鲢鱼降血压肽的酶解条件

以鲢鱼蛋白为原料,采用5种不同复合蛋白酶(P1,P2,P3,P4,P5)进行酶解,综合研究了复合生物酶酶量、酶解时间、pH、酶解温度等对鲢鱼蛋白酶解降血压肽ACE活性抑制效果的影响。结果表明,复合酶P1酶解降血压肽对ACE抑制率经体外检测为88.08%,具有最佳的酶解效果。对应最佳条件为:酶量25 IU/mL,酶解时间2 h,pH7.0,酶解温度60℃。     

鲢鱼降血压肽的酶解条件

以鲢鱼蛋白为原料，采用5种不同复合蛋白酶（P1，P2，P3，P4，P5）进行酶解。综合研究了复合生物酶酶量、酶解时间、pH、酶解温度等对鲢鱼蛋白酶解降血压肽ACE活性抑制效果的影响。结果表明。复合酶P1酶解降血压肽对ACE抑制率经体外检测为88．08％。具有最佳的酶解效果。对应最佳条件为：酶量25IU／mL，酶解时间2h，pH 7．0，酶解温度60℃。     

石斑鱼头酶解产物的制备及其功能性质

【目的】为了提高石斑鱼加工副产物的生物价值,对石斑鱼头中抗氧化活性物质进行分离与功能性质分析。【方法】以珍珠龙胆石斑鱼头为原料,采用生物酶解法,以水解度和DPPH自由基清除率为指标,通过单因素试验和响应面法优化酶解工艺;酶解产物经不同分子质量的超滤膜进行分离,对分离各级分的体外抗氧化活性与功能性质进行研究。【结果】石斑鱼头酶解产物的最佳制备条件是:风味蛋白酶添加量3 000 U/g,在pH 7.0,酶解温度为45℃、底物质量浓度300mg/mL下酶解4.5h,得到的石斑鱼头粗酶解产物的水解度为(20.27±1.28)%。分子质量3 ku的酶解产物(EHH-1)蛋白质质量分数为(13.18±1.17)%,抗氧化活性最显著,其对DPPH自由基、羟自由基、超氧阴离子自由基清除作用的IC_(50)值分别为0.35、4.60、0.46 mg/mL;在pH4.0～8.0之间均具有较好溶解性,氮溶指数达到(75.08～87.50)%,热稳定性达到(70.00～82.00)%,乳化活性为(23.52～37.45)m~2/g。【结论】石斑鱼头酶解产物分子质量3 ku的超滤级分具有较强的抗氧化性与良好的功能性质,在天然抗氧化剂和食品加工领域有潜在的利用价值。     

海参性腺酶解物制备及其体外清除自由基活性测定

【目的】酶解海参性腺并分析其体外清除自由基活性。【方法】采用外源性蛋白酶水解工艺制备海参性腺酶解产物,通过酶解效果比较分析,考察其体外清除DPPH自由基和羟自由基能力。【结果】确定木瓜蛋白酶为水解用酶,利用响应面优化实验确定酶解条件为酶解时间14 h、酶添加量3 000 U/g、料液比1∶40;酶解液灭酶后经离心、浓缩、干燥后得到粉状海参性腺酶解物,其主要成分质量分数分别为蛋白肽48.4%、矿物质16.8%、粗脂肪12.4%、水分9.8%;酶解产物清除DPPH自由基和羟自由基的IC50值分别为3.39、5.83 mg/mL。【结论】海参性腺酶解物具有良好的抗氧化活性。     

方格星虫酶解工艺优化及酶解物免疫活性

【目的】优化方格星虫(Sipunculus nudus L.)酶解工艺条件,探讨其酶解液对细胞免疫活性的影响。【方法】在确定实验用酶和单因素实验基础上,利用响应面设计建立数学模型,以水解度为响应值,进行3因素3水平的响应面分析,得到最适酶解条件。以小鼠脾淋巴细胞的增殖、小鼠腹腔巨噬细胞生成NO量和吞噬能力来评价酶解液对免疫细胞的促进作用。【结果】以动物水解蛋白酶和风味蛋白酶组合用酶效果最佳,方格星虫酶解的适宜条件为pH 7.0、加酶量5 000 U/g、液料比3∶1,55℃酶解5.2 h,其水解度为38.92%;以此条件得到的酶解液进行实验,酶解液质量浓度为0.9 mol/mL时,小鼠脾淋巴细胞的增殖率为30.57%,小鼠腹腔巨噬细胞生成NO能力为56.2μmol/L,吞噬能力D(540 nm)值均明显高于阳性对照组(P0.01)。【结论】方格星虫酶解液对细胞免疫具有促进作用。     

扁舵鲣抗氧化肽分离纯化及结构鉴定

【目的】从扁舵鲣(Auxis thazard)的木瓜蛋白酶酶解产物中分离鉴定具有较高1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除率的抗氧化肽。【方法】用木瓜蛋白酶水解扁舵鲣,以DPPH自由基清除能力为检测指标,通过超滤、凝胶过滤层析和反向高效液相色谱分离抗氧化肽,再经过超高效液相/三重四级杆飞行时间质谱(UPLC/Xevo G2-XS QTOF)进行结构鉴定。【结果】从酶解产物中获得3种抗氧化肽,其氨基酸序列分别为β-丙氨酸-1-甲基-L-组氨酸(241 u)、Gly-Ala-Gly-Gly-Pro(357 u)和Val-Glu(246 u)。【结论】扁舵鲣的木瓜蛋白酶酶解产物含有抗氧化活性的肽类,可为其抗氧化肽的开发提供理论依据。     

对虾蛋白自溶制备ACE抑制肽的工艺条件优化

以凡纳滨对虾为原料,以ACE抑制率为指标,利用响应面法对虾肉蛋白自溶制备ACE抑制肽的工艺条件进行了优化,即在酶解条件(pH值、温度、虾头虾肉质量比)和ACE(Angiotensin I-converting Enzyme,ACE)抑制率之间建立了数学模型Y=23.59-0.21X1+0.84X2+0.85X3-0.71X12-0.94X22-1.06X32+0.088X1X2-0.46X1X3-0.87X2X3。分析表明,在3个因素中,虾头与虾肉比例对ACE抑制率的影响最为显著。优化后的工艺参数为:pH7.35,温度57.2℃,虾头与虾肉比例为1∶1。根据回归方程的预测结果,反应时间为4 h,其ACE抑制率达41.9%。     

响应面法优化南极磷虾酶法制备DPP-IV抑制肽工艺条件的研究

采用动物蛋白水解酶酶解制备南极磷虾二肽基肽酶-IV(DPP-IV)抑制肽,以DPP-IV抑制率和可溶性蛋白含量为主要指标,考察温度、反应时间、pH、酶添加量对南极磷虾的酶解效果。在单因素的基础上,依据中心组合实验设计原理,运用4因素3水平的响应面分析法,建立动物蛋白水解酶酶解南极磷虾制备DPP-IV抑制肽的二次多项式数学模型,并以DPP-IV抑制率为响应值作响应面。结果表明:南极磷虾酶法制备DPP-IV抑制肽的最佳工艺参数为酶解时间3.85 h、温度45.02℃、pH值7.58、酶添加量237.55 U/g原料;在此条件下,酶解液DPP-IV抑制率的预测值为64.82%,验证值为64.78%,优化方法可行。     

牡蛎酶解工艺优化及其酶解产物对小鼠睾酮分泌的影响

【目的】优化太平洋牡蛎(Crassostreagigas)酶解工艺条件,并研究酶解产物及其超滤组分对体外培养的睾丸间质细胞增殖及睾酮分泌的影响。【方法】以多肽得率为参考标准,对木瓜蛋白酶酶解牡蛎的加酶量、初始pH、酶解时间、酶解温度进行单因素及正交实验优化;通过MTT法及酶联免疫吸附试验(ELISA)检测不同浓度牡蛎酶解产物及其超滤组分对TM3小鼠睾丸间质细胞增殖及睾酮分泌的影响。【结果】木瓜蛋白酶最佳酶解条件为加酶量3 000 U/g,初始pH 6. 5,酶解温度62.5℃,酶解时间4.5 h,在此条件下其多肽质量浓度为14.95 mg/mL;牡蛎酶解液及其超滤组分在0.25～1.00 mg/mL质量浓度范围内,与空白对照组相比,能够显著增加TM3细胞的增殖活性(P0.05),且具有浓度依赖性和时间依赖性。其中,5 ku和5～10 ku超滤组分细胞增殖活性及促进睾酮分泌活性均最强。【结论】牡蛎酶解产物及其超滤组分可有效增强小鼠睾丸间质细胞增殖活性并促进其分泌睾酮。     

酶解蓝圆鲹鱼肉蛋白制备小分子肽的工艺研究

以蓝圆鲹为原料,酶解蓝圆鲹鱼肉蛋白制备小分子肽。运用响应面试验设计优化蓝圆鲹蛋白酶解的工艺条件,探讨了酶种类和添加量、液固比、酶解温度、酶解时间对多肽提取率的影响。结果表明:选用木瓜蛋白酶,添加量为0.10%,液固比2∶1,酶解温度54.4℃,酶解时间3.18 h下,多肽提取率为32.35%。     

Enzymatic hydrolysis of defatted mackerel protein with low bitter taste

Ultrasound-assisted solvent extraction was confirmed as a novel,effective method for separating lipid from mackerel pro-tein,resulting in a degreasing rate (DR) of 95% and a nitrogen recovery (NR) of 88.6%.To obtain protein hydrolysates with high ni-trogen recovery and low bitter taste,enzymatic hydrolysis was performed using eight commercially available proteases.It turned out that the optimum enzyme was the ‘Mixed enzymes for animal proteolysis’.An enzyme dosage of 4%,a temperature of 50℃,and a hydrolysis time of 300 min were found to be the optimum conditions to obtain high NR (84.28%) and degree of hydrolysis (DH,16.18%) by orthogonal experiments.Glutamic acid was the most abundant amino acid of MDP (defatted mackerel protein) and MDPH (defatted mackerel protein hydrolysates).Compared with the FAO/WHO reference protein,the essential amino acid chemical scores (CS) were greater than 1.0 (1.0 1.7) in MDPH,which is reflective of high nutritional value.This,coupled with the light color and slight fishy odor,indicates that MDPH would potentially have a wide range of applications such as nutritional additives,functional ingredients,and so on.     

青鳞鱼蛋白复合酶控制水解动力学模型的研究

在假设复合酶恒温控制水解动力学遵循内切酶限制水解动力学历程的前提下 ,采用实验方法求出了复合酶恒温控制水解动力学模型。结果表明 ,复合酶对青鳞鱼蛋白进行控制水解的动力学模型为 :R =(0 .315 4e0 - 0 .0 186s0 )exp(- 0 .170DH) ,DH =5 .882ln[1+(0 .0 5 36 2e0 /s0 -0 .0 0 32 )t];其酶失活常数Kd=0 .0 5 36min- 1;水解反应能够顺利进行的条件是 :e0 /s0 >c0 ,常数c0 =5 .90× 10 - 2 。验证实验证明 ,根据复合酶恒温控制水解动力学模型得到的理论DH与实际DH基本吻合 ,该动力学模型具有很强的实用价值。     

青鳞鱼蛋白风味酶控制水解动力学模型

在假设风味酶恒温控制水解动力学遵循内切酶限制水解动力学历程的前提下,通过实验方法求出了风味酶恒温控制水解动力学模型。结果表明,风味酶对青鳞鱼蛋白进行控制水解的动力学模型为:R= ( 54 .782e0 -0 .045s0 )exp( -0 .398DH),DH=2 .513ln[1+(21 .803e0 /s0 -0. 0179)t];其酶失活常数kd=22. 155 6min-1;水解反应能够顺利进行的条件是:e0 /s0 >c0,常数c0 =8 .214×10-4。验证实验证明,根据风味酶恒温控制水解动力学模型得到的理论水解度与实际水解基本吻合。     

青鳞鱼HAP的试制

应用正交试验优选枯草杆菌中性蛋白酶和胃蛋白酶对青鳞鱼蛋白质的水解条件。结果表明：（１）枯草杆菌中性蛋白酶的最适条件为ｐＨ７．０、温度５０℃、时间３ｈ、酶浓度１０，０００μ／１００ｍｌ；（２）胃蛋白酶的最适条件为ｐＨ３．５、时间２．０ｈ、温度４０℃、酶浓度３００μ／１００ｍｌ；（３）以去内脏青鳞鱼为原料进行双酶水解，蛋白质水解度可达８３．８％。水解液经脱臭浓缩可制得ａ──氨基氮≥１．５ｇ／１００Ｉｎｌ的ＨＡＰ。它可作为海鲜食品的添加剂，也可制作海鲜汤料或作为膨化食品的辅料。     

DETERMINATION OF AMINO ACIDS IN TWO POLYSIPHONIA SPECIES AND STUDY OF ENZYMATIC HYDROLYSIS METHOD

The total content ot the nch amino acids in two common red algae, Potystpnoma urceolata and Polysiphonia japonica growing in the Qingdao seashore were determined. The algae powder was hydrolyzed by 6 mol/L HC1 at 110℃ for 48 h and determined by amino acid analyzer. The content was 25.35% and 24.16% , respectively, much higher than that of some other species. In addition, a nutritive liquid with abundant amino acids was prepared (by the enzymatic hydrolysis method using Polysiphonia urceolata ) as raw material for a kind of health beverage. The dried seaweed was decolored by 0.25% KMnO4 and 0.5% active carbon, then enzymalized. In the selection of enzymalizing condition, the orthogonal experimental design was used. Four factors including kinds of enzyme, quantity of enzyme, temperature and time were studied at 3 levels. According to the orthogonal design results, we can choose an optimal condition: hydrolyzing at 45℃ by neutral proteinase (0.25% , w/w) for 2 h, adjusting pH to 8.5, then adding trypsin (0.25% , w/w) and hydrolyzing for 2 h. Finally the above solution was alkalized by NaOH and neutralized by casein. After the hydrolyzed liquid was filtered and concentrated, suitable additives were added. The final products contain rich amino acids.     

The Hydrolyzation of Collagen by Fucoidan Oligosaccharide''s Complex with CeⅣ

Fucoidan is such a polysaccharide that its hydroxies are easy to combine with lanthanons ion (CeⅣ) to form complex. This work obtained the complexes of three fucoidan oligosaccharides with different molecular weights F1( ＞5 000), F2(1 000-5 000) and F3( ＜ 1 000) by hydrolyzing Oligosaccharide collagen with sulfuric acid. It is found that the fucoidan oligosaccharide F3 can form complex with more CeⅣ than F1 and F2. Hydrolyzing collagen with the complex was carried out to produce amino acid and peptides. All the three fucoidan oligosaccharide complexes with CeⅣ( F1, F2, F3) can catalyze by the artificial hydrolytic enzyme, and the activity of the complex of F3 is the highest.     

罗非鱼下脚料自溶条件的初步探讨

采用单因素和正交优化实验L9(43)确定酶法自溶罗非鱼下脚料的最适条件,结果表明,最适自溶条件为:料液质量比1∶3,反应温度50℃,pH7.5,反应时间6.5h;酶解结束蛋白质回收率为81.75%。     

Amino acid composition and functional properties of giant red sea cucumber (Parastichopus californicus) collagen hydrolysates

Giant red sea cucumber (Parastichopus californicus) is an under-utilized species due to its high tendency to autolysis.The aim of this study was to evaluate the functional properties of collagen hydrolysates from this species.The degree of hydrolysis (DH),amino acid composition,SDS-PAGE,emulsion activity index (EAI),emulsion stability index (ESI),foam expansion (FE),and foam stability (FS) of hydrolysates were investigated.The effects of pH on the EAI,ESI FE and FS of hydrolysates were also inves-tigated.The results indicated that the β and α1 chains of the collagen were effectively hydrolyzed by trypsin at 50℃ with an En-zyme/Substrate (E/S) ration of 1:20 (w:w).The DH of collagen was up to 17.3% after 3 h hydrolysis with trypsin.The hydrolysates had a molecular weight distribution of 1.1 17 kDa,and were abundant in glycine (Gly),proline (Pro),glutamic acid (Glu),alanine (Ala) and hydroxyproline (Hyp) residues.The hydrolysates were fractionated into three fractions (< 3 kDa,3 10 kDa,and > 10 kDa),and the fraction of 3 10 kDa exhibited a higher EAI value than the fraction of > 10 kDa (P<0.05).The fraction of > 10 kDa had higher FE and FS values than other fractions (P<0.05).The pH had an important effect on the EAI,ESI,FE and FS.All the fractions showed undesirable emulsion and forming properties at pH 4.0.Under pH 7.0 and pH 10.0,the 3 10 kDa fraction showed higher EAI value and the fraction of > 10 kDa showed higher FE value,respectively.They are hoped to be utilized as functional ingredients in food and nutraceutical industries.