海洋低温溶菌酶的制备及酶学性质

对一株海洋杆菌产低温溶菌酶的分离纯化及理化性质进行了研究。20℃下将菌株在含有葡萄糖、酵母膏、胰蛋白胨的标准海水培养基中震荡培养40h。获得活性大约为150U/ml的溶菌酶上清液。上清液经SMB-20生物型错流超滤系统浓缩、透析及CM-Sepharose-FF阳离子交换层析（     

产β-1,3-葡聚糖酶海洋细菌的筛选及其酶学性质研究

依据从海水中分离、筛选得到的β-1,3葡聚糖酶活较高的菌株SY-3之形态、生理及生化特性,初步鉴定为气单胞菌.该菌发酵产生的粗酶液经过硫酸铵沉淀、透析除盐后,对其酶学性质进行了初步研究.经测定,SY-3所产β-1,3葡聚糖酶的最适作用温度、pH分别为30 ℃、7.5.通过Lineweaver-Burk做图,得到其米氏常数(Km)为1.21 mg/ml.K+、Na+和Ca2+对酶促反应有促进作用,而Mg2+、Mn2+、Cu2+和Zn2+等有不同程度的抑制作用.     

壳聚糖固定化海洋低温碱性蛋白酶YS-80-122的研究

以壳聚糖为固定化载体,采用吸附交联法,以戊二醛为交联剂,对海洋低温碱性蛋白酶YS-80-122进行固定化。通过单因素和正交试验优化得到最佳固定化条件为pH7·0,戊二醛终浓度0·2%,吸附时间50min,酶载体比例40mg/g,交联时间16h,交联温度10℃,固定化酶酶活回收可达47·52%。固定化酶和游离酶的最适作用温度均为30℃,最适作用pH分别为9·0和10·0,相比游离酶,固定化酶可在更宽的温度和pH范围内发挥高活性,且其温度和pH稳定性得到显著提高。室温下经冷冻干燥的固定化酶贮存90d后活性基本保持不变,贮藏半衰期约为190d。以酪蛋白为底物,重复使用7次后酶活仍保留有79·2%,操作半衰期约为21次使用率。     

磁性壳聚糖微球固定化褐藻酸酶的研究

利用反相悬浮交联法制备磁性壳聚糖微球(magnetic chitosan microspheres, M-CS),并对褐藻酸酶进行固定化研究.结果表明,M-CS呈规则的圆球形,具有较好的磁响应性,可稳定地保存在弱酸和弱碱中.其弱碱交换量随着戊二醛用量的增加而减少,悬挂醛基则相应地增加.M-CS对褐藻酸酶的吸附动力学实验表明,M-CS容易吸附褐藻酸酶,但吸附的酶量受载体与酶的比例、溶液的离子浓度、戊二醛的用量、溶液pH的影响明显,而温度对吸附的酶量的影响则相对较弱.酶学性质研究表明,相对于游离的褐藻酸酶,固定化酶的最适温度略有升高,可明显改善其热稳定性和酸碱稳定性,与底物的亲和力也有所增强.     

海洋微生物酶的研究概况

海洋中蕴藏着丰富的微生物资源,海洋微生物对于维持海洋生态平衡发挥着重要作用。尽管海洋中的许多微生物推测是由陆地环境经河水、污水、雨水或尘埃等途径而来的,但特殊的海洋环境赋予海洋微生物以特殊的遗传结构和生活习性。海洋生态环境复杂,高盐度、高压力、低温及特殊的光     

不同载体固定化海洋微生物酯酶ETM-b的性能比较

分别采用海藻酸钠包埋法、明胶包埋交联法、壳聚糖吸附交联法制备固定化海洋芽孢杆菌酯酶ETM-b,并对其固定化条件进行了研究。结果发现,壳聚糖制备的固定化酶效果最好,壳聚糖2%、戊二醛浓度1%、小球与酶液4∶3(g/ml)时制备的固定化酶的活性回收最高,达到66%。壳聚糖制备的固定化酶使用10次,相对活性保留70%,具有良好的操作稳定性。固定化酶在非水介质中具有转化α-乙酸萘酯(α-Naphthyl acetate)的能力,在异辛烷、正辛烷、正己烷中活性表现最高。     

海洋微生物低温酶特性及其在工业中的潜在用途

依据近年来关于低温酶的研究资料，讨论了低温酶的研究现状，对产低温酶海洋细菌的种类、嗜冷特性、适应低浊的机制以及对其在工业上的应用作一评述。     

驼海燕内脏中蛋白酶的提取及其酶学性质的研究

以驼海燕为原料,用丙酮沉淀法制备蛋白酶粗酶,并对其酶学性质进行研究。试验结果表明,提取蛋白粗酶的最佳条件为:原酶液与丙酮的体积比为1∶1.6;蛋白酶反应的最适温度为45℃,最适pH值为8.1;pH为7～9时可保持较高的酶活力;蛋白酶的酶活力在20～50℃基本稳定。酶促反应结果表明,Na+、K+、Li+、Mg2+对蛋白酶活力具有促进作用,而Zn2+、Cu2+、Hg2+能抑制蛋白酶的活力,EDTA对蛋白酶活力的影响甚微。酶动力学方程表明,最大反应速度vmax=5.99U/mL,米氏常数Km=0.69g/mL。     

适用鱼油加工的Patatin酯酶固定化及其特性研究

Patatin (马铃薯糖蛋白)是一种酯水解酶,具有脂肪酶催化活性,可应用于脂肪的水解加工。针对游离Patatin酯酶稳定性差且工业生产中酶很难重复利用的缺点,利用ConA (刀豆蛋白A)耦联的纳米磁珠材料固定化Patatin酯酶,以提高其工业化应用的催化特性。通过研究,发现ConA耦联的纳米磁珠的平均吸附率为24.50%。同时筛选出适宜的Patain酯酶固定化材料为PAA (聚丙烯酸)-Fe₃O₄,其最优固定化条件为固定化时间47.2 min,固定化温度25.3℃,磁珠添加量为3.0 mg·mL^-1。Patain酯酶固定化纳米磁珠在40.0℃、pH 7.0时水解底物活性最高,且相比游离Patatin酯酶,其温度耐受性提高了123%左右,pH耐受性提高了47%左右,连续反应5次后仍保留56.60%酶活力,说明纳米磁珠固定化后提高了Patatin酯酶的酶催化特性,为其在水产品如鱼油中的加工应用提供了一种催化性能更强的新酶体系。     

固定化微生物技术及其在养殖水体中的应用

介绍了固定化微生物技术的定义,固定化微生物的制备方法,并重点介绍了固定化微生物技术在养殖水体中的应用以及存在的问题和展望。     

一株海洋过氧化氢酶高产菌的鉴定及产酶条件优化

对来自青岛近海海域底泥的一株产过氧化氢酶菌株YS0810进行形态学观察、16S rDNA序列同源性分析及生理生化特性的鉴定,在250 ml摇瓶中进行发酵产酶条件优化。初步确定该菌属于不动杆菌属Acinetobacter。发酵培养的最佳碳、氮源分别为蔗糖20 g/L和蛋白胨15 g/L,无机盐MgSO4·7H2O、NaCl、KH2PO4最佳浓度分别为0.9、5.0和1.0 g/L;菌株在培养基起始pH=7.0、4%接种量、50 ml装液量和25℃的条件下发酵24 h获得较高的酶产量。在最佳培养条件下酶产量为2469 U/ml,是优化前的5倍。     

一株海洋中性蛋白酶高产菌S-3685的鉴定及产酶条件

通过生理生化、分子生物学、单因子优化等方法研究了来自南海海域一株产中性蛋白酶菌株S-3685,并对其在250 ml摇瓶中的培养条件进行了优化。结果显示,该菌株初步确定为芽孢杆菌属(Bacillus),发酵培养的最佳碳、氮源分别为葡萄糖10 g/L和豆面10 g/L,牛肉膏5 g/L,酵母膏5 g/L。无机盐MgSO4·7H2O、Na2CO3、KH2PO4最佳浓度分别为0.2 g/L、2.0 g/L、1.0 g/L;菌株在培养基起始pH=7.0、4%接种量、15 ml/250 ml (v/v)装液量和30℃的条件下,发酵72 h获得较高的酶产量。在最佳培养条件下产酶量为4250 U/ml,是优化前的5倍。     

两株海洋微生物抗肿瘤代谢产物的筛选及其菌种鉴定

对5株海洋微生物进行发酵培养,发酵液离心后,收集上清液,采用饱和正丁醇萃取制备海洋微生物的代谢产物,采用MTT活性跟踪法,以BEL-7402、RKO、A549、U251和MCF-7等细胞系为模型,对代谢产物进行抗肿瘤活性的筛选。结果显示,菌株S-1和N16代谢产物的活性组分具有较好的抗肿瘤活性:菌株S-1对MCF-7、U251和BEL-7402三种肿瘤细胞的IC50值分别为44、102和82μg/ml;菌株N16对MCF-7和BEL-7402两种肿瘤细胞的IC50值分别为84、133μg/ml。对菌株S-1和N16进行了16S r RNA序列分析、生理生化特征鉴定,结果显示,菌株S-1为短芽孢杆菌属(Brevibacillus sp.),N16为芽孢杆菌属(Bacillus sp.)。通过显微镜观察了菌株Brevibacillus sp.S-1和Bacillus sp.N16代谢产物的活性组分对BEL-7402细胞形态的影响,表明两株菌的活性组分可以改变肝癌细胞BEL-7402的细胞形态,抑制细胞增殖。该研究为新型抗肿瘤先导化合物的发现提供了新的海洋微生物种质资源。     

海水桡足类的营养及在鱼、虾、蟹幼体培育中的应用

海水桡足类在海洋食物网和生态系统中具有重要的地位,是各种海洋经济鱼类和甲壳类的重要饵料,也是经济水产动物人工育苗阶段的优质活饵料源。研究海水桡足类营养和应用效果,对提高海水鱼、虾、蟹类幼体变态率、成活率、苗种质量和育苗成功率具有重要的意义。     

关于我国海水网箱养鱼发展现状与危机分析及可持续发展的探讨

我国海水网箱养鱼发展现状是以个体联合体生产经营为主体的低技术水平的数量规模化，众多传统港湾已形成了大范围的严重超负荷养殖状态，其对环境巨大的污染力引发鱼病猖獗和频受赤潮侵袭，已面临严峻的养殖环境危机。本文综合分析了我国海水网箱养鱼的发展现状与负面效应及其现状危机，并初步进行了危机根源和可持续发展的讨论分析。     

Functional Properties of the Marine Gastropod Molluscs Chicoreus ramosus and Babylonia spirata Collected from the Southern Coast of India

ABSTRACT

This study characterized the functional properties of ethyl acetate-methanol (EA-MeOH) and chloroform extracts of the muricid gastropod Chicoreus ramosus and buccinid Babylonia spirata. The EA-MeOH extract of B. spirata was a potent quencher of radical cation (IC₉₀ < 1 mg/mL), whereas that of C. ramosus held greater anti–inflammatory potential (IC₉₀ 5-lipoxygenase inhibition 1.74 mg/mL) than the buccinid gastropod. The EA-MeOH extract of C. ramosus displayed potent antidiabetic activities as deduced by its attenuation properties against carbolytic enzymes α-amylase and α-glycosidase (IC₉₀ 1.06 and 2.25 mg/mL, respectively) than those exhibited by B. spirata (IC₉₀ 2.32 and 4.50 mg/mL, respectively). The spectroscopic dereplication studies to determine probable chemical groups in the solvent extracts of the studied gastropods revealed the presence of functionalities, which might augment the electronic properties of the bioactive principles present within the extract, thereby enhancing their activity. Thus, the present study recognized that marine gastropods C. ramosus and B. spirata have potential functional implications against oxidative stress-induced diseases, such as diabetes and inflammation. This research supports further investigation of these previously uncharacterized marine gastropod species to isolate potential bioactive leads for use against various biological targets and to develop functional food formulations.     

Trophic mass balanced models and dynamic simulations of benthic communities from La Rinconada Marine Reserve off northern Chile: network properties and multispecies harvest scenario assessments

• 1. Mass balanced trophic models and dynamic simulations of two benthic ecological systems from La Rinconada Marine Reserve (Antofagasta Bay, SE Pacific) were constructed.
• 2. The scallop Argopecten purpuratus is the most important benthic resource in La Rinconada Marine Reserve, followed by the carnivorous snail Thais chocolata, and the filter‐feeder bivalves Tagelus dombeii and Transennella pannosa.
• 3. Information on biomass, P/B ratios, catches, food spectrum, consumption, and dynamics of commercial and non‐commercial species was obtained and examined using Ecopath with an Ecosim software package.
• 4. The bivalves A. purpuratus and T. dombeii represented the most abundant compartments in the studied subsystems. Of the carnivores, the snail T. chocolata was dominant, followed by the crabs Cancer spp. and the functional group of large epifauna.
• 5. The two subsystems presented similar values of system throughput. The mean trophic level of their fisheries also reached similar magnitudes (2.0), showing that the harvests in each system concentrated on secondary producers. Likewise, both subsystems presented similar A/C ratios (29.9 and 30.3), suggesting that they were immature.
• 6. The results obtained using mixed trophic impact (MTI) and Ecosim (increasing the fishing mortality F_i by four times) showed that only four species propagated the highest direct and indirect effects. Coincidentally, these species are the most economically important and the changes produced by the scallop A. purpuratus are noteworthy.
• 7. With regard to the system recovery time (SRT) estimates, only three species or functional groups presented the highest magnitudes, from highest to lowest: the sea star Luidia magallanica, the scallop A. purpuratus, and the crabs Cancer spp.
• 8. The topological keystone indexes of Jordán and Libralato had divergent results. According to Jordán's index, the keystone species were L. magallanica, Cancer spp., and detritus; whereas Libralato's index showed phytoplankton to be the keystone species.
• 9. Based on the results obtained, it is concluded that trophic mass balanced models and simulated management scenarios have considerable value for planning interventions and manipulations or for planning more sustainable management strategies in La Rinconada Marine Reserve. Copyright © 2009 John Wiley & Sons, Ltd.

     

Trypsin Enzyme from Viscera of Common Kilka (Clupeonella cultriventris caspia): Purification,Characterization, and Its Compatibility with Oxidants and Surfactants

The purification of trypsin from the common kilka (Clupeonella cultriventris caspia) viscera (pyloric caeca) resulted in a 28.3-fold increase and 12% yield by ammonium sulfate precipitation (30–60%), Sephadex G-75, and DEAE-cellulose chromatography. Trypsin showed a molecular weight of 23.2 kDa and appeared as a single band on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), native-PAGE, and zymography. The trypsin had optimal activity at pH 8.0 and 60°C for the hydrolysis of α-N-benzoyl-DL-arginine-ρ-nitroanilide hydrochloride (BAPNA) substrate. Trypsin was stable up to 50°C and at pH range of 7.0–10.0. Activity was significantly inhibited by soybean trypsin inhibitor (SBTI) and N-ρ-tosyl-L-lysine-chloromethylketone (TLCK) inhibitors (p < 0.05). The enzyme was relatively stable toward oxidizing agents, retaining 59.7 and 98.0% of its initial activity after 1 h incubation in the presence of 15% H₂O₂ and 1% sodium perborate, respectively. Trypsin was significantly activated by surfactants and Ca²⁺, Mg²⁺, and Mn²⁺ and inactivated by Fe²⁺, Zn²⁺, Cu²⁺, Al³⁺, Ba²⁺, and Co²⁺ (p < 0.05). Nevertheless, Na⁺ and K⁺ had no significant effect on trypsin activity (p > 0.05). The purified trypsin showed significantly higher catalytic efficiency (k_cat/K_m) than porcine pancreatic trypsin against BAPNA and N-α-p-Tosyl-L-arginine methyl ester hydrochloride (TAME) substrates (p < 0.05).