Characterization of proteases from Irpex lacteus grown on minimally denatured soybean meal.

BACKGROUND: Acid and thermal stabilities are important properties for the preparation of acidic protein beverage. It is an important method for enzymatic modification to improve the functional properties of protein. Irpex lacteus protease showed a selective hydrolysis to soy proteins. The purpose of this study was to investigate the mechanism of enzymatic hydrolysis and its effects on acid and thermal stabilities of soy proteins. RESULTS: The I. lacteus protease selectively hydrolyzed the α and α' subunits of the native soybean ß-conglycinin (7S globulin) to produce products that presented as the 55 kDa band upon sodium dodecyl sulfate polyacrylamide gel electrophoresis. The amino acid sequences of 55 kDa polypeptides were analyzed in gel multi-enzyme digestion followed by liquid chromatography-mass spectrometry. By matching the multi-enzyme digestion peptides with the published polypeptide chain sequences of the α and α' subunits, it was confirmed that the 55 kDa polypeptides were formed by eliminating amino acid residues on both sides of the N- and C-terminals. From the published protein structure database (https://www.uniprot.org/), it is known that the cleaved peptide bonds were in extension regions. Non-selective enzyme hydrolysis of both ß-conglycinin (7S globulin) and glycinin (11S globulin), with corresponding drastic increases in the degree of hydrolysis, was observed when the substrates were preheated to the denaturation degree of 40% and above. However, 55 kDa hydrolyzed products and B polypeptides showed some extent of resistance to the proteolysis by I. lacteus protease even if denaturation degree was 100%. Both selective and non-selective hydrolysis of soy proteins by I. lacteus protease improved the acid and heat stabilities under the same hydrolysis conditions (enzyme/substrate ratio, time, and temperature). CONCLUSION: Enzymatic hydrolysis of soybean proteins by the I. lacteus protease can effectively improve the acid and thermal stabilities of proteins. This discovery is significant to avoid aggregation during processing in the beverage industry. In the near future, the protease has potential application value for modification of other proteins. © 2022 Society of Chemical Industry.

Enzymatic preparation and potential applications of agar oligosaccharides: a review.

Oligosaccharides derived from agar, that is, agarooligosaccharides and neoagarooligosaccharides, have demonstrated various kinds of bioactivities which have been utilized in a variety of fields. Enzymatic hydrolysis is a feasible approach that principally allows for obtaining specific agar oligosaccharides in a sustainable way at an industrial scale. This review summarizes recent technologies employed to improve the properties of agarase. Additionally, the relationship between the degree of polymerization, bioactivities, and potential applications of agar-derived oligosaccharides for pharmaceutical, food, cosmetic, and agricultural industries are discussed. Engineered agarase exhibited general improvement of enzymatic performance, which is mostly achieved by truncation. Rational and semi-rational design assisted by computational methods present the latest strategy for agarase improvement with greatest potential to satisfy future industrial needs. Agarase immobilized on magnetic Fe3O4 nanoparticles via covalent bond formation showed characteristics well suited for industry. Additionally, albeit with the relationship between the degree of polymerization and versatile bioactivities like anti-oxidants, anti-inflammatory, anti-microbial agents, prebiotics and in skin care of agar-derived oligosaccharides are discussed here, further researches are still needed to unravel the complicated relationship between bioactivity and structure of the different oligosaccharides.

Study on the Performance of Liquid-Solid Contact Resistance Based on Magnetohydrodynamic Micro-Angular Vibration Sensor.

In this paper, the influence of the contact resistance between the conductive fluid and the metal electrode on the output characteristics of the magnetic fluid micro-angular vibration sensor (MHD sensor) is theoretically analyzed. The contact resistance models based on the solid-solid electric contact theory are established based on the resistivity, temperature, pressure and angular vibration of the materials between the conductive fluid and the metal electrode. The contact resistance was tested by setting up an experimental platform and making conductive fluid rings with electrode materials of Ag, Cu and Ti. The results show that the static contact resistance between the conductive fluid and the metal electrode is positively correlated with the material resistivity and temperature, and negatively correlated with the surface roughness and contact pressure of the metal electrode. The dynamic contact resistance fluctuation is proportional to the amplitude of the input voltage of the angle shaker and inversely proportional to the square of the input frequency. At the same time, reducing contact resistance can improve the MHD sensor's performance.

Single-Channel Blind Signal Separation of the MHD Linear Vibration Sensor Based on Singular Spectrum Analysis and Fast Independent Component Analysis.

An MHD vibration sensor, as a new type of sensor used for vibration measurements, meets the technical requirements for the low-noisy measurement of acceleration, velocity, and micro-vibration in spacecraft during their development, launch, and orbit operations. A linear vibration sensor with a runway type based on MHD was independently developed by a laboratory. In a practical test, its output signal was mixed with a large amount of noise, in which the continuous narrowband interference was particularly prominent, resulting in the inability to efficiently carry out the real-time detection of micro-vibration. Considering the high interference of narrowband noise in linear vibration signals, a single-channel blind signal separation method based on SSA and FastICA is proposed in this study, which provides a new strategy for linear vibration signals. Firstly, the singular spectrum of the linear vibration signal with noise was analyzed to suppress the narrowband interference in the collected signal. Then, a FastICA algorithm was used to separate the independent signal source. The experimental results show that the proposed method can effectively separate the useful linear vibration signals from the collected signals with low SNR, which is suitable for the separation of the MHD linear vibration sensor and other vibration measurement sensors. Compared with EEMD, VMD, and wavelet threshold denoising, the SNR of the separated signal is increased by 10 times on average. Through the verification of the actual acquisition of the linear vibration signal, this method has a good denoising effect.

Depth-Keeping Control for a Deep-Sea Self-Holding Intelligent Buoy System Based on Inversion Time Constraint Stability Strategy Optimization.

Based on the nonlinear disturbance observer (NDO), the inversion time-constraint stability strategy (ITCS) is designed to make the deep-sea self-holding intelligent buoy (DSIB) system hovered at an appointed depth within a specified time limit. However, it is very challenging to determine the optimal parameters of an ITCS depth controller. Firstly, a genetic algorithm based on quantum theory (QGA) is proposed to obtain the optimal parameter combination by using the individual expression form of quantum bit and the adjustment strategy of quantum rotary gate. To improve the speed and accuracy of global search in the QGA optimization process, taking the number of odd and even evolutions as the best combination point of the genetic and chaos particle swarm algorithm (GACPSO), an ITCS depth controller based on GACPSO strategy is proposed. Besides, the simulations and hardware-in-the-loop system experiments are conducted to examine the effectiveness and feasibility of the proposed QGA-ITCS and GACPSO-ITCS depth controller. The results show that the proposed GACPSO-ITCS depth controller provides higher stability with smaller steady-state error and less settling time in the depth-control process. The research of the proposed method can provide a stable operation condition for the marine sensors carried by the DSIB.

A Novel System for the Measurement of an Evaporation Duct Using the Magnetic Coupling Principle for Power Feeding and Data Transmission.

Since the evaporation duct height (EDH) only covers the antenna height of most shipborne microwave radars, mastering the EDH in advance has great significance in achieving long-range target detection. In this paper, a set of hydrological and meteorological sensors based on the gradient meteorological instrument (GMI) were built to monitor the evaporation duct of the South China Sea (SCS). However, the monitoring needed to be interrupted during the battery replacement of the sensor, which could result in the loss of some important data collection. On the basis of the inductively coupled power transfer (ICPT) technology, the resonance principle was used to compensate the inductive reactance on the closed steel ring (CSR), and the energy stored in the super capacitor was introduced for data collection and return. A novel measuring system for the detection of an evaporation duct was proposed. To avoid iterative calculation by setting the initial value of the current evaporation duct models in large-scale and multi time evaporation duct prediction and diagnosis, on the basis of the non-iterative air-sea flux (NAF) model, the EDH was obtained by introducing the K theoretical flux observation method into the atmospheric refractive index equation. Finally, preliminary experimental results are presented for the detection of evaporation duct to demonstrate the feasibility and effectiveness of the proposed system. The communication accuracy rate of the proposed system was 99.7%. The system transmission power reached 22.8 W. The research results of the NAF model adaptability showed that the mean value of the EDH was 8.7 m, which was lower than the mean EDH of the SCS. The EDH calculated by the NAF model in the unstable air-sea stratification state was slightly lower than that calculated by the NPS model. The diagnosis of the EDH by the NAF model was similar to that of the NPS model, but the calculation stability of the NAF model was better.

An On-Orbit Dynamic Calibration Method for an MHD Micro-Angular Vibration Sensor Using a Laser Interferometer.

The magnetohydrodynamic (MHD) micro-angular vibration sensor is a significant component of the MHD Inertial Reference Unit (MIRU) and measures micro-amplitude and wide frequency angular vibration. The MHD micro-angular vibration sensor must be calibrated in orbit since the ground calibration parameters may change after lift-off. An on-orbit dynamic calibration method for the MHD micro-angular vibration sensor is proposed to calibrate the complex sensitivity of the sensor in high frequency. An absolute calibration method that combines a homodyne laser interferometer and an angular retroreflector was developed. The sinusoidal approximation method was applied, and the calibration system was established and tested using a manufactured MHD sensor. Furthermore, the measurement principle and installation errors were analyzed, including the eccentric installation error of the retroreflector, the tilt installation error of the retroreflector, and the optical path tilt error. This method can be realized within a rotation range of ± 3 ∘ and effectively avoid the installation error caused by mechanical errors. The results indicate that the calibratable angular vibration frequency range is 25-800 Hz, and the angular velocity range is 0.076 -7590 mrad/s. The expanded uncertainties of the sensitivity amplitude and phase shift of the calibration system for the MHD micro-angular sensor are 0.04 % and 1.2 ∘ ( k = 2 ) .

Novel Effect of Zinc Nitrate/Vanadyl Oxalate for Selective Catalytic Oxidation of α-Hydroxy Esters to α-Keto Esters with Molecular Oxygen: An In Situ ATR-IR Study.

Selective oxidation of α-hydroxy esters is one of the most important methods to prepare high value-added α-keto esters. An efficient catalytic system consisting of Zn(NO3)2/VOC2O4 is reported for catalytic oxidation of α-hydroxy esters with molecular oxygen. Up to 99% conversion of methyl DL-mandelate or methyl lactate could be facilely obtained with high selectivity for its corresponding α-keto ester under mild reaction conditions. Zn(NO3)2 exhibited higher catalytic activity in combination with VOC2O4 compared with Fe(NO3)3 and different nitric oxidative gases were detected by situ attenuated total reflection infrared (ATR-IR) spectroscopy. UV-vis and ATR-IR results indicated that coordination complex formed in Zn(NO3)2 in CH3CN solution was quite different from Fe(NO3)3; it is proposed that the charge-transfer from Zn2+ to coordinated nitrate groups might account for the generation of different nitric oxidative gases. The XPS result indicate that nitric oxidative gas derived from the interaction of Zn(NO3)2 with VOC2O4 could be in favor of oxidizing VOC2O4 to generate active vanadium (V) species. It might account for different catalytic activity of Zn(NO3)2 or Fe(NO3)3 combined with VOC2O4. This work contributes to further development of efficient aerobic oxidation under mild reaction conditions.

Analysis of the Output Characteristics of a Novel Small-Angle Transducer Used in High-Precision Inertial Sensors.

This paper presents the design of a novel small-angle transducer characterized by a simple structure, fast response and very low reaction torque. A theoretical model is presented which describes the linear relationship between the output voltage and the angular displacement when the rotor rotates away from the null position. By analysis of the theoretical model, it is revealed that the small-angle transducer possesses a very high linearity within ±4° and a high sensitivity (approximately 0.34 V/°), and the parameters affecting output characteristics can be obtained. Furthermore, it is found that the transducer sensitivity can be improved by optimizing the load impedance and excitation frequency. These findings are verified by numerical evaluations. In addition, the established theoretical model and simulation analysis provide a quantitative method for analyzing the output characteristics of the novel small-angle transducer.

Error Analysis of Magnetohydrodynamic Angular Rate Sensor Combing with Coriolis Effect at Low Frequency.

The magnetohydrodynamic (MHD) angular rate sensor (ARS) with low noise level in ultra-wide bandwidth is developed in lasing and imaging applications, especially the line-of-sight (LOS) system. A modified MHD ARS combined with the Coriolis effect was studied in this paper to expand the sensor’s bandwidth at low frequency (<1 Hz), which is essential for precision LOS pointing and wide-bandwidth LOS jitter suppression. The model and the simulation method were constructed and a comprehensive solving method based on the magnetic and electric interaction methods was proposed. The numerical results on the Coriolis effect and the frequency response of the modified MHD ARS were detailed. In addition, according to the experimental results of the designed sensor consistent with the simulation results, an error analysis of model errors was discussed. Our study provides an error analysis method of MHD ARS combined with the Coriolis effect and offers a framework for future studies to minimize the error.

New insights into the humic acid fouling mechanism of ultrafiltration membranes for different Ca2+ dosage ranges: results from micro- and macro-level analyses.

To reveal the mechanisms of the influence of Ca2+ on membrane fouling with humic acid (HA), the adhesion forces of HA with both other HA molecules and the membrane, the HA fouling layer structure, HA fouling experiments, and the HA rejections at a wide range of Ca2+ dosages were investigated. The results indicated that the effect of Ca2+ on HA fouling can be divided into three stages. At lower ionic strength (IS) of CaCl2, the change in electrostatic forces is the main factor in controlling HA fouling behavior; i.e., increasing Ca2+ dosages resulted in more serious membrane fouling. When the IS of CaCl2 reached 10 mM, HA aggregates became the dominant factor in the fouling process, which could result in a porous fouling layer accompanied by less membrane fouling. Interestingly, much weaker membrane fouling was observed when the IS increased to 100 mM and the HA rejection began to decline. This was because a stronger hydration repulsion force was generated, which could weaken the compactness of the fouling layer and the adhesion forces of HA with both the membrane and HA, while enabling smaller-sized HA to pass more easily into the permeate, which led to less membrane fouling and a lower HA rejection.

Sol-gel encapsulation of pullulanase in the presence of hybrid magnetic (Fe3O4-chitosan) nanoparticles improves thermal and operational stability.

Pullulanase was sol-gel encapsulated in the presence of magnetic chitosan/Fe3O4 nanoparticles. The resulting immobilized pullulanase was characterized by scanning electron microscopy, vibrating sample magnetometry, Fourier transform infrared spectroscopy and thermogravimetric analysis. The results showed that the addition of pullulanase created a more regular surface on the sol-gel matrix and an enhanced magnetic response to an applied magnetic field. The maximal activity retention (83.9%) and specific activity (291.7 U/mg) of the immobilized pullulanase were observed under optimized conditions including an octyltriethoxysilane:tetraethoxysilane (OTES:TEOS) ratio of 1:2 and enzyme concentration of 0.484 mg/mL sol. The immobilized enzyme exhibited good thermal stability. When the temperature was above 60 °C, the immobilized pullulanase showed significantly higher activity than the free enzyme (p < 0.01); enzyme immobilized by simple sol-gel encapsulation and co-immobilized by crosslinking-encapsulation retained 52 and 69% of their initial activity after 5 h at 62 °C, respectively, compared to 11% for the free enzyme. Moreover, the stability of the pullulanase was improved by crosslinking-encapsulation, as the enzyme retained more than 85 and 81% of its original activity after 5 and 6 consecutive reuses, respectively, compared to 80 and 72% of its original activity for simple sol-gel encapsulated enzymes. This indicated the leakage of enzyme molecules through the pores of the gel was substantially abated by cross-linking. Such immobilized pullulanase provides high stability and ease of enzyme recovery, characteristics that are advantageous for applications in the food industry that involve continuous starch processing.

Theoretical and Experimental Study of Radial Velocity Generation for Extending Bandwidth of Magnetohydrodynamic Angular Rate Sensor at Low Frequency.

The magnetohydrodynamics angular rate sensor (MHD ARS) has received much attention for its ultra-low noise in ultra-broad bandwidth and its impact resistance in harsh environments; however, its poor performance at low frequency hinders its work in long time duration. The paper presents a modified MHD ARS combining Coriolis with MHD effect to extend the measurement scope throughout the whole bandwidth, in which an appropriate radial flow velocity should be provided to satisfy simplified model of the modified MHD ARS. A method that can generate radial velocity by an MHD pump in MHD ARS is proposed. A device is designed to study the radial flow velocity generated by the MHD pump. The influence of structure and physical parameters are studied by numerical simulation and experiment of the device. The analytic expression of the velocity generated by the energized current drawn from simulation and experiment are consistent, which demonstrates the effectiveness of the method generating radial velocity. The study can be applied to generate and control radial velocity in modified MHD ARS, which is essential for the two effects combination throughout the whole bandwidth.

Injected current reshaping in distance measurement by laser self-mixing interferometry.

Self-mixing interferometry (SMI), as an extremely simple and compact laser range finding technique, is especially appropriate to develop minitype sensors for narrow space and small precision parts. In order to enhance the distance resolution performance of this technique, we described the mechanism of nonlinearity in laser frequency under injected current tuning, and proposed a current reshaping method to linearize the laser frequency to attain higher resolution in the scheme of SMI. The proof of nonlinearity was obtained through numerical simulation by considering the change of temperature and carrier concentration and experiment by complex wavelet analysis. The current reshaping method, based on the experimental data of wavelength versus injected current, was proposed to suppress the nonlinearity and improve the distance resolution to better than 20 µm over the range of 2.4-20.4 cm. The influence of tuning parameters and other sources of error was discussed additionally.

The optical property of core-shell nanosensors and detection of atrazine based on localized surface plasmon resonance (LSPR) sensing.

Three different nanosensors with core-shell structures were fabricated by molecular self-assembly and evaporation techniques. Such closely packed nanoparticles exhibit fine optical properties which are useful for biochemical sensing. The refractive index sensitivity (RIS) of nanosensors was detected by varying the refractive index of the surrounding medium and the decay length of nanosensors was investigated using a layer-by-layer polyelectrolyte multilayer assembly. The results showed that the thickness of the Au shell plays an important role in determining the RIS and the decay length. A system based on localized surface plasmon resonances (LSPR) sensing was constructed in our study. The core-shell nanosensors can detect 10 ng/mL atrazine solutions and are suitable for pesticide residue detection.

A ß-Primeverosidase-like Enzyme in Soybean [Glycine max (L.) Merr] Hypocotyls: Specificity toward 1-Octen-3-yl and 3-Octanyl ß-Primeverosides.

A novel ß-primeverosidase-like enzyme, originating from the hypocotyl of soybeans, was isolated and characterized. This enzyme, with an estimated molecular weight of 44 kDa, was identified as a monomer and exhibited peak activity at 55 °C and pH 5.5. It demonstrated a specific and efficient hydrolysis of 1-octen-3-yl ß-primeveroside (1-octen-3-yl prim) and 3-octanyl ß-primeveroside (3-octanyl prim) but did not act on glucopyranosides. Mn2+ significantly enhanced its activity, while Zn2+, Cu2+, and Hg2+ exerted inhibitory effects. Kinetic analysis revealed a higher hydrolytic capacity toward 1-octen-3-yl prim. Partial amino acid sequences were determined and the N-terminal amino acid sequence was determined to be AIVAYAL ALSKRAIAAQ. The binding energy and binding free energy between the ß-primeverosidase enzyme and its substrates were observed to be higher than that of ß-glucosidase, thus validating its superior hydrolysis efficiency. Hydrogen bonds and hydrophobic interactions were the main types of interactions between ß-primeverosidase enzyme and 1-octen-3-yl prim and 3-octanyl prim, involving amino acid residues such as GLU-470, TRP-463, GLU-416, TRP-471, GLN-53, and GLN-477 (hydrogen bonds) and PHE-389, TYR-345, LEU-216, and TYR-275 (hydrophobic interactions). This study contributes to the application of a ß-primeverosidase-like enzyme in improving the release efficiency of glycosidically conjugated flavor substances.

Fermentation Characteristics, Antinutritional Factor Level and Flavor Compounds of Soybean Whey Yogurt.

Soybean whey contains high levels of off-flavors and anti-nutritional factors and is generally considered unsuitable for direct application in the food industry. In this work, to reduce beany off-flavors and anti-nutritional factors, and to improve its fermentation characteristics, soybean whey was treated with electrodialysis desalination, vacuum concentration and lactic acid bacteria (LAB) fermentation. The results showed that electrodialysis desalination increased the fermentation rate and the number of viable lactic acid bacteria of soybean whey yogurt. More than 90% of the antinutritional factor level (urease and trypsin inhibitory activity) was removed due to high-temperature denaturation inactivation and LAB degradation. Concentrated desalted soybean whey yogurt (CDSWY) possessed larger values for firmness and consistency, and a denser network microstructure compared with undesalted yogurt. Over 90% of off-flavors including hexanal, 1-octen-3-ol and 1-octen-3-one were removed after electrodialysis desalination and concentration treatment. Meanwhile, the newly generated ß-damascenone through carotenoid degradation and 2,3-butanedione improved the pleasant flavor and sensory quality of CDSWY, while the salty taste of CSWY lowered its sensory quality. This study provided a theoretical basis for better utilization of soybean whey to develop a plant-based yogurt like dairy yogurt.

Improving the thermal stability and branching efficiency of Pyrococcus horikoshii OT3 glycogen branching enzyme.

In practical applications, the gelatinisation temperature of starch is high. Most current glycogen branching enzymes (GBEs, EC 2.4.1.18) exhibit optimum activity at moderate or low temperatures and quickly lose their activity at higher temperatures, limiting the application of GBEs in starch modification. Therefore, we used the PROSS strategy combined with PDBePISA analysis of the dimer interface to further improve the heat resistance of hyperthermophilic bacteria Pyrococcus horikoshii OT3 GBE. The results showed that the melting temperature of mutant T508K increased by 3.1 °C compared to wild-type (WT), and the optimum reaction temperature increased by 10 °C for all mutants except V140I. WT almost completely lost its activity after incubation at 95 °C for 60 h, while all of the combined mutants maintained >40 % of their residual activity. Further, the content of the α-1,6 glycosidic bond of corn starch modified by H415W and V140I/H415W was approximately 2.68-fold and 1.92-fold higher than that of unmodified corn starch and corn starch modified by WT, respectively. Additionally, the glucan chains of DP < 13 were significantly increased in mutant modified corn starch. This method has potential for improving the thermal stability of GBE, which can be applied in starch branching in the food industry.

Advances in alginate lyases and the potential application of enzymatic prepared alginate oligosaccharides: A mini review.

Alginate is mainly a linear polysaccharide composed of randomly arranged ß-D-mannuronic acid and α-L-guluronic acid linked by α, ß-(1,4)-glycosidic bonds. Alginate lyases degrade alginate mainly adopting a ß-elimination mechanism, breaking the glycosidic bonds between the monomers and forming a double bond between the C4 and C5 sugar rings to produce alginate oligosaccharides consisting of 2-25 monomers, which have various physiological functions. Thus, it can be used for the continuous industrial production of alginate oligosaccharides with a specific degree of polymerization, in accordance with the requirements of green exploitation of marine resources. With the development of structural analysis, the quantity of characterized alginate lyase structures is progressively growing, leading to a concomitant improvement in understanding the catalytic mechanism. Additionally, the use of molecular modification methods including rational design, truncated expression of non-catalytic domains, and recombination of conserved domains can improve the catalytic properties of the original enzyme, enabling researchers to screen out the enzyme with the expected excellent performance with high success rate and less workload. This review presents the latest findings on the catalytic mechanism of alginate lyases and outlines the methods for molecular modifications. Moreover, it explores the connection between the degree of polymerization and the physiological functions of alginate oligosaccharides, providing a reference for enzymatic preparation development and utilization.

Identification, rapid screening, docking mechanism and in vitro digestion stability of novel DPP-4 inhibitory peptides from wheat gluten with ginger protease.

To better understand the hypoglycemic potential of wheat gluten (WG), we screened dipeptidyl peptidase IV (DPP-4) inhibitory active peptides from WG hydrolysates. WG hydrolysates prepared by ginger protease were found to have the highest DPP-4 inhibitory activity among the five enzymatic hydrolysates, from which a 1-3 kDa fraction was isolated by ultrafiltration. Further characterization of the fraction with nano-HPLC-MS/MS revealed 1133 peptides. Among them, peptides with P'2 (the second position of the N-terminal) and P2 (the second position of the C-terminal) as proline residues (Pro) accounted for 12.44% and 43.69%, respectively. The peptides including Pro-Pro-Phe-Ser (PPFS), Ala-Pro-Phe-Gly-Leu (APFGL), and Pro-Pro-Phe-Trp (PPFW) exhibited the most potent DPP-4 inhibitory activity with IC50 values of 56.63, 79.45, and 199.82 µM, respectively. The high inhibitory activity of PPFS, APFGL, and PPFW could be mainly attributed to their interaction with the S2 pocket (Glu205 and Glu206) and the catalytic triad (Ser630 and His740) of DPP-4, which adopted competitive, mixed, and mixed inhibitory modes, respectively. After comparative analysis of PPFS, PPFW, and PPF, Ser was found to be more conducive to enhancing the DPP-4 inhibitory activity. Interestingly, peptides with P2 as Pro also exhibited good DPP-4 inhibitory activity. Meanwhile, DPP-4 inhibitory peptides from WG showed excellent stability, suggesting a potential application in type 2 diabetes (T2DM) therapy or in the food industry as functional components.