Bioactive Peptides from Liquid Milk Protein Concentrate by Sequential Tryptic and Microbial Hydrolysis

Recently, bioactive peptides as a health-promoting agent have come to the forefront of health research; however, industrial production is limited, possibly due to the lack of the required technological knowledge. The objective of the investigation was to prepare bioactive peptides with hypoallergenic properties from liquid milk protein concentrate (LMPC), through sequential enzymatic and microbial hydrolysis. LMPC was produced from ultra-heat-treated (UHT) skimmed cow’s milk using a nanofiltration membrane. The effect of the concentration of trypsin (0.008–0.032 g·L−1) on the hydrolysis of LMPC was studied. Subsequently, the hydrolysis of tryptic-hydrolyzed LMPC (LMPC-T) with lactic acid bacteria was performed, and the effect of glucose in microbial hydrolysis was studied. Aquaphotomic analysis of the hydrolysis of LMPC was performed using the spectral range of 1300–1600 nm (near-infrared spectra). Changes in antioxidant capacity, anti-angiotensin-converting enzyme activity, and antibacterial activity against Bacillus cereus, Staphylococcus aureus and Listeria monocytogenes were noted after the sequential tryptic and microbial hydrolysis of LMPC. Allergenicity in LMPC was reduced, due to sequential hydrolysis with 0.016 g·L−1 of trypsin and lacteal acid bacteria. According to the aquaphotomic analysis result, there was a dissociation of hydrogen bonds in compounds during the initial period of fermentation and, subsequently, the formation of compounds with hydrogen bonds. The formation of compounds with a hydrogen bond was more noticeable when microbial hydrolysis was performed with glucose. This may support the belief that the results of the present investigation will be useful to scale up the process in the food and biopharmaceutical industries.

Optimization of Microbial Hydrolysis Parameters of Poultry By-Products Using Probiotic Microorganisms to Obtain Protein Hydrolysates

In connection with the active growth of poultry processing, the issue of rational use of poultry by-products is urgent. The paper proposes hydrolysis of the gizzards of broiler chickens and hens of the parent stock with bifidobacteria liquid concentrate (BLC) and Propionix liquid concentrated starter culture (Propionix LCSC). The effect of enzymatic treatment on changes in the structural components of the gizzards was studied using scanning electron microscopy (SEM) and determination of the dispersed composition. As a result of the research, the multiple regression equations and the response surfaces were obtained, which describe the optimal parameters of the gizzard´s hydrolysis process. The temperature and the time of hydrolysis are factors that have a significant effect on the degree of hydrolysis. The results of the structural and microscopic analysis confirm the high hydrolysability of the by-products by changing its structure and increasing the number of smaller protein particles. It was found experimentally that the gizzards of hens are more susceptible to the action of probiotic microorganisms’ enzymes compared to the gizzards of broiler chickens.

Human Saliva-Mediated Hydrolysis of Eugenyl-β-D-Glucoside and Fluorescein-di-β-D-Glucoside in In Vivo and In Vitro Models

Eugenyl-β-D-glucopyranoside, also referred to as Citrusin C, is a natural glucoside found among others in cloves, basil and cinnamon plants. Eugenol in a form of free aglycone is used in perfumeries, flavourings, essential oils and in medicinal products. Synthetic Citrusin C was incubated with human saliva in several in vitro models together with substrate-specific enzyme and antibiotics (clindamycin, ciprofloxacin, amoxicillin trihydrate and potassium clavulanate). Citrusin C was detected using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Citrusin C was completely degraded only when incubated with substrate-specific A. niger glucosidase E.C 3.2.1.21 (control sample) and when incubated with human saliva (tested sample). The addition of antibiotics to the above-described experimental setting, stopped Citrusin C degradation, indicating microbiologic origin of hydrolysis observed. Our results demonstrate that Citrusin C is subjected to complete degradation by salivary/oral cavity microorganisms. Extrapolation of our results allows to state that in the human oral cavity, virtually all β-D-glucosides would follow this type of hydrolysis. Additionally, a new method was developed for an in vivo rapid test of glucosidase activity in the human mouth on the tongue using fluorescein-di-β-D-glucoside as substrate. The results presented in this study serve as a proof of concept for the hypothesis that microbial hydrolysis path of β-D-glucosides begins immediately in the human mouth and releases the aglycone directly into the gastrointestinal tract.

Role of Microbial Hydrolysis in Anaerobic Digestion

There is a growing need of substrate flexibility for biobased production of energy and value-added products that allows the application of variable biodegradable residues within a circular economy. It can be used to balance fluctuating energy provision of other renewable sources. Hydrolysis presents one of the biggest limitations during anaerobic digestion. Methods to improve it will result in broader process applicability and improved integration into regional material cycles. Recently, one focus of anaerobic digestion research has been directed to systems with a separate hydrolysis–acidogenesis stage as it might be promised to improve process performance. Conditions can be adjusted to each class of microorganisms individually without harming methanogenic microorganisms. Extensive research of separate biomass pretreatment via biological, chemical, physical or mixed methods has been conducted. Nevertheless, several methods lack economic efficiency, have a high environmental impact or focus on specific substrates. Pretreatment via a separate hydrolysis stage as cell-driven biotransformation in a suspension might be an alternative that enables high yields, flexible feeding and production, and a better process control. In this review, we summarize existing technologies for microbial hydrolytic biotransformation in a separate reactor stage and the impacts of substrate, operational parameters, combined methods and process design as well as remaining challenges.

Comparative Studies on Production of Bioethanol from Rice Straw Using Bacillus subtilis and Trichoderma viride as Hydrolyzing Agents

Ethanol production by S. cerevisiae was carried out using rice straw as substrate and B. subtilis and T. viride as hydrolyzing agents. The aim of this research is to compare the potential of rice straw (non-edible waste material) for bioethanol production using Bacillus subtilis and Trichoderma viride as cellulose hydrolyzing agents. The sample was dried and ground; and was subjected to chemical pretreatment and microbial hydrolysis to maximize sugar production. Standard methods were used to carry out isolation, identification and analysis of sample which includes proximate, mineral and physicochemical analysis. The sample was fermented for seven days during which ethanol yield was determined. Cellulose hydrolysis screening carried out on each of the two organisms revealed T. viride having the higher clearance zone of 1.8 cm, while B. subtilis had 1.5 cm. Proximate analysis obtained from the samples showed that the pretreatment method was relatively effective giving an increase in the cellulose and decrease in the hemicellulose and lignin contents of the samples. This showed rice straw having a cellulose content of 51.33 ± 0.17% after pretreatment. Potassium content was relatively high (17.96 mg/g), Hydrolysis using T. viride gave higher reducing sugar yield than that obtained using B. subtilis with 26.6 g and 12.21 g respectively. The pH was observed to decrease during fermentation while total titratable acidity observed showed an increase. Highest ethanol yield of 16.21 g/100 g was obtained using T. viride as hydrolyzing agent.

Comparative analysis of functionality of spray dried whey protein hydrolysates obtained by enzymatic and microbial hydrolysis

The aim of this study was to examine the bioactive potential of hydrolysate powders produced by enzymatic and microbial hydrolysis of whey proteins followed by spray drying, in order to reveal which one of these processes result in a product with significantly improved functional properties. Hydrolysate powders produced by the two different biotechnological processes were compared based on their antioxidant (DPPH and FTC), antibacterial as well as erythrocyte membrane stabilizing activities. The performed tests revealed that the concentration of at least 178.4 mg mL-1 of the whey protein hydrolysate powder, produced by tryptic digestion, could inhibit the process of lipid peroxidation by 50 %, suppress the microbial contamination caused by S. aureus ATCC25923, B. cereus ATCC 11778 and L. monocytogenes, and provide the antioxidant and membrane stabilizing activities greater than 50 %. On the other hand, the hydrolysate powder obtained by whey fermentation at the concentration of at least 811.5 mg mL-1 achieved 50 % of all tested bioactivities, with the emphasis on the significantly more pronounced antibacterial activity against all tested strains. In that sense, tryptic hydrolysis could be highlighted as an optimal process that provides production of the whey hydrolysate with pronounced bioactive properties that could be considered as a very promising natural food supplement.