Changes of enzymatic activity during larval development of carp, Catla catla (Ham.)

Quantitative assays of trypsin, amylase and alkaline phosphatases were made in relation to age and food during the larval development of the Indian major carp Catla catla. The responses of all the test enzymes to age and food were identical. No enzymes were detected from the fertilized eggs. Detectable amount of enzymes were first observed in the first day old hatchlings. All the test enzymes in the group fed normal feed tended to rise gradually with advancement of age till day 22 after which an asymptotic level was attained. Absence of food throughout the rearing period caused the enzymatic activity of the larva to remain at the lowest level throughout. When starvation was followed by feeding, enzymatic activity in the former group was consistently higher than that of latter, suggesting that feeding activity was primarily responsible in maintaining the enzymatic activity of carp larva. The enzymatic activity of zooplankton was significantly higher than carp larva till day 6 to 12 after which the latter exceeded the former implying that carp larva during development utilizes the exogenous enzymes of zooplankton.

CHARACTERISTICS OF FATTY ACID METHYL ESTERS (FAMEs) AND ENZYMATIC ACTIVITIES IN SEDIMENTS OF TWO EUTROPHIC LAKES

Enzymatic activities and fatty acid methyl esters (FAMEs) in the sediments of two eutrophic lakes in Wuhan city were investigated. The results showed phosphatase and dehydrogenase activities in the lotus zone and plant floating bed zone were significantly lower than those in other sites, and urease activity was the highest where microorganism agents were put in. Fatty acid group compositions indicated the predominance of aerobic bacteria in the surface sediments in shallow lakes. The ratios of FAMEs specific for bacteria and Gram-positive bacteria exibited significant differences between the two lakes. The results of trans to cis indicated that the microorganisms in Lake Yuehu could adapt themselves to environmental stress better. The enzymatic activities and FAMEs showed differences in different sites, indicating that ecological restoration measures and environmental conditions could affect lake sediment to some extent. But the monitoring, work would be done in series to exactly evaluate the effect of the remediation measures.

Enzymatic activities in constructed wetlands and di-n-butyl phthalate (DBP) biodegradation

The bioaccumulation of phthalate acid esters (PAEs) from industrial products and their mutagenic action has been suggested to be a potential threat to human health. The effects of the most frequently identified PAE, Di-n-butyl phthalate (DBP), and its biodegradation, were examined by comparison of two small scale plots (SSP) of integrated vertical-flow constructed wetlands. The influent DBP concentration was 9.84 mg l(-1) in the treatment plot and the control plot received no DBP. Soil enzymatic activities of dehydrogenase, catalase, protease, phosphatase, urease, cellulase, beta-glucosidase, were measured in the two SSP after DBP application for 1 month and 2 months, and 1 month after the final application. Both treatment and control had significantly higher enzyme activity in the surface soil than in the subsurface soil (P < 0.001) and greater enzyme activity in the down-flow chamber than in the up-flow chamber (P < 0.05). In the constructed wetlands, DBP enhanced the activities of dehydrogenase, catalase, protease, phosphatase and inhibited the activities of urease, cellulase and beta-glucosidase. However, urease, cellulase, beta-glucosidase activities were restored 1 month following the final DBP addition. Degradation of DBP was greater in the surface soil and was reduced in sterile soil, indicating that this process may be mediated by aerobic microorgansims. DBP degradation fitted a first-order model, and the kinetic equation showed that the rate constant was 0.50 and 0.17 d(-1), the half-life was 1.39 and 4.02 d, and the r(2) was 0.99 and 0.98, in surface and subsurface soil, respectively. These results indicate that constructed wetlands are able to biodegrade organic PA-Es such as DBP. (c) 2005 Elsevier Ltd. All rights reserved.

酸化油固定床酶法合成生物柴油研究

酸化油是油脂工业中以皂脚、油脚经酸化处理得到的产品。它的主要成分是游离脂肪酸及中性油，是生产脂肪酸的重要原料，但生产过程中有水解废水的产生，若将其直接排放，既污染了环境又浪费了资源。生物柴油的主要成分是脂肪酸甲酯（fatty acid methyl ester，FAME）。它具有原料丰富而且可再生、可生物降解、无毒、不含芳香烃、二氧化硫等污染物、燃烧排放低、闪点高、运输储存安全等特点。作为石化柴油的潜在替代能源，生物柴油因其独特的优越性和现实的需求越来越受到关注。利用酸化油生产生物柴油不仅可以缓解生物柴油原料不足问题，还可解决酸化油所带来的环境问题。

The convertion of acid oil to biodiesel by use of immobilized Candida lipase absorbed on textile cloth was studied in a fixed bed reactor, which can not only reduce the environmental pollution of acid oil, but also produce a substitute for petroleum diesel. The acid oil mixed with methanol was pumped into three fixed bed reactors in series, and the methanol was added with the molar flow rate same as the acid oil in each reactor. The effects of enzyme content, solvent content, water content, flow rate of reactant and temperature on the enzymatic reaction were analyzed. The result of orthogonal experiments indicates that the optimal transesterification can be performed under the following conditions： immobilized lipase content in acid oil, 20% ; hexane content in acid oil, 10% ; water content in acid oil, 10%, reaction temperature, 50 ℃ ; and flow rate of reactant, 0.08 g/rain. Under these conditions, the FAME content of 90.18% in the product is obtained. The immobilized lipase can be reused with relatively stable activity after glycerol being removed from the surface. By refining, most of the chemical and physical properties of biodiesel will meet the American and Germany biodiesel standards and exceed the Chinese standard of 0^# petroleum diesel except for carbon residue, density and kinematic viscosity.

Kinetic Model Study on Enzymatic Hydrolysis of Cellulose Using Artificial Neural Networks

Enzymatic hydrolysis of cellulose was highly complex because of the unclear enzymatic mechanism and many factors that affect the heterogeneous system. Therefore, it is difficult to build a theoretical model to study cellulose hydrolysis by cellulase. Artificial neural network (ANN) was used to simulate and predict this enzymatic reaction and compared with the response surface model (RSM). The independent variables were cellulase amount X-1, substrate concentration X-2, and reaction time X-3, and the response variables were reducing sugar concentration Y-1 and transformation rate of the raw material Y-2. The experimental results showed that ANN was much more suitable for studying the kinetics of the enzymatic hydrolysis than RSM. During the simulation process, relative errors produced by the ANN model were apparently smaller than that by RSM except one and the central experimental points. During the prediction process, values produced by the ANN model were much closer to the experimental values than that produced by RSM. These showed that ANN is a persuasive tool that can be used for studying the kinetics of cellulose hydrolysis catalyzed by cellulase.

Enzymatic reaction of the immobilized enzyme on porous silicon studied by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry

Desorption/ionization on silicon mass spectrometry (DIOS-MS) is a matrix-free technique that allows for the direct desorption/ionization of low-molecular-weight compounds with little or no fragmentation of analytes. This technique has a relatively high tolerance for contaminants commonly found in biological samples. DIOS-MS has been applied to determine the activity of immobilized enzymes on the porous silicon surface. Enzyme activities were also monitored with the addition of a competitive inhibitor in the substrate solution. It is demonstrated that this method can be applied to the screening of enzyme inhibitors. Furthermore, a method for peptide mapping analysis by in situ digestion of proteins on the porous silicon surface modified by trypsin, combined with matrix-assisted laser desorption/ionization-time of flight-MS has been developed.