Optimization Of Fermentation Process Research Articles

Real-time monitoring of multiparameter in 1,3-propanediol production process with near-infrared spectroscopy

1,3-propanediol (1,3-PDO) is a significant product of fermentation, with glycerol serving as the primary substrate in most cases. Bioprocess control based on real-time information of feedstock and main products is crucial for reducing the cost of production. However, rapid quantification of 1,3-PDO and glycerol remains challenging due to their highly similar molecular structures. In this study, the feasibility of near-infrared (NIR) spectroscopy to monitor 1,3-PDO, glycerol, acetate, and butyrate concentrations in the fermentation process using strain Clostridium pasteurianum was evaluated. NIR spectra were acquired through at-line measurement involving sampling and ex-situ analysis or on-line measurement with a fiber optic probe immersed in fermentation broth, integrated with Partial Least Squares (PLS) regression to establish calibration models on a laboratory-scale and pilot-scale. The best PLS regressions of 1,3-PDO, glycerol, acetate, and butyrate with two measurement approaches provided excellent performance, with the root-mean-squared errors of prediction (RMSEP) of 1.656g/L, 1.502g/L, 0.746g/L, and 0.557g/L in at-line measurement and 1.113g/L, 1.581g/L, 0.415g/L, and 0.526g/L in on-line measurement. The cross-scale application performance of at-line measurement was evaluated by an external fermentation trial and an acceptable result was achieved. At-line measurement technique represents a superior choice for the optimization of fermentation process since the robustness across varying fermentation scales and its applicability in multiple bioreactors. Thus, a calibration model developed for one bioreactor is likely to be used in other bioreactors, which enables the reduction of modeling costs. On-line measurement technique, owing to its automated operation and frequent data acquisition, enables real-time monitoring and precise control of the fermentation process, thereby reducing cost and improving production efficiency.

Multiscale modelling of bioprocess dynamics and cellular growth

BackgroundFermentation processes are essential for the production of small molecules, heterologous proteins and other commercially important products. Traditional bioprocess optimisation relies on phenomenological models that focus on macroscale variables like biomass growth and protein yield. However, these models often fail to consider the crucial link between macroscale dynamics and the intracellular activities that drive metabolism and proteins synthesis.ResultsWe introduce a multiscale model that not only captures batch bioreactor dynamics but also incorporates a coarse-grained approach to key intracellular processes such as gene expression, ribosome allocation and growth. Our model accurately fits biomass and substrate data across various Escherichia coli strains, effectively predicts acetate dynamics and evaluates the expression of heterologous proteins. By integrating construct-specific parameters like promoter strength and ribosomal binding sites, our model reveals several key interdependencies between gene expression parameters and outputs such as protein yield and acetate secretion.ConclusionsThis study presents a computational model that, with proper parameterisation, allows for the in silico analysis of genetic constructs. The focus is on combinations of ribosomal binding site (RBS) strength and promoters, assessing their impact on production. In this way, the ability to predict bioreactor dynamics from these genetic constructs can pave the way for more efficient design and optimisation of microbial fermentation processes, enhancing the production of valuable bioproducts.

Optimisation of the Ethanol Fermentation Process Using Hydrothermal Pretreatment of Cellulose Waste-Effect of Fermentation Pattern and Strain.

Suitable fermentation substrates and fermentation modes can effectively improve the fermentation ethanol yield. In this study, we optimised the hydrothermal pretreatment conditions by orthogonal optimisation using waste tissue paper as substrate. These conditions consisted of 50 min duration in a high-pressure reactor with pure water as solvent at a temperature of 160 °C. The biomass to water ratio was maintained at a constant level. The cellulose content of the pretreated TP was 81.19 ± 4.06%, which was an increase of 21.59% compared to the blank control. The 72 h reducing sugar yield of pretreated TP was 0.61 g sugar/g paper, which was 38.64% higher than that of untreated TP. Subsequently, the pretreated TP was fermented under optimal conditions. The mixed group of Saccharomyces cerevisiae and Candida shehatae (SC) showed a distributed saccharification fermentation pattern, with an ethanol yield of 28.11 g/L in 72 h. On the other hand, the single Saccharomyces cerevisiae (S) exhibited a homobloc saccharification fermentation pattern, with an ethanol yield of 35.15 g/L in 72 h.

Investigation of the Fermentation Process of Moringa oleifera Leaves and Its Effects on the Growth Performance, Antioxidant Capacity, and Intestinal Microbiome of Procambarus clarkii.

Moringa oleifera is renowned for its high antioxidant activity. However, few studies have been conducted on its effects on aquatic animals. The aim of this experiment was to investigate the optimal fermentation process of M. oleifera leaves and to evaluate the effects of fermented M. oleifera leaves on crayfish (9.11 ± 0.3 g) in terms of growth performance, antioxidant capacity, and gut microbiological parameters. By optimizing the fermenting material/water ratio, fermentation time, temperature, and strain, the optimal fermentation conditions of a 10% water ratio + 48 h + 30 °C + inoculation with 2% B. amyloliquefaciens (107 CFU mL-1) were obtained. These conditions resulted in notable increases in the contents of the total protein, total phenols, flavonoids, and amino acids (p < 0.05) while also leading to a notable decrease in the content of tannins in contrast to those of unfermented M. oleifera leaves (p < 0.05). The fermented M. oleifera (FMO) leaves were incorporated at five concentrations, including 0% (control (CT)), 0.25% (0.25FMO), 0.5% (0.5FMO), 1% (1FMO), and 2% (2FMO). The results showed that the 1FMO group performed better in terms of the final body weight (FBW), weight gain rate (WGR), and specific weight gain rate (SGR) compared with the CT group (p < 0.05). In addition, amylase and lipase activities were significantly higher in the 1FMO and 2FMO groups compared with the other groups (p < 0.05). The fermented M. oleifera leaves significantly increased the catalase (CAT) activity in the crayfish (p < 0.05). The superoxide dismutase (SOD) activity was significantly increased in the 0.25FMO, 1FMO, and 2FMO groups, and the malondialdehyde (MDA) content was significantly decreased while the glutathione peroxidase (GSH-Px) content was significantly increased in the 0.5FMO, 1FMO, and 2FMO groups (p < 0.05). Furthermore, the 1FMO group was observed to significantly increase the abundance of Firmicutes while simultaneously reducing the abundance of Aeromonas (p < 0.05) and adjusting the structure of the intestinal microbiome. In conclusion, this study established the optimal fermentation conditions for M. oleifera and obtained a product with high nutrient and low tannin contents. Furthermore, the incorporation of 1% FMO was demonstrated to facilitate growth, enhance the antioxidant capacity, and optimize the gut microbiology in crayfish.

Elucidating Bile Acid Tolerance in Saccharomyces cerevisiae: Effects on Sterol Biosynthesis and Transport Protein Expression.

The tolerance of Saccharomyces cerevisiae to high concentrations of bile acids is intricately linked to its potential as a probiotic. While the survival of yeast under high concentrations of bile acids has been demonstrated, the specific mechanisms of tolerance remain inadequately elucidated. This study aims to elucidate the tolerance mechanisms of S. cerevisiae CEN.PK2-1C under conditions of elevated bile acid concentrations. Through growth curve analyses and scanning electron microscopy (SEM), we examined the impact of high bile acid concentrations on yeast growth and cellular morphology. Additionally, transcriptomic sequencing and molecular docking analyses were employed to explore differentially expressed genes under high bile acid conditions, with particular emphasis on ATP-binding cassette (ABC) transporters and steroid hormone biosynthesis. Our findings indicate that high concentrations of bile acids induce significant alterations in the sterol synthesis pathway and transporter protein expression in S. cerevisiae. These alterations primarily function to regulate sterol synthesis pathways to maintain cellular structure and sustain growth, while enhanced expression of transport proteins improves tolerance to elevated bile acid levels. This study elucidates the tolerance mechanisms of S. cerevisiae under high bile acid conditions and provides a theoretical foundation for optimizing fermentation processes and process control.

Investigation into optimizing fermentation processes to enhance uric acid degradation by probiotics

To enhance the activity of Lactiplantibacillus plantarum FS4722 (L. plantarum FS4722) in reducing uric acid, this study optimized the culture medium components and fermentation conditions using response surface methodology. The results indicated that the optimal culture medium comprised glucose at 25.61 g/L, yeast extract powder (YEP) at 26.86 g/L, and nucleoside addition at 0.48 g/L. The optimal fermentation conditions were: a fermentation time of 10 h, an initial pH of 6.5, a fermentation temperature of 33 °C, and an inoculum size of 1.0 %. Under these conditions, the nucleoside degradation ability of L. plantarum FS4722 increased by 41.9 times, enhancing its potential applications in reducing uric acid and developing anti-gout foods, health supplements, and pharmaceuticals.

Raman‐Enabled Predictions of Protein Content and Metabolites in Biopharmaceutical Saccharomyces cerevisiae Fermentations

ABSTRACTRaman spectroscopy, a robust and non‐invasive analytical method, has demonstrated significant potential for monitoring biopharmaceutical production processes. Its ability to provide detailed information about molecular vibrations makes it ideal for the detection and quantification of therapeutic proteins and critical control parameters in complex biopharmaceutical mixtures. However, its application in Saccharomyces cerevisiae fermentations has been hindered by the inherent strong fluorescence background from the cells. This fluorescence interferes with Raman signals, compromising spectral data accuracy. In this study, we present an approach that mitigates this issue by deploying Raman spectroscopy on cell‐free media samples, combined with advanced chemometric modeling. This method enables accurate prediction of protein concentration and key process parameters, fundamental for the control and optimization of biopharmaceutical fermentation processes. Utilizing variable importance in projection (VIP) further enhances model robustness, leading to lower relative root mean squared error of prediction (RMSEP) values across the six targets studied. Our findings highlight the potential of Raman spectroscopy for real‐time, on‐line monitoring and control of complex microbial fermentations, thereby significantly enhancing the efficiency and quality of S. cerevisiae‐based biopharmaceutical production.

Optimisation of fermentation condition for production of hight monacolin k (mk) yield by monascus-strains

Monacolin K (MK), produced by Monascus fermentation, has significant hypolipidemic effects. In this study, under solid-state fermentation (SSF) quinoa was used as substrate to investigate the effect of the MK production by Monascus species quinoa. The results of the single factor experiment showed that the amount of the substrate loaded, addition of water to the substrate, and fermentation temperature change time have a significant effect on the MK production of Monascus-fermented quinoa. It was found that neither M. pilosus-fermented quinoa nor M. ruber-fermented quinoa produced citrinin. Consequently, these three factors were selected for the response surface optimization to determine the optimal fermentation process. The results showed that the optimal fermentation process of M. pilosus-fermented quinoa was as follows: quinoa loading amount of 30 g/300 ml, addition of 20 ml water to substrate, seed liquid inoculation volume of 10%, cultured at 30°C for 2 days, followed by a temperature change to 25°C and continued cultivation until 12 days. Under this fermentation process, the yield of MK reached 2.51 mg/g and without production of any citrinin. The optimal fermentation process of M. ruber-fermented quinoa was as follows: quinoa loading amount of 31 g/300 ml, water addition of substrate of 20 ml, seed liquid inoculation volume of 20%, cultured at 30°C for 2 days, followed by a temperature change to 25°C and continued cultivation until 12 days. Under this fermentation process, the yield of MK reached 3.22 mg/g and without any citrinin production. Bangladesh J. Bot. 53(3): 717-729, 2024 (September) Special

Xylitol production from passion fruit peel hydrolysate: Optimization of hydrolysis and fermentation processes

The passion fruit peel (PFP) has a high cellulose and hemicellulose content, which can be used to produce fermentable sugars. In this context, this study aims to optimize the release of xylose and the production of xylitol from PFP. The optimized conditions were 0.71 M dilute sulfuric acid and a 21.84-minute treatment, yielding 19.03 g/L of xylose (PFP-1). Different PFP hydrolysates were evaluated to improve xylitol production by the yeast Kluyveromyces marxianus ATCC 36907: PFP-2 (PFP1 treated with Ca(OH)2), PFP-3 (PFP-1 treated with Ca(OH)2 and activated carbon), PFP-4 (PFP-3 with biological elimination of glucose with S. cerevisiae, and concentrated at different xylose concentrations). The applied methods resulted in higher xylitol production (14.97 g/L), when PFP hydrolysate was detoxified with Ca(OH)2, treated with activated charcoal for 1 h, biotreated for glucose removal, and concentrated to 40 g/L of xylose.

Waste to wealth: Polyhydroxyalkanoates (PHA) production from food waste for a sustainable packaging paradigm

The growing demand for sustainable food packaging and the increasing concerns regarding environmental pollution have driven interest in biodegradable materials. This paper presents an in-depth review of the production of Polyhydroxyalkanoates (PHA), a biodegradable polymer, from food waste. PHA-based bioplastics, particularly when derived from low-cost carbon sources such as volatile fatty acids (VFAs) and waste oils, offer a promising solution for reducing plastic waste and enhancing food packaging sustainability. Through optimization of microbial fermentation processes, PHA production can achieve significant efficiency improvements, with yields reaching up to 87 % PHA content under ideal conditions. This review highlights the technical advancements in using PHA for food packaging, emphasizing its biodegradability, biocompatibility, and potential to serve as a biodegradable alternative to petroleum-based plastics. However, challenges such as high production costs, mechanical limitations, and the need for scalability remain barriers to industrial adoption. The future of PHA in food packaging hinges on overcoming these challenges through further research and innovation in production techniques, material properties, and cost reduction strategies, along with necessary legislative support to promote widespread use.

Exploring Calcium Alginate-Based Gels for Encapsulation of Lacticaseibacillus paracasei to Enhance Stability in Functional Breadmaking.

This study focuses on evaluating the efficiency of acid-tolerant Lacticaseibacillus paracasei bacteria encapsulated in an alginate-based gel matrix during repeated sourdough fermentation cycles, as well as their preservation during storage and throughout baking at high temperature. A double-coating procedure was applied, involving the encapsulation of bacterial cells in calcium alginate, which was further coated with chitosan. The encapsulation efficiency (EE) did not show significant difference between alginate and alginate-chitosan (97.97 and 96.71%, respectively). The higher number of L. paracasei bacteria was preserved in double-coated microbeads, with survivability rates of 89.51% and 96.90% in wet and dried microbeads, respectively. Encapsulated bacteria demonstrated effective fermentation ability, while double gel-coated cells exhibited slower acidification during sourdough fermentation, maintaining higher efficiency in the second fermentation cycle. The addition of freeze-dried, alginate-based gel-encapsulated bacteria (2-4%, w/w flour) significantly (p < 0.05) improved bread quality and extended its shelf life. A double-layer coating (alginate-chitosan) can be introduced as an innovative strategy for regulating the release of lactic acid bacteria and optimizing fermentation processes. Powdered alginate or alginate-chitosan gel-based L. paracasei microcapsules, at appropriate concentrations, can be used in the production of baked goods with acceptable quality and sensory properties, achieving a lactic acid bacteria count of approximately 106 CFU/g in the crumb, thereby meeting the standard criteria for probiotic bakery products.

Parametric adjustment of two different kinetic models with constant and variable yield coefficients. Butanol production from glycerol fermentation by Clostridium pasteurianum

The study of bioreactors, for their design and operation, is crucial for optimizing fermentation processes. This can be achieved through computational simulations based on unstructured kinetic models. Generally, unstructured kinetic models are established with yields associated with the concentrations of biomass, substrate, and product of constant type. It has been reported that variable polynomial yields can better describe these processes. Therefore, this work proposes the study of a glycerol fermentation system to produce butanol in the presence of Clostridium pasteurianum. Parametric identification of two unstructured kinetic models (Moser-Boulton and Moser-Levespiel) was performed. Three mass balances were proposed for a batch-operated reactor (biomass, glycerol, and butanol). Constant and variable yields of first, second, and third order were considered. Finally, the simulated data were validated using correlation coefficients with the corresponding experimental data.

Production of Amylase by Solid State Fermentation Using Agricultural Waste

This study presents a comprehensive investigation into the production of amylase, a crucial enzyme with wide-ranging industrial applications, using locally sourced substrates from Kachchh, Gujarat. The research employed the Bacillus licheniformis strain and substrates such as coconut, rice husk, wheat bran, paddy straw, and maize straw. The study found paddy straw to be the most promising substrate for amylase production. The research also systematically optimized various process parameters for amylase production in Solid-State Fermentation (SSF) using the One Variable at a Time (OVAT) method. These parameters included incubation period, temperature, inoculum level, additional carbon sources, starch concentrations, additional nitrogen sources, initial pH, different mineral salt ions, initial moisture level, and surfactants. The results showed that the optimal conditions for maximum amylase yield were an incubation period of 48 hours, an incubation temperature of 35°C, an inoculum level of 10%, starch as the additional carbon source, a starch concentration of 2.5%, yeast extract as the additional nitrogen source, an initial pH of 7, NaCl as the mineral salt, an initial moisture level of 75%, and Tween 80 as the surfactant. This research provides a reliable and sustainable approach to enzyme production, offering valuable insights for the optimization of the solid-state fermentation process for maximum amylase production.

Effects of tomato straw fermentation on nutrients and bacterial community structure

Unsustainable straw treatment methods detrimentally affect the environment and ecology. Aerobic fermentation (AE) and anaerobic fermentation (AN) are environmentally friendly treatments that better utilise straw resources. In this study, high-throughput sequencing was used to investigate the effects of AE and AN on nutrient content and microbial community structure during tomato straw fermentation. Nitrate nitrogen, available phosphorus, available potassium, and fulvic acid contents following AE were 1250.04 mg/kg, 80.34 %, 161.39 %, and 49.31 %, respectively, which were higher than those following AN. Ammonium nitrogen, humic acid, and humic substance levels following AN were 309.07 %, 31.18 %, and 17.38 %, respectively, which were higher than those following AE. Firmicutes (24.76 %) and Actinobacteria (12.93 %) were more abundant following AE, whereas Proteobacteria (33.82 %) and Bacteroidetes (33.82 %) exhibited higher abundance following AN. AE more effectively eliminated pathogenic bacteria (22.01%–0.26 %) and encouraged stronger interactions between dominant bacterial genera. Redundancy and Mantel test analyses revealed that electrical conductivity and temperature were the most important environmental factors affecting bacterial communities in AE and AN, respectively. AE had a stronger effect on effective nutrient release from tomato straw, implying its greater application potential as a fertiliser. Overall, our study provides a theoretical basis for the optimisation of fermentation methods and processes.

Utilization of citrus, date, and jujube substrates for anaerobic digestion processes

AbstractThis research explores the potential for generating biogas and clean energy by processing organic waste, a process that can become a sustainable solution to Syria's energy needs. Focusing on agricultural residues generated from citrus fruit orange, date, and jujube cultivation in Syria, this study evaluates the potential for anaerobic digestion of these residues for biogas production. It highlights the influence of substrate composition and the optimization of fermentation processes on biogas and methane production. The study focuses on evaluating the anaerobic digestion process by examining various dosages ranging from 20% to 50% dry matter for citrus orange waste, and different types of substrate with a fixed ratio of 20% substrate dry matter. It specifically discusses the factors influencing the inhibitory effect of anaerobic digestion, giving particular consideration to orange waste, a significant byproduct of the citrus industry. The biogas produced maintained a stable methane content when a citrus‐to‐inoculum ratio of 30:70 was used. Jujube waste, characterized by a composition rich in cellulose and hemicellulose, exhibited a higher potential for biogas and methane generation among the fruit waste investigated, particularly when combined with the inoculum in a 20:80 ratio. The research findings underscore the potential of using Syrian agricultural residues, including orange citrus peel, date, and jujube fruit, for the production of biogas through anaerobic digestion.

Impact of Bacillus subtilis on Chinese yellow rice wine (Huangjiu) fermentation: Method variations and flavor analysis

This investigation explores the impact of various fermentation techniques and the inoculation of Bacillus subtilis spores on the physicochemical properties and principal flavor profiles of Huangjiu. Employing sensory analysis, headspace solid-phase microextraction, gas chromatography-tandem mass spectrometry (HS-SPME-GC–MS), and orthogonal partial least squares discriminant analysis (OPLS-DA), we observed that these variables significantly alter the physicochemical attributes of Huangjiu. Our analysis, integrating volatile organic compounds (VOCs) with odor activity values (OAV), revealed that while B. subtilis inoculation modifies the concentrations of key flavor compounds, it does not affect their types. Notably, the inoculation enhances the concentrations of 13 primary flavor compounds, thereby enriching floral and fruity notes while reducing higher alcohol levels. These findings contribute valuable insights into the flavor formation mechanisms of Huangjiu and guide the optimization of fermentation processes.

A Preliminary Study on the Effect of Adding Sugarcane Syrup on the Flavor of Barley Lager Fermentation.

This study focuses on the diversified utilization of the sugarcane industry, and sugarcane syrup, as a by-product of the sugarcane industry, is a good raw material for fermentation. Bringing sugarcane syrup into beer is conducive to the enrichment of the sugar industry, and it can improve the flavor of beer and make it more aromatic. This study determined the optimal fermentation process for beer. By analyzing the consumption rate of the carbon and nitrogen sources of raw materials, the nutrient utilization of yeast, and the causes of differences in flavor substances, the flavor composition and flavor stability of beer were determined by SPME-HS-GC-MS technology. The results showed that beer brewed with sugarcane syrup as an auxiliary raw material met the basic specifications of beer. The addition of sugarcane syrup to the wort base increased the utilization of amino acids by the yeast, and LS (lager with added cane syrup) increased the nine flavor compounds of the beer, which constituted the basic flavor of the beer, bringing new flavor compounds compared with the normal all-barley beer. Forced aging experiments showed that LS produced fewer aging compounds than OWBL. Various experiments have shown that it is feasible to ferment beer with sugarcane syrup instead of partial wort.

Online Monitoring of the Temperature and Relative Humidity of Recycled Bedding for Dairy Cows on Dairy Farms

In large-scale dairy farming, the use of high-temperature-fermented dairy manure bedding instead of rice husk-based bedding and other commercial types of bedding is widely favored. Strip-stacking aerobic fermentation is the main production method of dairy manure bedding, but it has problems including unstable fermentation and the secondary breeding of pathogens. In this work, a multi-probe, integrated, online monitoring system for temperature and relative humidity was used for fermentation process optimization. The effects of the temporal and spatial distribution of fermentation temperature and relative humidity on the nutrient content curve and the moisture and ash content of manure bedding materials were systematically studied. The effect of the fermentation process on the retention rate of effective bedding materials (cellulose, hemicellulose, and lignin) was analyzed. The experiments proved that high-quality bedding material can be obtained through reasonable stacking fermentation. The fabricated bedding material has a total dry base content consisting of cellulose, hemicellulose, and lignin of 78%, an ash content of 6%, and a nutrient content of 17%. The obtained bedding material was produced to increase the bed rest rate and continuously inhibit the bedding bacteria content, keeping it at a low level for 5 days. This study proves that temperature and humidity monitoring can guide the optimization of the strip-stacking fermentation process of dairy manure and that it can be applied to large-scale farms to improve fermentation parameters.

Enhancing the yield of Xenocoumacin 1 in Xenorhabdus nematophila YL001 by optimizing the fermentation process

Xenocoumacin 1 (Xcn 1), antibiotic discovered from secondary metabolites of Xenorhabdus nematophila, had the potential to develop into a new pesticide due to its excellent activity against bacteria, oomycetes and fungi. However, the current low yield of Xcn1 limits its development and utilization. To improve the yield of Xcn1, response surface methodology was used to determine the optimal composition of fermentation medium and one factor at a time approach was utilized to optimize the fermentation process. The optimal medium composed of in g/L: proteose peptone 20.8; maltose 12.74; K2HPO4 3.77. The optimal fermentation conditions were that 25 °C, initial pH 7.0, inoculum size 10%, culture medium 75 mL in a 250 mL shake flask with an agitation rate of 150 rpm for 48 h. Xenorhabdus nematophila YL001 was produced the highest Xcn1 yield (173.99 mg/L) when arginine was added to the broth with 3 mmol/L at the 12th h. Compared with Tryptic Soy Broth medium, the optimized fermentation process resulted in a 243.38% increase in Xcn1 production. The obtained results confirmed that optimizing fermentation technology led to an increase in Xcn1 yield. This work would be helpful for efficient Xcn1 production and lay a foundation for its industrial production.

Isolation, purification, characterization, and analysis of the antioxidant activity of antioxidant peptides from walnut milk fermented with Lactobacillus paracasei SMN-LBK

Microbial fermentation is an important method to obtain antioxidant peptides. In this study, Lactobacillus paracasei SMN-LBK was used to ferment biprotein (walnut milk) to enhance the antioxidant capacity of the active peptide. The optimal fermentation process of 5% inoculum, 1:7 plant protein ratio, 37 °C and 12 h resulted in 77.63% ± 0.36% DPPH scavenging, 93.31% ± 0.25% ABTS scavenging and 79.37% ± 0.02% metal ion chelating capacity. The hydrolysis products were classified into three molecular weight classes using an ultrafiltration membrane system. DPAP-3 with a molecular weight of less than 3 kDa had the highest antioxidant activity. DPAP-3 was further separated into three subclasses using gel filtration chromatography. These subclasses were characterized using nano-liquid chromatography-tandem mass spectrometry (NanoLC-MS/MS) and screened by searching https://biochemia.uwm.edu. Four antioxidant peptides were identified using a combination of hydrophobic amino acids, positive ion-charge interactions, and numerical values. Two of these peptides were from animals and one from plants. Their amino acid sequences are as follows PCRGVLLR, IHIPLPRWV, QSKVLPVPQK, and KVLPVPQKA, with molecular docking binding energies of −7.8, −8.3, −7.4, and −7.7 kcal/mol, respectively. The results suggest that fermentation of plant-animal proteins with Lactobacillus paracasei can produce antioxidant peptides that can be used as functional ingredients in food products. This has the potential to conserve animal protein resources.