Chemical and Biochemical Hydrolysis of Persian Sturgeon (Acipenser persicus) Visceral Protein

Chemical (pH 3.3, 70 °C, 85 °C; pH 12, 70 °C, 85 °C) and biochemical (Alcalase, Protamex, Neutrase, Flavourzyme, and Trypsin) hydrolysis of Persian sturgeon (Acipenser persicus) visceral protein was investigated. The results of this study revealed that there are significant differences between enzymes in terms of degree of hydrolysis (DH%; P < 0.05). Alcalase-hydrolyzed fish protein had the highest DH% (50.13%), and Trypsin-hydrolyzed fish protein had the minimum DH% (14.21%). The highest DH% in chemical hydrolysis was related to pH 3.3 at 85 °C (68.87%). The highest protein recovery (83.64%) and protein content (73.34) were related to enzymatic hydrolysis by Alcalase. The results of current study showed the significant effect of hydrolysis conditions on fish protein hydrolysate properties. Microbial enzymes could produce fish hydrolysates with higher degree of hydrolysis when compared to animal enzyme. Also, in chemical hydrolysis it is clear that hydrolysis at the lower pH and at higher temperature causes to more protein recovery and degree of hydrolysis.     

Functional properties of hydrolysates from Phaseolus lunatus seeds

Use of low degree of hydrolysis (DH < 10%) with enzymatic treatment can produce protein hydrolysates with functional properties superior to the raw material. Suspensions of Phaseolus lunatus protein isolate (PPI) were treated with one of two commercial enzymes (Alcalase or Flavourzyme) at 50 °C and pH 8.0. DH with Alcalase was greater than Flavourzyme at 5 or 15 min of reaction. Alcalase-prepared hydrolysates had more peptides than those prepared with Flavourzyme. All the hydrolysates had higher solubility than the PPI, the highest being for the Alcalase-prepared hydrolysate at 15 min reaction time. Overall, the Alcalase-prepared hydrolysates had better solubility characteristics, whereas the Flavourzyme-prepared hydrolysates had better film properties (maximum emulsifying capacity and the highest foam formation values). This is probably because of the greater ease of movement toward the interface as shown by their high surface hydrophobicity values. The Alcalase-prepared hydrolysates had generally low or nonexistent film properties.     

高底物浓度玉米蛋白的双酶复合水解效果研究

利用碱性蛋白酶(Alcalase)、风味蛋白酶(Flavourzyme)和复合蛋白酶(Protamex)对高底物浓度(135g/L)玉米蛋白进行双酶复合水解,研究复合水解对水解物的水解度、可溶性蛋白质含量和抗氧化活性的影响,并对双酶酶解效果较好的酶解液进行了分子量分布测定。结果表明,Flavourzyme和Alcalase、Flavourzyme和Protamex、Protamex和Alcalase顺次水解玉米黄粉,总水解度分别为27.11%、26.95%和19.76%,可溶性蛋白质含量分别为50.33、40.32、48.85mg/ml,抗氧化活性分别为634.35、576.79和593.21 U/ml。多肽分子量主要分布在5 801.170～238.962u,与单酶水解相比均有显著提高。     

两种酶水解鲢鱼蛋白产物功能性质研究

采用Alcalase 2.4L和Flavourzyme 500L蛋白酶对鲢鱼肉进行水解,研究了水解产物的水解度与体系的pH值对其功能性质的影响。水解产物的溶解度在pH4时最低,碱性时具有较高的溶解度;浊度随pH值变化趋势与溶解度相反,浊度越大,溶解度越小。随水解度的增加,水解产物的乳化活性指数、乳化稳定性、起泡性和泡沫稳定性减小(p<0.05);在水解度相同情况下,水解产物的功能性质取决于所用酶的种类。结果表明,鲢鱼肉蛋白水解产物的功能性质受其水解度和所用酶种类的影响。     

Calcium-binding peptides derived from tilapia (Oreochromis niloticus) protein hydrolysate

Calcium-binding peptide was derived from protein hydrolysates. In this study, tilapia protein at a concentration of 2 % (w/v) was hydrolyzed using various proteases including Alcalase 2.4 L, Flavourzyme 1,000L, Protease GN, and papain at 50 °C, pH 8 for 6 h. It was found that the degree of hydrolysis increased with the time of the incubation in all cases. The highest calcium-binding capacity of the hydrolysate was 65 mg/g protein at 27.7 % degree of hydrolysis by Alcalase 2.4 L. The molecular weight of the calcium-binding peptides characterized by gel-filtration chromatography on a Sephadex G-25 was 1.2 kDa. The calcium-binding motif of the hydrolyzed peptides identified by the automated Edman degradation was a short peptide (Trp-Glu-Trp-Leu-His-Tyr-Trp). The results of this study suggested that tilapia protein is a good source for calcium-binding peptides.     

蚕蛹多肽制备工艺及其体外抗氧化活性

采用碱性蛋白酶、风味蛋白酶、复合蛋白酶、木瓜蛋白酶、胰蛋白酶水解蚕蛹蛋白,以水解度和清除DPPH·能力为指标对酶解过程进行分析,并研究水解产物多肽的体外抗氧化活性。结果表明,碱性蛋白酶对蚕蛹蛋白具有较好的水解效果,其水解产物有较高抗氧化活性,对DPPH·、超氧阴离子自由基(O2·)和羟自由基(·OH)都具有较强的清除能力。     

Hydrolysis conditions for antioxidant peptides derived from enzymatic hydrolysates of sandfish (<Emphasis Type="Italic">Arctoscopus japonicus</Emphasis>)

Sandfish (Arctoscopus japonicus) meat and roe were used as natural materials for the preparation of antioxidant peptides using enzymatic hydrolysis. Meat and roe were hydrolyzed using Alcalase 2.4 L and Collupulin MG, respectively. Optimal hydrolysis conditions were determined through the effects of pH, temperature, enzyme concentration, and hydrolysis time on the radical scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH). The optimal hydrolysis conditions for meat hydrolysate (MHA) obtained via Alcalase 2.4 L treatment were a pH of 6.0, temperature of 70 °C, enzyme concentration of 5% (w/w), and a hydrolysis time of 3 h. The optimal hydrolysis conditions for roe hydrolysate (RHC) obtained via Collupulin MG treatment were pH 9.0, 60 °C temperature, 5% (w/w) enzyme concentration, and 1 h hydrolysis time. Under the optimal conditions, the DPPH radical scavenging activities of MHA and RHC were 60.04 and 79.65%, respectively. These results provide fundamental data for the production of antioxidant peptides derived from sandfish hydrolysates.     

双酶法水解猪皮工艺的优化

以猪皮为原料,以水解度(DH)为评价指标,在用Flavourzyme,Protamex,Neutrase,Alcalase,Papain进行单酶、双酶水解的基础上,筛选出了Flavourzyme和Protamex两种水解效果较好的酶,并采用Box-Benhnken响应面分析法,设计了3因素3水平实验,得到了双酶法水解猪皮的优化条件为:pH5.85,温度48.37℃,加酶量2.61%,底物浓度15%,时间6h,在此条件下,猪皮水解度最高,可达到19.25%。     

Alcalase 2.4L碱性内切酶和Flavourzyme风味酶酶解大豆分离蛋白及脱苦工艺优化

以大豆分离蛋白为原料,选用Alcalase 2.4L碱性内切酶和Flavourzyme风味蛋白酶对大豆分离蛋白进行酶法水解及脱苦工艺研究。以水解度和苦味分值为考察值,对酶解工艺进行优化,确定最佳条件。结果表明:Alcalase2.4L碱性内切酶最佳酶解条件为加酶量14 000 U/g、酶解温度60℃、酶解pH8.5、底物质量分数5%,酶解时间2h,最终水解度为45.34%,此时水解液苦味值为4。Flavourzyme风味蛋白酶对水解液进行二次水解的最优酶解条件为加酶量300 U/g、酶解温度55℃、酶解pH 7.0、酶解时间3 h,此条件下大豆分离蛋白水解液苦味值最低为1.2。Alcalase2.4L碱性内切酶和Flavourzyme风味蛋白酶水解大豆分离蛋白使水解度得到较大提高的同时也解决了水解液的苦味问题。     

Physicochemical and bitterness properties of enzymatic pea protein hydrolysates

ABSTRACT:  The effects of different proteolytic treatments on the physiochemical and bitterness properties of pea protein hydrolysates were investigated. A commercial pea protein isolate was digested using each of 5 different proteases to produce protein hydrolysates with varying properties. After 4 h of enzyme digestion, samples were clarified by centrifugation followed by desalting of the supernatant with a 1000 Da membrane; the retentates were then freeze-dried. Alcalase and Flavourzyme™ produced protein hydrolysates with significantly higher ( P < 0.05) degree of hydrolysis when compared to the other proteases. Flavourzyme, papain, and alcalase produced hydrolysates that contained the highest levels of aromatic amino acids, while trypsin hydrolysate had the highest levels of lysine and arginine. Papain hydrolysate contained high molecular weight peptides (10 to 178 kDa) while hydrolysates from the other 4 proteases contained predominantly low molecular weight peptides (≤ 23 kDa). DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical scavenging activity of the Flavourzyme hydrolysate was significantly ( P < 0.05) the highest while alcalase and trypsin hydrolysates were the lowest. Inhibition of angiotensin converting enzyme (ACE) activity was significantly higher ( P < 0.05) for papain hydrolysate while Flavourzyme hydrolysate had the least inhibitory activity. Sensory analysis showed that the alcalase hydrolysate was the most bitter while papain and α-chymotrypsin hydrolysates were the least. Among the 5 enzymes used in this study, papain and α-chymotrypsin appear to be the most desirable for producing high quality pea protein hydrolysates because of the low bitterness scores combined with a high level of angiotensin converting enzyme inhibition and moderate free radical scavenging activity.     

双酶法酶解猪瘦肉蛋白工艺优化研究

为制备腊味香精提供更丰富的前体物,以猪瘦肉为原料,水解度为指标,选用Alcalase和Flavourzyme为试验用酶,通正交试验优化双酶法水解猪瘦肉蛋白最佳工艺条件。结果表明:Alcalase和Flavourzyme水解瘦肉蛋白其最佳条件为,pH 9时,用1 000U/g Alcalase于60℃下水解6h,再用2 500U/g Flavourzyme于50℃、pH 4.5继续水解4h。此时,水解度达到34.82%,比Alcalase和Flavourzyme单酶水解度分别提高34.12%和27.48%,且酶解产物还含有丰富的制备腊味香精的前提物。     

Antioxidant properties and protein compositions of porcine haemoglobin hydrolysates

Porcine haemoglobin hydrolysates were prepared through hydrolysis by Alcalase followed by Flavourzyme, and their protein compositions were analyzed using Sephadex G-50 gel filtration chromatography. The antioxidant activities, including reducing power, ferrous ion chelating ability, and DPPH radical scavenging activity, of the hydrolysates were evaluated. The results showed that the hydrolysates of haemoglobin exhibited low reducing powers, but high ferrous ion chelating abilities and DPPH radical scavenging activities. The hydrolysate, obtained through hydrolysis by 2% Alcalase for 4 h and followed by 1% Flavourzyme for 6 h, had the highest ferrous ion chelating ability of 63.54% at a concentration of 5.0 mg/mL. The hydrolysate, obtained through hydrolysis by 2% Alcalase for 4 hrs, had the highest DPPH radical scavenging activity of 41.94% at a concentration of 5.0 mg/mL. According to the results of protein composition analysis, we divided the hydrolysates into three groups, including high molecular weight (MW) group (Group I), medium MW group (Group II), and low MW group (Group III). The reducing power and ferrous ion chelating ability of the hydrolysates were significantly and positively correlated to the relative amount of Group I, and negatively correlated to the relative amount of Group III. This study revealed that the antioxidant activities of porcine haemoglobin hydrolysates were dependent on their protein compositions. The high MW protein fraction (Group I) was responsible for the high reducing power and ferrous ion chelating ability of the hydrolysate.     

大豆活性肽酶解制备的工艺条件研究

以大豆分离蛋白为原料,选用碱性蛋白酶和风味蛋白酶,分别从酶解pH 值、酶解温度、酶用量和底物质量分数因素研究其对单酶酶解大豆分离蛋白的影响。并通过Minitab 软件,利用响应曲面试验优化双酶酶解工艺条件。结果表明,其最佳酶解条件为酶解pH7.7、碱性蛋白酶用量为110mg/g 底物、风味蛋白酶用量为90mg/g底物、酶解温度56℃、底物质量分数8%、酶解时间7h,所得的大豆活性肽的分子量主要集中在1000D 以下。     

Enzymatic hydrolysis of heated whey: iron-binding ability of peptides and antigenic protein fractions

This study evaluated the influence of various enzymes on the hydrolysis of whey protein concentrate (WPC) to reduce its antigenic fractions and to quantify the peptides having iron-binding ability in its hydrolysates. Heated (for 10 min at 100°C) WPC (2% protein solution) was incubated with 2% each of Alcalase, Flavourzyme, papain, and trypsin for 30, 60, 90, 120, 150, 180, and 240 min at 50°C. The highest hydrolysis of WPC was observed after 240 min of incubation with Alcalase (12.4%), followed by Flavourzyme (12.0%), trypsin (10.4%), and papain (8.53%). The nonprotein nitrogen contents of WPC hydrolysate followed the hydrolytic pattern of whey. The major antigenic fractions (β-lactoglobulin) in WPC were degraded within 60 min of its incubation with Alcalase, Flavourzyme, or papain. Chromatograms of enzymatic hydrolysates of heated WPC also indicated complete degradation of β-lactoglobulin, α-lactalbumin, and BSA. The highest iron solubility was noticed in hydrolysates derived with Alcalase (95%), followed by those produced with trypsin (90%), papain (87%), and Flavourzyme (81%). Eluted fraction 1 (F-1) and fraction 2 (F-2) were the respective peaks for the 0.25 and 0.5 M NaCl chromatographic step gradient for analysis of hydrolysates. Iron-binding ability was noticeably higher in F-1 than in F-2 of all hydrolysates of WPC. The highest iron contents in F-1 were observed in WPC hydrolysates derived with Alcalase (0.2 mg/kg), followed by hydrolysates derived with Flavourzyme (0.14 mg/kg), trypsin (0.14 mg/kg), and papain (0.08 mg/kg). Iron concentrations in the F-2 fraction of all enzymatic hydrolysates of WPC were low and ranged from 0.03 to 0.05 mg/kg. Fraction 1 may describe a new class of iron chelates based on the reaction of FeSO₄·7H₂O with a mixture of peptides obtained by the enzymatic hydrolysis of WPC. The chromatogram of Alcalase F-1 indicated numerous small peaks of shorter wavelengths, which probably indicated a variety of new peptides with greater ability to bind with iron. Alcalase F-1 had higher Ala (18.38%), Lys (17.97%), and Phe (16.58%) concentrations, whereas the presence of Pro, Gly, and Tyr was not detected. Alcalase was more effective than other enzymes at producing a hydrolysate for the separation of iron-binding peptides derived from WPC.     

不同蛋白酶对小麦蛋白酶解物抗氧化活性的影响

小麦蛋白是小麦淀粉加工的副产物,酶解是提高小麦蛋白溶解性和功能性的有效方式,而酶解用酶种类可能对酶解产物的功能性如抗氧化活性有一定影响。采用碱性蛋白酶、中性蛋白酶、胃蛋白酶、风味蛋白酶、胰蛋白酶、木瓜蛋白酶6种常用的蛋白酶分别对小麦蛋白进行酶解,并对酶解4 h后酶解物的多肽得率、分子质量分布、1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率、超氧阴离子自由基(O_2~-·)清除率、羟自由基(·OH)清除率等反映水解程度和抗氧化能力的主要指标进行评价。结果表明,风味蛋白酶酶解物中多肽得率最高,达91.44%,且分子质量小于3 000 D的多肽含量达76.9%;酶解物质量浓度为3 mg/m L时,木瓜蛋白酶酶解物对DPPH自由基清除作用最好,清除率为65.12%(P0.01),其次是风味蛋白酶(58.43%)和碱性蛋白酶(55.29%);碱性蛋白酶酶解物对O_2~-·清除率效果最好,清除率为58.68%(P0.01),其次是风味蛋白酶(49.25%);碱性蛋白酶和木瓜蛋白酶酶解物对·OH清除效果最佳,清除率分别为59.23%和58.16%。结果说明,蛋白酶种类对小麦蛋白酶解物抗氧化活性影响显著,风味蛋白酶对提高蛋白水解程度和生成小分子质量多肽的作用明显,而碱性蛋白酶、木瓜蛋白酶和风味蛋白酶对提高酶解产物抗氧化活性效果较好。     

杏仁蛋白水解物对血管紧张素转化酶抑制作用的研究

分别采用Protamex、Alcalase、Neutrase、Flavourzyme、Proleather FG-F、木瓜蛋白酶水解杏仁蛋白,利用高效液相色谱法测定水解物对血管紧张素转化酶(ACE)的抑制活性,以水解度(DH)和水解产物对ACE的抑制率为指标对酶解过程进行分析,并研究水解物的体外消化稳定性。结果表明,Proleather FG-F 和Alcalase 对杏仁蛋白有较好的水解效果,其水解物对ACE 抑制率较高,IC50 分别为1.24mg/ml 和0.98mg/ml。模拟胃肠消化实验结果表明,在消化酶的作用下杏仁蛋白水解物仍具有较强的ACE 抑制活性。     

Antioxidant activity and water-holding capacity of canola protein hydrolysates

Canola protein hydrolysates were prepared using commercial enzymes, namely Alcalase, an endo-peptidase and Flavourzyme with both endo- and exo-peptidase activities. The hydrolysates so prepared were effective as antioxidants in model systems, mainly by scavenging of free radicals and acting as reducing agents. This effect was concentration-dependent and also influenced by the type of enzyme employed in the process. The hydrolysate prepared using flavourzyme showed the highest antioxidant activity among all samples, whereas the hydrolysates prepared by combination of Alcalase and Flavourzyme did not differ significantly (P > 0.05) in antioxidant effectiveness from that produced by Alcalase alone. The hydrolysates were also found to be effective in enhancing water-holding capacity and cooking yield in a meat model system. Their capability in improving the cooking yield of meat was in the order of Flavourzyme hydrolysates > combination hydrolysates > Alcalase hydrolysates. These results suggest that canola protein hydrolysates can be useful in terms of their functionality and as functional food ingredients and that their composition determines their functional properties and thus their potential application in the food and feed industries.     

Quality of Fish Protein Hydrolysates from Herring (Clupea harengus)

Fish protein hydrolysates were prepared, using minced fillets and Al-calase and papain, from raw herring and from herring defatted by ethanol extraction, cooking and pressing. Physicochemical, sensory and storage properties as well as the molecular weight sizes in spraydried hydrolysates were evaluated. Fat extraction before hydrolysis reduced degree of hydrolysis. Alcalase hydrolyzed samples to a higher degree than papain. Ethanol extraction reduced fishy odor to barely detectable levels. Papain hydrolysates were more bitter than those made with Alcalase. Color and nonenzymic browning measurements indicated darkening during 3 mo storage at room temperature (～20°C).     

Enzymic modification of extruded soy protein concentrates as a method of obtaining new functional food components

Response surface methodology was used to determine optimal conditions for limited hydrolysis of the extruded soy protein concentrate with Alcalase and Esperase, which would allow one to obtain new products with desirable functional properties. The best results can be obtained by conducting the process at 60°C with water addition 13 g/g protein, enzyme addition 8 × 10⁻³ Anson Units/g protein, as well as pH 8.3 and 8.6 and time 120 and 60 min using Alcalase and Esperase, respectively. It was shown that hydrolysates obtained from extruded soy protein concentrate differ from those from non texturized raw material in the content of sulfhydryl groups, disulphide bonds and surface hydrophobicity. Also, as was shown by SDS-PAGE, protein breakdown in them was less selective.     

Enzymatic extraction of protein from toasted and not toasted soybean meal

The soybean is one import font of vegetable protein, that contains whole the essential amino acids to the human organism. The enzymatic process is a alternative for the protein extraction and offer advantages. In this work, two commercial proteases, Alcalase and Flavourzyme, are used to extract protein from toasted and not toasted degreased soybean meal. The tests (duplicate) are carried out in 40 °C, initial pH of 7,0, enzyme concentration of 1% (protein/protein) and times from 1 up to 24 hours. At the end of the hydrolysis the suspension pH is measured and again corrected for 7,0.