黑豆蛋白肽果汁复合饮料的研制

黑豆是种皮为黑色的大豆，其蛋白质含量高达50%，是极具开发潜力的植物蛋白资源，但黑豆蛋白质分子高度压缩折叠、不易消化，使其加工利用受到了限制。该试验采用2709碱性蛋白酶对黑豆蛋白进行有控制的水解，制备酸溶性好、易消化吸收的黑豆蛋白肽，经水解度与酸溶性蛋白肽得率测定，确定了酶解工艺条件：底物浓度为每100 mL黑豆浆含3.75 g蛋白质，酶-底物比为8 000 u/g，pH值9.0，温度50℃，酶解时间2.0 h，水解度为10%，酸溶性蛋白肽得率达85%。2709蛋白酶对黑豆蛋白具有较好的水解效果，1 kg黑豆经磨浆、酶解与离心去渣脱苦制得20 L黑豆蛋白肽混合液，将其与天然澄清苹果汁按80∶20(V/V)混合调配制备黑豆蛋白肽果汁复合饮料，产品色泽棕红清亮，具天然果汁与黑豆清香风味，蛋白质与肽类物质含量(m/V)≥1.5%，酸度(以柠檬酸计) ≥0.4 g/100 mL。     

酶解膜分离两步分离乳清中β-乳球蛋白的研究

为研究乳清中β-乳球蛋白环保、温和、高效的分离工艺,该文对采用蛋白酶选择性水解然后超滤分离乳清水解液中的β-乳球蛋白进行研究.试验对乳清水解物进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析,比较了胃蛋白酶、胰蛋白酶、木瓜蛋白酶和中性蛋白酶的选择性,并分别采用分子截留量10000的聚砜膜(PS-10)和聚醚膜(PES-10)对胃蛋白酶解液进行超滤处理.结果显示,β-乳球蛋白对胃蛋白酶耐受性较好,α-乳白蛋白对木瓜蛋白酶抗性较佳,但二者均对中性蛋白酶较敏感.较优的水解分离条件如下:乳清蛋白质量浓度8%,胃蛋白酶添加量为乳蛋白质量的0.3%,pH值2.1,温度37℃,水解2h,α-乳白蛋白近似完全水解而β-乳球蛋白几乎不被降解.水解液用PS-10膜超滤分离,β-乳球蛋白的纯度和产率达到了较高,分别为94.6%,75.6%.因此,采用选择性酶解处理乳清然后超滤分离β-乳球蛋白是可行的.     

酶解膜分离两步分离乳清中β-乳球蛋白的研究(简报)

为研究乳清中β-乳球蛋白环保、温和、高效的分离工艺,该文对采用蛋白酶选择性水解然后超滤分离乳清水解液中的β-乳球蛋白进行研究。试验对乳清水解物进行十二烷基硫酸钠－聚丙烯酰胺凝胶电泳（SDS-PAGE）分析,比较了胃蛋白酶、胰蛋白酶、木瓜蛋白酶和中性蛋白酶的选择性,并分别采用分子截留量10000的聚砜膜(PS-10)和聚醚膜(PES-10)对胃蛋白酶解液进行超滤处理。结果显示,β-乳球蛋白对胃蛋白酶耐受性较好,α-乳白蛋白对木瓜蛋白酶抗性较佳,但二者均对中性蛋白酶较敏感。较优的水解分离条件如下：乳清蛋白质量浓度8%, 胃蛋白酶添加量为乳蛋白质量的0.3%,pH值2.1,温度37℃,水解2 h,α-乳白蛋白近似完全水解而β-乳球蛋白几乎不被降解。水解液用PS-10膜超滤分离,β-乳球蛋白的纯度和产率达到了较高,分别为94.6%,75.6%。因此,采用选择性酶解处理乳清然后超滤分离β-乳球蛋白是可行的。     

酶解法制备豆粕水解肽工艺优化

为制备苦味低且生物活性高的豆粕水解肽,本试验在综合考虑水解酶酶切位点后选取7种酶组合对低温豆粕进行水解,以蛋白质转化率、水解度、分子量分布及智能感官评价等作为水解肽的评价指标,比较不同酶组合的作用效果。结果表明,优化得到豆粕水解肽的最佳酶法制备工艺条件:复合酶的最佳组合为碱性蛋白酶+中性蛋白酶+脱苦蛋白酶,酶解pH值依次为8.5、7.5、7.5,反应温度保持55℃,反应时间分别为3、2、2 h,经最佳酶组合制得的水解肽水解度为16%,分子量≤1 000 Da的水解肽可达89.8%,制备的水解肽理化性质及口感更优。本研究结果为豆粕资源开发及功能性肽产品的应用奠定了理论基础。     

碱性蛋白酶(alcalase)水解菜籽清蛋白的工艺优化

采用响应曲面法对碱性蛋白酶(alcalase)水解菜籽清蛋白工艺进行系统地研究。确定最佳水解条件如下：温度50℃、酶浓度0.38 AU/g、底物浓度4.87%。同时,葡聚糖凝胶(Sephadex G-25)柱层析显示水解物较原清蛋白分子量变小。氨基酸组成分析结果表明菜籽清蛋白水解物可作为一种营养丰富的食品添加剂加以广泛利用。     

大米降压肽酶法制备工艺及其性质研究

该文利用碱性蛋白酶水解大米蛋白制备大米降压肽，确定碱性蛋白酶水解终点为水解度18.17%。采用9种大孔吸附树脂吸附大米降压肽，结果显示NKA型树脂对大米降压肽的吸附效果较好，大米降压肽中灰分含量由脱盐前的16.64%减少到脱盐后的2.32%。对大米降压肽的相对分子质量、溶解性进行分析，大米降压肽的相对分子质量在138～1461之间；在pH 3～11的范围内，大米降压肽的溶解度在97.6%左右。     

三文鱼皮四种蛋白酶酶解物抗氧化活性及功能特性的差异研究

为进一步合理开发利用三文鱼皮,本研究对碱性蛋白酶、中性蛋白酶、风味蛋白酶和复合蛋白酶水解所制备三文鱼皮酶解物的抗氧化活性与功能特性进行了比较。结果表明,三文鱼皮碱性蛋白酶酶解物的水解度(20.18%)和三氯乙酸可溶性氮得率(40.14%)最高,小分子肽含量高达99.97%,其抗氧化活性显著优于其他组(P<0.05);且碱性蛋白酶酶解物的溶解性和持水性最高,分别为87.21%和26.92%;中性蛋白酶酶解物的持油性和乳化性能最强,分别为4.67%和14.69%;而风味蛋白酶酶解物的乳化稳定性显著优于其他组(P<0.05)。综上所述,碱性蛋白酶为制备三文鱼皮蛋白抗氧化肽的最优蛋白酶。本研究为三文鱼皮副产物的高值化利用提供了数据支持和理论基础。     

白果活性蛋白的酶法水解及抗氧化活性研究

白果活性蛋白(GAP)是华中农业大学天然产物化学实验室首次从白果中分离得到的一种以清蛋白为主的且具有显著生物活性的蛋白,为扩大该蛋白在食品中的用途,提高其生物活性,研究了酶法水解制备白果肽段的方法及其抗氧化活性的变化。通过单因素分析、正交试验以及对试验结果的分析,确定了2709碱性蛋白酶水解白果活性蛋白的最佳水解条件为：底物浓度1%,pH值8.5,水解温度45℃,酶浓度7000 U/g,反应时间6 h。以Fenton体系和邻苯三酚体系检验其抗氧化活性,结果表明白果肽段较酶解前的活性蛋白,抗氧化活性有不同程度的提高,但两者均具有较强的抗氧化活性。     

大米肽的酶法制备工艺及其特性的研究

该文利用蛋白酶水解大米蛋白制备得到大米肽。比较碱性蛋白酶、中性蛋白酶、复合蛋白酶和风味酶水解大米蛋白的进程曲线，结果显示碱性蛋白酶的水解效果最好，其较佳作用条件为：底物浓度10%、pH值9.0、温度45℃、酶与底物比48 AU/kg、时间150 min。在此条件下，大米肽的得率为46.8%，纯度为71.3%。大米肽具有溶解性较好和黏度较低的特性，可以在食品中广泛应用。     

抑制玉米淀粉回生的面粉蛋白酶解液筛选

为防止玉米淀粉食品会因回生而降低品质,该文利用酸性、中性和碱性3种蛋白酶水解小麦面粉中的球蛋白、谷蛋白和醇溶蛋白,研究酶解物中混合多肽对玉米淀粉回生的影响。研究结果表明,酸性蛋白酶水解谷蛋白所得多肽混合液能强烈抑制玉米淀粉回生,使玉米淀粉回生率由14.0%降低到8.0%。其他2种水解物促进玉米淀粉回生,促进最多的是碱性蛋白酶水解醇溶蛋白,使玉米淀粉回生率由14.0%升高到19.5%。通过红外和核磁分析了混合多肽抑制或促进玉米淀粉回生的可能机理。研究成果为控制淀粉回生提供一条全新的途径。     

Angiotensin I converting enzyme-inhibitory peptides from commercial wet- and dry-milled corn germ

Bioprocesses were developed to enhance the value of proteins from deoiled corn germ. Proteins were hydrolyzed with trypsin, thermolysin, GC 106, or Flavourzyme to generate the bioactive peptide sequences. At an enzyme to substrate ratio of 1:100, protein hydrolysis of wet-milled germ was greatest using thermolysin followed by trypsin, GC 106, and Flavourzyme. For the dry-milled corn germ, protein hydrolysis was greatest for GC 106 and least for Flavourzyme. Electrophoretic patterns indicated that the hydrolysis conditions used were adequate for generating low molecular weight peptides for both germs. Unhydrolyzed dry- and wet-milled corn germ did not appear to contain angiotensin I converting enzyme (ACE)-inhibitory peptides. After hydrolysis with trypsin, thermolysin, and GC 106 but not Flavourzyme, ACE inhibition was observed. ACE inhibition was greatest for the GC 106 hydrolysate for both wet- and dry-milled corn germ. Denaturing the protein with urea before hydrolysis, in general, increased the amount of ACE-inhibitory peptides found in the hydrolysate. Membrane fractionations of both the wet- and dry-milled hydrolysates indicated that most of the ACE-inhibitory peptides were in the <1 kDa fraction. Examination of the control total protein extracts (before treatment with proteases) from wet- and dry-milled germ revealed that neither had ACE-inhibitory properties. However, when both total corn germ control protein extracts were fractionated, the <1 kDa fraction of wet-milled corn germ proteins exhibited ACE inhibition, whereas the comparable low molecular weight fraction from dry-milled corn germ did not.     

Preparation of whey protein hydrolysates using a single- and two-stage enzymatic membrane reactor and their immunological and antioxidant properties: characterization by multivariate data analysis

An initial 5% (w/v), followed thereafter with replacement aliquots of 3% (w/v), whey protein isolate (WPI) (ca. 86.98% Kjeldahl N x 6.38), was hydrolyzed using Protease N Amano G (IUB 3.4.24.28, Bacillus subtilis) in an enzymatic membrane reactor (EMR) fitted with either a 10 or 3 kDa nominal molecular weight cutoff (NMWCO) tangential flow filter (TFF) membrane. The hydrolysates were desalted by adsorption onto a styrene-based macroporous adsorption resin (MAR) and washed with deionized water to remove the alkali, and the peptides were desorbed with 25, 50, and 95% (v/v) ethyl alcohol. The desalted hydrolysates were analyzed for antibody binding, free radical scavenging, and molecular mass analysis as well as total and free amino acids (FAA). For the first time a quantity called IC50, the concentration of peptides causing 50% inhibition of the available antibody, is introduced to quantify inhibition enzyme-linked immunosorbent assay (ELISA) properties. Principal component analysis (PCA) was used for data reduction. The hydrolysate molecular mass provided the most prominent influence (PC1 = 57.35%), followed by inhibition ELISA (PC2 = 18.90%) and the antioxidant properties (PC3 = 10.43%). Ash was significantly reduced in the desalted fractions; the protein adsorption recoveries were high, whereas desorption with alcohol was prominently influenced by the hydrophobic/ hydrophilic amino acid balance. After hydrolysis, some hydrolysates showed increased ELISA reactivity compared with the native WPI.     

Isolation and characterization of antioxidative peptides from gelatin hydrolysate of Alaska pollack skin

Gelatin extracted from Alaska pollack skin was hydrolyzed with serial digestions in the order of Alcalase, Pronase E, and collagenase using a three-step recycling membrane reactor. The fraction from the second step, which was hydrolyzed with Pronase E, was composed of peptides ranging from 1.5 to 4.5 kDa and showed high antioxidative activity. Two different peptides showing strong antioxidative activity were isolated from the hydrolysate using consecutive chromatographic methods including gel filtration on a Sephadex G-25 column, ion-exchange chromatography on a SP-Sephadex C-25 column, and high-performance liquid chromatography on an ODS column. The isolated peptides, P1 and P2, were composed of 13 and 16 amino acid residues, respectively; and both peptides contained a Gly residue at the C-terminus and the repeating motif Gly-Pro-Hyp. The antioxidative activities of the purified peptides were measured using the thiobarbituric acid method, and the cell viability was measured with MTT assay. The results showed that P2 had potent antioxidative activity on peroxidation of linoleic acid. Moreover, the cell viability of cultured liver cells was significantly enhanced by addition of the peptide. These results indicate that the purified peptide, P2, from gelatin hydrolysate of Alaska pollack skin is a natural antioxidant which has potent antioxidative activity.     

beta-lactoglobulin hydrolysis. 2. Peptide identification, SH/SS exchange, and functional properties of hydrolysate fractions formed by the action of plasmin.

beta-Lactoglobulin (betaLg) was hydrolyzed by plasmin to a degree of hydrolysis of 4%. The hydrolysate was fractionated by ion-exchange chromatography and subsequent hydrophobic-interaction chromatography. The betaLg peptide fraction consisting of smaller peptides (mostly <2 kDa) had poor foam- and emulsion-forming and -stabilizing properties. Most of the betaLg peptides were identified (in either the nonreduced or reduced form) by mass spectrometry on the basis of the known primary structure of the intact protein and the specificity of the enzyme. The peptides formed during betaLg/plasmin-hydrolysis were (1) peptides lacking a cysteyl residue, (2) peptides composed of a single amino acid chain containing intramolecular disulfide bonds, and (3) peptides composed of two amino acid chains linked by an intermolecular disulfide bond. It appeared that significant SH/SS-exchange had taken place during hydrolysis. Many of the peptides present in the peptide fraction that exhibited good functional properties were disulfide-linked fragments.     

Enzymatic hydrolysis, grease permeation, and water barrier properties of zein isolate coated paper

An inexpensive zein-lipid mixture was isolated from yellow dent, dry-milled corn. Grease permeation through zein isolate applied to brown Kraft paper was found to be independent of loading levels at zein isolate levels above 30 mg/16 in.(2). The data shows that water vapor transmission rates depended on the amount of coating applied. Triacylglycerols were the most abundant lipid in milled corn but were absent in the zein isolate (perhaps due to hydrolysis by lipases). Zein from the paper was hydrolyzed enzymatically and the hydrolysis monitored by SDS-capillary electrophoresis. At an E:S ratio of 1:100 no further increase in the hydrolysate peak occurred after 10 and 30 min for alpha-chymotrypsin and pancreatin 8 x; however, zein and lipid were still present 1 h after hydrolysis by pancreatin 1 x.     

Emulsion properties of casein and whey protein hydrolysates and the relation with other hydrolysate characteristics

Casein and whey protein were hydrolyzed using 11 different commercially available enzyme preparations. Emulsion-forming ability and emulsion stability of the digests were measured as well as biochemical properties with the objective to study the relations between hydrolysate characteristics and emulsion properties. All whey protein hydrolysates formed emulsions with bimodal droplet size distributions, signifying poor emulsion-forming ability. Emulsion-forming ability of some casein hydrolysates was comparable to that of intact casein. Emulsion instability was caused by creaming and coalescence. Creaming occurred mainly in whey hydrolysate emulsions and in casein hydrolysate emulsions containing large emulsion droplets. Coalescence was dominant in casein emulsions with a broad particle size distribution. Emulsion instability due to coalescence was related to apparent molecular weight distribution of hydrolysates; a relative high amount of peptides larger than 2 kDa positively influences emulsion stability.     

Correlations between biochemical characteristics and foam-forming and -stabilizing ability of whey and casein hydrolysates

Whey protein and casein were hydrolyzed with 11 commercially available enzymes. Foam properties of 44 samples were measured and were related to biochemical properties of the hydrolysates using statistical data analysis. All casein hydrolysates formed high initial foam levels, whereas whey hydrolysates differed in their foam-forming abilities. Regression analysis using the molecular weight distribution of whey hydrolysates as predictors showed that the hydrolysate fraction containing peptides of 3-5 kDa was most strongly related to foam formation. Foam stability of whey hydrolysates and of most casein hydrolysates was inferior to that of the intact proteins. The foam stability of casein hydrolysate foams was correlated to the molecular weight distribution of the hydrolysates; a high proportion of peptides >7 kDa, composed of both intact casein and high molecular weight peptides, was positively related to foam stability.     

Characterization of squid-processing byproduct hydrolysate and its potential as aquaculture feed ingredient

The squid (Loligo pealei) byproduct composed of heads, viscera, skin, fins, and small tubes was subjected to hydrolysis at 55 degrees C and natural pH (6.8) using endogenous proteases. Squid hydrolysate was characterized during the course of hydrolysis for changes in the degree of hydrolysis, viscosity, electrophoretic pattern of proteins and peptides, and amino acid and fatty acid profiles. The change in viscosity can be used to monitor the progress of protein hydrolysis up to the molecular mass of 26.63 kDa. The 2 h hydrolysis resulted in a 2-fold increase in the total free amino acids and yielded hydrolysate with protein molecular mass of < or =45 kDa having feed attractability and good amino acid and fatty acid profiles with high contents of essential amino acids and fatty acids. Such hydrolysis-induced changes can make squid byproduct hydrolysate a good source of aquaculture feed ingredient, especially for a starter diet for larval fish.     

Investigation of enzymatic hydrolysis conditions on the properties of protein hydrolysate from fish muscle (Collichthys niveatus) and evaluation of its functional properties

This study was carried out to investigate the enzymatic hydrolysis conditions on the properties of protein hydrolysate from fish muscle of the marine fish species Collichthys niveatus. About 160 fish samples were tested, and the analyzed fish species was found to be a lean fish with low fat (1.77 ± 0.01%) and high protein (16.76 ± 1.21%). Fish muscle of C. niveatus was carefully collected and hydrolyzed with four commercial enzymes: Alcalase, Neutrase, Protamex, and Flavourzyme under the conditions recommended by the manufacturers. Among the tested proteases, Neutrase catalyzed the hydrolysis process most effectively since the hydrolysate generated by Neutrase has the highest content of sweet and umami taste amino acids (SUA). The effect of hydrolysis conditions was further optimized using response surface methodology (RSM), and the optimum values for temperature, pH, and enzyme/substrate ratio (E/S ratio) were found to be 40.7 °C, 7.68, and 0.84%, respectively. Finally, the amino acid composition of the hydrolysate was analyzed by AccQ·Tag derivatization and HPLC-PDA determination. Major amino acids of the muscle of C. niveatus were threonine, glutamic acid, phenyalanine, tryptophan, and lysine, accounting for respectively 10.92%, 10.85%, 10.79%, 9.86%, and 9.76% of total amino acid content. The total content of essential amino acids was 970.7 ng·mL(-1), while that of nonessential amino acids was 709.1 ng·mL(-1). The results suggest that the fish muscle and its protein hydrolysate from C. niveatus provide a versatile supply of the benefits and can be incorporated as supplements in health-care foods.