Enzyme Dosage Research Articles

Effect of different pretreatments on the hydrolysis efficiency and flavor of squid viscera (Dosidicus gigas)

In order to better utilize the proteins in squid viscera (Dosidicus gigas), the effects of different pretreatments on the degree of hydrolysis (DH) and volatile compounds of hydrolysates were studied. The DH was used as an indicator to select alkaline protease as the optimal protease from 8 types of proteases. The hydrolysis conditions of alkaline protease and the flavor of the hydrolysates were investigated through single-factor, response surface methodology, and pretreatment-assisted hydrolysis experiments. The results showed that alkaline protease had the highest DH (42.08 ± 0.58%), with enzyme dosage of 1.5%, hydrolysis temperature of 45 °C, hydrolysis time of 5 h, and liquid-solid ratio of 4:1. The DH of the ultrasound group was significantly higher than other groups, with the highest DH (45.67 ± 0.34%) after ultrasound for 40 min (U40). The proportion of molecular weight < 1 kDa and the content of free amino acids (1613.64 mg/100 g) were the highest at U40. The electronic nose results indicated that the composition of organic sulfur and aliphatic compounds in U40 showed significant differences compared to other products. The U40 sample had the highest type and content of volatile compounds (especially benzaldehyde), which was more conducive to the volatilization of flavor substances. In conclusion, U40 pretreatment can effectively improve the DH and flavor of hydrolysates.

Effects of Phytase Enzyme Supplementation on Protein Digestibility, Efficiency of Feed Utilization, Growth and Carcass Protein Content of Tambaqui (Colossoma macropomum)

The success of intensive cultivation of tambaqui (Colossoma macropomum) is largely determined by the availability of high-quality feed. High-quality fish diets consist of fish meal produced from animal-based proteins in order to provide fish with the protein requirement. In intensive farming systems, feed costs account for 60–70% of total production costs. Due to fish meal's increasing cost; hence, less expensive ingredients are required to substitute fish meal. Soybean meal can be used as an alternative source of protein to substitute for fish meal due to its lower price and high protein content in fish diet. Phytate acid, one of the antinutritional materials contained within soybean meal, may decrease the nutrient content of feed by affecting with the fish's ability to absorb nutrients. The research aimed to examine the effect of adding the phytase enzyme to diet feed on increasing protein digestibility, efficiency of feed utilization and growth of tambaqui fish. The fingerlings of tambaqui used as test fish had an average weight of 3.85 ± 0.08 g/fish. The diet treatments in this study were phytase enzyme addition in diet with different doses, namely: A (0 FTU/kg feed), B (250 FTU/kg feed), C (500 FTU/kg feed), D (750 FTU/kg feed) and E (1000 FTU/kg feed). The results showed that phytase enzyme addition in diet significantly increased protein digestibility, efficiency of feed utilization, growth, and carcass protein content of tambaqui fish. The most effectiveness phytase enzyme dose for tambaqui fish was 1000 FTU/kg feed, which resulted in highest values of ADCp, PER, FCR, and RGR at 68.46±0.28%, 2.27±0.05, 1.53±0.04, 74.59±0.45%, and 6.01±0.19%/day, respectively.

Alginate-laccase beads in the decolourization of indigo carmine

The aim of the study was to assess the feasibility of utilizing sodium alginate biopolymer as animmobilization carrier for laccase in the removal of indigo carmine (IC), an anionic dye. The main goal of this work was to optimize the decolourization process by selecting the appropriate immobilized enzyme dose per 1 mg of dye, as well as the process temperature. The effective immobilization of laccase using sodium alginate as a carrier was confirmed by Raman spectroscopy. An analysis of the size and geometric parameters of the alginate beads was also carried out. Tests of IC decolourization using alginate-laccase beads were conducted. Applying the most effective dose of the enzyme (320 mg of enzyme/1 mg of IC) made it possible to remove 92.5% of the dye over 40 days. The optimal temperature for the IC decolourization process, using laccase immobilized on sodium alginate, was established at 30-40ºC. The obtained results indicate that laccase from Trametes versicolor immobilized on sodium alginate was capable of decolourizing the tested dye primarily based on mechanism of biocatalysis.

Enhanced bio-extraction of medicinally important plants using xylano-pectinolytic enzymes produced concurrently by Bacillus pumilus AJK

Medicinal plant extracts obtained by chemical methods have drawbacks such as high chemical, time and energy consumption, byproduct formation, lower quality and efficiency, high toxicity and lead to degradation of thermo-sensitive bioactive compounds. The purpose of this study was to investigate the ability of crude xylano-pectinolytic enzymes concoction in bio-extraction of medicinally important plant extracts, in order to reduce the use of chemical solvents for extraction. In this regard, various conditions including material to liquid ratio (MLR), enzyme dose, treatment time and stirring speed were optimized and several physico-chemical parameters including pH, total soluble solid (TSS), total dissolved solid (TDS), clarity, reducing sugar, polyphenolic content, viscosity and filterability were studied. The bio-extraction efficiency was found to be best at material-to-liquid ratio (MLR) 1:5-1:6 (g/mL), enzyme dose range between 15:60 to 20:80 (IU/g), treatment time range between 180- 240 min and stirring speed range between 50-60 rpm. Enzymatic extraction of medicinal plants resulted in 7-17% increase in clarity, 27-50% increase in TSS, 3-42% increase in reducing sugars, 20-42% increase in polyphenolic content, 13-53% increase in TDS, 15-22% increase in filterability along with decrease of 3-5% in pH and 18-43% in viscosity. Hence, this eco-friendly bio-extraction strategy could reduce the chemical usage, along with improvement of the physico-chemical properties of medicinally important extracts. The technique also enables the application of greener chemistry in the pharmaceuticals to optimize cleaner methods of extraction. This is the first report manifesting bio-extraction of medicinally important plant extracts using crude xylano-pectinolytic enzymatic concoction produced by a single bacterial isolate.

Application of Thermomyces lanuginosus polygalacturonase produced in Komagataella phaffii in biomass hydrolysis and textile bioscouring

In this work, the polygalacturonase (TL-PG1) from the thermophilic fungus Thermomyces lanuginosus was heterologously produced for the first time in the yeast Komagataella phaffii. The TL-PG1 was successfully expressed under the control of the AOX1 promoter and sequentially purified by His-tag affinity. The purified recombinant pectinase exhibited an activity of 462.6 U/mL toward polygalacturonic acid under optimal conditions (pH 6 and 55 ˚C) with a 2.83 mg/mL and 0.063 μmol/minute for Km and Vmax, respectively. When used as supplementation for biomass hydrolysis, TL-PG1 demonstrated synergy with the enzymatic cocktail Ctec3 to depolymerize orange citrus pulp, releasing 1.43 mg/mL of reducing sugar. In addition, TL-PG1 exhibited efficiency in fabric bioscouring, showing potential usage in the textile industry. Applying a protein dosage of 7 mg/mL, the time for the fabric to absorb water was 19.77 seconds (ten times faster than the control). Adding the surfactant Triton to the treatment allowed the reduction of the enzyme dosage by 50% and the water absorption time to 6.38 seconds. Altogether, this work describes a new versatile polygalacturonase from T. lanuginosus with the potential to be employed in the hydrolysis of lignocellulosic biomass and bioscouring.

Exocrine pancreatic insufficiency and fat malabsorption related to pancreatectomy and other gastrointestinal surgery: A narrative review.

Surgical resection is the mainstay of treatment for patients with tumors of the pancreas. There are a number of well-recognized complications that account for the significant morbidity associated with the operation, including exocrine pancreatic insufficiency (EPI). Patients with pancreatic cancer commonly have evidence of EPI prior to surgery, and this is exacerbated by an operation, the extent of the insult being dependent on the indication for surgery and the operation performed. There are accumulating data to demonstrate that treatment of EPI with pancreatic enzyme replacement (PERT) enhances clinical outcomes after surgery by reducing critical complications; this in turn may enhance oncological outcomes. Data would indicate that quality of life (QoL) is also improved after surgery when enzymes are prescribed. To date, many surgeons and clinicians have not appreciated the need for PERT or the benefits it may bring to their patients; therefore, education of clinicians remains a significant opportunity. In turn, patient education about consumption of the correct dose of enzymes at the appropriate time is key to an optimal outcome. In addition, because of the complex nature of the regulation of pancreatic exocrine function, there is evidence to support the presence of EPI following operations performed on other gastrointestinal (GI) organs, including the esophagus, stomach, and small intestine. The aim of this review is to document the existing published evidence in relation to EPI and its treatment with PERT following GI surgery.

Changes in the Main Physicochemical Properties and Electrochemical Fingerprints in the Production of Sea Buckthorn Juice by Pectinase Treatment.

Enzymatic hydrolysis using pectinase is critical for producing high-yield and quality sea buckthorn juice. This study determined the optimal temperature, time, and enzyme dosage combinations to guide manufacturers. A temperature of 60 °C, hydrolysis time of 3 h, and 0.3% enzyme dosage gave 64.1% juice yield-25% higher than without enzymes. Furthermore, monitoring physicochemical properties reveals enzyme impacts on composition. Higher dosages increase soluble solids up to 15% and soluble fiber content by 35% through cell wall breakdown. However, excessive amounts over 0.3% decrease yields. Pectin concentration also declines dose-dependently, falling by 91% at 0.4%, improving juice stability but needing modulation to retain viscosity. Electrochemical fingerprinting successfully differentiates process conditions, offering a rapid quality control tool. Its potential for commercial inline use during enzymatic treatment requires exploration. Overall, connecting optimized parameters to measured effects provides actionable insights for manufacturers to boost yields, determine enzyme impacts on nutrition/functionality, and introduce novel process analytical technology. Further investigations of health properties using these conditions could expand sea buckthorn juice functionality.

Case study of chemical and enzymatic degumming processes in soybean oil production at an industrial plant

The vegetable oil degumming process plays a critical role in refining edible oil. Phospholipids (PL) removal from crude extracted soybean oil (SBO) by the enzymatic degumming process has been investigated in this work. Enzymatic degumming of extracted SBO with microbial phospholipase A1 PLA-1 Quara LowP and Lecitase Ultra enzymes have also been studied comparatively. The main novelty of our work is the use of the enzymatic degumming process on an industrial scale (600 tons a day). Many parameters have been discussed to understand in detail the factors affecting oil losses during the degumming process. The factors such as chemical conditioning (CC) by phosphoric acid 85%, the enzyme dosage mg/kg (feedstock dependent), the enzymatic degumming reaction time, and the characteristics of the plant-processed SBO have been discussed in detail. As a main point, the degummed oil with a phosphorus content of < 10 mg/kg increases yield. Quara LowP and Lecitase Ultra enzymes are not specific for certain phospholipids PL; however, the conversion rate depends on the SBO phospholipid composition. After 4 h, over 99% of Phospholipids were degraded to their lysophospholipid LPL (lysolecithin). The results showed a significant effect of operating parameters and characteristics of different origins of SBO, fatty acids FFA content, Phosphorus content and total divalent metals (Calcium Ca, Magnesium Mg and Iron Fe mg/kg) content on the oil loss. The benefit of using enzymatic degumming of vegetable oils rather than traditional chemical refining is that the enzymatic degumming process reduces total oil loss. This decrease is known as enzymatic yield. The enzymatic degumming also decreases wastewater and used chemicals and running costs; moreover, it enables physical refining by lowering the residue phosphorus to < 10 mg/kg.

Ozone Exposure Controls Oxidative Stress and the Inflammatory Process of Hepatocytes in Murine Models.

(1) Background: Ozone exposure is a promising tool for treating liver damage since it is known to control the release of free radicals and increase the expression of antioxidant enzymes. The objective is to investigate the main intracellular pathways activated after exposure to ozone, considering the dosage of antioxidant enzymes and markers of oxidative stress. (2) Methods: This systematic review was performed based on the PRISMA guidelines and using a structured search in MEDLINE (PubMed), Scopus, and Web of Science. Bias analysis and methodological quality assessments were examined using the SYRCLE Risk of Bias tool. (3) Results: Nineteen studies were selected. The results showed that the exposure to ozone has a protective effect on liver tissue, promoting a decrease in inflammatory markers and a reduction in oxidative stress in liver tissue. In addition, ozone exposure also promoted an increase in antioxidant enzymes. The morphological consequences of controlling these intracellular pathways were reducing the tissue inflammatory process and reducing areas of degeneration and necrosis. (4) Conclusions: Ozone exposure has a beneficial effect on models of liver injury through the decrease in oxidative stress in tissue and inflammatory markers. In addition, it regulates the Nrf2/ARE antioxidant pathway and blocks the NF-κB inflammatory pathway.

Three cases of scombroid syndrome in Italy: clinical and preventive considerations.

Foodborne diseases are common sources of morbidity and mortality worldwide. Scombroid syndrome represents a particular condition since it is not directly related to the ingestion of spoiled food but is determined by high levels of histamine, a chemical mediator naturally produced within the human body under particular conditions. In these cases, histamine is formed as a result of the bacterial activity from histidine, an amino acid present at high levels in some fish species. The resulting symptomatology can range from mild symptoms such as headache and skin rash to more severe manifestations such as hypotension and coronary spasms. Reference regulations in Italy set maximum levels of histamine in food at 200 mg/kg. The cases described involve a family of three who, following the ingestion of a tuna dish, started to exhibit symptoms typical of an allergic reaction. In one case, hypotension, tachycardia, and electrocardiographic changes in the ST-tract suggestive of myocardial ischemia also appeared with negative myocardionecrosis enzyme dosage. All three cases experienced complete remission of symptoms in the absence of sequelae. Histamine concentrations in fish sampled three days later were 169 mg/kg. The cases described emphasize the importance of proper differential diagnosis as well as the importance of implementing specific controls in food hygiene.

Comparative study of chemical and enzymatic pre-treatments of Kitchen waste (KW) to generate fermentable sugars for the production of polyhydroxyalkanoates (PHA)

Kitchen waste (KW) production is anticipated to rise because of accelerated urban and economic growth, particularly in Asian nations. Therefore, applications/recycling of KW should be explored for its effective utilization to generate a sustainable source of energy that will ultimately avoid massive economic and energy losses. The main goal of this study was to investigate the combinational effect of thermochemical and enzymatic pre-treatment on synthetic KW to enhance the release of fermentable sugars for the synthesis of value-added products such as polyhydroxyalkanoates (PHAs). The thermochemical pre-treatment of KW samples was performed using 4 N HCl, 4 N H2SO4, and 4 N NaOH at a concentration of 0–30% (v/v), 30–100 °C for 0–120 min. Furthermore, to enhance the total fermentable sugars, variable enzyme dosages were added after acidic or alkaline pre-treatments (0–1% v/v α-amylase and 0–1% v/v amyloglucosidase-AMG) while maintaining a constant pH 5, temperature 50 °C, and time 30 min. Pre-treatment of KW with acid (15% v/v 4 N HCl in water) at temperature 100 °C, followed by enzymatic hydrolysis, i.e., 0.6% (v/v) α-amylase and 0.4% (v/v) amyloglucosidase (at pH 5, 50 0C and 60 min) resulted in the maximum concentration of fermentable reducing sugars (46.13 g/L) in the hydrolysate.

Comparative study for enzymatic hydrolysis of sugarcane bagasse using free and nanoparticle immobilized holocellulolytic enzyme cocktail

This study aimed at a comparative analysis of enzymatic hydrolysis of sugarcane bagasse biomass by using a free holocellulase cocktail of an enzyme (Cellic CTec2) and immobilized enzyme system on IOMNP@SiO2-NH2 support to make the process cost-effective. The immobilization was substantiated by examining the nano biocatalysts using FTIR, TEM, and DLS analysis. IOMNP@SiO2 –NH2 and immobilized enzyme system have average particle sizes of 81.1, and 455.9 nm confirmed by DLS. Response surface methodology (RSM) based on the Box Behnken design was employed to optimize the saccharification process conditions. The free cellulase cocktail showed a maximum hydrolysis yield of 70.71 % at 12.5FPU/gm enzyme dose, while the immobilized enzyme at a dose of 20 FPU/gm achieved a conversion rate of 74.19 % under optimized parameters likes hydrolysis time, surfactant and substrate concentration. Pretreated sugarcane bagasse substrate was used to evaluate the reusability potential of magnetically immobilized cellulase enzyme compared to recovered free enzyme up to 5 cycles. In the second and third cycles, the conversion rates of cellulose to glucose were determined to be 68.21 %, and 52.6 %, respectively. The findings of this study demonstrate that the immobilized Cellic CTec2 enzyme cocktail on functionalized IOMNP is effective for hydrolyzing sugarcane bagasse which also provides the advantages of easy recovery and reusability of enzyme for multiple hydrolysis cycles.

Sustainable cellobionic acid biosynthesis from starch via artificial in vitro synthetic enzymatic biosystem

Cellobionic acid (CBA), a kind of aldobionic acid, offers potential applications in the fields of pharmaceutical, cosmetic, food, and chemical industry. To tackle the high cost of the substrate cellobiose in CBA production using quinoprotein glucose dehydrogenase, this study developed a coenzyme-free and phosphate-balanced in vitro synthetic enzymatic biosystem (ivSEBS) to enable the sustainable CBA synthesis from cost-effective starch in one-pot via the CBA-synthesis module and gluconic acid-supply module. The metabolic fluxes of this artificial biosystem were strengthened using design−build−test−analysis strategy, which involved exquisite pathway design, meticulous enzyme selection, module validation and integration, and optimization of the key enzyme dosage. Under the optimized conditions, a remarkable concentration of 6.2 g/L CBA was achieved from initial 10 g/L maltodextrin with a starch-to-CBA molar conversion rate of 60 %. Taking into account that the biosystem simultaneously accumulated 3.6 g/L of gluconic acid, the maltodextrin utilization rate was calculated to be 93.3 %. Furthermore, a straightforward scaling-up process was performed to evaluate the industrial potential of this enzymatic biosystem, resulting in a yield of 21.2 g/L CBA from 50 g/L maltodextrin. This study presents an artificial ivSEBS for sustainable production of CBA from inexpensive starch, demonstrating an alternative paradigm for biomanufacturing of other aldobionic acids.

Production of xylooligosaccharides from different pretreated Camellia oleifera shell with crude enzyme from Aspergillus fumigatus

Production of xylooligosaccharides (XOS) from Camellia oleifera shell (COS) using the crude enzyme from Aspergillus fumigatus (A. fumigatus) was investigated. COS was subjected to different pretreatments, namely thermal autoclaving, combination of diluted acid, diluted alkali or concentrated alkali with thermal autoclaving, microwave-assited ethanol pretreatment. The results showed that the combination of diluted alkali with thermal autoclaving is the best pretreatment for XOS production. The optimal conditions for enzymatic xylooligosaccharides production from the best pretreatment are: temperature 50 °C, pH 4.5, substrate concentration 8% (m/V), enzymatic hydrolysis time 9 h, and enzyme dose of 40 IU/g substrate. A maximum yield of 17.08 ± 0.67 g/L of XOS was obtained under the above optimum conditions for the best pretreatment.

An ecofriendly and sustainable method for efficient cleaning of raw cashmere fiber with enzymes in SCF-CO2

An efficient and ecofriendly method to remove impurities from raw cashmere fiber was constructed by employing various biological enzymes (lipase, cellulase, pectinase and protease) to form reverse microemulsion in supercritical carbon dioxide. An investigation and optimization of various parameters, such as the dosage of biological enzymes, cleaning temperature, pressure and duration for this method were performed according to the whiteness and weight loss of the treated samples. Then the proposed method was further validated by energy dispersive X-ray spectrometry (EDS), scanning electron microscopy (SEM) analysis to verify the impurity removal from cashmere fiber. Moreover, the aggregation structure and properties of cleaned cashmere fiber were also investigated by X-ray powder diffraction (XRD), strength measurement, moisture absorption testing and thermal analysis (TG-DTG). The results declare that, via forming a type of (W+enzymes)/SCF-CO2 microemulsion to transfer the biological enzymes into supercritical medium, various impurities from raw cashmere fiber could be efficiently decreased and removed, which cleaning performance was better than a conventional control process. The cleaning efficiency for various impurities was notably influenced by the dosages of the applied enzymes and system parameters, which could be improved at an appropriate condition in SCF-CO2. Furthermore, the cleaning feasibility, reliability and efficiency of the enzymatically catalyzed method for raw cashmere fiber was successfully verified by SEM, EDS, XRD, TG-DTG, tensile strength analysis, and a supercritical cleaning mechanism for raw cashmere was further proposed.

Modelling and optimization of simultaneous saccharification and fermentation of agro-food residues

In terms of productivity, bioethanol production from agri-food residues can be improved by carrying out simultaneously the two involved stages (enzyme hydrolysis and alcohol fermentation). This is because the shorter the process duration, the higher the overall productivity achieved. In this work, two kinetic models previously developed by the authors have been combined to obtain a general kinetic model for the simultaneous saccharification and fermentation of lignocellulosic materials. The validity of the combined model has been tested using three different agri-food residues: wheat straw, rice husks and exhausted sugar beet pulp. Later, the model has been used to calculate productivity under many different conditions and to optimize the industrial process. After this study, it is concluded that some important operating variables, such as the enzyme dose and the inoculum strength, must be well coupled to obtain the maximal yield from residues. Thus, the overall productivity obtained in the laboratory experiments was under 5 mg of ethanol per gram of solid waste and per hour, but the simulations in the optimal operating conditions predict productivities of up to twice that.

Long-Term Outcome of Infantile Onset Pompe Disease Patients Treated with Enzyme Replacement Therapy - Data from a German-Austrian Cohort.

Enzyme replacement therapy (ERT) with recombinant human alglucosidase alfa (rhGAA) was approved in Europe in 2006. Nevertheless, data on the long-term outcome of infantile onset Pompe disease (IOPD) patients at school age is still limited. We analyzed in detail cardiac, respiratory, motor, and cognitive function of 15 German-speaking patients aged 7 and older who started ERT at a median age of 5 months. Starting dose was 20 mg/kg biweekly in 12 patients, 20 mg/kg weekly in 2, and 40 mg/kg weekly in one patient. CRIM-status was positive in 13 patients (86.7%) and negative or unknown in one patient each (6.7%). Three patients (20%) received immunomodulation. Median age at last assessment was 9.1 (7.0-19.5) years. At last follow-up 1 patient (6.7%) had mild cardiac hypertrophy, 6 (42.9%) had cardiac arrhythmias, and 7 (46.7%) required assisted ventilation. Seven patients (46.7%) achieved the ability to walk independently and 5 (33.3%) were still ambulatory at last follow-up. Six patients (40%) were able to sit without support, while the remaining 4 (26.7%) were tetraplegic. Eleven patients underwent cognitive testing (Culture Fair Intelligence Test), while 4 were unable to meet the requirements for cognitive testing. Intelligence quotients (IQs) ranged from normal (IQ 117, 102, 96, 94) in 4 patients (36.4%) to mild developmental delay (IQ 81) in one patient (9.1%) to intellectual disability (IQ 69, 63, 61, 3x <55) in 6 patients (54.5%). White matter abnormalities were present in 10 out of 12 cerebral MRIs from 7 patients. Substantial motor, cardiac, respiratory, and cognitive deficits are frequent in IOPD long-term survivors who started ERT before 2016. The findings of this study can be valuable as comparative data when evaluating the impact of newer treatment strategies including higher enzyme dosage, immunomodulation, modified enzymes, or early start of treatment following newborn screening.

Produção de queijo fresco usando papaína liofilizada como coagulante vegetal

Abstract The study examined the efficacy of freeze-dried papain enzyme obtained from three Vasconcellea species (V. pubescens, V. chachapoyensis, V. heilbornii) as a natural coagulant in cheese making. Notably, the enzyme V. pubescens demonstrated the most promising results when concentrations of 2 g/L, 4 g/L, and 6 g/L were used to produce fresh cheese, while other enzyme species exhibited lower efficacy. The optimal yield of fresh cheese with minimal residual enzyme was achieved when a 2 g/L dose of papain enzyme was employed at a coagulation temperature of 30 °C, resulting in physicochemical and organoleptic characteristics comparable to those produced with commercial Hansen’s rennet. Nevertheless, an increase in the coagulation temperature (42 °C) and a higher dose of papain enzyme (4 g/L) resulted in a reduction in the yield of fresh cheese and; consequently, the residual enzyme increased. Further studies are required to determine the purity of freeze-dried papain and the most effective dosage to increase profitability for producers and consumers. Such findings could facilitate the ecological application of this alternative in producing of fresh cheese.

Study on extraction of antioxidant polypeptides from byproducts of Procambarus clarkii

This article primarily focuses on studying the peptide extraction method and antioxidant properties of crayfish by-products using alkaline protease. Taking the degree of protein hydrolysis as a measurement indicator, and considering enzyme dosage, solid-liquid ratio, and hydrolysis time as optimization factors, we utilized single-factor and orthogonal experiments to refine the extraction process. Additionally, we assessed its antioxidant activity using the DPPH method. The findings revealed that the optimal process conditions were as follows: an enzyme dosage of 2% alkaline protease, a solid-liquid ratio of 1:20, and an enzymatic hydrolysis time of 2 hours. The measured protein hydrolysis degree was 21.08%, and its DPPH free radical scavenging rate reached 70.55%. When compared to vitamin C, it demonstrated superior antioxidant capabilities in scavenging free radicals.

Extraction of Polysaccharides from Shiitake Mushroom Based on Pineapple Enzymatic Technology

The pineapple enzymatic technique was used to extract polysaccharides from shiitake mushrooms, with glucose serving as the standard for calculating the yield of the polysaccharides. The effects of acidity, alkalinity, extraction time, extraction temperature, enzyme dosage, and number of extractions on the extraction rate of mushroom polysaccharides were investigated under the control of a single factor. Orthogonal tests showed that the optimal process combination for the extraction of mushroom polysaccharides under the influence of multiple factors was pH 5, an extraction time of 1.5 hours, extraction temperature of 45°C, enzyme dosage of 0.8/100 g, and the number of extraction times of 3. The yield of mushroom polysaccharides under optimal processing conditions could reach 6.47%. The method is easy to operate, accurate, highly efficient, and it is a non-polluting and cost-effective process for extracting shiitake polysaccharides.