Studies on the digestive lipase of the surf clam Spisula solidissima

Crude molluscan lipase was prepared by homogenizing in tris buffer either fresh or lyophilized crystalline styles of Spisula solidissima. Both preparations showed specific activities less than 2% of standard hog pancreatic lipase, and pH and temperature optima of 8.0 and 20°C, respectively. When compared with mammalian hog pancreatic lipase, the clam lipase was found to be the more versatile enzyme on primary wax esters as well as methyl esters. Neither lipase was capable of hydrolyzing the oleate or palmitate esters of the secondary alcohols of glycerol, cholesterol or isopropyl alcohol; however, both showed activity on the oleate ester of isobutyl alcohol, a primary alcohol. S. solidissima lipase, hog pancreatic lipase, and a lipase isolated from the pancreas of the skate Raja erinacea, all demonstrated an inability to hydrolyze the methyl esters of eicosatetraenoic, eicosapentaenoic and docosahexaenoic acids. The lipase-resistant polyunsaturated fatty acid esters are discussed in terms of their role in the lipid biochemistry of marine organisms.     

多环芳烃生物修复中的表面活性剂

由于其致癌、致突变和致畸性，多环芳烃(PAHs)成为环境中一类重要的有机污染物。生物修复是一种经济和有效的修复污染土壤的方法。由于PAHs低的水溶性、强的吸附性，使其生物可利用性降低，不利于生物修复。添加表面活性剂是一种常见的加强PAHs生物利用性的方法。文章概述了近年来在多环芳烃生物修复中关于表面活性剂的研究进展。     

Controlling microbiological interfacial behaviors of hydrophobic organic compounds by surfactants in biodegradation process

Bioremediation of hydrophobic organic compounds （HOCs） contanlinated soils involves several physicochemical and microbiological interracial processes among the soil-water-microorganism interfaces. The participation of surfactants facilitates the mass transport of HOCs in both the physicochemical and microbiological interfaces by reducing the interfacial tension. The effects and underlying mechanisms of surfactants on the physi-cochemical desorption of soil-sorbed HOCs have been widely studied. This paper reviewed the progress made in understanding the effects of surfactant on microbiological interlhcial transport of HOCs and the underlying mechanisms, which is vital for a better understanding and control of the mass transfer of HOCs in the biodegradation process. In summary, surfactants affect the microbiological interfacial behaviors of HOCs during three consecutive processes： the soil solution-microorganism sorption, the transmembrane process, and the intracellular metabolism. Surfactant could promote cell sorption of HOCs depending on the compatibility of surfactant hydrophile hydrophilic balance （HLB） with cell surface properties; while the dose ratio between surfactant and biologic mass （membrane lipids） determined the transmembrane processes. Although surfactants cannot easily directly affect the intracellular enzymatic metabolism of HOCs due to the steric hindrace, the presence of surfactants can indirectly enhanced the metabolism by increasing the substrate concentrations.     

Response of gill ATPase and liver esterase of pseudorasobora parva to a two month exposure to glyphosate and metsulfuron methyl

Responses of gill ATPase and liver esterase of topmouth gudgeon, Pseudorasobora parva under sublethal exposure to glyphosate (WSC 41% as isopropyl amine salt) at 1.0, 5.0 and 25?mg/L and metsulfuron methyl (technical 98.2%) at 0.0095, 0.085 and 0.85?mg/L of water were measured at 8th, 16th, 24th and 65th days of exposure. Two way analysis of variance (ANOVA) followed by least significant difference (LSD) test and parameter estimates indicated a significant inhibitory effect on gill ATPase activity (max. 57%) by metsulfuron methyl and liver esterase activity by glyphosate (max. 43%) as compared to the control, but the differences in the residual activities among the concentration levels of the herbicides were not significant. The activities changed significantly with the sampling times except gill ATPase activity under metsulfuron methyl exposure. In most of the cases, the maximum inhibitory effect on the enzymes was recorded on the 8th day and over-recovering appeared with time.     

Synthesis of butyl butyrate in 93 % yield by <Emphasis Type="Italic">Thermomyces lanuginosus</Emphasis> lipase on waste eggshells

There is a high demand for immobilized enzymes in the industry because it is a green technology, it improves the efficiency of the crude enzyme and it minimizes the step of product recovery. Selection of a suitable carrier for enzyme immobilization is crucial. Eggshells are abundantly available as an organic waste. Here, lipase from Thermomyces lanuginosus was immobilized on eggshell to produce butyl butyrate. We evaluated the effect of lipase loading, from 0.5 to 3.0 %, size of eggshells, lower than 1, 1–2 and 2–4 mm, and temperature, from 30 to 50 °C. Results show that the optimum conditions calculated by response surface methodology analysis for butyl butyrate synthesis were 40 % w/v of immobilized lipase loading and 0.2 M of substrate concentration at 38 °C during 24 h. The predicted yield was 92.78 %, and the yield obtained during synthesis was 93.48 %.     

Effect of surfactants on phosphatase level of fresh water fish Labeo rohita

Alterations in the activity of enzymes Acid phosphatase (E.C.3.1.3.2) and Alkaline phosphatase (EC 3.1.3.1) in organs such as liver, gills and muscle of rohu following its exposure to surfactants viz. CTAB, SDS and Triton X-100 were analyzed. Different levels of exposure were given depending on the LC50 value of the surfactant used. Also, the influence of age and weight of the organisms was tested simultaneously. The activity of ACP in the tissues of fish exposed to all the three surfactants showed marked enhancement after exposure; the effect being highest in the liver followed by gill and muscle. Activity levels of ALP in different tissues of the fish exposed to the surfactants also showed an increase. Maximum increase was found in the liver followed by muscle, and gill. The increase in the levels of these enzymes indicates a stressful condition of the fish.     

Effects of fertilization and development of the common mussel Mytilus edulis after long-term exposure to a nonionic surfactant

Common mussels (Mytilus edulis L.) were exposed over a 5-month period to low-level concentrations (0.5 to 1.5 ppm) of a nonionic surfactant. Upon maturation at the end of this period, spawning ability was examined. Fertilization occurred at low-level concentrations (0.1 to 2.0 ppm) of the surfactant, and was most successful for gametes from the long-term controls and the highest long-term concentration (1.5 ppm). Inhibited or delayed larval development was observed, related to the concentration gradient of the short-term exposures. Gametes from mussels long-term exposed to the surfactant were more sensitive than those from the long-term control.     

Surfactants in sludge-amended agricultural soils: a review

Surfactants are included in different detergent formulations and are one of the most ubiquitous and important families of organic compounds. Although the generic term applies to a great number of products, 80% of their demand is covered by only ten types of compounds. The global surfactant market volume size is more than 18 million tons per year. Large quantities of surfactants are continuously released into the environment, where they can or cannot be degraded depending on their structure. The alkylbenzenesulphonate (LAS) is the most widely used surfactant. LAS can be degraded under aerobic conditions but is persistent in the environment under anaerobic conditions. Surfactants may enter the terrestrial environment through several routes, with the use of sewage sludge as fertiliser on agricultural land being by far the most important. High concentrations of surfactants and their degradation products may affect the biota. On the other hand, due to their amphiphilic nature, surfactants may interact both with inorganic as well as organic contaminants affecting their bioavailability.     

表面活性剂对菲的增溶动力学及质量增溶比（WSR）与亲水亲油平衡值（HLB）的关系

采用振荡实验方法,比较研究了2种非离子表面活性剂（Triton X-100和Tween80）和2种阴离子表面活性剂（SDS和SDBS）对菲的最佳增溶振荡时间及其对菲的增溶能力,同时也比较分析了表面活性剂对菲的质量增溶比（WSR）与其亲水亲油平衡值（HLB）的关系.结果表明,Triton X-100、Tween 80、S...     

Biological effects of surface active agents on marine animals

The biological effects of 5 surface active agents (the anionic ABS, LAS and LES 3 EO and the nonionic NP 10 EO and TAE 10 EO) on marine fishes, crustaceans and bivalves have been tested in, continuous-flow systems. Concentrations from 100 to 0.5 ppm were normally used. Fishes were found to be more susceptible (96 hLC 50 range: 0.8 to 6.5 ppm) than bivalves (96 hLC50 range: 5 to >100 ppm) while crustaceans were considerably more resistant (96 h LC 50 range: 25 to > 100 ppm). Within each of these 3 systematic groups, more active species were found to be more sensitive than less active species. Developmental stages were also more sensitive than adults. The resistance of crustaceans to surfactants decreases immediately after moulting. The most toxic agent for fishes and decapods was the soft anionic LAS, and for bivalves and barnacles the hard nonionic NP 10 EO. Ability to recover normal behaviour after exposure decreases with increasing concentration and time, and ceases earlier in anionic than in nonionic surfactants. The first reaction to the surfactants is increased activity (avoidance reaction of mobile species), followed successively by inactivation, immobilization and death. Nonionic surfactants affect the equilibrium in fishes. Sublethal effects appear as impaired locomotory activity and breathing rate in fishes and crustaceans, impaired byssus activity and shell closure in Mytilus edulis, burrowing in Cardium edule, Astarte montagui and Astarte sulcata. Siphon retraction is affected in Mya arenaria and Cardium edule, as is also the response to food of the Leander spp.     

Agricultural waste from the tequila industry as substrate for the production of commercially important enzymes

Approximately 1 million tons of Agave tequilana plants are processed annually by the Mexican Tequila industry generating vast amounts of agricultural waste. The aim of this study was to investigate the potential use of Agave tequilana waste as substrate for the production of commercially important enzymes. Two strains of Aspergillus niger (CH-A-2010 and CH-A-2016), isolated from agave fields, were found to grow and propagate in submerged cultures using Agave tequilana waste as substrate. Isolates showed simultaneous extracellular inulinase, xylanase, pectinase, and cellulase activities. Aspergillus CH-A-2010 showed the highest production of inulinase activity (1.48 U/ml), whereas Aspergillus niger CH-A-2016 produced the highest xylanase (1.52 U/ml) and endo-pectinase (2.7U/ml) activities. In both cases production of enzyme activities was significantly higher on Agave tequilana waste than that observed on lemon peel and specific polymeric carbohydrates. Enzymatic hydrolysis of raw A. tequilana stems and leaves, by enzymes secreted by the isolates yielded maximum concentrations of reducing sugars of 28.2 g/l, and 9.9 g/l respectively. In conclusion, Agave tequilana waste can be utilized as substrate for the production of important biotechnological enzymes.     

混合表面活性剂对菲和芘的增溶作用

比较了非离子表面活性剂 (Tween2 0 ,Tween40 ,Tween60 ,Tween80 ,Brij35和Brij5 8)与SDBS混合表面活性剂对菲和芘的增溶作用 .结果表明 ,在临界胶束浓度 (CMC)以上 ,表面活性剂对菲和芘有显著的增溶作用 ,菲的增溶顺序为 :Tween40 >Tween60 >Tween2 0 >Tween80 >Brij5 8>Brij35 ;芘的增溶顺序为 :Tween60 >Tween80 >Tween40 >Brij5 8>Tween2 0 >Brij35 .阴 非离子混合表面活性剂溶液的CMC值降低 ,胶束 水中溶质的分配系数Kmc增大 ,由此对菲和芘产生协同增溶作用 ,其顺序均为 :SDBS Tween60 >SDBS Tween80 >SDBS Tween40 >SDBS Brij5 8>SDBS Tween2 0 >SDBS Brij35 ,协同增溶程度在 1 1 7%— 65 8%之间 .     

Spore inhibition-based enzyme substrate assay for monitoring of aflatoxin M1 in milk

A spore germination-based concept and its transformation into a field level prototype for monitoring aflatoxin M₁ (AFM₁) in milk was developed. Initially, 15 strains of Bacillus spp. procured from different culture collection were screened for AFM₁ sensitivity using spot assay and marker strain showing inhibition at 0.5 ppb was selected based upon maximum zone of inhibition. The selected strain B. megaterium 2949 was further screened for different enzymes activities and subsequently its spores were produced to an extent of 73.13% ± 3.197% in newly developed sporulation medium containing beef extract (0.0075% ± 0.0004%), yeast extract (0.015% ± 0.001%), peptone (0.0375% ± 0.0016%), and sodium chloride (0.0375% ± 0.0018%). A spore germination-based concept/ assay was optimized by immobilizing spores in eppendorf with pretreated milk (80°C/15 min) containing germinant and chromogenic substrate followed by incubation at 37°C. The appearance of sky blue color within real time of 45 min indicated spores germination and release of specific marker enzyme such as acetyl esterase and its specific action on chromogenic substrate which demonstrates absence of AFM₁ in milk. However, if there was no color change, presence of AFM₁ at 0.5 ppb MRL was denoted by Codex. The developed concept on AFM₁ detection was validated and a correlation of 0.97 was established with AOAC approved Charm 6602 and ELISA at Codex MRL with minimal false positive and negative results. The cost effective test has potential application in dairy farms, manufacturing, and R&D units for routine monitoring of AFM₁ in milk.     

Ontogenetic changes in digestive enzyme activity of the spiny lobster,<Emphasis Type="Italic"> Jasus edwardsii</Emphasis> (Decapoda; Palinuridae)

Digestive enzyme profiles of puerulus, post-puerulus, juvenile and adult stages of the spiny lobster Jasus edwardsii Hutton were determined in order to identify ontogenetic changes in digestive capabilities and assess capacity to use dietary components and to exploit diets to meet nutritional requirements. Juvenile and adult lobsters exhibited a diverse range of enzymes, suggesting they can exploit varied diets. Proteolytic activity of trypsin, chymotrypsin and carboxypeptidase A (1.86–3.70 U mg^–1 digestive gland protein) was significantly higher than carbohydrase activity of amylase, -glucosidase, cellulase and chitinase (0.0014–0.38 U mg^–1 digestive gland protein), thus showing that dietary protein was more important than carbohydrate and that the lobster is carnivorous. These conclusions are consistent with other studies that found spiny lobsters to feed predominantly on crabs, bivalves, ophiuroids and sponges. Lipase activity (0.371 U mg^–1 digestive gland protein) was also relatively high, thus showing the importance of dietary lipid. Total activities (units per digestive gland) of every enzyme assayed increased significantly with lobster carapace length. There were few significant ontogenetic trends in specific enzyme activity (U mg^–1 digestive gland protein). Amylase and laminarinase specific activity was significantly higher in small lobsters than large lobsters (regression analyses, respectively, F_(1,23)=9.84, P=0.005; F_(1,11)=19.65, P=0.001), suggesting that carbohydrates including laminarin are more important in the diet of small juveniles. Trypsin, amylase and lipase activities were detected in all non-feeding puerulus stages, suggesting that feeding is not a cue for digestive enzyme production in J. edwardsii. Significant variations in total and specific activities of amylase (F_(1,3)=14.2, P=0.00; F_(1,3)=14.2, P=0.00) and trypsin (F_(1,3)=8.8, P=0.00; F_(1,3)=21.41, P=0.00) and a declining trend in lipase total activity between non-feeding puerulus stages suggests that they may be hydrolysing endogenous energy reserves to sustain their onshore swimming activity prior to settlement. Profiles suggest carbohydrate and lipid are utilised first, followed by protein. Consistently high levels of lipase in all puerulus stages (0.24–0.7 U digestive gland^–1; P>0.05) confirm the importance of lipid as a major energy substrate.Communicated by G.F. Humphrey, Sydney     

白眉蝮蛇（Agkistrodon halys ussuriensis）蛇毒精氨酸酯酶对小鼠的生殖毒性及F1子代的影响

研究了白眉蝮蛇蛇毒中分离的纯精氨酸酯酶对小鼠的一般生殖毒性及对F1代小鼠的影响 .0 .36U/kg剂量使雄性F0 小鼠体重增长缓慢 ,并使睾丸系数增大 (P <0 .0 5 ) ,但病理学检验睾丸和附睾正常 ;该剂量处理孕鼠 ,减轻了胎鼠和新生F1代小鼠的体重 (P <0 .0 5 ) ,但对F0 雌性小鼠的生殖功能及F1代小鼠的一般体征、中枢神经系统发育及生殖机能均无影响 (P >0 .0 5 ) .纯精氨酸酯酶对小鼠生殖无毒剂量为 0 .18U/kg .表 2参 9     

Simultaneous copper bioaccumulation,growth and lipase production of Rhizopus delemar in molasses medium: optimisation of environmental conditions using RSM

In this study, the bioaccumulation of Cu(II) ions by R. delemar and their influence on growth of the microorganism were assessed, and the impact of Cu(II) ions on the activity of lipase enzyme degradation of lipids in wastewaters by enzymatic hydrolysis was received for consideration. Optimisation of Cu(II) bioaccumulation process was performed by varying three independent parameters (pH, temperature and Cu(II) concentration) using a central composite design (CCD) under response surface methodology (RSM). For the maximum Cu(II) bioaccumulation and lipase production at a desired level of the biomass concentration level, a total of 20 experimental runs were set and the experimental data fitted to the empirical second-order polynomial models. The adequacies of the predicted models for the Cu(II) bioaccumulation, lipase activity and biomass concentration were confirmed by the coefficients of multiple regression (R²), which were adjusted to be 99.6%, 94.5%, 93.4%, respectively indicating reasonably good models for practical implementation. A maximum Cu(II) bioaccumulation of 36.81 mg/L was obtained at pH 6.0, at 32.6°C, and at 50 mg/L initial Cu(II) concentration. The lipase activity of R. delemar decreased only from 505 U/L in Cu(II)-free fermentation medium to 476 U/L in the presence of Cu(II) giving 5.74% decrease in lipase activity.     

Effervescence assisted dispersive liquid–liquid microextraction followed by microvolume UV-Vis spectrophotometric determination of surfactants in water

The novel and simple methods for the sensitive determination of cationic and anionic surfactants in water based on effervescence assisted dispersive liquid–liquid microextraction and microvolume UV-Vis spectrophotometry have been developed. The method involves ion-pair extraction of cationic and anionic surfactants with organic dyes (methyl orange and azure A, respectively) during the dispersion of extraction solvent (CHCl₃) by CO₂ bubbles which are formed by the injection of a mixture of the extraction solvent and proton donor solvent into the sample solution which contains carbonate-ions as effervescency agent. The analytical performance of the proposed procedure was compared with the conventional dispersive liquid–liquid microextraction. Appropriate experimental conditions for both methods were investigated. The absorbances of the colored extracts at wavelengths of 440 and 625 nm obey Beer's law within the range of 0.1–5.0 mg/L for both cationic and anionic surfactants. The limit of detection (LOD) obtained using the effervescence assisted extraction method was 30 µg/L for cationic and anionic surfactants. The proposed methods were successfully applied to determination of surfactants in water samples.     

Effect of dietary factors and environmental chemicals on intestinal drug metabolizing enzymes

The liver is the major site of metabolism of foreign chemicals, but contribution from intestinal biotransformation can influence the overall disposition of chemicals. The intestinal drug‐metabolizing enzymes are located in the endoplasmic reticulum and they possess biochemical properties similar to those of the hepatic system. In general, the rate of in vitro metabolism of drug substrates in intestines is lower than in the liver. There seem to be differences in regulation and induction of hepatic and intestinal drug‐metabolizing enzymes. The in vitro intestinal metabolism of foreign chemicals is affected by several factors including the nutritional status of the animal. Dietary components such as vitamins, lipids and vegetables can modify the activities of intestinal drug‐metabolizing enzymes. Phenobarbital or 3‐methyl‐cholanthrene (3‐MC) pre‐treatment induces the hepatic metabolism of a wide variety of drug substrates in a number of species, while the stimulation of intestinal enzymes is both substrate and species specific. Rabbit intestinal drug‐metabolizing enzymes seem to be resistant to induction by foreign chemicals.     

Cu2+或表面活性剂AE对黄鳝肝损伤的超微结构观察

利用透射电镜研究了亚致死浓度的Cu^2 或表面活性剂AE(脂肪醇聚氧乙烯醚)对黄鳝肝细胞结构的损伤作用．观察发现，在ρ(Cu^2 )=4．5mg／L的污染环境中，黄鳝肝脏细胞核变形，染色质凝集，内质网囊泡化，细胞质中出现较多的溶酶体和过氧化物酶体；在ρ(AE)=4．0mg／L的表面活性剂污染下，黄鳝肝脏细胞核变形，内质网呈线状，溶酶体、过氧化物酶体体积大、数量多，部分细胞质解体，细胞中出现空腔甚至死亡．图版1参10     

全氟烷基磺酸盐(PFOS)及其替代品对两栖类胚胎的发育毒性

用非洲爪蟾胚胎致畸实验和黑斑蛙胚胎发育毒性实验,比较研究了PFOS及其4种替代品对两栖动物胚胎的发育毒性。结果发现,用调聚法合成的织物三防整理剂和50%的全氟丁基有机铵盐阳离子表面活性剂同PFOS一样对非洲爪蟾胚胎有明显毒性,且织物三防整理剂和表面活性剂的毒性强于PFOS;电解氟化法合成的C4织物及C6织物三防整理剂(以下简称C4及C6织物三防整理剂)对非洲爪蟾胚胎没有明显毒性。与非洲爪蟾不同,PFOS和4种替代品对黑斑蛙胚胎没有明显毒性。结果显示,从生物安全性的角度分析,C4和C6织物三防整理剂可作为PFOS的替代品使用,而织物三防整理剂和表面活性剂的毒性比PFOS大,作为PFOS替代品使用需要慎重考虑。