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1.
选择胰蛋白酶、弹性蛋白酶、枯草杆菌蛋白酶、胰凝乳蛋白酶及其特异底物,建立比色法检测,对沙塘鳢、鲫鱼、草鱼、乌鳢鱼卵的丝氨酸蛋白酶抑制活性进行了比较。试验结果表明,胰蛋白酶抑制活性依次为乌鳢草鱼沙塘鳢鲫鱼;弹性蛋白酶抑制活性依次为沙塘鳢鲫鱼草鱼乌鳢;枯草杆菌蛋白酶抑制活性依次为沙塘鳢鲫鱼草鱼乌鳢,胰凝乳蛋白酶活性只受乌鳢抑制。研究表明这可能与抑制剂P1位点的残基种类有关。此外还对鱼卵中的抑制剂成分进行热稳定性与酸碱稳定性的检测,结果表明其具有很强的热稳定与酸碱稳定性。  相似文献   

2.
以罗非鱼肠为原料,采用超声波辅助提取技术、电泳技术和层析技术等对罗非鱼肠道蛋白酶进行研究,结果表明:将鱼肠匀浆经超声波提取的粗酶液,用30%~70%的硫酸铵盐进行盐析、HitrapTM Q FF阴离子交换柱纯化及Sephadex G-100凝胶柱分离纯化,得到罗非鱼肠蛋白酶纯品,其比活为335U/mg,得率为32.8%;SDS-PAGE电泳为单一蛋白酶带,分子量为28ku。该酶最适pH为8.0~8.5,在pH7.0~9.0的条件下稳定;最适温度为37~42℃,热稳定性好;该酶的Km值和Vmax值分别为0.605g/L和9.407μg/min。金属离子Ag+、Pb2+对蛋白酶有完全抑制作用,Na+、K+对该酶无抑制作用。丝氨酸蛋白酶抑制剂能完全抑制该酶活性,胃蛋白酶抑制剂和脲素对该酶有一定抑制作用,EDTA没有明显抑制作用,DTT能激活该酶活性,该酶为丝氨酸蛋白酶。  相似文献   

3.
草鱼肠道、肝胰脏对饲料蛋白质酶解速度的比较   总被引:14,自引:0,他引:14       下载免费PDF全文
草鱼样品体重(1543±213)g。采用离体消化培养和茚三酮比色方法,以草鱼前肠、中肠、后肠和肝胰脏的粗酶液作为酶源,对鱼粉、豆粕、菜粕和棉粕进行酶解,并测定酶解14h内,豆粕酶解液中氨基酸生成量随时间的变化,以及0至4h,内4种饲料蛋白质酶解液中氨基酸的生成量。结果表明,(1)在0至4h内,4种饲料酶解液OD570随时间变化的线性关系较好。(2)对于同种蛋白质饲料原料,以中肠组织粗酶液的酶解速度最大,其余依次为前肠、后肠及肝胰脏粗酶液。表明草鱼中肠对饲料蛋白质的酶解能力强于前肠和后肠,同时,肠道酶液的酶解速度均大于肝胰脏。(3)对于4种不同的蛋白质饲料原料,以鱼粉酶解氨基酸生成速度最大,其他依次为豆粕、菜粕、棉粕。(4)草鱼肝胰脏粗酶液对豆粕的酶解氨基酸生成速度最大,表明草鱼对豆粕进行酶解消化的能力较强。实验证实,饲料原料种类的差异和饲料蛋白质组成与性质的差异使其酶解氨基酸生成量和生成速度有差异。  相似文献   

4.
草鱼和银鲫肠道产消化酶细菌的研究   总被引:3,自引:1,他引:3  
检测了分别从草鱼(Ctenopharyngodon idellus)和银鲫(Carassius auratus gibelio)肠道中分离的180株细菌的蛋白酶、脂肪酶、淀粉酶和纤维素酶的产酶能力。结果显示,两种鱼肠道内可分泌胞外消化酶的细菌包括Aero-monas(气单胞菌属,Aer.)、Vibrio(弧菌属,Vib.)、Bacillus(芽孢杆菌属,Bac.)、Pseudomonas(假单胞菌属,Pse.)四个种属的细菌,Aer.在其中占主要优势,45.71%的Aer.可分泌胞外消化酶。草鱼可分泌上述四种胞外消化酶的菌株共有33株,占肠道菌总数的36.67%;银鲫43株,占47.78%。产酶菌的分布上,草鱼中肠内产消化酶细菌数量显著多于前肠和后肠(P<0.05),前、中、后肠分别是6株、20株和7株;银鲫中肠和后肠数量差异不显著,前肠分布最少。草鱼分泌蛋白酶、脂肪酶、淀粉酶和纤维素酶的菌株分别有21株(23.33%)、10株(11.11%)、30株(33.33%)和16株(17.78%)。银鲫肠道内未检测到可分泌纤维素酶的细菌,蛋白酶、脂肪酶和淀粉酶菌株的数量分别是21株、37株和17株。可见鱼类肠道细菌对食饵消化有重要作用。  相似文献   

5.
沸石粉作草鱼饲料矿物添加剂的研究   总被引:1,自引:1,他引:1  
在草鱼饲料中添加5%的沸石粉,使草鱼成活率提高,生长加快,饵料消耗降低。草鱼肠道蛋白酶活性提高27%,而肝胰脏蛋白酶活性不受影响。对血液中 BUN、GPT、CR、TG 等生化指标及草鱼肌肉的品质无不良影响。  相似文献   

6.
应用对比试验和正交试验方法对草鱼进行酶解技术研究。试验结果表明,胃蛋白酶酶解草鱼酶用量为(E/S)0.6%,温度55℃,pH2.0,酶解5h氨基态氮含量较高,为0.0190g/L;枯草杆菌蛋白酶酶解草鱼酶用量为(E/S)0.01%,温度45℃,pH6.5,酶解4h氨基态氮含量较高,为0.0417g/L;胰蛋白酶酶解草鱼酶用量为(E/S)0.64%,温度50℃,pH8.0,酶解5h氨基态氮含量较高,为0.0472g/L。通过对3种蛋白酶在最优条件下酶解草鱼肉的比较可知,枯草杆菌蛋白酶和胰蛋白酶酶解效果较好。为草鱼深加工提供了一定的科学依据。  相似文献   

7.
徐凯慧  陈云峰  章玉贵  李小勤  冷向军 《水产学报》2023,47(10):109603-109603
为了探究饲料中补充不同水平丁酸钠对草鱼生长性能、肠道消化酶活性、血清生化指标和肠道组织形态的影响,分别添加0 mg/kg (SB0)、1 000 mg/kg (SB1)、2 000 mg/kg(SB2)和3 000 mg/kg (SB3)的微囊丁酸钠(含50%丁酸钠)至基础饲料中,设计4组等氮等脂的实验饲料。选取300尾体质健康的草鱼(10.0±0.1) g,分成4组,每组3个重复,每个重复25尾鱼,放置在水泥池中养殖8周。结果显示,各组草鱼增重率为273.7%~279.9%,饲料系数为1.55~1.60,饲料中补充丁酸钠对草鱼生长性能和体成分无显著影响。与对照组相比,添加不同水平的丁酸钠均显著提高了肠道蛋白酶活性和血清超氧化物歧化酶(SOD)活性,降低了血清谷草转氨酶(AST)活性,SB1组血清碱性磷酸酶(AKP)活性显著升高,而SB2和SB3组血清谷丙转氨酶(ALT)活性显著降低。各组肠道淀粉酶活性、血清酸性磷酸酶(ACP)、甘油三酯(TG)、葡萄糖(GLU)、总蛋白(TP)和D-乳酸(D-LA)无显著差异。在肠道组织学方面,各丁酸钠补充组肠道绒毛高度和SB1、SB2组肌层厚度显著...  相似文献   

8.
冯健 《水产学报》2004,28(5):505-509
为了研究草鱼肠道完整吸收小肽与肠道中小肽的关系,用6.25%酶解酪蛋白、酸解酪蛋白、酪蛋白溶液和生理盐水进行草鱼肠道灌注试验(1mL·100g-1体重)。20min后尾静脉采血制备血浆,高效液相色谱分析结果表明:灌注酶解酪蛋白溶液的草鱼血液循环中总肽量和某些肽量显著(P<0.05)高于灌注生理盐水、酸解酪蛋白、酪蛋白溶液;较灌注生理盐水草鱼血液循环中总肽量提高18.73%。肠道灌注酸解酪蛋白、酪蛋白溶液对血液循环中总肽量影响不大,无显著性差异(P>0.05)。草鱼血浆中肽量的增加与肠道提供的肽种类和数量有关。实验结果表明,草鱼肠道能够完整地吸收某些小肽进入血液循环。  相似文献   

9.
七种常见养殖鱼血清胰酶抑制活性的比较   总被引:3,自引:0,他引:3       下载免费PDF全文
黄素文 《水产学报》2003,27(5):415-419
α2巨球蛋白能结合并抑制蛋白酶的活性,但不抑制该蛋白酶对人工合成的小分子底物的酶活。利用这一特性,选择胰酶的一种有色小分子底物BAPNA,建立BAPNA比色检测法,对7种常见养殖鱼:草鱼、鲫、鲢、胡子鲇、鳙、加州鲈和黄鳝的血清胰酶抑制活性进行了比较,胰酶抑制活性由高到低依次为:草鱼>鲫>黄鳝>鳙>鲢>胡子鲇>加州鲈;α2巨球蛋白活性由高到低依次为:黄鳝>草鱼>胡子鲇>加州鲈>鲢>鳙>鲫;黄鳝和草鱼的α2巨球蛋白活性明显高于其他几种鱼的α2巨球蛋白活性。  相似文献   

10.
草鱼和青鱼消化酶的分布特性   总被引:4,自引:0,他引:4  
刘忠义  王璋 《淡水渔业》2006,36(1):14-18
选择草鱼(C tenopharyngodon idellus)和青鱼(Mylopharyngodon piceus)作试验鱼,对它们消化道的酸性蛋白酶、碱性蛋白酶、淀粉酶以及三酰基甘油脂酶沿消化道及肝脏和胰腺中的活性分布进行了比较研究。研究表明:这些酶在消化道中呈现不同的分布规律,在肝脏和胰腺中酶的活性高于在消化道中相应酶的活性;同种鱼不同消化酶的分布有所差别;两种鱼消化组织中的各种酶活性分布规律也有差别;青鱼和草鱼的主要消化酶在不同消化器官中的分布有较大差别。  相似文献   

11.
Vermiculated sailfin catfish (Pterygoplichthys disjunctivus, Weber, 1991), a member of the Loricariidae family and an invasive species of several inland waters around the world, possess an enormous digestive tract representing about 10% of fish weight. Thus, the aim of this study was to partially characterize proteases from their digestive tracts. Azocasein digestion of the crude extract of intestine at different pH values and temperatures revealed the presence of alkaline proteases with optimum activities at pH 9.0 and 50°C. Incubation assays of the crude extract with inhibitors such as phenyl methyl sulfonyl fluoride, N-α-p-tosyl-l-lysine chloromethyl ketone, N-tosyl-phenyalanine chloromethyl ketone, benzamidine, pepstatin A and ethylenediamine tetra-acetic acid showed that trypsin and chymotrypsin are the main alkaline proteinases present. Zymography showed that the crude extract of Pterygoplichthys disjunctivus viscera contained proteases with molecular masses ranging from 21.5 to 116 kDa. Trypsin and chymotrypsin were inhibited by the following ions in decreasing order: Hg2+, Fe2+, Cu2+, Li+, Mg2+, K+, while Mn2+, and Ca2+ had no effect. Activities decreased continuously as the NaCl concentration increased from 0 to 30%. These results constitute important background information for future studies and for the potential biotechnological use of the crude digestive extract from this invasive species.  相似文献   

12.
Digestive enzymes of freshwater fish Labeo rohita (family Cyprinidae; class Actinopterygii; infraclass Teleostei; order Cypriniformes) were studied during ontogenic development. Amylase, protease and lipase activities showed a polynomial relationship with age of rohu, whereas trypsin, chymotrypsin and lipase activities exhibited exponential trends with the age of rohu larvae. SDS‐PAGE of crude enzyme extract revealed that the proteases of higher molecular weight (MW) appeared during early ontogeny, whereas low MW proteases were observed at a later stage. Substrate SDS‐PAGE supported the quantitative study of protease activities as evidenced with the increasing number and intensity of activity bands with the age of fish. The number of protease activity bands observed in 4, 10, 12 and 24 DAH (days after hatching) larvae were 5, 7, 8 and 9, respectively. Inhibition of protease activities with soybean trypsin inhibitor (SBTI) (58.6–81.2%), phenyl methyl sulphonyl fluoride (PMSF) (55.6–70%), Nαp‐tosyl‐l ‐lysine chloromethyl ketone (TLCK) (41.1–52.3%) and N‐tosyl‐l ‐phenylalanine chloromethyl ketone (TPCK) (27.9–44.5%) indicated the presence of serine proteases, trypsin‐ and chymotrypsin‐like enzymes in rohu larvae. Inhibition with SBTI and TPCK showed power, TLCK and ethylenediaminetetraacetic acid showed exponential, whereas PMSF showed polynomial relationships during the study period.  相似文献   

13.
Digestive alkaline proteinases from golden grey mullet (Liza aurata) were extracted and characterized. The crude alkaline protease showed optimum activity at pH 8.0 and 60°C, and it was highly stable over a wide range of pH from 4.0 to 10.0, retaining more than 80% activity after incubation for 1 h at 4°C. The alkaline proteases showed extreme stability toward nonionic and anionic surfactants after preincubation for 1 h at 25°C and relative stability toward oxidizing agents. Additionally, the crude enzyme showed excellent stability and compatibility with various solid and liquid detergents. Further, proteases from golden grey mullet viscera were found to be effective in the deproteinization of shrimp wastes. The protein removal after 3 h at 45°C with an enzyme/substrate (E/S) ratio of 10 U/mg protein was about 76%. The golden grey mullet proteases were also shown to be efficient in the production of antioxidant protein hydrolysate.  相似文献   

14.
The monoclonal antibody (MAb-001), which was produced against a surface membrane glycoprotein on C. salmositica , significantly inhibited the activities of the intracellular proteases of the parasite. The total activity in the partially purified metallo-protease, and about 80% of activity in the partially purified cysteine protease, were inhibited by the antibody (at 10 μg protein ml–1). The inhibitory effects of the antibody on the proteases were also demonstrated using haemoglobin (substrate)-SDS-PAGE. The activities of the metallo-protease band (200 kDa) and the three cysteine protease bands (66, 70 and 97 kDa) were inhibited by MAb-001, but the activity of the fourth cysteine protease band (49 kDa) was not affected. Similar inhibitory effects of the antibody were also found in the crude protease extract (parasite lysate), except that more antibody was required to obtain the same level of inhibition. The metallo-protease band was more sensitive than the cysteine protease bands to the antibody.  相似文献   

15.
We have investigated the antioxidant activities of eight hydrolysates from cuttlefish by-products obtained by treatment with various gastrointestinal proteases (chymotrypsin, trypsin, and crude alkaline enzyme extracts from cuttlefish and sardinelle) and bacterial proteases (Alcalase and crude enzymes from Bacillus pumilus A1, Bacillus mojavensis A21, and Bacillus cereus BG1). The antioxidant activities of the cuttlefish by-products protein hydrolysates (CPHs) were evaluated using various in vitro antioxidant assays, such as 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity, reducing power, and total antioxidant capacity. All hydrolysates showed different degrees of hydrolysis (DH) and varying degrees of antioxidant activity. Among the different hydrolysates, cuttlefish crude enzyme hydrolysate exhibited the highest antioxidant activity, followed by sardinelle crude enzyme and Alcalase hydrolysates. Further, CPHs with different degrees of hydrolysis were prepared by treatment with proteolytic enzymes from cuttlefish, sardinelle, and B. mojavensis A21. All hydrolysates showed a greater antioxidative activity as indicated by all the methods considered. In addition, antioxidant activity in hydrolysates was positively correlated with the increase of DH. The results of this study indicated that CPHs might be a good candidate for further investigation in developing new antioxidants.  相似文献   

16.
This work provides a comparative study of the inhibitory effect of several plant protein sources on digestive proteases of two snappers: yellow snapper (Lutjanus argentiventris) and dog snapper (Lutjanus novemfasciatus). Seed extracts did not affect gastric proteases whereas they significantly inhibit intestinal proteases. Inhibition of alkaline proteases showed that pancreatic proteases of L. argentiventris were more sensitive to seed protease inhibitors than those of L. novemfasciatus. Legume seeds showed the highest inhibitory capacity on alkaline proteases causing inhibition higher than 50% in total proteolytic activity. Protease inhibition on digestive extracts was assessed using different relative concentration of seed extracts and represented by constructing dose response curves. In order to reduce the inhibitory effect, seed extracts were acid-treated before the inhibition assay. Results showed that acid treatment did not affect the inhibitory capacity of seeds on alkaline proteases in both species. However, when the action of gastric enzymes was simulated on seed extracts, the inhibitory capacity was reduced significantly, mainly in the case of L. novemfasciatus. The responses of fish enzymes to heat-treated seed extracts were also tested. Only higher temperatures were capable of reducing the inhibitory capacity of seed, with the specific response to the snapper species. The use of biochemical assays allows us to quantify the action of inhibitors on total proteolytic activity. In addition, zymograms obtained by substrate-SDS-PAGE provided qualitative information about the number and type of proteases affected by each inhibitor. Each seed extract produces a characteristic profile of inhibition on alkaline protease. The results obtained are important for future formulation of feeds for these snapper species.  相似文献   

17.
为了对扇贝裙边进行综合利用,本文选择由中性蛋白酶、胰蛋白酶和复合蛋白酶3种酶组合成3对混合酶对扇贝裙边进行酶水解实验,以水解液中游离氨基酸态氮为考察指标,采用正交实验设计研究了加酶量、酶解时间、酶解温度及酶的种类对酶解效果的影响,从而优化了扇贝裙边的酶法水解工艺条件。以最佳水解工艺条件获得的酶解液经喷雾干燥制成的氨基酸营养粉,经检测,其粗蛋白含量为82.89%,粗脂肪含量为1.55%,总糖含量为6.82%。该氨基酸营养粉中氨基酸种类齐全,每100g粗蛋白中氨基酸总量达79.06%,其中游离氨基酸总量达51.53%。  相似文献   

18.
以毛蚶(Arco subcrenat)、紫贻贝(Mytilus edulis)、脉红螺(Rapana venosa)和四角蛤蜊(Mactra veneriformis)的软体部分为原料,经水煮法粗提牛磺酸和酶解法粗提糖胺聚糖,并比较了4种贝类中牛磺酸和糖胺聚糖的含量。结果表明,四角蛤蜊、毛蚶、脉红螺中的牛磺酸平均含量分别为2.708、3.532、3.662g/kg,以脉红螺中牛磺酸含量最为丰富;四角蛤蜊、紫贻贝、毛蚶中糖胺聚糖平均含量为8.27%、4.53%、3.87%,以四角蛤蜊的含量最高。  相似文献   

19.
华南沿海5种养殖贝类营养成分的比较分析   总被引:16,自引:0,他引:16  
对华南沿海养殖的方斑东风螺、近江牡蛎、波纹巴非蛤、管角螺和九孔鲍软体部的常规营养成分、氨基酸、脂肪酸和矿物元素含量进行了比较研究。结果表明:小规格东风螺[平均体重(6.62±0.23)g]的粗蛋白质含量高于大规格东风螺[平均体重(29.74±3.26)g]和其他几种贝类;波纹巴非蛤软体部所含氨基酸总量和人体必需氨基酸的总量(未包括色氨酸)最高;呈味氨基酸含量以管角螺最高;方斑东风螺支/芳比和多不饱和脂肪酸含量最高。5种贝类软体部常量元素含量差异不大,微量元素含量差异较大。  相似文献   

20.
Nymphoides hydrophylla (NH), a lesser known aquatic vegetable, is a popular dish among the Hakka community in Taiwan. This study aimed to examine the proximate and polyphenolic compositions of NH and evaluate its antioxidant activities using various antioxidant assays. Results showed that the moisture, crude protein, crude fat, carbohydrate (crude fiber), and ash contents of fresh NH were 89.46, 0.46, 0.47, 4.11 (3.65), and 5.50%, respectively. In the antioxidant studies, the NH ethanolic extract (NH-E) exhibited higher antioxidant activities than the aqueous extract (NH-H) in the antilipid peroxidation, reducing power, metal chelation, and 2,2?-azino-bis(3-ethylbenthiazoline-6-sulfonic acid) or ABTS radical scavenging assays, while NH-H possessed more potent activity in the superoxide and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays. Compared with NH-H (6.86 mg/g for total flavonoids and 48.83 mg/g for total phenolics), NH-E showed a higher content of total flavonoids (197.21 mg/g) and total phenolics (79.43 mg/g). High-performance liquid chromatography (HPLC) analysis showed that the main difference between NH-H and NH-E polyphenolic contents was derived from the concentrations of kaempferol and quercetin.  相似文献   

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