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1.
胡昂  段蕊  刘志东  林娜  张俊杰  李磊  蒋玫 《海洋渔业》2020,42(1):98-109
采用响应面法优化中性蛋白酶酶解海湾扇贝(Argopecten irradias)副产物蛋白制备抗氧化酶解物工艺。以DPPH自由基清除率和还原能力为响应值,在底物浓度、温度、时间、酶与底物比4个单因素实验基础上,优化确定3个显著性因素。获得实验条件下最佳制备工艺为:底物浓度25 mg·mL^-1、温度50℃、时间3.8 h、酶与底物比1.5%。在此条件下,实验验证酶解海湾扇贝副产物蛋白制备抗氧化酶解物的DPPH清除率为81.93%(预测值为83.19%),还原能力为58.12%(预测值为58.92%),DPPH自由基清除率和还原能力的IC 50值分别为0.264 mg·mL^-1和3.136 mg·mL^-1,且表现出较强的抗氧化活性。研究表明,优化后的回归模型用于预测中性蛋白酶酶解海湾扇贝副产物蛋白制备抗氧化酶解物是可行的。  相似文献   

2.
琼胶寡糖的制备及其_13_C_NMR研究   总被引:4,自引:0,他引:4  
毛文君 《水产学报》2001,25(6):582-584
天然多糖酶降解是制备寡聚糖的一个重要途径。酶降解由于具有特异性 ,可选择性地酶解切断特定链 ,从而制得特定的寡聚糖 ,并且反应条件温和 ,降解过程易于控制 ,在多糖降解中运用日益增多。在对多种微生物筛选研究基础上 ,从一种海洋微生物中分离获得了对琼胶具有较高降解活性的琼胶酶 ,并将此琼胶酶应用于琼胶糖降解修饰研究。本文研究了琼胶寡糖的制备 ,并运用波谱分析对琼胶寡糖的结构进行研究 ,为琼胶的构效关系研究提供基础实验资料。1 材料与方法1 .1 琼胶糖的酶解取 5g琼胶糖 ,加入 2 5 0mL蒸馏水并加热至 10 0℃摇动使其溶解 ,…  相似文献   

3.
罗非鱼鱼鳞经水洗、酸浸、碱浸处理后,采用热水法提取胶原蛋白并对其进行酶解,以酶解液水解度为指标筛选适合的酶,研究酶添加量、酶解时间、底物浓度和酶解温度对水解度的影响。在单因素实验的基础上,通过正交实验优化鱼鳞胶原蛋白的酶解条件。实验结果表明:当酶添加量6 000 U/g、酶解时间8 h、底物浓度5%、酶解温度50℃时,鱼鳞胶原蛋白酶解液水解度最大,达到24.23%。此条件下制备的分子量小于1 000 Da胶原寡肽占82.40%。  相似文献   

4.
异枝麒麟菜酶法降解工艺的优化研究   总被引:1,自引:0,他引:1  
以施氏假单胞菌分泌的卡拉胶酶为工具,采用二次回归正交旋转组合设计,对异枝麒麟菜的酶法降解工艺进行了研究,探讨pH、温度和卡拉胶酶添加量对还原糖生成量的影响。pH、温度和卡拉胶酶添加量与还原糖生成量的数学回归模型为:Y=0.3310 0.0107Z1-0.0848Z2 0.0451Z3-0.0012Z31-0.0055Z32 0.0186Z21-0.0940Z21-0.0635Z22-0.0125Z23;最佳的酶解参数为:pH 6.27,温度39.6℃,加酶量74.4 U,底物质量浓度为0.7595 g/L(总糖含量),酶解8 h。  相似文献   

5.
以壳聚糖为载体固定化胰蛋白酶,制备牡蛎肽,然后以肽得率为指标,分别研究了pH值,酶加量,酶解时间和温度等单因子对固定化胰蛋白酶酶解牡蛎蛋白质的影响。实验结果表明,酶解牡蛎蛋白的最佳工艺条件是:底物浓度为200μg/mL,加固定化胰蛋白酶量7mg/3mL,酶解时间3h,pH 8.0,温度40℃。在此条件下水解牡蛎蛋白,肽得率可达到20.39%。  相似文献   

6.
鲣鳔蛋白抗氧化酶解物制备工艺   总被引:3,自引:1,他引:2       下载免费PDF全文
为有效提高鲣鳔蛋白的附加值,研究以DPPH自由基清除率为抗氧化活性评价指标,采用蛋白酶酶解制备活性多肽的工艺,选用菠萝蛋白酶、复合蛋白酶、碱性蛋白酶、木瓜蛋白酶、胃蛋白酶、胰蛋白酶、中性蛋白酶7种酶在各自最适的条件下酶解,筛选出复合蛋白酶为最适用酶,通过单因素实验分别研究加酶量、溶液初始p H、酶解温度和时间对酶解物抗氧化活性的影响,在此基础上,根据响应面法优化鲣鳔抗氧化酶解物的制备工艺。结果显示,最佳酶解工艺条件为加酶量8.53 U/mg,p H 5.54,温度50.03°C,时间5.07 h。此外,利用超滤法对最佳条件下制备的酶解物进行初步分级,得到分子质量分别为大于10 000 u、3000~10 000 u和小于3000 u的3段组分,且这3段组分对DPPH自由基的半抑制浓度IC50值分别为0.64、0.52和0.37 mg/m L。研究表明,最优条件下制备的酶解物的DPPH清除率达72.00%,与模型预测值71.60%接近,且其中小于3000 u的组分具有较强的DPPH自由基清除活性。  相似文献   

7.
紫菜低聚糖的制备技术研究   总被引:1,自引:0,他引:1  
姚兴存  商勇  刘芳 《水产科学》2005,24(5):38-40
通过正交试验确定紫菜酸解的最佳条件为盐酸浓度0 05mol/L,水解时间120min,底物浓度3%,温度80℃,在此条件下低聚糖的提取率为11 30%。  相似文献   

8.
为确定酶解法提取菲律宾蛤仔(Ruditapes philippinarum)抗肿瘤活性多肽的最佳工艺条件,选用5种蛋白酶在相同的料液比、酶解时间和酶添加量、各自最适温度和pH的条件下酶解菲律宾蛤仔。通过甲醛电位滴定法和Cell counting kit-8(CCK-8)方法分别测定了菲律宾蛤仔酶解液氨基态氮含量和酶解多肽的抗肿瘤活性,分析了氨基态氮含量与A549抑制率之间的相关性。在单因素试验的基础上,以人非小细胞肺癌A549抑制率为指标,正交分析优化酶解工艺条件。结果显示,菲律宾蛤仔的5种酶解产物中,风味蛋白酶的酶解液中氨基态氮含量最高,而胃蛋白酶获得的酶解多肽抗肿瘤活性最高。胃蛋白酶酶解菲律宾蛤仔正交实验确定酶解最佳工艺条件为:料液比(w/w)1∶1,pH 2.0,酶解时间1 h,酶添加量2 000 U/g,酶解温度37℃。当设定最佳酶解工艺获得的酶解多肽浓度为20 mg/m L时,其对A549的抑制率为99.83%。研究表明,酶的种类对菲律宾蛤仔酶解液氨基态氮含量和酶解多肽的抗肿瘤活性有较大影响,菲律宾蛤仔酶解液中氨基态氮含量与A549抑制率没有直接的正相关性。  相似文献   

9.
大鲵肉酶解产物的制备及其抗氧化性的研究   总被引:1,自引:0,他引:1  
以大鲵肉为原料,利用Aspergillus sp.酸性蛋白酶进行酶解,研究其最佳的酶解条件以及酶解产物的抗氧化作用。结果表明,大鲵肉酶解的最佳工艺条件为Aspergillus sp.酸性蛋白酶加酶量为0.4%(质量比)及底物浓度为0.1g/mL时,酶解时间5.5h,pH 2.0,温度45℃。时间飞行质谱表明酶解产物的分子量小于2 000,苯酚硫酸法测定糖含量为2%,Folin-酚试剂法测蛋白含量为93%。大鲵酶解产物清除羟基自由基(.OH)和DPPH自由基的能力随浓度升高而增强。  相似文献   

10.
龟类不仅营养价值高,而且具有潜在的药用价值,通过研究龟肉酶解产物的功能特性,可以科学认识其营养保健功能.以黄缘盒龟肉为原料,羟自由基清除率为指标,碱性蛋白酶为水解酶,通过正交试验L9(34)得到制备抗氧化肽的最佳酶解条件为pH值8.0、酶解温度55℃、料液比1.5∶20 g/mL、加酶量(酶/底物,E/S)3.0%、酶解时间3h,酶解液对羟自由基清除率达到82.08%.酶解产物体外抗氧化活性的结果表明,酶解产物对羟自由基、超氧阴离子自由基、DPPH自由基和过氧化氢均有较好的清除作用,还具一定的还原能力、亚铁离子螯合能力和亚油酸自氧化抑制能力.总体而言,黄缘盒龟酶解产物在不同的体外抗氧化体系中均表现出一定的抗氧化效果,具有良好的开发利用前景.  相似文献   

11.
非淀粉多糖酶对蛋白酶体外酶解棉籽粕效果的影响研究   总被引:1,自引:0,他引:1  
采用二步酶解法,先单独使用纤维素酶和果胶酶或二者合用,再用中性蛋白酶(中性蛋白酶500U/g酶解时间5h)酶解棉籽粕,以还原糖生成量、蛋白质水解度(degreeofhydrolysis,DH)和三氯乙酸氮溶指数(trichloroaceticacid-nitrogen solution index,TCA-NSI)为标识,研究NSP酶水平对还原糖生成量、DH和TCA-NSI的影响。结果为:在先用非淀粉多糖(non-starch polysaccharides,NSP)酶水解,然后用中性蛋白酶500U/g水解5h时,单独使用纤维素酶和果胶酶均可提高还原糖生成量、DH和TCA-NSI,适宜水平分别为:纤维素酶350U/g,果胶酶250U/g;同时使用纤维素酶和果胶酶,对提高还原糖生成量、DH具有协同效应,适宜水平为:纤维素酶350U/g和果胶酶250U/g。  相似文献   

12.
Bacterial cellulase can be a promising biotechnological approach in seaweed degradation to facilitate extraction of bioactive molecules. In the present study, bacterial cultures were isolated from marine and terrestrial samples and screened for cellulase production. The selected cultures were grouped by random amplification of polymorphic DNA (RAPD) profiling, and the cultures showing a distinct banding pattern were identified by 16S rDNA sequence analysis. Cellulase from Bacillus megaterium (CB-sw1-I), a native isolate, was purified by ion-exchange chromatography with a purification fold of 6.91 and specific activity of 1.66 U. The purified enzyme with molecular weight of 43 kDa was optimally active at pH 6.0 and temperature of 60°C. The potentiality of enzyme in degradation of Sargassum thallus was evident with an increase in reducing sugar. Morphology of the thallus after 10 days digestion observed by scanning electron microscope also provided additional evidence for degrading ability of the enzyme. This enzyme could subsequently be used in hydrolysis of seaweed for extraction of bioactive molecules.  相似文献   

13.
ABSTRACT:   This study was conducted to optimize the cultivation conditions for Bacillus subtilis to produce proteases, amylases and cellulase, and to further investigate the hydrolysis ability against mackerel and asparagus. The extracellular enzymes from B. subtilis after 2 and 4 days incubation in a modified medium, containing 1% skim milk, 1% soya meal, 0.25% starch, 0.25% K2HPO4, 0.5% NaCl and 0.05% MgCl2 were collected for the hydrolysis of asparagus and minced mackerel, respectively. Except for the α,α-diphenyl-β-picrylhydrazyl (DPPH) scavenging ability of the hydrolyzed asparagus, the trolox equivalent antioxidation capacity and DPPH scavenging ability of both samples increased significantly ( P  < 0.05) after 1 h hydrolysis and further increased during elongated hydrolysis at 50°C. Sodium dodecylsulfate–polyacrylamide gel electrophoresis indicated severe degradation of muscle proteins during hydrolysis. Changes in reducing sugar, soluble proteins and peptides before/after hydrolysis suggested the extracellular enzymes from B. subtilis could effectively hydrolyze the mackerel or asparagus, and subsequently improve their antioxidation ability.  相似文献   

14.
Guan-James  WU  Guo-Jane  TSAI 《Fisheries Science》2004,70(6):1113-1120
ABSTRACT:   Samples of colloidal chitin and chitosans with deacetylation degrees (DD) of 50, 65, 75 and 95% (DD50, DD65, DD75 and DD95, respectively) were prepared from shrimp chitin. The hydrolytic activity of cellulase on these samples increased with the increasing DD of chitosan, with DD95 being the most easily hydrolyzed. Colloidal chitin was hardly hydrolyzed. The optimal reaction temperature and pH for cellulase digestion of chitosan were 55°C and pH 5.2, respectively. During cellulase digestion of chitosan, the 9-h hydrolysate had the highest enhancing effect on the proliferation of a human hybridoma cell, HB4C5. This hydrolysate is composed of low-molecular-weight chitosan (LMWC), with a molecular mass of 20.0 kDa, and chitooligosaccharides, which are composed of sugars with a degree of polymerization of 1–6. In vitro , the 9-h hydrolysate increased both cell proliferation and immunoglobulin (Ig)M secretion of HB4C5 because of the presence of chitooligosaccharides for the former activity and LMWC for the latter activity. In vivo , samples of the chitosan hydrolysate, chitooligosaccharide mixture and LMWC significantly increased the levels of serum IgG and IgM, and enhanced concanavalin A- and lipopolysaccharide-induced proliferation of mouse lymphocytes.  相似文献   

15.
构建了枯草芽孢杆菌CH_2菌株(Bacillus subtilis CH_2)壳聚糖酶基因的真核表达载体p PIC9K-CH2-Cns,在毕赤酵母GS115(Pichia pastoris)中进行重组表达,获得了有生物学活性的重组壳聚糖酶,对重组壳聚糖酶的酶学特性及酶解产物进行了分析。结果显示,重组壳聚糖酶的分子量为29 k D,粗酶的比酶活达到133.60 U/mg,纯化后重组蛋白的比酶活为338.08 U/mg;该酶的最适作用温度为50℃,最适pH为4.5,酶动力学常数Vmax=24.39(μmol/mg)·min~(-1),Km=5.48 mg/m L;Fe2+、K+等对其酶活力有一定的激活作用,其中Mn~(2+)能使酶活力提升2.4倍,Ag~+、Mg~(2+)、Hg~(2+)、EDTA、EGTA和SDS等存在时则对其酶活力有强烈抑制作用。利用重组壳聚糖酶酶解壳聚糖,酶解产物主要为聚合度2~10的壳寡糖,且分布均匀。以上结果表明,枯草芽孢杆菌CH_2菌株壳聚糖酶基因在毕赤酵母GS115中成功表达,获得了一种内切型的高酶活力重组壳聚糖酶,该重组酶具有反应条件温和、酶解产物均一等特点,可被应用于海洋甲壳质加工应用中。  相似文献   

16.
The aim of this work was to study the effect of adding sugar during proteolysis to promote the Maillard reaction and mask the initial fish odor and off-flavors generated. An experimental design, based on the Doehlert plan, was used to study the influence of hydrolysis conditions (time, temperature, sugar, and antioxidant addition) on the odor characteristics of hydrolysates, soluble protein levels, and amino acid content. Results showed that the lowest level of sugar (10 g of D-xylose added to 1 kg of by-products) was enough to develop a grilled odor in hydrolysates. In the hydrolysis conditions used—i.e., enzyme inactivation at 95°C for 30 min—hydrolysis temperature had no effect on grilled odor production but significantly affected the soluble protein fraction, as did hydrolysis time. Soluble protein content and essential amino acid content increased with the enzymatic reaction but were not modified by adding sugar. Hydrolysis conditions that promote Maillard reactions while keeping a nutritional balance have been identified.  相似文献   

17.
朱思咪  张萍  陈丽娇  程艳 《福建水产》2012,34(2):134-140
为探讨罗非鱼碎鱼肉酶法制备蛋白胨的加工工艺,采用正交试验的方法,分别研究了木瓜蛋白酶、AS.1398中性蛋白酶、复合蛋白酶水解罗非鱼碎肉制备蛋白胨的工艺条件。结果表明:木瓜蛋白酶的最佳水解工艺条件为pH6.5,温度65℃,水解时间4h,加酶量1250 U/g,蛋白胨得率达12.63%;AS.1398中性蛋白酶最佳水解工艺条件为pH 7.5,温度55℃,水解时间4 h,加酶量750 U/g,蛋白胨得率达13.25%;复合蛋白酶的最佳水解工艺条件为pH 7.0,温度50℃,水解时间4.5 h,加酶量850 U/g,蛋白胨得率达11.43%。  相似文献   

18.
扇贝裙边富含蛋白质、脂质等营养成分。为了高值化利用扇贝裙边,本研究选用中性蛋白酶、动物蛋白酶、风味蛋白酶、木瓜蛋白酶和酸性蛋白酶5种酶,以酶解液中游离氨基酸态氮为考察指标,对扇贝裙边进行酶解工艺条件探讨。首先,将5种酶制成复合蛋白酶进行正交实验,确定最佳酶解时间、温度、pH及加酶量,经检验,氨基酸转化率为77%;然后,通过实验确定CaCl2为最适钙源,以贝壳为原料,通过水飞法和酸法转化可制得贝壳源CaCl2。将扇贝裙边酶解液中复合氨基酸与来源于贝壳的钙螯合制备复合氨基酸螯合钙,以正交实验筛选出最佳螯合条件。经检验,该螯合反应螯合率达92%。  相似文献   

19.
We isolated a cellulase from the digestive organs of the short-spined sea urchin Strogylocentrotus intermedius using a combination of ion-exchange chromatography and gel filtration together with an assay for carboxymethylcellulase activity. The isolated cellulase was stained as a single band by Congo red. The molecular weight of the isolated cellulase, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, was 59?kDa. The isolated cellulase exhibited hydrolytic activity toward carboxymethyl cellulose, with an optimum temperature and pH of 30?°C and pH 8.0, respectively. The thermal stability of the enzyme was characterized by determining the temperature at which activity decreased by 50?% with treatment for 30?min at pH 7.0 and found to be 32?°C. Cellulase activity remained at a high level at 5?C20?°C, which is the growth temperature of the short-spined sea urchin. These results confirm that the short-spined sea urchin should preferably be reared at a water temperature of <20?°C.  相似文献   

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