This study investigated the synergistic effect of combining flavourzyme, a natural enzyme, and floating electrode-dielectric barrier discharge (FE-DBD) plasma (1.1 kV, 43 kHz, N2 1.5 m/s) treatment, a non-thermal decontamination technology, against Escherichia coli biofilms in squid. E. coli (ATCC 35150 and ATCC 14301) biofilms were formed on the surface of squid and treated with different minimum inhibitory concentrations (MICs) of flavourzyme (1/8; 31.25 μL/mL, 1/4; 62.5 μL/mL, 2/4; 125 μL/mL, and 3/4 MIC; 250 μL/mL) and FE-DBD plasma (5, 10, 30, and 60 min). Independently, flavourzyme and FE-DBD plasma treatment decreased by 0.26-1.71 and 0.19-1.03 log CFU/cm2, respectively. The most effective synergistic combination against E. coli biofilms was observed at 3/4 MIC flavourzyme + 60 min FE-DBD plasma exposure, resulting in a reduction of 1.55 log CFU/cm2. Furthermore, the combined treatment exhibited higher efficacy in E. coli biofilm inactivation in squid compared to individual treatments. The pH values of the synergistic combinations were not significantly different from those of the untreated samples. The outcomes indicate that the combined treatment with flavourzyme and FE-DBD plasma can effectively provide effective control of E. coli biofilms without causing pH changes in squid. Therefore, our study suggests a new microbial control method for microbial safety in the seafood industry.

Calcium peptide chelates are developed as efficient supplements for preventing calcium deficiency. Spent hen meat (SHM) contains a high percentage of proteins but is generally wasted due to the disadvantages such as hard texture. We chose the underutilized SHM to produce peptides to bind calcium by proteolysis and aimed to investigate chelation between calcium and peptides in hydrolysate for a sustainable purpose. The optimized proteolysis conditions calculated from the result of response surface methodology for two-step hydrolysis were 0.30% (wenzyme/wmeat) for papain with a hydrolysis time of 3.5 h and 0.18% (wenzyme/wmeat) for flavourzyme with a hydrolysis time of 2.8 h. The enzymatic hydrolysate (EH) showed a binding capacity of 63.8 ± 1.8 mg calcium/g protein. Ethanol separation for EH improved the capacity up to a higher value of 68.6 ± 0.6 mg calcium/g protein with a high association constant of 420 M-1 (25°C) indicating high stability. The separated fraction with a higher amount of Glu, Asp, Lys, and Arg had higher calcium-binding capacity, which was related to the number of ─COOH and ─NH2 groups in peptide side chains according to the result from amino acid analysis and Fourier transform infrared spectroscopy. Two-step enzymatic hydrolysis and ethanol separation were an efficient combination to produce peptide mixtures derived from SHM with high calcium-binding capacity. The high percentage of hydrophilic amino acids in the separated fraction was concluded to increase calcium-binding capacity. This work provides foundations for increasing spent hen utilization and developing calcium peptide chelates based on underutilized meat.

The interest in incorporating potatoes into wheat dough is increasing. However, potatoes exhibit significant viscosity during thermal processing, affecting product processing and quality. This study aims to find an effective method to reduce the viscosity of mashed potatoes. We aimed to compare the effects of different enzymes (α-amylase, β-amylase, and flavourzyme) and concentrations (0.01%, 0.05%, and 0.1%) on the micromorphology and rheological properties of mashed potatoes and potato-wheat dough. The impact of flavourzyme was the most significant (p<0.05). When enzyme concentration increased, viscosity decreased, and the degree of structural damage, indicated by increased porosity. Notably, the addition of flavourzyme can increase the content of sweet and umami free amino acids, improving the flavor of mashed potatoes. The scanning electron microscopy and confocal laser scanning microscopy images of potato-wheat dough revealed that enzyme-hydrolyzed mashed potatoes had improved homogeneity, reestablished the dough continuity, and strengthened the three-dimensional structure comprising proteins and starch. Notably, flavourzyme demonstrated the most significant effect on enhancing the protein-starch network structure. This was attributed to the exposure of functional groups resulting from protein hydrolysis, facilitating interaction with starch molecules. Our findings indicate that the addition of 0.1% flavourzyme (500 LAPU/g, pH 5.5, 55 ± 2°C, 30 min treated) was the most effective in reducing viscosity and reconstructing the gluten network. Enzymatic hydrolysis plays a vital role in the production of high-quality potato products, with particular importance in the baking industry, where flavourzyme exhibits significant potential. PRACTICAL APPLICATION: Enzymatic hydrolysis plays a vital role in the production of high-quality potato products, with particular importance in the baking industry, where flavourzyme exhibits significant potential.

Enzymatic protein hydrolysis is a well-established method for improving the quality of dietary proteins, including edible insects. Finding effective enzymes from natural sources is becoming increasingly important. This study used nuruk extract concentrate (NEC), an enzyme-rich fermentation starter, to produce protein hydrolysate from defatted Tenebrio molitor (also called mealworm, MW). The nutritional, functional, and sensorial properties of the hydrolysate were then compared to those obtained using commercial proteases (alcalase and flavourzyme). The protease activities of the crude nuruk extract (CNE), NEC, alcalase, and flavourzyme were 6.78, 12.71, 11.07, and 12.45 units/mL, respectively. The degree of hydrolysis and yield of MW hydrolysis by NEC were 15.10 and 35.92% (w/w), respectively. MW hydrolysate was obtained using NEC and had a significantly higher free amino acid content (90.37 mg/g) than alcalase (53.01 mg/g) and flavourzyme (79.64 mg/g) hydrolysates. Furthermore, the NEC hydrolysis of MW increased the antioxidant and angiotensin-converting enzyme inhibitory activity, with IC50 values of 3.07 and 0.15 mg/mL, respectively. The enzymatic hydrolysis also improved sensory properties, including umaminess, sweetness, and saltiness. Overall, this study found that the NEC hydrolysis of MW outperformed commercial proteases regarding nutritional quality, sensory attributes, and biological activity. Therefore, nuruk could potentially replace commercial proteases, lowering the cost of enzymatic protein hydrolysis.

Spent hen meat is considered as a category of waste generated by the poultry sector which can lead to serious environmental concerns if not disposed and utilized properly. In this work, spent hen meat was hydrolysed by 2% Flavourzyme (6.5 pH, 55 °C) followed by ultrafiltration to produce three peptide fractions with molecular weights > 10 kDa, 5-10 kDa and < 5 kDa. These fractions were evaluated for antioxidant potential, SDS PAGE and amino acid profile. The SDS PAGE profile demonstrated bands in the low molecular weight (< 10 kDa) region. Peptide fractions of < 5 kDa exhibited highest antioxidant activity and, essential as well as hydrophobic amino acid composition than whole hydrolysate and other peptide fractions. Incorporation of the identified hydrolysate fraction in food could improve its shelf stability while serving as a preventive component against human degenerative diseases.

Application of non-thermal treatment to proteins prior to enzymatic hydrolysis can facilitate the release of novel bioactive peptides (BPs) with unique biological activities. In this study, lupin protein isolate was pre-treated with ultrasound and hydrolysed using alcalase and flavourzyme to produce alcalase hydrolysate (ACT) and flavourzyme hydrolysate(FCT). These hydrolysates were fractionated into 1, 5, and 10 kDa molecular weight fractions using a membrane ultrafiltration technique. The in vitro angiotensin-converting enzyme (ACE) studies revealed that unfractionated ACT (IC50 = 3.21 mg mL-1) and FCT (IC50 = 3.32 mg mL-1) were more active inhibitors of ACE in comparison to their ultrafiltrated fractions with IC50 values ranging from 6.09 to 7.45 mg mL-1. Molecular docking analysis predicted three unique peptides from ACT (AIPPGIPY, SVPGCT, and QGAGG) and FCT (AIPINNPGKL, SGNQGP, and PPGIP) as potential ACE inhibitors. Thus, unique BPs with ACE inhibitory effects might be generated from ultrasonicated lupin protein.

The present study aimed to investigate the change in physicochemical and safety properties of grass carp during fermentation with flavourzyme and Lactobacillus plantarum (FLF). The natural fermentation (NF) and fermentation with Lactobacillus plantarum (LF) samples were used as control. The results showed that with increasing fermentation time, the pH and water activity (Aw) in each fermented grass carp sample gradually decreased, while the a-amino nitrogen (ANN), thiobarbituric acid reactive substance (TBARS), total volatile base nitrogen (TVB-N), biogenic amines (BAs), and harmful microbial gradually increased. Besides, compared with NF samples at each fermentation time, significantly lower pH, Aw, TBARS, TVB-N, BAs, and harmful microbial presented in LF and FLF samples. However, FLF samples have a higher AAN content than that of NF and LF samples during fermentation. Overall, the fermentation with Lactobacillus plantarum and flavourzyme could contribute to fermented grass carp products with better physicochemical and safety properties.

The physicochemical features of mung bean protein (MBP) and adzuki bean protein (ABP) hydrolysates derived from Alcalase (MBPHA, ABPHA) and Flavourzyme (MBPHF, ABPHF) were assessed using FTIR, hydrophobicity, emulsion activity, zeta potential, and health-promoting activities. The results proved that the choice of peptidase and substrate both have a significant effect on the hydrolysates in different physicochemical, structural and functional properties. Size exclusion-HPLC was used to fractionate the MBP and ABP hydrolysates. The results demonstrated that Alcalase hydrolysates included smaller peptides than Flavourzyme hydrolysates, and the chromatogram patterns of the two peptidases were similar. The peptides with the most potent antioxidant and ACE-inhibitory properties were identified using MALDI-TOF-MS. The fraction (F4) of MBPHA exhibited the highest levels of metal chelating activity. The Flavourzyme hydrolysates fraction (F2) and the ABPHA fraction (F2) showed the highest ABTS radical scavenging activity and ACE-inhibitory activity, respectively. Pro-Pro was identified in peptide sequences with ABTS radical scavenging activity as an active component while Pro-Gln was identified in peptide sequences with ACE-inhibitory activity. As a result, Pro-Pro and Pro-Gln, respectively, are likely-one of the characteristics of antioxidant and ACE-inhibitory peptides from MBP and ABP. Compared to mung bean and adzuki bean protein as substrate, Alcalase and Flavourzyme as peptidases significant impacted the development of distinct functionalities and biological activities.

This study aimed to investigate the effects of flavourzyme addition on protein degradation and flavor formation in grass carp during fermentation. The related results showed that the addition of flavourzyme reduced the moisture content and accelerated the hydrolysis of protein and generation of water-soluble flavor substances (e.g., TCA-soluble peptides, α-amino nitrogen, and free amino acids), thereby contributing to fermented grass carp products with a better taste quality. Besides, radar map results of electronic tongue and electronic nose showed that flavourzyme addition gives fermented products a more intense umami taste and odor. Meanwhile, sensory evaluation results also further confirmed that the addition of flavourzyme significantly improved the sensory attributes of fermented grass carp products, especially the taste and odor attributes. Overall, flavourzyme addition may be an effective way to shorten fermentation time and improve the flavor quality of fermented grass carp products during fermentation. PRACTICAL APPLICATIONS: In this study, to study the effects of flavourzyme addition on protein degradation and flavor formation in grass carp during fermentation, the related indicators include the moisture content, total nitrogen, non-protein nitrogen, protein degradation index, TCA-soluble peptides, α-amino nitrogen, free amino acids, electronic tongue, electronic nose, and sensory attributes were analyzed. This study may provide some useful information for the improvement of fermentation methods and the production of high-quality fermented grass carp products.

This study aimed to carefully investigate the effect of hydrolysis using Flavourzyme on meat quality, antioxidative status, and taste-related compounds in breast of Samgyetang that was supplemented with black garlic (BG). Four different treatment groups were compared: (1) conventional Samgyetang (control), (2) Samgyetang hydrolyzed with Flavourzyme (1%, v/w) (FS), (3) Samgyetang made with the BG extract without hydrolysis (NBG), and (4) BG samgyetang pre-treated with Flavourzyme (1%, v/w) in a water bath at 55°C for 2.5 h and hydrolyzed before being processed (HBG). All the treatment groups were cooked by retorting at conditions 121°C and 1.5 kg/cm2 for 1 h. Improved umami profiles through the increase of umami-related nucleotides (5\'-GMP, 5\'-IMP) and free amino acids-aspartic acid and glumtamic acid, in Samgyetang breast was recorded following hydrolysis. The HBG group tended to impart stronger scavenging activity toward free radicals compared with the other two groups, while not differing with NBG group regarding suppressing malondialdehyde. Textural properties were improved through hydrolysis, wherein the shear force value decreased from 2.29 kgf in the control to 1.19 and 1.25 kgf in the FS and HBG group. Moisture percentages were highly retained, with the redness score increasing and the lightness color decreasing following hydrolysis. In conclusion, the results of this study can be a preliminary information of the effect of hydrolysis pre-treatment for BG samgyetang. Further experiments are required to compare various enzymes along with its organoleptic acceptances.