Optimization Of Enzymatic Hydrolysis Research Articles

Optimization of Enzymatic Hydrolysis and Fermentation Processing for Set-Type Oat Yogurt with Favorable Acidity and Coagulated Texture.

The key role of enzymatic hydrolysis and fermentation in the sensory quality of set yogurt made from whole oats was demonstrated. The optimal process was established by the orthogonal and response surface methodology based on the acidity, textural, and rheological properties. The results indicated that the enzymatic hydrolysis appropriately consisted of liquefaction with 12 U/mL α-amylase at 70 °C and pH 6.5 for 60 min, followed by saccharification with 400 U/mL α-1,4-glucan glucohydrolase at 60 °C and pH 4.5 for 60 min. The Streptococcus thermophilus ST15 and Lactobacillus bulgaricus 20249 strains were the most efficacious strains, with a 0.1% inoculation for the fermentation at 42 °C for 16 h. So, a soft semisolid oat yogurt formed with an 8% solid-liquid ratio, which exhibited an acidity of 73.17 °T, a cohesiveness ratio of 0.51, and a maximum apparent viscosity of 1902.67 Pa·s. The coagulated texture of the oat yogurt was closely associated with the exopolysaccharide (EPS) yield up to 304.99 mg/L. These findings supported the optimal processing of oat yogurt, especially its correlation with the high capacity of EPS production by strains. It is an innovative and feasible way to improve the properties of set-type oat yogurt, especially the utilization of the whole oat.

Optimization of enzymatic hydrolysis of cellulose extracted from bamboo culm for bioethanol production by Saccharomyces cerevisiae modified via CRISPR/Cas9

Optimization of enzymatic hydrolysis of cellulose extracted from bamboo culm for bioethanol production by Saccharomyces cerevisiae modified via CRISPR/Cas9

Optimization of enzymatic hydrolysis and fermentation of bleached garlic straw for bioethanol production

Optimization of enzymatic hydrolysis and fermentation of bleached garlic straw for bioethanol production

Sequential Enzymatic Hydrolysis and Ultrasound Pretreatment of Pork Liver for the Generation of Bioactive and Taste-Related Hydrolyzates.

In the study of protein-rich byproducts, enzymatic hydrolysis stands as a prominent technique, generating bioactive peptides. Combining exo- and endopeptidases could enhance both biological and sensory properties. Ultrasound pretreatment is one of the most promising techniques for the optimization of enzymatic hydrolysis. This research aimed to create tasteful and biologically active pork liver hydrolyzates by using sequential hydrolysis with two types of enzymes and two types of ultrasound pretreatments. Sequential hydrolyzates exhibited a higher degree of hydrolysis than single ones. Protana Prime hydrolyzates yielded the largest amount of taste-related amino acids, enhancing sweet, bittersweet, and umami amino acids according to the Taste Activity Value (TAV). These hydrolyzates also displayed significantly higher antioxidant activity. Among sequential hydrolyzates, Flavourzyme and Protana Prime hydrolyzates pretreated with ultrasound showed the highest ferrous ion chelating activity. Overall, employing both Alcalase and Protana Prime on porcine livers pretreated with ultrasound proved to be highly effective in obtaining potentially tasteful and biologically active hydrolyzates.

Development of an LC-MS/MS method for the determination of five psychoactive drugs in postmortem urine by optimization of enzymatic hydrolysis of glucuronide conjugates.

Toxicological analyses of biological samples play important roles in forensic and clinical investigations. Ingested drugs are excreted in urine as conjugates with endogenous substances such as glucuronic acid; hydrolyzing these conjugates improves the determination of target drugs by liquid chromatography-tandem mass spectrometry (LC-MS/MS). In this study, we sought to improve the enzymatic hydrolysis of glucuronide conjugates of five psychoactive drugs (11-nor-9-carboxy-Δ9-tetrahydrocannabinol, oxazepam, lorazepam, temazepam, and amitriptyline). The efficiency of enzymatic hydrolysis of glucuronide conjugates in urine was optimized by varying temperature, enzyme volume, and reaction time. The hydrolysis was performed directly on extraction columns. This analysis method using LC-MS/MS was applied to forensic autopsy samples after thorough validation. We found that the recombinant β-glucuronidase B-One® quantitatively hydrolyzed these conjugates within 3min at room temperature directly on extraction columns. This on-column method saved time and eliminated the loss of valuable samples during transfer to the extraction column. LC-MS/MS-based calibration curves processed with this method showed good linearity, with r2 values exceeding 0.998. The intra- and inter-day accuracies and precisions of the method were 93.0-109.7% and 0.8-8.8%, respectively. The recovery efficiencies were in the range of 56.1-104.5%. Matrix effects were between 78.9 and 126.9%. We have established an LC-MS/MS method for five psychoactive drugs in urine after enzymatic hydrolysis of glucuronide conjugates directly on extraction columns. The method was successfully applied to forensic autopsy samples. The established method will have broad applications, including forensic and clinical toxicological investigations.

Enzymatic Hydrolysis Optimization of Yak Whey Protein Concentrates and Bioactivity Evaluation of the Ultrafiltered Peptide Fractions.

Yak whey protein concentrates (YWPCs) have good functional properties, but there is still a gap in the study of their peptides. In this study, peptides were obtained by enzymatic hydrolysis, and the bioactivity of each ultrafiltration fraction was evaluated using an optimal process. YWPCs were isolated and purified from yak milk as the raw material. Alkaline protease, trypsin, and papain were used to hydrolyze YWPCs. The protease with the highest degree of hydrolysis (DH) and peptide concentration was selected as the most suitable enzyme. The effects of pH, temperature, time, and the enzyme-to-substrate ratio (E/S) on the DH and peptide concentration were investigated, and response surface methodology was utilized to optimize the hydrolysis process. The hydrolysate was separated using ultrafiltration membranes with molecular weight cut-offs of 10 kDa, 5 kDa, 3 kDa, and 1 kDa. The bioactivity of each ultrafiltration component was analyzed, including the inhibition rates of α-amylase and xanthine oxidase (XOD) activities and the scavenging rates of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) cation radicals. The results indicated that alkaline protease was the best enzyme for hydrolyzing YWPCs. The peptide concentration in the YWPC hydrolysate was the highest (17.21 mg/mL) at a pH of 8 and a concentration of 7500 U/g, after 2.5 h at 62 °C. The enzymatic hydrolysate was ultrafiltered to yield four peptide fractions, of which the <1 kDa peptides exhibited the highest α-amylase inhibitory activity (22.06%), XOD inhibitory activity (17.15%), and ABTS cationic free radical scavenging rate (69.55%). This demonstrates the potential of YWPC hydrolyzed peptides for hypoglycemic, uric acid-lowering, and antioxidant applications, providing a theoretical basis for the high-value utilization of YWPCs.

Use of an inexpensive carbon source for the production of a cellulase enzyme complex from Penicillium ucsense S1M29 and enzymatic hydrolysis optimization

AbstractThe high cost of cellulolytic enzyme complexes (CECs) has been a significant impediment to the commercial production of bioproducts from lignocellulose biomass. This study aimed to develop a cost‐effective CEC derived from Penicillium ucsense (former Penicillium echinulatum), utilizing diverse forms of pretreated sugarcane bagasse as the primary carbon/inductor source. Among the different pretreatments used, the hydrothermal pretreatment followed by NaOH delignification (BHD) produced higher FPase and xylanase activities (4.5 FPU mL–1 and 120 IU mL–1) in bioreactor experiments at 20 g BHD L–1 initial concentration. A batch‐mode assay conducted across a range of initial carbon source (5 to 60 g L–1) confirmed the highest FPase activity (4.0 to 5.0 FPU mL–1 at 120 h), in the range of 20–40 g BHD L–1. During these assays the agitation rate, controlled by dissolved O2, tended to stabilize at lower levels, indicating substrate limitation. Conversely, higher initial carbon source concentrations led to an excess of glucose, likely triggering carbon catabolite repression and inhibiting cellulase production. This insight prompted the development of a controlled pulsed fed‐batch strategy, resulting in FPase activity of 11 FPU mL–1 at 220 h using 90 g L–1 BHD controlled fed into the bioreactor. An enzymatic hydrolysis procedure using the generated CEC was also optimized using a central composite rotational design (CCRD). The optimized enzyme hydrolysis conditions achieved a reducing sugar concentration of 80.9 g L–1 in 48 h using 170 g L–1 of BHD as the substrate at a ratio of 15 FPU of enzyme substrate per g of BHD. A preliminary economic assessment demonstrated that, for a first‐ and second‐generation (1G + 2G) ethanol biorefinery, the cost contribution of enzymes would be about US$0.2/L of biofuel. In conclusion, an efficient and highly productive procedure was developed successfully for the production of a CEC. It was particularly effective for the enzymatic hydrolysis of pretreated sugarcane bagasse.

Pretreatment and enzymatic hydrolysis optimization of lignocellulosic biomass for ethanol, xylitol, and phenylacetylcarbinol co-production using Candida magnoliae.

Cellulosic bioethanol production generally has a higher operating cost due to relatively expensive pretreatment strategies and low efficiency of enzymatic hydrolysis. The production of other high-value chemicals such as xylitol and phenylacetylcarbinol (PAC) is, thus, necessary to offset the cost and promote economic viability. The optimal conditions of diluted sulfuric acid pretreatment under boiling water at 95°C and subsequent enzymatic hydrolysis steps for sugarcane bagasse (SCB), rice straw (RS), and corn cob (CC) were optimized using the response surface methodology via a central composite design to simplify the process on the large-scale production. The optimal pretreatment conditions (diluted sulfuric acid concentration (% w/v), treatment time (min)) for SCB (3.36, 113), RS (3.77, 109), and CC (3.89, 112) and the optimal enzymatic hydrolysis conditions (pretreated solid concentration (% w/v), hydrolysis time (h)) for SCB (12.1, 93), RS (10.9, 61), and CC (12.0, 90) were achieved. CC xylose-rich and CC glucose-rich hydrolysates obtained from the respective optimal condition of pretreatment and enzymatic hydrolysis steps were used for xylitol and ethanol production. The statistically significant highest (p ≤ 0.05) xylitol and ethanol yields were 65% ± 1% and 86% ± 2% using Candida magnoliae TISTR 5664. C. magnoliae could statistically significantly degrade (p ≤ 0.05) the inhibitors previously formed during the pretreatment step, including up to 97% w/w hydroxymethylfurfural, 76% w/w furfural, and completely degraded acetic acid during the xylitol production. This study was the first report using the mixed whole cells harvested from xylitol and ethanol production as a biocatalyst in PAC biotransformation under a two-phase emulsion system (vegetable oil/1M phosphate (Pi) buffer). PAC concentration could be improved by 2-fold compared to a single-phase emulsion system using only 1M Pi buffer.

Optimization of Enzymatic Hydrolysis by Protease Produced from Bacillus subtilis MTCC 2423 to Improve the Functional Properties of Wheat Gluten Hydrolysates

This study is aimed at investigating the reutilizing of gluten protein from the wheat processing industry by Bacillus subtilis MTCC 2423 protease to obtain gluten hydrolysates with high added value. Gluten protein hydrolysis using protease achieved a 34.07% degree of hydrolysis with 5% gluten protein, at a hydrolysis time of 2 h for 1000 U/mL at pH 8.0 and temperature of 40°C. Compared to the wheat gluten, the obtained hydrolysates exhibited enhanced functional attributes, including heightened solubility (43%), increased emulsifying activity (93.08 m2/g), and improved radical scavenging properties. Furthermore, these hydrolysates demonstrated enhanced antioxidant potential, as evidenced by elevated ABTS (2,2 ′‐azino‐bis‐(3‐ethylbenzothiazoline‐6‐sulfonic acid) of 81.25% and DPPH (2,2‐diphenyl‐1‐picrylhydrazyl) of 56.46% radical scavenging activities and also exhibited a higher α‐amylase inhibitory effect of 33.98%. The enhancement in functional characteristics of wheat gluten hydrolysates was observed by Fourier transform infrared spectroscopy. The percentage of free amino acids obtained by protease‐mediated hydrolysates increased significantly compared to the unhydrolyzed wheat, which was observed by high‐performance liquid chromatography. These findings suggest that wheat gluten hydrolysates hold promising potential as functional and nutritional food ingredients in the food industry, owing to their enhanced functionalities and potential antioxidant and antidiabetic properties.

Optimization of Enzymatic Hydrolysis of Mackerel (Rastrelliger sp.) Muscle Protein Hydrolysate Using Response Surface Methodology

Mackerel (Rastrelliger sp.) is a widely distributed epipelagic species in South East Asia. Mackerel has a high amount nutrient such as protein (20.83 %) and fat (1.03 %). The high amount of protein and low amount of fat will allow it to be used as a material to produce a good protein hydrolysate. The aim of this study is to determine the optimal enzymatic hydrolysis conditions (time, temperature, and pH) using Response Surface Methodology (RSM). Mackerel Protein Hydrolysate (MPH) was prepared using commercial Flavourzyme. Optimization of MPH was performed by employing Box Behnken Design method of RSM. SN-TCA method was used to calculate the degree of hydrolysis (DH) which is the key parameter in hydrolysis reaction. Optimum hydrolysis conditions were obtained at pH 7, temperature 55°C and 60 min of process. Under these conditions the DH obtained was 17.7293 % with 4% enzyme to substrate ratio. The suggested model for the hydrolysis process is quadratic with the desirability factor of 1. The MPH was further assessed for its amino acid composition using High Performance Liquid Chromatography (HPLC). The hydrolysis process increases the amino acid amounts namely L-Glutamic Acid (19.77%), L-Valin (14.20%), L-Aspartic Acid (11.42%), Glycine (11.04%), L-Alanin (14.20%), L-Prolin (16.80%), and L-Histidin (27.06%). The study suggested that mackerel muscle can be considered to be utilized as fish protein hydrolysis materials.

Optimization and kinetic analysis of untreated brewers’ spent grain saccharification process via enzymatic hydrolysis

Optimization and kinetic analysis of untreated brewers’ spent grain saccharification process via enzymatic hydrolysis

Enzymatic Hydrolysis Optimization for Preparation of Sea Cucumber (Holothuria scabra) Hydrolysate with an Antiproliferative Effect on the HepG2 Liver Cancer Cell Line and Antioxidant Properties.

The sea cucumber body wall was subjected to enzymatic hydrolysis using papain. The relationship between the enzyme concentration (1-5% w/w protein weight) and hydrolysis time (60-360 min) and the degree of hydrolysis (DH), yield, antioxidant activities, and antiproliferative activity in a HepG2 liver cancer cell line was determined. The surface response methodology showed that the optimum conditions for the enzymatic hydrolysis of sea cucumber were a hydrolysis time of 360 min and 4.3% papain. Under these conditions, a 12.1% yield, 74.52% DH, 89.74% DPPH scavenging activity, 74.92% ABTS scavenging activity, 39.42% H2O2 scavenging activity, 88.71% hydroxyl radical scavenging activity, and 9.89% HepG2 liver cancer cell viability were obtained. The hydrolysate was produced under optimum conditions and characterized in terms of its antiproliferative effect on the HepG2 liver cancer cell line.

Isolation of Fatty Acids from the Enzymatic Hydrolysis of Capsaicinoids and Their Use in Enzymatic Acidolysis of Coconut Oil

Herein we report the optimization of enzymatic hydrolysis of a mixture of capsaicinoids, capsaicin and dihydrocapsaicin obtained from chili peppers, and the utilization of the isolated fatty acids for the modification of coconut oil using enzyme catalyzed acidolysis. This work was carried out as the fatty acids that can be isolated from capsaicinoid hydrolysis have been shown to possess interesting biological properties. These biological properties could be better exploited by incorporating the fatty acids into a suitable delivery vehicle. The enzymatic hydrolysis of the mixture of capsaicin and dihydrocapsaicin was carried out using Novozym® 435 in phosphate buffer (pH 7.0) at 50℃. The enzyme catalyst could be reused in multiple cycles of the hydrolysis reaction. The desired 8-methyl-6-trans-nonenoic acid and 8-methylnonanoic acid were isolated from the hydrolysis reaction mixture using a simple extraction procedure with a 47.8% yield. This was carried out by first extracting the reaction mixture at pH 10 with ethyl acetate to remove any dissolved capsaicinoids and vanillyl amine side product. The fatty acids were isolated after adjustment of the pH of the reaction mixture to 5 and second extraction with ethyl acetate. The acidolysis of coconut oil with the obtained fatty acids was performed using Lipozyme® TL IM. The performance of the acidolysis reaction was evaluated using 1H-NMR spectroscopy and verified in selected cases using gas chromatography. The best performing conditions involved carrying out the acidolysis reaction at 60℃ with a 1.2 w/w ratio of the fatty acids to coconut oil and 10% enzyme loading for 72 h. This resulted in the incorporation of 26.61% and 9.86% of 8-methyl-6-trans-nonenoic acid and 8-methylnonanoic acid, respectively, into the modified coconut oil product. This product can act as a potential delivery vehicle for these interesting compounds.

Response surface optimization of enzymatic hydrolysis of germinated brown rice for higher reducing sugar production

The hydrolysis of germ rice by the use of &amp;alpha;-amylase and glucoamylase enzymes will&amp;nbsp;help increase the reducing sugar content, reduce viscosity, and improve the yield of milk&amp;nbsp;solution compared to the traditional extraction method. The liquefaction experiment was&amp;nbsp;arranged with two factors, which are substrate ratio: &amp;alpha;-amylase concentration and &amp;alpha;-amylase&amp;nbsp;concentration: different hydrolysis time. The saccharification experiment was carried out&amp;nbsp;based on a multivariate model according to the Central Composite Design method. As a&amp;nbsp;result, a 1 : 2 substrate ratio, 0.5% &amp;alpha;-amylase concentration (approx. 11U/g starch) and 50&amp;nbsp;minutes hydrolysis time were selected as the basis for the next experiment. Analysis of&amp;nbsp;variance in the regression model showed that the quadratic model was significant (p &amp;lt;&amp;nbsp;0.0001). Lack of fit (p &amp;gt; 0.05) this indicates that the model is suitable for all data. The&amp;nbsp;reliability of the model R2 = 0.993 shows that the built regression model fits the data set&amp;nbsp;99.3%. CV = 1.19% indicated a better precision and reliability of the experiments carried&amp;nbsp;out. Optimal conditions for hydrolysis of glucoamylase concentration of 0.399% (approx.&amp;nbsp;119.863U/g starch), temperature of 59.813&amp;deg;C and hydrolysis time of 160.468 minutes gave&amp;nbsp;the highest DE content at 25.245% and higher than the non-enzymatic method (DE = 8.985&amp;nbsp;&amp;plusmn; 0.062). &amp;nbsp;

Optimization of enzymatic hydrolysis by alcalase and flavourzyme to enhance the antioxidant properties of jasmine rice bran protein hydrolysate

This study aimed to optimize the hydrolysis conditions for producing jasmine rice bran protein hydrolysate (JBH) using response surface methodology (RSM). The independent variables were the ratio of flavourzyme to alcalase (Fl:Al; 0: 100 to 15: 85; 2.84% enzyme concentration) and hydrolysis time (60–540 min). The optimum hydrolysate was obtained at an Fl:Al ratio of 9.81: 90.19 for 60 min, since it enabled high amounts of protein, high antioxidant activity and more low molecular weight proteins. The experimental values obtained were a degree of hydrolysis (DH) of 7.18%, a protein content of 41.73%, an IC50 for DPPH of 6.59 mg/mL, an IC50 for ABTS of 0.99 mg/mL, FRAP of 724.81 mmol FeSO4/100 g, and 322.35 and 479.05 mAU*s for peptides with a molecular weight of < 3 and 3–5 kDa, respectively. Using a mixture of enzymes revealed the potential of mixed enzymes to produce JBH containing more small peptides and high antioxidant activity.

Optimization of enzymatic hydrolysis of boso fish (Oxyeleotris marmorata) protein based on the degree of hydrolysis and the physical properties of the resultant hydrolysates

Fish protein hydrolysates (FPHs) were produced from boso fish (Oxyeleotris marmorata) through enzymatic hydrolysis using papain as the hydrolysis agent. Papain concentration (0.2–0.4%), temperature (45–55°C), and time (4–12 hrs) were chosen as the independent factors while the degree of hydrolysis (DH), viscosity, and turbidity of the resultant FPHs were recorded as the responses. This study aimed to optimize the enzymatic hydrolysis of boso fish protein using the Box-Behnken Response Surface Methodology (RSM) design. The optimum condition for the boso fish protein hydrolysis based on the DH was predicted at 0.2% of papain concentration, 55°C, and 12 hrs, which would give a DH of 18.63% and an R2 of 90.90%. The FPHs prepared with moderate hydrolysis conditions (0.3% of papain concentration at 50°C for 8 hrs) were spray-dried with the yield ranging from 5.37% to 16.70% (w/w). The spray-dried FPH displayed a globular structure with a diameter of 1–7 μm as observed under the scanning electron microscope (SEM). The total colour difference of the dried FPH was 10.15 compared to the standard white tile. The particle size analysis in distilled water exhibited some distinct peaks which were distributed within 200–1,100 nm with an intensity of 18.1%. This study showed the potential of boso fish for FPH production by using the papain enzyme.

Response surface optimization of enzymatic hydrolysis and ROS scavenging activity of silk sericin hydrolysates

Context Sericin, a protein found in wastewater from the silk industry, was shown to contain a variety of biological activities, including antioxidant. The enzymatic conditions have been continuously modified to improve antioxidant effect and scavenging capacity against various free radicals of silk sericin protein. Objective Variables in enzymatic reactions, including pH, temperature and enzyme/substrate ratio were analysed to discover the optimum conditions for antioxidant activity of sericin hydrolysates. Materials and methods Hydrolysis reaction catalysed by Alcalase® was optimized through response surface methodology (RSM) in order to generate sericin hydrolysates possessing potency for % inhibition on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals, ferric-reducing power and peroxyl scavenging capacity. Flow cytometry was performed to evaluate cellular ROS level in human HaCaT keratinocytes and melanin-generating MNT1 cells pre-treated either with 20 mg/mL RSM-optimized sericin hydrolysates or 5 mM N-acetyl cysteine (NAC) for 60 min prior exposure with 1 mM hydrogen peroxide (H2O2). Results Among these three variables, response surface plots demonstrate the major role of temperature on scavenging capacity of sericin hydrolysates. Sericin hydrolysates prepared by using Alcalase® at RSM-optimized condition (enzyme/substrate ratio: 1.5, pH: 7.5, temperature: 70 °C) possessed % inhibition against H2O2 at 99.11 ± 0.54% and 73.25 ± 8.32% in HaCaT and MNT1 cells, respectively, while pre-treatment with NAC indicated the % inhibition only at 30.26 ± 7.62% in HaCaT and 51.05 ± 7.14% in MNT1 cells. Discussion and conclusions The acquired RSM information would be of benefit for further developing antioxidant peptide from diverse resources, especially the recycling of waste products from silk industry.

Optimization of Enzymatic Hydrolysis of Perilla Meal Protein for Hydrolysate with High Hydrolysis Degree and Antioxidant Activity.

Botanical oils are staple consumer goods globally, but as a by-product of oil crops, meal is of low utilization value and prone to causing environmental problems. The development of proteins in meal into bioactive peptides, such as Perilla peptide, through biotechnology can not only solve environmental problems, but also create more valuable nutritional additives. In the present work, the hydrolysis process of Perilla meal protein suitable for industrial application was optimized with the response surface methodology (RSM) on the basis of single-factor experiments. Alcalase was firstly selected as the best-performing among four proteases. Then, based on Alcalase, the optimal hydrolysis conditions were as follows: enzyme concentration of 7%, hydrolysis temperature of 61.4 °C, liquid-solid ratio of 22.33:1 (mL/g) and hydrolysis time of 4 h. Under these conditions, the degree of hydrolysis (DH) of Perilla meal protein was 26.23 ± 0.83% and the DPPH scavenging capacity of hydrolysate was 94.15 ± 1.12%. The soluble peptide or protein concentration of Perilla meal protein hydrolysate rose up to 5.24 ± 0.05 mg/mL, the ideal yield of which was estimated to be 17.9%. SDS-PAGE indicated that a large proportion of new bands in hydrolysate with small molecular weights appeared, which was different from the original Perilla meal protein. The present data contributed to further, more specific research on the separation, purification and identification of antioxidant peptide from the hydrolysate of Perilla meal protein. The results showed that the hydrolysis of Perilla meal protein could yield peptides with high antioxidant activity and potential applications as natural antioxidants in the food industry.

Response surface optimisation of enzymatic hydrolysis of cassava peels without chemical and hydrothermal pretreatment

Response surface optimisation of enzymatic hydrolysis of cassava peels without chemical and hydrothermal pretreatment

Optimization of enzymatic hydrolysis of Pleurotus ostreatus derived proteins through RSM and evaluation of nutritional and functional qualities of mushroom protein hydrolysates

Abstract Pleurotus ostreatus (Jacq.) P. Kumm., the second most widely cultivated oyster mushroom was grown on paddy straw, which is cheap and readily available waste material. After harvesting and drying, nutritional, and antinutritional composition of P. ostreatus were estimated using the standard assay methods. Tannin and phytic acid were present in very negligible amount (0.095 ± 0.027 mg/g and 0.150 ± 0.083 mg/g, respectively), whereas oxalate and cyanide were absent in whole mushroom. In fact, P. ostreatus was hydrolysed with commercially available proteinase K, pepsin and trypsin with different concentrations of the enzymes (0.05%, 0.10% and 0.15%), at different temperatures (30 °C, 40 °C and 50 °C) for different time periods (60, 90 and 120 min) to get the mushroom protein hydrolysates. Degree of hydrolysis and protein content varied from 4.29 ± 1.12% to 99.42 ± 0.02% and from 0.25 ± 0.07 mg/mL to 3.22 ± 0.12 mg/mL, respectively. Maximum degree of hydrolysis and the highest protein content of protein hydrolysate was obtained when using 0.15% proteinase K, at 50 °C for 120 minutes. Mushroom protein hydrolysates thus obtained exhibited improved functional characteristics such as foaming capacity, foaming stability and emulsifying property than the unhydrolysed mushroom. Based on the result of the present study, the mushroom protein hydrolysates could be served as useful ingredient for food and nutraceutical applications.