Functionalities and antioxidant properties of protein hydrolysates from the muscle of ornate threadfin bream treated with pepsin from skipjack tuna

Functional properties and antioxidant activities of protein hydrolysates prepared from ornate threadfin bream (Nemipterus hexodon) muscle, using skipjack tuna pepsin, with different degrees of hydrolysis (DH: 10%, 20% and 30%), were evaluated. Emulsifying and foaming properties of hydrolysates were governed by their DH and concentrations used. Hydrolysates with 20% DH had the highest scavenging activities for ABTS and DPPH radicals. However, chelating activity of hydrolysates for ferrous ion increased as DH increased. Size exclusion chromatography of the hydrolysate with 20% DH using Sephadex G-25 revealed that antioxidative peptides with molecular weight of approximately 1.3 kDa exhibited the highest ABTS radical-scavenging activity. In vitro simulated gastrointestinal digestion indicated that ABTS radical-scavenging activity of the antioxidative peptides was not affected by pepsin hydrolysis, whilst further digestion by pancreatin enhanced the activity. Therefore, protein hydrolysate from the muscle of ornate threadfin bream produced by skipjack tuna pepsin can be used as a promising source of functional peptides with antioxidant properties.     

Proteolysis characteristics of sarcoplasmic, myofibrillar, and stromal proteins separated from grass carp and antioxidant properties of their hydrolysates

Proteolysis of grass carp sarcoplasmic, myofibrillar, and stromal proteins by 5 commercial proteases were studied. Sarcoplasmic and myofibrillar protein could be well hydrolyzed by Alcalase 2.4 L to reach high protein recoveries (PR) (71.86±2.46 and 80.77±3.05%, respectively), while the maximum PR for stromal protein was only 42.83±2.84%. However, stromal hydrolysates, containing mostly 6–10 kDa fraction, exhibited higher ·OH scavenging activities due to its high content of antioxidant-assisting amino acids. Alcalase 2.4 L and pancreatin 6.0, which produced hydrolysates with relative high degree of hydrolysis (DH), were used for further hydrolysis of whole grass carp protein with the assistance of response surface methodology (RSM). The results showed that serine proteases (Alcalase 2.4 L and pancreatin 6.0) could produce sarcoplasmic, myofibrillar, or stromal hydrolysates with relatively high PR, DH, and strong ·OH scavenging activity, which may be used to prepare antioxidant hydrolysates from grass carp.     

Angiotensin I‐converting enzyme inhibitory activity of protein hydrolysates prepared from three freshwater carps (Catla catla,Labeo rohita and Cirrhinus mrigala) using Flavorzyme

Fish protein hydrolysates from three freshwater carps, Catla catla, Labeo rohita and Cirrhinus mrigala with different degree of hydrolysis (DH) (5%, 10%, 15% and 20%), were prepared using Flavorzyme enzyme and designated as HCF, HRF and HMF, respectively. The angiotensin I‐converting enzyme (ACE) inhibitory activity of hydrolysates was found to vary from 43 ± 2% to 71 ± 3%. Based on ACE inhibitory activity, HRF with DH‐15% was taken up for further study. The mode of ACE activity inhibition by HRF‐DH 15% was mixed type as revealed by Lineweaver–Burk plot. Sequential digestion of HRF‐DH 15% using pepsin and pancreatin decreased the ACE inhibitory activity from 76% to 63%. Partial purification of HRF‐DH 15% by size exclusion chromatography gave three different fractions designated as F‐1, F‐2 and F‐3 with the molecular mass in the range of 6456–407 Da. Fraction 2 had significantly higher ACE inhibitory activity than the other fractions.     

Improving antioxidant activities of whey protein hydrolysates obtained by thermal preheat treatment of pepsin,trypsin, alcalase and flavourzyme

Antioxidant activity of whey protein concentrate (WPC) hydrolysates was evaluated. Hydrolysates were obtained by pepsin, trypsin, alcalase and flavourzyme enzymatic reaction and preheat treatment of 95 °C for 5 or 10 min. The degree of hydrolysis (DH) was determined by 2,4,6‐trinitrobenzene sulphonic acid method, and antioxidant properties were determined by three spectrophotometric methods: ferricyanide method, ferric reducing/antioxidant power assay and diphenyl‐picryl hydrazinyl radical‐scavenging activity. For all the enzymes, briefly preheat treatment (95 °C/5 min) increased DH of WPC. Alcalase hydrolysates showed the highest antioxidant activity by three methods. The changes in antioxidant activity was coincidental with the changes in DH (R² = 0.988). Hydrolysates analysed by polyacrylamide gel electrophoresis and high performance liquid chromatography indicated that the α‐La was hydrolysed completely by pepsin, trypsin and alcalase and was resistant to flavourzyme to some extent; β‐lactoglobulin was only completely hydrolysed by trypsin and alcalase. Results indicated that antioxidant activity of hydrolysates was greatly related to the exposure of amino acid residues.     

Angiotensin‐I converting enzyme inhibitory and antioxidant activities of peptide fractions extracted by ultrafiltration of cowpea Vigna unguiculata hydrolysates

BACKGROUND: Enzymatic proteolysis of food proteins is used to produce peptide fractions with the potential to act as physiological modulators. Fractionation of these proteins by ultrafiltration results in fractions rich in small peptides with the potential to act as functional food ingredients. The present study investigated the angiotensin‐I converting enzyme (ACE‐I) inhibitory and antioxidant activities for hydrolysates produced by hydrolyzing Vigna unguiculata protein extract as well as ultrafiltered peptide fractions from these hydrolysates. RESULTS: Alcalase^®, Flavourzyme^® and pepsin–pancreatin were used to produce extensively hydrolyzed V. unguiculata protein extract. Degree of hydrolysis (DH) differed between the enzymatic systems and ranged from 35.7% to 58.8%. Fractionation increased in vitro biological activities in the peptide fractions, with IC₅₀ (hydrolysate concentration in µg protein mL^?1 required to produce 50% ACE inhibition) value ranges of 24.3–123 (Alcalase hydrolysate, AH), 0.04–170.6 (Flavourzyme hydrolysate; FH) and 44.7–112 (pepsin–pancreatin hydrolysate, PPH) µg mL^?1, and TEAC (Trolox equivalent antioxidant coefficient) value ranges of 303.2–1457 (AH), 357.4–10 211 (FH) and 267.1–2830.4 (PPH) mmol L^?1 mg^?1 protein. CONCLUSION: The results indicate the possibility of obtaining bioactive peptides from V. unguiculata proteins by means of a controlled protein hydrolysis using Alcalase^®, Flavourzyme^® and pepsin–pancreatin. The V. unguiculata protein hydrolysates and their corresponding ultrafiltered peptide fractions might be utilized for physiologically functional foods with antihypertensive and antioxidant activities. Copyright © 2010 Society of Chemical Industry     

Peptic and tryptic hydrolysis of native and heated whey protein to reduce its antigenicity

This study examined the effects of enzymes on the production and antigenicity of native and heated whey protein concentrate (WPC) hydrolysates. Native and heated (10 min at 100°C) WPC (2% protein solution) were incubated at 50°C for 30, 60, 90, and 120 min with 0.1, 0.5, and 1% pepsin and then with 0.1, 0.5, and 1% trypsin on a protein-equivalent basis. A greater degree of hydrolysis was achieved and greater nonprotein nitrogen concentrations were obtained in heated WPC than in native WPC at all incubation times. Hydrolysis of WPC was increased with an increasing level of enzymes and higher incubation times. The highest hydrolysis (25.23%) was observed in heated WPC incubated with 1% pepsin and then with 1% trypsin for 120 min. High molecular weight bands, such as BSA, were completely eliminated from sodium dodecyl sulfate-PAGE of both native and heated WPC hydrolysates produced with pepsin for the 30-min incubation. The α-lactalbumin in native WPC was slightly degraded when incubated with 0.1% pepsin and then with 0.1% trypsin; however, it was almost completely hydrolyzed within 60 min of incubation with 0.5% pepsin and then with 0.5% trypsin. Incubation of native WPC with 1% pepsin and then with 1% trypsin for 30 min completely removed the BSA and α-lactalbumin. The β-lactoglobulin in native WPC was not affected by the pepsin and trypsin treatments. The β-lactoglobulin in heated WPC was partially hydrolyzed by the 0.1 and 0.5% pepsin and trypsin treatments and was completely degraded by the 1% pepsin and trypsin treatment. Antigenicity reversibly mimicked the hydrolysis of WPC and the removal of β-lactoglobulin from hydrolysates. Antigenicity in heated and native WPC was reduced with an increasing level of enzymes. A low antigenic response was observed in heated WPC compared with native WPC. The lowest antigenicity was observed when heated WPC was incubated with 1% pepsin and then with 1% trypsin. These results suggested that incubation of heated WPC with 1% pepsin and then with 1% trypsin was the most effective for producing low-antigenic hydrolysates by WPC hydrolysis and obtaining low molecular weight small peptides. Further research is warranted to identify the low molecular weight small peptides in the WPC hydrolysates produced by pepsin and trypsin, which may enhance the use of whey.     

Functional Properties and Bioactivities of Pine Nut (Pinus gerardiana) Protein Isolates and Its Enzymatic Hydrolysates

The functional properties and bioactivities of the pine nut protein isolates (PPI) and its enzymatic hydrolysates (PPH) prepared with Alcalase at 5 %, 10 %, 15 % and 25 % degree of hydrolysis (DH) were studied. The solubility of PPH significantly increased (p?<?0.05) with the increase of the DH, while the foaming capacity of PPH was only improved at a low DH. However, enzymatic hydrolysis reduced the emulsifying capacity of PPH. The DPPH radical scavenging and inhibition of linoleic acid oxidation activities of PPH were significantly improved by a low DH (5 %) compared with those of PPH with a higher DH and the original PPI (p?<?0.05). The reducing power of PPH at all DH decreased in comparison to that of the original PPI. Potent angiotensin-converting enzyme (ACE) inhibitory peptides could be generated by hydrolysis with Alcalase, and the ACE inhibitory activity of PPH increased (p?<?0.05) with the DH. These results revealed that a low degree of enzymatic hydrolysis was appropriate to obtain PPH with improved functional properties and good antioxidant activities, while a high degree of hydrolysis was essential to obtain highly potent ACE inhibitory peptides from PPI. These results suggest that the control of the DH may be an effective strategy to modify specific functional and bioactive properties of PPH, and PPH has potential as a functional food ingredient for related functional and health benefits.     

Antioxidant properties of porcine liver proteins hydrolyzed using <Emphasis Type="Italic">Monascus purpureus</Emphasis>

In this study, the antioxidant activities of porcine liver proteins, hydrolyzed using Alcalase^®, papain, pepsin, or a microbial suspension of Monascus purpureus (APLH, PaPLH, PePLH, and MPLH, respectively), were investigated. The results indicated that the yield and degree of hydrolysis (DH) of hydrolysates increased with hydrolysis time. The highest yield and peptide content were obtained from APLH, whereas the DH of PaPLH was higher than that of the others. MPLH exhibited the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and reducing power, whereas APLH and PaPLH exhibited the higher ferrous ion-chelating ability than that of the MPLH. The molecular weights of all the hydrolysates were <10 kDa. The PaPLH exhibited the highest contents of total amino acids and hydrophobic amino acids. Fifteen antioxidant fractions obtained from MPLH contained one or more of the following amino acids in their sequences: Tyr, Trp, Ala, Pro, Met, Lys, Asp, Cys, Val, Leu, and His.     

Enzymatic hydrolysis of by‐products from the fish‐filleting industry; chemical characterisation and nutritional evaluation

Fish frames without heads from Atlantic cod and Atlantic salmon were proteolysed with the industrial enzymes neutrase®, alcalase® and pepsin for 1, 15, 30, 45, 60, 90 and 120 min. After 120 min of hydrolysis, salmon treated with alcalase and cod treated with pepsin yielded significantly (p < 0.05) higher protein recoveries (67.6 and 64% respectively) as compared to salmon treated with neutrase or pepsin and cod treated with neutrase or alcalase (53–62%). To minimise bitterness in the fish hydrolysates, kojizyme™ was added after 120 min of pre‐hydrolysis with alcalase, and the hydrolysis was run for additional times of 120, 240, 360, 480, 600 and 720 min. Protein recovery did not change significantly during the hydrolysis with kojizyme, but the degree of hydrolysis increased significantly (p < 0.01) in both the cod and salmon hydrolysates. A hydrolysate from cod treated with alcalase (150 min) followed by treatment with kojizyme (510 min) was produced. The final hydrolysate was freeze‐dried to a fish protein hydrolysate (FPH) and chemically characterised. The nutritional value of the FPH was established in an experiment with rats. Inclusion of 10% FPH‐N showed significantly (p < 0.05) higher nutritional value as compared to rats fed higher inclusion levels of FPH. © 2000 Society of Chemical Industry     

Solubility and Emulsifying Properties of Soy Protein Isolates Modified by Pancreatin

Soy protein isolates (SPI) with varying degrees of hydrolysis (DH of 7, 11, 15, 17%) were produced using pancreatin. The surface hydrophobicity indices of pancreatin hydrolyzed SPI (PSPI) (34.5, 34.9, 39. 1, and 40.7 for 7, 11, 15, 17% DH, respectively) were higher than that of SPI (10.5) and control SPI (CSPI) (12.5, 11.9, 12.9, and 12.6 for 10, 60, 120, and 180 min incubation, respectively). The solubilities of PSPI at pH 4.5 were 2.7, 9.1, 11.9, and 18.7%, for 7, 11, 15, and 17% DH, respectively, while the solubilities of SPI and CSPI at the same pH were about 1. 6%. Solubilities of PSPI at pH 7. 0 were > 90% for all DHs tested, while those of SPI and CSPI were 85%. The emulsifying activity index (EAI) of PSPI increased with increasing DH. PSPI with 15% DH had highest EAI (1. 122) which was higher (P < 0. 05) than those of SPI (0.550) and CSPI after 120 min incubation without enzyme (0.568). These results suggest that PSPI could be used as an ingredient for emulsified products and where high solubility at low pH is required.     

Enzymatic hydrolysis of fish gelatin under high pressure treatment

The effect of high pressure treatment on the hydrolysis of fish skin gelatin and the antioxidant properties of the hydrolysates was evaluated. Hydrolysis was performed by Alcalase, collagenase, trypsin and pepsin both at atmospheric pressure and under high pressure (100 MPa/15 and 30 min, 200 MPa/15 and 30 min, 300 MPa/15 min). The degree of hydrolysis (DH) was determined according to the base consumption via pH‐stat as well as the percentage of soluble nitrogen in trichloroacetic acid (TCA). About 16% of nitrogen in the gelatin was still soluble in 10% TCA, indicative of a significant amount of low molecular weight components. The high pressure treatment increased the DH with all the enzymes used between 5% and 10%. However, in comparison with the hydrolysates obtained at atmospheric pressure after 3 h of digestion under controlled conditions (using a pH‐stat), the radical scavenging capacity of the pressured hydrolysates was only significantly enhanced when Alcalase or collagenase were used. High pressure may be a useful tool for the quick obtaining of gelatin hydrolysates with antioxidant capacity.     

Preparation and Functional Properties of Enzymatically Deamidated Soy Proteins

Heat-denatured soy protein was hydrolyzed by Alcalase to 2.0% or 4.0% degree of hydrolysis (DH), heated again at 100°C and deamidated with B. circulans peptidoglutaminase. The extent of deamidation was 6.0% and 8.2% for 2.0 DH hydrolysates and 12.8% and 16.0% for 4.0 DH hydrolysates heated for 15 and 30 min, respectively. Deamidation increased protein solubility and substantially enhanced emulsifying activity under mildly acidic (pH 4–6) as well as alkaline conditions. Deamidation improved emulsion stability and foaming power of heat-denatured hydrolysed soy proteins. Enzymatically deamidated soy protein hydrolysates had improved functional properties compared to nondeamidated hydrolysates and the native soy protein.     

琥珀酸脱酰胺对小麦面筋蛋白酶解产物抗氧化性的影响

研究琥珀酸不同脱酰胺程度对小麦面筋蛋白胰酶的酶解产物抗氧化性的影响。在相同水解度下,以DPPH.清除率、.OH清除率、ORAC值为抗氧化指标,结果表明低脱酰胺程度(22.4%)的小麦面筋蛋白酶解产物的抗氧化性有较大幅度的提高,高脱酰胺程度(60.4%)的小麦面筋蛋白酶解产物的抗氧化性降低。水解度15%的酶解产物的肽分子量分布表明,低脱酰胺的小麦面筋蛋白酶解产物中分子量小于3000u的肽段增加,高脱酰胺的小麦面筋蛋白酶解产物中分子量小于3000u的肽段减少;在相同水解度下的氨基酸分析结果表明,脱酰胺后的小麦面筋蛋白酶解生成的肽的疏水性和抗氧化性的氨基酸含量提高。因此,低脱酰胺的小麦面筋蛋白酶解产物抗氧化性提高的主要原因是产物中含疏水性和抗氧化性氨基酸的小分子肽含量提高。     

Controlled enzymatic hydrolysis on characteristic and antioxidant properties of soybean protein isolate-maltodextrin conjugates

This study aimed to investigate the effect of limited hydrolysis on conformational and antioxidant properties of soy protein isolate-maltodextrin (SPI-Md) conjugates. Extrinsic fluorescence analysis showed unfolding of the protein molecule and exposure of hydrophobic groups in SPI-Md conjugate hydrolysates. Free amino acid analysis showed that, the contents of hydrophobic amino acids in SPI-Md conjugates increased after hydrolysis. The contents of leucine, isoleucine, phenylalanine increased from 0.32, 0.30 and 0.54 to 1.36, 1.86 and 2.60, respectively, when the hydrolysis degree (DH) gradually increased from 0% to 5.7%. The FT-IR spectrum showed that C = O absorption of the amide group formed by glycosylation continued unabated after limited hydrolysis (DH 2.9%). The glycated SPI products showed good reducing power and superior resistance to lipid oxidation (34%, 12 mg mL^?1), whereas the limit hydrolysates (DH 2.9%) of SPI-Md conjugates showed more ef?cient radical-scavenging capacity (89.5%, at 12 mg mL^?1) and iron-chelation activity (91.3%, at 12 mg mL^?1). Results of this study indicated that, slight enzymatic hydrolysis (DH 0–2.9%) could help partially unfolding the globular structure of SPI-Md conjugates without deteriorating amide bonds and had a positive effect on their antioxidant properties.     

Effect of Enzymatic Digestion, pH and Molecular Weight on the Iron Solubilizing Properties of Chicken Muscle

The effects of a simulated gastrointestinal pH, enzymatic digestion and molecular weight (MW) on the iron solubilizing properties of a heated dilute salt insoluble fraction of chicken muscle were examined. The solubility of 50 ppm added FeCl₃ increased linearly from 0–260 min during pepsin digestion. The total soluble iron reached a maximum concentration following a 120 min pepsin-30 min pancreatin digestion, with pepsin digestion products ranging in MW from 6200–2500. Solubilization capacity, defined as an in vitro measure of total iron bioavailability, did not correlate to binding by free sulfhydryl groups. The soluble low molecular weight iron chelates found may explain, in part, the mechanism by which the “meat factor” enhances iron bioavailability.     

Purification and characterisation of antioxidative peptides from unfractionated rice bran protein hydrolysates

Crude rice bran protein (CRBP) was prepared by alkaline extraction and then treated with 0.6 m HCl to remove phytic acid. The phytate‐free rice bran protein (PFRBP) was hydrolysed with proteases M, N, S, P and pepsin under optimal conditions. Hydrolysates obtained from various hydrolysis periods were subjected to analysis for their degree of hydrolysis (DH) and functional properties. The hydrolysates were fractionated by reversed‐phase column chromatography on Kaseigel ODS resin (120–140 μm) using a stepwise gradient of aqueous ethanol, and their activities were measured. The 40% ethanol fraction of protease P 4 h‐hydrolysate was separated by successive reversed‐phase high‐performance liquid chromatography and the amino acid sequences of isolated antioxidative peptides were determined by a protein sequencer and matrix‐assisted laser desorption ionisation‐time of flight mass spectrometry. Crude rice bran protein had higher antioxidative activity than PFRBP, due to the presence of phytic acid. Phytate contents of rice bran, CRBP and PFRBP were 2.5%, 1.42% and 0%, respectively. The activity of PFRBP increased upon protease digestion. Protease M hydrolysates showed the highest DH, but the lowest antioxidative activity. Hydrolysates with DH below 10% had higher antioxidative activity than those above 20%. This result indicates that the antioxidative activity of the hydrolysates is inherent to their characteristics amino acid sequences of peptides depending on the protease specificities.     

Compositions and antioxidant properties of protein hydrolysates from the skins of four carp species

Compositions and antioxidant properties of protein hydrolysates prepared from four carp skins: black carp, grass carp, silver carp and bighead carp, using pepsin, with a degree of hydrolysis (DH) of 6–15%, were investigated. The yield of freeze‐dried hydrolysates was in the range of 54–62 g/100 g (dry skin). The content of protein and ash in four freeze‐dried hydrolysates was 72–81% and 8–17%, respectively. All hydrolysates contained high amount of hydrophobic amino acid residues (389–480 residues/1000 residues). Meanwhile, their antioxidant properties were evaluated by in vitro assays. The results revealed that all hydrolysates possessed potent antioxidant activities and showed dose dependency as the activity increased with sample concentration, capable of scavenging 72–88% of DPPH and 61–69% of hydroxyl radicals, respectively, at the highest tested concentration. The hydrolysates exhibited high reducing power and β‐carotene–linoleic acid oxidation inhibition. Among the four hydrolysates, the hydrolysate derived from bighead carp skin was superior to others in terms of yield, DH and antioxidant activities.     

The effect of lactic acid bacteria fermentation on the antioxidant activity of wheat gluten pancreatin hydrolysates

The antioxidant activities of the fermented wheat gluten hydrolysates with different fermentation times were investigated to elucidate the impact of lactic acid bacteria (LAB) fermentation on the wheat gluten hydrolysates. Prior to LAB fermentation, wheat gluten was deamidated by hydrochloric acid and then hydrolysed by pancreatin to 12 and 24 h, respectively. Results showed that LAB fermentation had significant impacts on the enzymatic efficiency and antioxidant activities of wheat gluten. The degree of hydrolysis and protein recovery of hydrolysates gradually increased and then reached maximum values, respectively, when fermenting with LAB for 36 h. The hydrolysis degree and protein recovery of fermented pancreatin 24‐h hydrolysates were larger than that of the fermented pancreatin 12‐h hydrolysates during the whole fermentation. The antioxidant activity analysis revealed a marked increase and improvement in the scavenging activities of 1,1‐Diphenyl‐2‐picrylhydrazyl·radicals, hydroxyl radicals and oxygen radical absorbance capacity, while the scavenging activities of ABTS⁺ radical decreased as the fermentation time extended. The antioxidant activities of pancreatin 24‐h hydrolysates were higher than that of the pancreatin 12‐h hydrolysates during the whole LAB fermentation.     

Impact of enzyme preparation and degree of hydrolysis on peptide profile and nitrogen solubility of sodium caseinate hydrolysates

The solubility of casein ingredients is limited under certain pH conditions. Therefore, sodium caseinate (NaCN) was hydrolysed at laboratory scale with four enzyme preparations (Alcalase, Prolyve, FlavorPro Whey and pepsin) yielding hydrolysates having degree of hydrolysis (DH) values between 0.19 ± 0.11 and 19.25 ± 0.73%. The nitrogen solubility index (NSI) over the pH range 2.0–8.0 was affected by (i) the specificity of the enzyme preparation, (ii) the DH and (iii) the presence of unhydrolysed NaCN within the hydrolysate. The trends observed at laboratory scale (<5 L) were also seen with the semi‐pilot‐scale (300 L) hydrolysates generated with Alcalase and FlavorPro Whey. Removal of unhydrolysed NaCN from the FlavorPro Whey hydrolysate using a 5‐kDa ultrafiltration (UF) membrane increased NSI. This study has highlighted the importance of carefully selecting the proteolytic preparation along with the DH to design casein‐based ingredients with enhanced technofunctional properties.     

大豆蛋白酶解肽的分子量分布及抑制ACE活性关系研究

研究不同蛋白酶作用的酶解肽表现在血管紧张素转化酶(ACE)活性抑制差异,酶解产物的水解度、分子量分布与ACE抑制率的相互关系。用胰蛋白酶、胃蛋白酶、中性蛋白酶、木瓜蛋白酶、碱性蛋白酶等五种蛋白酶对大豆分离蛋白酶解,进行了多肽增量、水解度、超滤膜分离及其ACE抑制率对比等实验。结果表明,碱性蛋白酶的多肽增量最大,胃蛋白酶次之,依次为木瓜蛋白酶和中性蛋白酶,胰蛋白酶则出现反常;水解度随着酶解的时间而增加,碱性蛋白酶的最大水解度可达到21%,依次为胃蛋白酶、木瓜蛋白酶和中性蛋白酶,最低为胰蛋白酶仅为9%左右;与此对应的碱性蛋白酶的酶解物的ACE活性抑制率为最高(44.9%),胃蛋白酶次之(43.5%)。分子量范围在1000Da以下组分对ACE的抑制效果最高,碱性蛋白酶作用获得的小分子肽组成为71.25%,胃蛋白酶的为69.35%,但其对应的ACE抑制率却为64.57%和78.49%。中性蛋白酶、木瓜蛋白酶和胰蛋白酶作用获得的小分子肽的ACE抑制率分别为45.7%、47.3%和29.6%。胃蛋白酶的降压肽制备效果为最好,其次为碱性蛋白酶、木瓜蛋白酶、中性蛋白酶和胰蛋白酶。