. Bennell Dei G. Huws 《Marine Georesources & Geotechnology》2001,19(4):221-243
The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths. 相似文献
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Geoacoustic Inversion via Genetic Algorithm and Its Application to Manganese Sediment Identification
An acoustic inversion method using a wide-band signal and two near field receivers is proposed and applied to multiple layered seabed models including a manganese sediment. The inversion problem can be formulated into a probabilistic model comprised of signals, a forward model, and additive noise. The forward model simulates wide-band signals, such as chirp signals, and is chosen to be the source-waveletconvolution plane wave modeling method. The wavelet matching technique, using weighted least-squares fitting, estimates the sediment sound-speed and thickness on which determination of the possible numerical ranges for a priori uniform distribution is based. The genetic algorithm is applied to a global optimization problem to find a maximum a posteriori solution for determined a priori search space. Here the object function is defined by an L 2 norm of the difference between measured and modeled signals. Not only the marginal pdf but also its statistics are calculated by numerical evaluation of integrals using the samples selected during importance sampling process of the genetic algorithm. 相似文献
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海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明：土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。 相似文献
17.
The advanced piston corer (APC) has been used by the Ocean Drilling Program since 1985 for recovering soft sediments from the ocean floor. The pullout force measured on extracting the core barrel from the sediment is shown to correlate with the average shear strength of the sediment core measured in the ship's laboratory. A simple rule of thumb is derived relating the shear strength of the sediment to the pullout force. Multiple APC holes at individual sites allow the consistency of the pullout measurements to be assessed. The effects of different operational procedures during APC coring are also explored. Although generally applicable, the correlation between pullout force and laboratory measurements of shear strength breaks down for some APC holes, possibly because of the disturbance of some sediment types during the APC coring process. A better understanding of the physical process of APC coring, and its effect on the properties of the sediment both inside and immediately outside the core barrel, would indicate what confidence can be put on the measurement of pullout force as a way of evaluating the in situ shear strength of deep sea sediments. 相似文献
18.
Shigehisa Nakamura 《Marine Geodesy》2000,23(1):55-61
Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone. 相似文献
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