Effect of residual chymotryptic activity in a trypsin preparation on peptide aggregation in a β-lactoglobulin hydrolysate

Peptide composition and peptide aggregation in β-lactoglobulin (β-LG) hydrolysate were studied as related to residual chymotryptic activity in a commercial trypsin (CT) preparation. Residual chymotryptic activity produced smaller and more hydrophobic peptides in tryptic hydrolysate of β-LG, which enhanced peptide aggregation, mainly at acidic pH. The contribution of the chymotryptic peptide β-LG 15–20 to this aggregation process appeared to be very important, but other peptides (i.e., β-LG 41/43−60, 1–8 and 61−69/70+149−162) and residual α-LA may also decrease peptide solubility. When using CT mixtures in the preparation of whey protein hydrolysates, the impact of residual chymotryptic activity should not be neglected because of its influence on peptide–peptide interactions and on the resulting solubility of the hydrolyzed product.     

Chemical characterisation and determination of sensory attributes of hydrolysates produced by enzymatic hydrolysis of whey proteins following a novel integrative process

The overall aim of this work was to characterise the major angiotensin-converting enzyme (ACE) inhibitory peptides produced by enzymatic hydrolysis of whey proteins, through the application of a novel integrative process. This process consisted of the combination of adsorption and microfiltration within a stirred cell unit for the selective immobilisation of β-lactoglobulin and casein-derived peptides (CDP) from whey. The adsorbed proteins were hydrolysed in situ, which resulted in the separation of peptide products from the substrate and fractionation of peptides. Two different hydrolysates were produced: (i) from CDP (IC₅₀ = 287 μg/mL) and (ii) from β-lactoglobulin (IC₅₀ = 128 μg/mL). The well-known antihypertensive peptide IPP and several novel peptides that have structural similarities with reported ACE inhibitory peptides were identified and characterised in both hydrolysates. Furthermore, the hydrolysates were assessed for bitterness. No significant difference was found between the bitterness of the control (milk with no hydrolysate) and hydrolysate samples at different concentrations (at, below and above the IC₅₀).     

Interaction of whey protein with polyphenols from salal fruits (Gaultheria shallon) and the effects on protein structure and hydrolysis pattern by Flavourzyme®

Milk whey can interact with polyphenols leading to the formation of complexes. In this research, whey protein was fortified with salal fruits (SB) extract and the effect on protein structure was investigated. Particle size and tertiary structure analysis indicates α-lactalbumin–ligand interactions when whey is supplemented with SB extract. Circular dichroism spectroscopy suggests conformational changes of α-Lac to a partially unfolded state as indicated by the decrease in α-helix structures. Enzymatic treatment of whey protein mixed with SB revealed differences in the hydrolysis pattern. LC-MS/MS data analysis indicates that a higher number of peptides are released when whey is mixed with SB. Peptides of known bioactivity were identified in all hydrolysates. The supplementation of whey protein with SB extract can influence protein hydrolysis and the release of peptides following enzymatic treatment with commercial proteases which may affect the functional and health-related properties of the hydrolysate.     

Effect of enzymatic hydrolysis and polysaccharide addition on the β-lactoglobulin adsorption at the air-water interface

The effect of enzymatic hydrolysis and polysaccharide addition on the interfacial adsorption of β-lactoglobulin (β-LG) was investigated in this work. The enzymatic treatment was performed in the hydrolysis degree (HD) range of 0.0-5.0% using bovine α-chymotrypsin II immobilized on agarose beads. Anionic non-surface active polysaccharides (PS), sodium alginate (SA) and λ-carrageenan (λ-C) were studied in the concentration range of 0.0-0.5 wt.%. The adsorption process at the air-water interface was evaluated by means of tensiometry and surface dilatational rheology. Biopolymer interactions in solution were analyzed by extrinsic fluorescence spectroscopy. The enzymatic hydrolysis improved β-LG interfacial properties. On the other hand, at low HD (1.0%), PS addition enhanced surface and elastic properties of β-LG hydrolysate films probably due to a higher repulsion between biopolymers in solution. However, at high HD (3.0-5.0%), SA addition caused a deterioration of surface and elastic properties of β-LG hydrolysate films probably due to the segregation and hydrolysate aggregation in solution, whereas λ-C addition could promote the formation of soluble complexes leading to a better control of elastic properties of β-LG hydrolysate films.     

Identification of dipeptidyl peptidase-IV inhibitory peptides from mare whey protein hydrolysates

Inhibition of dipeptidyl peptidase-IV (DPP-IV) activity is a promising strategy for treatment of type 2 diabetes. In the current study, DPP-IV inhibitory peptides were identi?ed from mare whey protein hydrolysates obtained by papain. The results showed that all the mare whey protein hydrolysates obtained at various hydrolysis durations possessed more potent DPP-IV inhibitory activity compared with intact whey protein. The 4-h hydrolysates showed the greatest DPP-IV inhibitory activity with half-maximal inhibitory concentration of 0.18 mg/mL. The 2 novel peptides from 4-h hydrolysate fractions separated by successive chromatographic steps were characterized by liquid chromatography–electrospray ionization tandem mass spectrometry. The novel peptides Asn-Leu-Glu-Ile-Ile-Leu-Arg and Thr-Gln-Met-Val-Asp-Glu-Glu-Ile-Met-Glu-Lys-Phe-Arg, which corresponded to β-lactoglobulin 1 f(71–77) and β-lactoglobulin 1 f(143–155), demonstrated DPP-IV inhibitory activity with half-maximal inhibitory concentrations of 86.34 and 69.84 μM, respectively. The DPP-IV inhibitory activity of the 2 peptides was retained or even improved after simulated gastrointestinal digestion in vitro. Our findings indicate that mare whey protein-derived peptides may possess potential as functional food ingredients in the management of type 2 diabetes.     

Calcium binding of peptides derived from enzymatic hydrolysates of whey protein concentrate

This study was carried out to investigate the peptides derived from enzymatic hydrolysates of whey protein concentrate. The physiological activity of peptides in whey protein may be used in food additives to promote the absorption of calcium and prevent bone disorders. The whey protein was hydrolysed by trypsin, and the separation of peptides, the properties of hydrolysates and the analysis of the ability to inhibit the formation of calcium phosphates were then investigated. Calcium-binding peptides were produced by tryptic hydrolysis of whey protein concentrate and further purified by precipitation and chromatography on DEAE-cellulose. The hydrolysates were loaded onto an ion-exchange column, followed by stepwise elution with 0, 0.25, 0.5, and 0.75 m NaCl in equilibration buffer to separate the peptides. Trypsin hydrolysates were shown to peak with 0.25 m NaCl and 0.5 m NaCl. The results of SDS-PAGE analysis showed that the peptides with a small molecular weight of about 1.4 to 3.4 kDa were present in the fraction resulting from 0.25 m and 0.5 m NaCl stepwise elution by ion-exchange chromatography of tryptic hydrolysates. The results of this study show that the whey protein hydrolysates produced by the action of trypsin have the ability to inhibit the formation of calcium phosphates.     

PARTIAL CHARACTERIZATION OF A WHEY PROTEIN CONCENTRATE AND ITS ENZYME HYDROLYSATES

A whey protein concentrate (WPC) was produced from fresh whey by ultrafiltration (MW cut off 10 kDa) and lyophilization. Enzymatic hydrolysis was performed with three enzyme systems: pancreatin (PA), protamex (PR) or alcalase 0.6L (AL) to produce hydrolysates with 20% degree of hydrolysis (DH). The peptide profiles of the hydroly sates were determined by high performance capillary electrophoresis (HPCE). The relationship between enzyme system and preferential protein substrate could be established. The alcalase hydrolysate (ALH) differed from the other two hydrolysates, and the enzyme showed the lowest specificity for β‐lactaglobulin. Considering the protein content from WPC the pancreatin hydrolytic system was the most efficient leaving only 4.69% unhydrolyzed protein in the final hydrolysate (PAH). For 20% degree of hydrolysis alcalase left 7.98% unhydrolyzed protein, while protamex left 9.81% unhydrolyzed protein in the final hydrolysate.     

Separation of iron-binding protein from whey through enzymatic hydrolysis

Whey protein concentrate (WPC) was hydrolyzed by nine proteolytic enzymes to examine the effectiveness of the hydrolysates to bind iron. Degree of WPC hydrolysis was higher with pancreatin (13.91%), alcalase (13.60%), and flavourzyme (12.80%) compared with other enzymes (esperase, neutrase, papain, pepsin, protease and trypsin). Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis and reverse-phase high performance liquid chromatography analyses revealed maximum hydrolysis of α-LA and β-LG with alcalase. Molecular masses of peptides derived from alcalase hydrolysate were smaller than 6.5 kDa. Iron-binding by alcalase hydrolysate was the highest (97.6%) of all other hydrolysates. Using ion-exchange chromatography alcalase hydrolysate was eluted at a 0.25 m NaCl gradient concentration with higher iron-binding ability. This eluted fraction had higher Lys (18.09%), Ala (17.24%), and Phe (16.58%) contents. Alcalase showed noticeably better effectiveness than other enzymes to produce a hydrolysate for the separation of iron-binding peptides derived from WPC.     

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.     

Short communication: Tryptic β-casein hydrolysate modulates enteric nervous system development in primary culture

The intestinal tract of the newborn is particularly sensitive to gastrointestinal disorders, such as infantile diarrhea or necrotizing colitis. Perinatal development of the gut also encompasses the maturation of the enteric nervous system (ENS), a main regulator of intestinal motility and barrier functions. It was recently shown that ENS maturation can be enhanced by nutritional factors to improve intestinal maturation. Bioactivity of milk proteins is often latent, requiring the release of bioactive peptides from inactive native proteins. Several casein-derived hydrolysates presenting immunomodulatory properties have been described recently. Furthermore, accumulating data indicate that milk-derived hydrolysate can enhance gut maturation and enrichment of milk formula with such hydrolysates has recently been proposed. However, the capability of milk-derived bioactive hydrolysate to target ENS maturation has not been analyzed so far. We, therefore, investigated the potential of a recently described tryptic β-casein hydrolysate to modulate ENS growth parameters in an in vitro model of rat primary culture of ENS. Rat primary cultures of ENS were incubated with a bioactive tryptic β-casein hydrolysate and compared with untreated controls or to cultures treated with native β-casein or a Prolyve β-casein hydrolysate (Lyven, Colombelles, France). Differentiation of enteric neurons and enteric glial cells, and establishment of enteric neural network were analyzed using immunohistochemistry and quantitative PCR. Effect of tryptic β-casein hydrolysate on bone morphogenetic proteins (BMP)/Smad pathway, an essential regulator of ENS development, was further assessed using quantitative PCR and immunochemistry. Tryptic β-casein hydrolysate stimulated neurite outgrowth and simultaneously modulated the formation of enteric ganglia-like structures, whereas native β-casein or Prolyve β-casein hydrolysate did not. Additionally, treatment with tryptic bioactive β-casein hydrolysate increased the expression of the glial marker glial fibrillary acidic protein and induced profound modifications of enteric glial cells morphology. Finally, expression of BMP2 and BMP4 and activation of Smad1/5 was altered after treatment with tryptic bioactive β-casein hydrolysate. Our data suggests that this milk-derived bioactive hydrolysate modulates ENS maturation through the regulation of BMP/Smad-signaling pathway. This study supports the need for further investigation on the influence of milk-derived bioactive peptides on ENS and intestinal maturation in vivo.     

Emulsifying Property of Whey Peptide Fractions as a Function of pH and ionic Strength

Solubility and emulsifying properties of whey protein concentrate (WPC), heat-treated WPC (90°C, pH 2.5, 10 min), and their tryptic or chymotryptic peptide fractions obtained by ultrafiltration were measured from pH 3 to 9 at two ionic strengths (uμ=0 and 0.6). There was no correlation between solubility and emulsifying capacity (EC). Heat treatment induced some conformational changes in WPC, resulting in better EC. Some peptide fractions had better EC than proteins at pH 7 and 9. Treatments resulting in more hydrophobic peptides or a higheT content of larger peptides produced fractions with better emulsifying properties. pH and ionic strength affected conformation and emulsifying properties of proteins and peptides.     

大黄鱼蛋白肽-钙螯合物的制备及性质研究

目的 制备大黄鱼蛋白肽-钙螯合物,并研究其结构性质。方法 以大黄鱼鱼糜为原料,通过胃蛋白酶、胰蛋白酶、中性蛋白酶进行酶解制备多肽,将肽与钙进行螯合,探究大黄鱼蛋白肽及其肽-钙螯合物的性质功能。结果 3种酶解肽中,胰酶肽钙螯合含量最高,胰酶肽的粒径最小,肽-钙螯合物相比于肽的粒径都有所减小,说明肽-钙螯合物是一种致密的纳米颗粒并呈现出折叠多孔的网络结构。这些变化可归因于肽和钙离子之间的相互作用。紫外吸收光谱、荧光光谱、红外图谱和圆二色光谱的结果表明大黄鱼蛋白肽与钙离子螯合,生成了新的肽-钙螯合物。结论 3种酶解肽中,胰蛋白酶水解物的钙离子结合能力在3种蛋白酶获得的水解物中最高。因此,在本研究中胰蛋白酶被认为是制备大黄鱼蛋白水解物钙离子结合活性肽的最优酶制剂,为大黄鱼的高值化利用和新型钙制剂的开发提供了理论依据和实验参考。     

Rheological and structural characterization of gels from whey protein hydrolysates/locust bean gum mixed systems

The gelling ability of whey proteins can be changed by limited hydrolysis and by the addition of other components such as polysaccharides. In this work the effect of the concentration of locust bean gum (LBG) on the heat-set gelation of aqueous whey protein hydrolysates (10% w/w) from pepsin and trypsin was assessed at pH 7.0. Whey protein concentrate (WPC) mild hydrolysis (up to 2.5% in the case of pepsin and 1.0% in the case of trypsin) ameliorates the gelling ability. The WPC synergism with LBG is affected by the protein hydrolysis. For a WPC concentration of 10% (w/w), no maximum value was found in the G′ dependence on LBG content in the case of the hydrolysates, unlike the intact WPC. However, for higher protein concentrations, the behaviour of gels from whey proteins or whey protein hydrolysates towards the presence of LBG becomes very similar. In this case, a small amount of LBG in the presence of salt leads to a big enhancement in the gel strength. Further increases in the LBG concentration led to a decrease in the gel strength.     

Innovative applications of high-intensity ultrasound in the development of functional food ingredients: Production of protein hydrolysates and bioactive peptides

In recent years, the research on functional peptide generation for the development of functional foods has focused, among other issues, on the enhancement of enzymatic hydrolysis by means of high-intensity ultrasound (HIU) application. It has been suggested that the use of HIU in pretreatment and during the hydrolysis process can modify protein conformation by affecting hydrogen bonds and hydrophobic interactions, disrupting the quaternary and/or tertiary structure of proteins due to the effects of cavitation. Therefore, these structural modifications may expose more hydrolysis sites to be accessible by the enzyme, causing an increase in degree of hydrolysis and bioactivity. The main objective of this work was to review recent advances in food science and technology on the application of HIU for the enhancement of enzymatic hydrolysis of proteins of both animal and plant origin in order to generate novel protein hydrolysates and bioactive peptides which could be used as functional-food ingredients.     

Characteristic Property of Low Bitterness in Protein Hydrolysates by a Novel Soybean Protease D3

ABSTRACT:  Enzymatic hydrolysis is 1 means of improving the functional properties of food protein; however, in most cases, bitter peptides are generated by such treatment, and the resulting product is therefore not acceptable as a food ingredient. We have already reported a novel cysteine protease, D3, purified from germinating soybean cotyledons. Because of its substrate specificities, most hydrophobic amino acid residues in the hydrolysate are presumed not to be located at the peptide termini. It was therefore expected that protein hydrolysate by protease D3 would taste less bitter than other enzymatic hydrolysates. The objective of this study was to demonstrate the low bitterness of protein hydrolysates by protease D3. For that purpose, soy protein and casein hydrolysates were prepared with treatment of protease D3, subtilisin, pepsin, trypsin, and thermolysin, respectively. The bitterness of these hydrolysates was evaluated by measuring points of subjective equality (PSE). The PSE value demonstrated that the protein hydrolysates by protease D3 were significantly less bitter than the other enzymatic hydrolysates, indicating that the products had a taste mild enough to be acceptable as a less-bitter peptide food ingredient. These results suggested that a prominent feature of protease D3 was its capacity to produce less-bitter peptides. Therefore, it is thought that protease D3 could be applied to produce protein hydrolysates for use as ingredients in a variety of food products.     

Influence of enzymatic hydrolysis,pH and storage temperature on the emulsifying properties of canola protein isolate and hydrolysates

The aim of this work was to enhance emulsification properties of canola proteins through enzymatic proteolysis and pH variaton. Canola protein isolate (CPI) and hydrolysates (CPHs) were used to form emulsions at pH 4.0, 7.0 and 9.0 followed by storage at 4 or 25 °C for 7 days. Controlled enzymatic hydrolysis led to increased peptide bond cleavage with time (0.23 g/100 g in CPI to 7.18 g/100 g after 24‐h Alcalase hydrolysis). Generally, oil droplet sizes were smaller for emulsions made at pH 9.0, which suggest better quality than those made at pH 4.0 and 7.0. Trypsin hydrolysate emulsions were the most physically stable at pH 7.0 and 9.0; in contrast, the pepsin hydrolysate emulsions were unstable at all conditions. The results suggest that selective enzymatic hydrolysis could play an important role in enhancing successful incorporation of canola proteins and peptides into food systems as protein emulsifiers.     

Foaming characteristics of β-lactoglobulin as affected by enzymatic hydrolysis and polysaccharide addition: Relationships with the bulk and interfacial properties

The objective of this work was to study the effect of enzymatic hydrolysis and polysaccharide addition on the foaming characteristics of β-lactoglobulin (β-LG). Enzymatic treatment was performed in the hydrolysis degree (HD) range of 0.0–5.0% using bovine α-chymotrypsin II immobilized on agarose microbeads. Anionic non-surface active polysaccharides (PS), sodium alginate (SA) and λ-carrageenan (λ-C) were studied in the concentration range of 0.0–0.5 wt.%. Foaming characteristics were determined by conductimetric and optical methods and were linked to protein diffusion kinetics, film mechanical properties and biopolymer molecular dynamics in solution. Experiments were performed at constant temperature (20 °C), pH 7 and ionic strength 0.05 M. Limited hydrolysis improved the formation and stability of β-LG foam possibly due to an increased protein diffusion rate and film dilatational elasticity. Furthermore, PS addition caused different effects on β-LG foaming characteristics depending on the PS type, their relative concentration and extent of enzymatic treatment (HD). Diffusion rate and interfacial rheological behavior of mixed systems could exert a decisive role in foaming characteristics of β-LG and its hydrolysates in close connection with biopolymer interactions in solution, e.g., macromolecule repulsion, protein segregation/aggregation and soluble complexes formation.     

Preparation of whey protein hydrolysates with ACE-inhibitory activity using cysteine peptidases from Bromelia hieronymi Mez. (Bromeliaceae)

A proteolytic extract from fruits of Bromelia hieronymi was used to hydrolyse bovine whey proteins. The peptide profile obtained exhibited a gradual fading of the main whey proteins and a decrease in the content of hydrophobic peptides. The 180-min hydrolysate showed a hydrolysis degree of 15.2% and inhibited the angiotensin-converting enzyme (ACE), showing an IC₅₀ of 0.17 mg/mL. By bioinformatics analysis, several theoretical sequences of possible ACE-inhibitory peptides were deduced. These results showed that proteolytic extracts from B. hieronymi can be used to prepare whey hydrolysates that would serve as a potential bioactive ingredient of functional foods.     

Interactions of milk proteins during heat and high hydrostatic pressure treatments — A Review

Pressure treatment of β-lactoglobulin (β-LG), whey protein concentrate (WPC), whey protein isolate and skim milk has been explored by many groups using a wide range of techniques. In general terms, heat treatment and pressure treatment have similar effects: denaturing and aggregating the whey proteins and diminishing the number of viable microorganisms. However, there are significant differences between the effects of the two treatments on protein unfolding and the subsequent thiol-catalysed disulfide-bond interchanges that lead to different structures and product characteristics. Application of a range of techniques has given insight into the subtle differences between the pathways from native proteins to the final product mix. This review covers some of the techniques used and their strengths, and the probable pathways from native protein to the final products. β-LG is one of the most pressure-sensitive proteins and α-lactalbumin (α-LA) is one of the most pressure resistant. In a heated WPC system, bovine serum albumin is very sensitive and β-LG is more resistant. In a heated milk system, β-LG reacts with κ-casein (κ-CN) and not with α_S2-CN, but, in pressure-treated milk, β-LG forms adducts with either κ-CN or α_S2-CN. In both treatments, the role of β-LG is central to the ongoing reactions, involving α-LA and κ-CN in heated systems but involving κ-CN, α_S2-CN and α-LA in pressurized systems.Industrial relevanceHigh hydrostatic pressure (HHP) processing, as opposed to heat treatment, has received much attention recently as a means of processing milk proteins. This review examines the differences in the denaturation pathways that give rise to different final products.     

分步酶解小麦面筋蛋白制备低苦味肽粉的研究

目的 研究分步酶解小麦面筋蛋白(wheat gluten, WG)制备低苦味肽粉的工艺。方法 选用中性蛋白酶、木瓜蛋白酶、胃蛋白酶水解WG至8%水解度,接着用风味蛋白酶对水解产物进行脱苦处理,对不同酶解产物中苦味肽的特性进行系统研究,探究苦味肽含量、氨基酸组成、分子量分布、表面疏水性等指标变化对WG酶解物苦味值的影响,对比风味蛋白酶对不同单酶酶解物的脱苦效果差异,分析风味蛋白酶对WG酶解物脱苦的内在机理,进而确定制备低苦味小麦蛋白肽粉的最佳酶解工艺。结果 中性蛋白酶的酶解产物经风味蛋白酶作用后,脱苦效果最显著,苦味肽苦味值从4.08降至2.25,酶解产物的苦味值可下降56.42%。木瓜蛋白酶的酶解产物经风味蛋白酶作用4 h后,酶解产物的苦味值最低,制备出苦味值为1.28的WG低苦味肽粉。结论 经分步酶解作用后,酶解产物中苦味肽的含量下降;疏水性氨基酸比例的下降和游离氨基酸含量的升高引起苦味肽苦味阈值的增大,共同导致酶解产物苦味值显著降低,该研究为酶解脱苦技术的快速发展和WG活性肽工业化生产提供新的参考。