Hydrolysis of Soybean Milk Protein by Papain: Antioxidant, Anti-Angiotensin, Antigenic and Digestibility Perspectives.

The objective of the investigation was to understand the biochemical activities of hydrolysate of soybean milk protein (SMP). Hydrolysis was carried out by different concentrations of papain (0.008 g·L-1, 0.016 g·L-1, 0.032 g·L-1 and 0.064 g·L-1). The antioxidant capacity was measured by the ferric-reducing ability of plasma (FRAP) and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assays. The anti-angiotensin activity of hydrolysate was measured by the recombinant angiotensin converting enzyme and substrate Abz-FRK(Dnp)-P. The contributions of the Kunitz trypsin inhibitor (KTI) and Bowman-Birk inhibitor (BBI) on antigenicity, and the in vitro digestion of papain-hydrolyzed SMP were studied. Rabbit polyclonal anti-KTI and anti-BBI antibodies together with peroxidase-labelled goat anti-Rb IgG secondary antibody were used to identify the antigenicity of KTI and BBI in unhydrolyzed and papain-hydrolyzed SMP. The antioxidant capacity and anti-angiotensin activity of SMP were increased after the papain hydrolysis of SMP. The KTI- and BBI-specific antigenicity were reduced in SMP by increasing the concentration of papain. However, there was interaction between papain-hydrolyzed SMP and trypsin in native gel, while interaction with chymotrypsin was absent. The interaction between trypsin and SMP was reduced due to the hydrolysis of papain in a concentration-dependent manner. According to the in vitro gastrointestinal digestion simulation protocol (Infogest), the digestibility of SMP was not statistically increased.

Effect of Combination of Time and Temperature on Quality Characteristics of Sous Vide Chicken Breast.

The use of minimal thermal processing techniques such as sous vide technology to improve the quality of meat-based foods has gained a special focus in recent years. A proper combination of temperature and time parameters in sous vide processing plays an important role in the water-holding capacity, texture properties, and juiciness of the meat. The present study aimed to assess the impact of the one-step and two-step sous vide processing on different quality properties of chicken breast with special emphasis on the cooking loss, color, texture properties, protein solubility, and lipid oxidation. According to the results, chicken breast treated with a two-step temperature (50 and 60 °C) showed improved texture parameters (shear force, hardness, chewiness, and gumminess), lower cooking loss, acceptable redness values, and decreased lipid oxidation levels than the chicken breast treated with the one-step temperature of 60 °C. Moreover, the two-step sous vide technique revealed significantly higher total protein solubility of the chicken breast than the one-step sous vide. Based on pasteurization values, the two-step sous vide technique was equally safe as the one-step sous vide technique for vegetative cells' inactivation for the main pathogens of interest (C. perfringens and L. monocytogenes).

Effect of α-Terpineol on Chicken Meat Quality during Refrigerated Conditions.

The present study was designed to evaluate the in vitro antimicrobial properties of nine bioactive compounds (BACs). Applying the disc paper and minimum inhibitory concentration (MIC) assays, we found that the BACs with the widest spectrum of in vitro antibacterial activity against the studied bacteria were carvacrol and α-terpineol (αTPN). Subsequently, αTPN was selected and applied at different concentrations into the fresh minced chicken meat. The meat was then vacuum packaged and stored for 14 days at 4 °C. Physicochemical properties, lipid oxidation (thiobarbituric acid reactive substances, TBARS), electronic-nose-based smell detection, and microbiological characteristics were monitored. At day 14, meat treated with higher concentrations of αTPN (MIC-2 and MIC-4) exhibited a significantly increased pH and lightness (L*), increased yellowness (b*), decreased redness (a*), caused a significant decrease in water holding capacity (WHC), and decreased lipid oxidation by keeping TBARS scores lower than the control. Although αTPN showed perceptibly of overlapped aroma profiles, the E-nose was able to distinguish the odor accumulation of αTPN between the different meat groups. During the 2-week storage period, αTPN, particularly MIC-4, showed 5.3 log CFU/g reduction in aerobic mesophilic counts, causing total inhibition to the Pseudomonas lundessis, Listeria monocytogenes, and Salmonella Typhimurium. These promising results highlight that αTPN is exploitable to improve the shelf life and enhance the safety of meat and meat products.

Antioxidant and Antibacterial Peptides from Soybean Milk through Enzymatic- and Membrane-Based Technologies.

Enzymatic hydrolysis of soybean milk proteins with cysteine protease papain was performed in an advanced bioreactor, operated with batch mode. In soybean milk protein hydrolysis reaction, enzyme and substrate ratio and reaction temperature were varied, ranging from 0.029:100-0.457:100 and 30-60 °C, respectively. The degree of hydrolysis of soybean milk proteins was increased with increase of enzyme and substrate (soybean milk protein) ratio. However, the degree of hydrolysis was increased due to change of reaction temperature from 30 °C to 60 °C with enzyme and substrate ratio 0.229:100 and was reduced when hydrolysis reaction was performed with enzyme and substrate ratio 0.11:100 at hydrolysis temperature 60 °C. Antioxidant capacity of enzyme-treated milk had a similar trend with degree of hydrolysis. In a later exercise, a membrane bioreactor was adopted for continuous production of antioxidant and antibacterial peptides from soybean milk. The membrane bioreactor was operated for 12 h with constant feeding. Ceramic-made tubular membrane with a pore size 20 nm was used. Application of static turbulence promoter in a membrane separation process was investigated and its positive effects, with respect to higher permeate flux and lower energy consumption in filtration process, were proven. Antioxidant capacity and antibacterial activity against Bacillus cereus of enzyme-hydrolyzed milk and permeate from membrane were confirmed.

Production of Hypoallergenic Antibacterial Peptides from Defatted Soybean Meal in Membrane Bioreactor: A Bioprocess Engineering Study with Comprehensive Product Characterization.

Hypoallergenic antibacterial low-molecular-mass peptides were produced from defatted soybean meal in a membrane bioreactor. In the first step, soybean meal proteins were digested with trypsin in the bioreactor, operated in batch mode. For the tryptic digestion of soybean meal protein, optimum initial soybean meal concentration of 75 g/L, temperature of 40 °C and pH=9.0 were determined. After enzymatic digestion, low-molecular-mass peptides were purified with cross-flow flat sheet membrane (pore size 100 µm) and then with tubular ceramic ultrafiltration membrane (molecular mass cut-off 5 kDa). Effects of transmembrane pressure and the use of a static turbulence promoter to reduce the concentration polarization near the ultrafiltration membrane surface were examined and their positive effects were proven. For the filtration with ultrafiltration membrane, transmembrane pressure of 3·105 Pa with 3-stage discontinuous diafiltration was found optimal. The molecular mass distribution of purified peptides using ultrafiltration membrane was determined by a liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry setup. More than 96% of the peptides (calculated as relative frequency) from the ultrafiltration membrane permeate had the molecular mass M≤1.7 kDa and the highest molecular mass was found to be 3.1 kDa. The decrease of allergenic property due to the tryptic digestion and membrane filtration was determined by an enzyme-linked immunosorbent assay and it was found to exceed 99.9%. It was also found that the peptides purified in the ultrafiltration membrane promoted the growth of Pediococcus acidilactici HA6111-2 and they possessed antibacterial activity against Bacillus cereus.