罗非鱼鱼鳞胶原寡肽制备

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%。     

紫菜活性肽复合酶法制备与清除羟自由基作用

以条斑紫菜为原料,利用胰蛋白酶和木瓜蛋白酶组成的复合酶水解紫菜蛋白制备生物活性肽并研究清除羟自由基作用。采用单因素试验考察胰蛋白酶与木瓜蛋白酶的比例、底物质量浓度、酶用量、水解温度、水解时间、pH对羟自由基清除率及水解度的影响,响应面法优化酶解条件,SephadexG-15凝胶层析法测定活性肽的分子质量分布。试验结果表明,复合酶最佳酶解工艺条件为胰蛋白酶与木瓜蛋白酶复合酶比例1.67,底物质量浓度20mg/mL,酶用量1.67%,pH 7.0,温度55℃,酶解4h,紫菜活性肽对羟自由基的清除率为80.6%,清除率为50%时的质量浓度为0.744mg/mL,分子质量大多分布于500～1500ku。     

酶解罗非鱼鱼皮胶制备降血压肽的研究

为探讨罗非鱼鱼皮胶制备降血压肽的酶解工艺,研究选用Neutrase中性蛋白酶、Alcalase碱性内切蛋白酶、胰蛋白酶、木瓜蛋白酶和复合蛋白酶分别对罗非鱼鱼皮胶进行酶解,对其酶解液的水解度进行比较。结果表明复合蛋白酶的酶解效果最好。对复合蛋白酶的酶解工艺进行单因素优化,确定其最佳酶解工艺为:温度50℃、pH值7.0、料液比1∶5、酶量为底物量的1.5%。在此条件下水解6h时,水解度为60%,酶解产物对血管紧张素转化酶(ACE)的抑制率最高,达到78.96%。     

木瓜蛋白酶对罗非鱼下脚料的水解作用

试验结果表明,木瓜蛋白酶水解罗非鱼下脚料蛋白的适宜条件为:底物浓度1∶1.5,温度60℃,加酶量1100 IU/g原料,初始pH,酶解3 h;最佳水解条件下的酶解液氨基态氮含量达1.25 g/L,蛋白质含量5.19%,氮回收率为69.88%。酶活力下降是酶解反应减缓的主要原因之一。     

响应面法优化末水坛紫菜蛋白酶解工艺及其酶解液抗氧化活性研究

为实现末水坛紫菜(Porphyra haitanensis)的高值化利用,研究了末水坛紫菜的蛋白酶解工艺及其酶解液的抗氧化活性。以酶解产物水解度和还原力为指标,分别采用单因素和响应面优化实验筛选出最适蛋白酶和最佳酶解工艺参数;通过测定酶解液还原力对1,1-二苯基-三硝基苯肼(DPPH)自由基、羟自由基(·OH)和超氧阴离子自由基(O_2~-·)的清除作用,研究了最高水解度下的酶解液的抗氧化性活性。结果表明,中性蛋白酶是6种蛋白酶中的最适用酶;最佳酶解条件为:底物质量浓度35 g·L~(-1)、加酶量31 200 U·g~(-1)、温度45℃、pH 7.6、酶解时间5 h,在此条件下坛紫菜水解度达31.37%;酶解液还原力为2.2,对DPPH、·OH和O_2~-·自由基清除率分别为56.26%、85.84%和72.73%。结果表明,中性蛋白酶可以有效水解末水坛紫菜,水解后的酶解产物具有较好的抗氧化能力和应用前景。     

酶水解美洲帘蛤蛋白质及产物抗氧化活性初探

采用二苯基苦基苯肼(DPPH)自由基法和改进的邻苯三酚法检测了胃蛋白酶对美洲帘蛤的水解作用及其水解液的抗氧化活性,探讨了酶质量分数、底物质量分数、pH、水解时间、温度对美洲帘蛤蛋白质水解度的影响.试验结果表明,酶质量分数为5%,底物质量分数为10%.pH 1.0,酶解3 h.温度为50℃时可以获得较高的水解度,同时水解液对超氧阴离子自由基和DPPH自由基有较高的清除率.     

鳕鱼下颌肉水解蛋白制备工艺优化

通过单因素实验(蛋白酶种类、加酶量、料液比、水解时间、温度和pH值),对鳕鱼下颌肉进行酶解分析,以水解度(DH)为指标,确定最适蛋白酶为复合风味蛋白酶,其单因子最佳水解条件为:加酶量为1 400 U/g(相对于鱼糜质量)、料液比为1:3、水解时间为5 h、温度为43℃、pH 7.6。经正交试验验证,复合风味蛋白酶的最佳酶解工艺条件为:pH7.0,料液比1∶2,加酶量为1 400 U/g,水解8 h,DH可达42.62%。     

乌贼墨黑色素粗提物的提取工艺优化及其抗氧化能力测定

本文对乌贼墨粉进行超声波辅助蛋白酶水解研究,以水解度为考察指标,筛选最适蛋白酶,并通过单因素试验和L_(16)(4~5)正交试验优化酶解条件。试验结果表明,酸性蛋白酶为墨粉水解的最佳试验用酶,最佳酶解条件为温度50℃、水解时间2 h、加酶量3.0%、反应pH 3.5、料液比1∶15,在此最优条件下对墨粉进行水解试验,制得水解后墨黑色素,并测得水解度为11.45%。水解后墨黑色素氧化产物2,3,5-三羧酸吡咯(PTCA)含量为17.621μg/mg,水解后墨黑色素总抗氧化能力为18.9 U/g,总抗氧化能力保持率为99%。     

壳聚糖固定化胰蛋白酶制备牡蛎肽的研究

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%。     

基于美拉德反应的狭鳕鱼排复配酶解工艺优化

为提高水产品加工副产物中蛋白质资源的综合利用率,采用复合蛋白酶和风味蛋白酶对阿拉斯加狭鳕鱼排进行复配酶解。以水解度为指标,考察初始p H、温度、料液比、加酶量、复配酶比例、时间对酶解液水解度的影响。在单因素试验基础上,采用Box-Behnken中心组合设计和响应面分析法,确定最佳的酶解工艺条件为:初始p H 7.0,温度47℃,料液比1∶2,加酶量3.0%,复合蛋白酶与风味蛋白酶比例1∶1,酶解时间3 h,此时酶解液水解度可达到38.74%。进一步的美拉德反应试验研究表明,在持续酶解9 h后,酶解液水解度、游离氨基酸含量增加趋势均趋于平缓,同时对美拉德反应产物的风味评价差异不显著,因此最终确定酶解时间为9 h。该研究可为利用狭鳕鱼排酶解液进行美拉德反应制备不同肉风味香料提供理论依据。     

酶法提取鱿鱼皮胶原蛋白工艺条件的研究

研究了酶法提取鱿鱼皮胶原蛋白的工艺条件。根据5种蛋白酶水解液中羟脯氨酸（HYP）的含量,确定了胰蛋白酶和木瓜蛋白酶这2种提取率高的酶为水解用酶,采用L16（4^5）正交试验确定了这2种酶水解鱿鱼皮以制备胶原蛋白的最佳酶解条件。结果表明,胰蛋白酶和木瓜蛋白酶水解鱿鱼皮以制备胶原蛋白的最佳温度、加酶量、底物浓度、pH、时间分别为55℃、1200U·g^-1、1：20、pH8.0、4h和50℃、3200U·g^-1、1：20、pH6.0、6h。2种蛋白酶提取的胶原蛋白含量分别为11.08%和11.36%,提取率分别为95.16%和97.56%。     

Optimization of Enzymatic Protein Hydrolysis from Cobia (Rachycentron canadum) Frame Using Alcalase®

The combined effects of hydrolysis time, temperature, pH, and ratio of enzyme to substrate on the degree of hydrolysis (DH) of cobia frame were determined using Response Surface Methodology. The effects of these factors were employed using a three-level factors face-centered central composite design. The proximate compositions of cobia frame and cobia hydrolysate powder were determined as well. The generated model equation gave a quadratic fit with experimental data. It is suggested that hydrolysis conditions for obtaining the optimum DH using Alcalase® were: temperature of 58°C, hydrolysis time of 134 min, pH of substrate at 9.4, and an enzyme concentration of 8.3%. Proximate analyses revealed that cobia frame contained 47.0% protein, 27.6% fat, and 24.8% ash; whereas cobia frame hydrolysate powder contained 88.8% protein, 0.58% fat, and 5.05% ash. The high protein content indicated that cobia frame hydrolysate is a potential ingredient for food and feed.     

木瓜蛋白酶在制取斑鰶鱼蛋白中的应用

为确定木瓜蛋白酶酶解斑鰶鱼的条件。研究了酶解温度、酶浓度、酶解时间、加水量、pH值对斑鰶鱼氨基氮含量和总氮得率的影响。结果表明：温度小于55℃。随着酶解温度的提高、酶浓度的增加、酶解时间的延长斑鰶鱼酶解液氨基氮含量和总氮得率均增加；pH值对水解效果及氮回收率影响不大。综合考虑，最佳水解条件为：温度55℃。加酶量6．0‰。酶解时间1．5h．原料：水=1：2．5，pH值6．8。此时。氨基氮生成率达26．7％。总氮得率2．28％。     

鲢鱼骨胶原多肽螯合钙的制备研究

文章探讨了以鲢（Hypophthalmichthysmolitrix）鱼骨为原料,采用风味蛋白酶酶解制备骨胶原多肽水解液,剩余骨渣经乳酸提取制备骨粉酸解液（钙液）,研究其通过螯合制备鲢鱼骨胶原多肽螯合钙的工艺条件。分析了酶解时间对鲢鱼骨蛋白酶解产物产品品质和功能特性（溶解性和热稳定性）的影响以及采用乳酸提取鲢鱼骨中钙时提取温度对钙溶出率的影响,并以螯合率为指标,考察pH、时间、温度和多肽液与钙液体积比对螯合反应的影响。结果表明,在温度50℃,按原料鱼排质量0．09％加入风味蛋白酶酶解30min制备出的鲢鱼骨胶原多肽具有良好的产品品质和功能特性;乳酸提钙时提取温度对钙溶出率没有显著性影响（P〉0．05）;在pH8、温度25℃条件下,多肽液与钙液体积比1：1反应40min得到的鲢鱼骨胶原蛋白螯合钙螯合率最高（85．24％）。     

金乌贼肌肉中三甲胺脱甲基酶的分离纯化及酶学性质

为了提取纯化金乌贼肌肉中的三甲胺脱甲基酶(TMAOase),本研究采用含有0.1 mol/L NaCl、pH 7.0、浓度为20 mmol/L的三羟甲基氨基甲烷(Tris)-醋酸缓冲液提取粗酶液,经透析、浓缩处理后,通过DEAE-52阴离子交换柱层析和Sephacryl S-300柱层析得到了纯化的TMAOase,并对其酶学性质进行了研究。结果显示,经Sephacryl S-300柱层析的TMAOase相比粗酶纯化了209.54倍;粗酶和纯化酶的最适温度分别为55和50°C,当温度高于最适温度时,酶活性开始出现显著下降,粗酶在80°C仍残留21.9%的活性;而纯化酶在80°C时,几乎检测不到酶活;粗酶和纯化酶的最适p H均为7.0,中性条件下表现稳定,在酸性和碱性条件下稳定性下降,p H为9.0时,粗酶残留60.7%的活性,而纯化酶的活性仅为20.5%。以双倒数作图法(Lineweaver-Burk法)测得纯化的TMAOase的Km值为22.8 mmol/L;经SDS-PAGE电泳分析,测得其分子量为21.3 ku;在化学物质中,柠檬酸和CaCl_2对酶活性具有显著的促进作用,H_2O_2和Na_2S对TMAOase活性有显著抑制作用。     

Influence of Degree of Hydrolysis on Chemical Composition,Functional Properties,and Antioxidant Activities of Chinese Sturgeon (Acipenser sinensis) Hydrolysates Obtained by Using Alcalase 2.4L

Protein hydrolysates from Chinese sturgeon were prepared using Alcalase 2.4L enzyme. Under the optimum conditions (enzyme–substrate ratio of 3.5%, pH of 8.5, and temperature of 55°C), the degree of hydrolysis (DH) was 13.8%, 16.7%, and 19.1% after 1, 3, and 6 h, respectively. The contents of crude protein and amino acid increased at DH of 19.1% to 86.97% and 78.29%, respectively. There was an obvious increase in the low-molecular-weight peptides, which could enhance the hydrolysate’s functional properties such as solubility, representing more than 90% at different pH levels. The obtained protein hydrolysates revealed good emulsification properties and high oil absorption. Furthermore, good antioxidant activities such as 1, 1-diphenyl-2-picrylhydrazyl (DPPH) inhibition, ferric reducing power, and ferrous ion (Fe²⁺⁾ chelating ability were attained depending on the solution concentration. The findings indicate that the functional and antioxidant properties of protein hydrolysates could be useful in many applications of the food industry.     

Partial characterization and activity distribution of proteases along the intestine of grass carp, Ctenopharyngodon idella (Val.)

A series of studies based on biochemical assays and electrophoretical observations has been conducted in order to investigate activity distributions and partially characterize various types of proteinases in the digestive tract of grass carp, Ctenopharyngodon idella (Val.). The casein digestion assays revealed that the presence of acidic proteinase had the highest activity at pH 2.5–3.0 and the alkaline proteinases at pH 10.0. The acidic proteinase activity distribution was found to decrease gradually from the oesophagus to the anus. Pepstatin A and EDTA inhibited the acidic proteinases activity. The SDS‐substrate‐PAGE showed that crude extraction of grass carp intestine contained an acidic proteinase active component with molecular mass of 28.5 ku. The substrate‐PAGE at neutral pH condition showed the presence of two acidic proteinase active components. The activity distribution of alkaline proteinase was found to slightly fluctuate along the intestine. And the whole intestine had very high activity. The inhibition assays and substrate specificity assays showed that trypsin was the main active component of the alkaline proteinases. The SDS‐substrate‐PAGE further showed that the crude extraction of grass carp intestine had four types of alkaline proteinase with molecular mass of 26.4, 30.8, 43.0 and 105.0 ku respectively. They were characterized to be trypsin (26.4, 30.8 and 43.0 ku) and un‐serine proteinase (105.0 ku) respectively. No chymotrypsin was detected.     

Phospholipase A<Subscript>1</Subscript> activity of crude enzyme extracted from the ovaries of skipjack tuna

The phospholipid class composition, fatty acid composition and phospholipase A₁ (PLA₁) activity from the ovaries of skipjack tuna were compared with those of six other species of marine fish. In the skipjack ovaries, the lysophosphatidylcholine (LPC) proportion for the phospholipid, the docosahexaenoic acid (DHA) percentage for the total fatty acids of the phospholipids and the PLA₁ activity of the crude enzyme were the highest among those of the seven species. The optimum pH and temperature for the PLA₁ activity of the crude enzyme from the skipjack ovaries were in the range of pH 6–7 and 20–30°C, respectively, and calcium ions were not required. As a substrate, phosphatidylcholine was more easily hydrolyzed than phosphatidylethanolamine by this enzyme, and the plasmalogen-type phospholipid was much lower than the acyl-type phospholipid. After a 6-h hydrolysis reaction of the purified phospholipid extracted from the mixed ovaries of skipjack and yellowfin tuna by this enzyme, the LPC ratio of the phospholipid increased from 20 to 72.6% and the percentage of DHA for the total fatty acids of the phospholipid also increased. Thus, skipjack ovaries might possibly be used as a source of PLA₁.     

罗非鱼碎肉酶法制备蛋白胨的工艺研究

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%。