Optimal Hydrolysis Conditions Research Articles

Combined Effect of Protease, Hemicellulase and Pectinase on the Quality of Hemp Seed Oil (Cannabis sativa L.) Obtained by Aqueous Enzymatic Extraction as an Eco-friendly Method.

The objective of this research was to evaluate the efficiency of aqueous enzymatic extraction (AEE) to obtain oil from hemp seeds (Cannabis sativa L.) grown in northern Morocco. Optimisation of AEE extraction parameters, including pH, enzyme concentration (hemicellulase, protease and pectinase), temperature and incubation time, to maximize oil yield was achieved using response surface methodology with a central composite design. For comparison, the solvent extraction (Soxhlet) (SE) method was also used. Optimized hydrolysis conditions involved incubation for 4 hours at 60°C with a pH of 6.5, using a multi-enzyme preparation comprising protease, hemicellulase and pectinase at concentrations of 55, 202.5 and 234 U/mg, respectively. Referring to the conventional Soxhlet extraction (SE), Aqueous Enzymatic Extraction (AEE) achieved a 30.65% oil recovery rate under the optimized parameters mentioned above. The use of enzymes produced an oil that was more stable against oxidation than the solvent-extracted oil, with a peroxide value (PV) of 19.54 and 47.87 meq O 2 /kg, respectively. Furthermore, HPLC-DAD analysis of tocopherol content indicated a higher total tocopherol content (547.2 mg/kg) in Aqueous Enzymatic Extraction (AEE) compared to Soxhlet Extraction (SE) (513.51 mg/kg), with γ-tocopherol being the predominant form. No significant differences in fatty acid composition were observed between the two extraction methods with linoleic acid and alpha-linolenic acid being the predominant constituents.

Preparation, Identification, and Digestive Stability of Antioxidant Peptides From Chlorella vulgaris

Chlorella, a single‐celled green algae rich in proteins, holds promise as a source of bioactive peptides. This research aimed to isolate an antioxidant peptide from Chlorella, examining its stability through digestion. Using Response Surface Methodology (RSM), we identified optimal hydrolysis conditions for Chlorella Protein Hydrolysate (CPH): 55°C, pH 8.75, and an enzyme‐to‐substrate ratio of 2.15%. These conditions led to CPH with notable radical scavenging activity against 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) (53.19 ± 0.99%), 2,2′‐azino‐bis (3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) (76.86 ± 0.28%), hydroxyl radical (53.39 ± 1.74%), and Fe2+ chelating rates of 22.56 ± 2.97%. Subsequently, peptides under 3 kDa isolated by ultrafication proved particularly stable and antioxidative after in vitro digestion. In addition, further purify to obtain component C1 with DPPH radical scavenging ability of 59.62 ± 1.29% and an Fe2+ chelating ability of 49.78 ± 1.55%. Subsequent LC‐MS/MS identification and virtual screening led to the discovery of a stable and antioxidative peptide, Ser‐Gly‐His‐His‐Lys‐Pro‐Leu (SGHHKPL). The study provides the theoretical basis for the development and utilization of chlorella.

Optimization, Scale-Up, and Economic Analysis of the Ethanol Production Process Using Sargassum horneri

Recently, the extensive spread of some algae along coastlines has surged into unmanageable thick decomposition layers. This study aimed to demonstrate the use of Sargassum horneri as a biomass resource for ethanol production through the continuous hydrolysis, enzymatic saccharification, and fermentation process. Sugars from S. horneri were obtained using a combination of thermal acid hydrolysis and enzymatic saccharification. The optimal conditions for thermal acid hydrolysis involved a 10% (w/v) S. horneri slurry treated with 100 mM H2SO4 at 121 °C for 60 min; enzymatic saccharification using 16 U/mL Cellic CTec2 further boosted the monosaccharide concentration to 23.53 g/L. Fermentation experiments were conducted with mannitol-adapted Saccharomyces cerevisiae BY4741 using S. horneri hydrolysate. Enhanced ethanol production was observed in the hydrolysate, particularly with mannitol-adapted S. cerevisiae BY4741, which yielded 10.06 g/L ethanol. Non-adapted S. cerevisiae produced 8.12 g/L ethanol, as it primarily utilized glucose and not mannitol. Regarding ethanol fermentation using 5 L- and 500 L-scale fermenters, the ethanol concentrations reached 10.56 g/L and 7.88 g/L with yields of 0.51 and 0.45, respectively, at 48 h. This study confirmed the economic viability of ethanol production using waste seaweed with optimized pretreatment conditions and the adaptive evolution of S. cerevisiae to mannitol.

Optimization of ultrasound-assisted enzymatic hydrolysis of channel catfish (Ictalurus punctatus) frames yield high-calcium and high-amino acid hydrolysates

ABSTRACT In this study, a high-calcium and high-amino acid fish frame hydrolyzate (FFH) was prepared from channel catfish (Ictalurus punetaus) through ultrasound-assisted enzymatic hydrolysis (UAEH). A 4-factor and 3-level Box-Behnken design of response surface methodology was employed to optimize the hydrolysis conditions, where the independent variables were sonication power (W), sonication time (min), enzyme activity (U) and water-substrate ratio (v/w). The degree of hydrolysis (DH) and soluble calcium content (SCC) were used as response factors. The optimal hydrolysis conditions were found to be sonication power (X1) = 492.91 W, sonication time (X2) = 15.01 min, enzyme activity (X3) = 6677.88 U, and water-to-substrate ratio (X4) = 3.56 v/w. Under these conditions, UAEH effectively disintegrated fish frame collagen and converted bound calcium to soluble calcium. Nevertheless, this study investigated the pattern of collagen transformation into short peptides and free amino acids throughout the degradation process. These findings highlight the potential of the FFH from channel catfish to serve as a foundation for creating highly nutritious food additive. The outcomes of this study further enhance the food industry’s ability to capitalize on the nutritional benefits offered by fish frame hydrolyzates. This expansion of health-focused product options and endorsement of the sustainable use of fish by-products harmonizes seamlessly with the industry’s evolving goals of waste reduction and value optimization.

Reutilization of food industrial waste for lutein production with heterotrophic microalgae Chlorella sorokiniana MB-1-M12

BackgroundLutein is a xanthophyll carotenoid commonly found in photosynthetic organisms, such as microalgae and plants. It serves as a photosynthetic accessory pigment and possesses antioxidant properties. Lutein is recognized for its prophylactic use in preventing Age-Related Macular Degeneration (AMD) and is widely employed in health supplements. The demand for lutein is steadily increasing. This study utilized Chlorella sorokiniana MB-1-M12 for heterotrophic growth and lutein production, employing molasses as a model of food industrial waste as an alternative carbon source. MethodsMolasses was hydrolyzed using acids, and the optimal hydrolysis conditions were determined by varying the types and concentrations of the acids. The resulting hydrolysate, which contains reducing sugars, was then recovered and used to support the heterotrophic growth of C. sorokiniana MB-1-M12 for lutein production. Significant FindingsThe best efficiency in molasses hydrolysis was achieved using 0.15 M sulfuric acid, resulting in a maximum sugar recovery of 0.721 gs of reducing sugar per gram of molasses. When the molasses hydrolysate reached a sugar concentration of 10 g/L, it yielded a biomass of 2.57 g/L and a lutein content of 2.1 mg/g. These findings indicate that molasses can effectively serve as a substitute for glucose as a carbon source, although it cannot provide all the essential micronutrients required for biomass growth. Consequently, the addition of nutrients in BG-11 medium became necessary. This study successfully demonstrates the upcycling of waste resources, such as molasses, into a high-value product like lutein, all achieved in a cost-effective and resource-efficient manner.

Optimization of the Hydrolysis Process of Microalgae Porphyridium cruentum Biomass with Variations of Hydrochloric Acid Concentration, Temperature, and Time using Response Surface Methodology (RSM)

Microalgae Porphyridium cruentum has potential as a raw material for bioethanol production because it has a high carbohydrate content. These carbohydrates can be broken down into reducing sugars through the hydrolysis process. The reducing sugar obtained will be used as a substrate in the production of bioethanol. This study aimed to produce a substrate with the best-reducing sugar indicator and to determine the optimum conditions for hydrolysis of P. cruentum microalgae biomass following the Box-Behnken statistical experimental design, using Response Surface Methodology (RSM). The parameters of the optimized hydrolysis process were HCl concentration (2 - 0.2 N), temperature (60 -120 ˚C), and hydrolysis time (30-180 min). The optimum conditions that have been achieved using RSM are an HCl concentration of 1.91 N, a temperature of 60 °C, and a hydrolysis time of 180 min were predicted a maximum total reducing sugar production of 810 mg/L. The experimental result of total reducing sugar obtained at optimum conditions was 895 mg/L, which was well close to the predicted value, verifying the appropriateness of the model.Abstract is informed about the statements of the problem, methods, scientific finding results and conclusion concisely

Extraction, optimization, and functional quality evaluation of carotenoproteins from shrimp processing side streams through enzymatic process.

The study aimed to develop an effective and eco-friendly enzymatic process to extract carotenoproteins from shrimp waste. The optimization of enzymatic hydrolysis conditions to maximize the degree of deproteinization (DDP) of carotenoprotein from shrimp head waste (SHW) and shrimp shell waste (SSW) was conducted separately using the Box-Behnken design of response surface methodology (RSM). To achieve a maximum DDP of 92.32% for SSW and 96.72% for SHW, the optimal hydrolysis conditions were determined as follows: temperature (SSW: 53.13°C; SHW: 45.90°C), pH (SSW: 7.13; SHW: 6.78), time (SSW: 90min; SHW: 61.18min), and enzyme/substrate ratio (SSW: 2g/100g; SHW: 1.18g/100g). The carotenoprotein effluent obtained was subjected to spray drying and subsequently assessed for color, nutritional, and functional characteristics. The carotenoprotein from shrimp shell (CpSS) contained a higher essential amino acid score than carotenoprotein from shrimp head (CpSH). CpSS had a higher whiteness index of 82.05, while CpSH had 64.04. Both CpSS and CpSH showed good functional properties viz solubility, emulsion, and foaming properties. The maximum solubility of CpSH and CpSS was determined to be 92.94% and 96.48% at pH 10.0, respectively. The highest emulsion capacity (CpSH: 81.33%, CpSS: 70.13%) and stability (CpSH: 57.06%, CpSS: 63.05%) were observed at 3% carotenoprotein concentration. Similarly, the highest values of foaming capacity (CpSH: 27.66%, CpSS: 105.5%) and stability (CpSH: 23.83%, CpSS: 105.33%) were also found at the same 3% carotenoprotein concentration. In conclusion, the carotenoproteins obtained from shrimp waste showed favorable attributes in terms of color, amino acid composition, and functional properties. These findings strongly suggest the potential applicability of CpSS and CpSH as valuable resources in various domains. CpSS, with its higher whiteness index, greater amino acid content, and superior functional characteristics, may find suitability as functional ingredients in human food products. Conversely, CpSH could be considered for incorporation into animal feed formulations.

Optimization of thermochemical hydrolysis of potassium hydroxide-delignified Cocoa (Theobroma cacao L) pod husks under low combined severity factors (CSF) conditions

This study identified the optimal conditions for acid hydrolysis of cocoa pod husks (CPH) pretreated and then hydrolyzed under combined low gravity conditions. Pre-treatment was carried out using 1% potassium hydroxide (KOH), using a biomass to solvent ratio of 9.143% (w/v), for 4 h at 30 °C, then the mixture is autoclaved for 15 min at 128 °C. Lignocellulosic analyzes were carried out on raw and pretreated husk. In order to determine the structural changes operated on the pretreated pod, Fourier transform infrared (FTIR) analyzes were also carried out. These analyzes confirmed a 93.87% removal of lignin after delignification. Furthermore, scanning electron microscopy/energy dispersive X‐ray spectrometry (SEM/EDX) revealed several changes within the pretreated matrix; showing a dominant presence of the elements carbon, oxygen and potassium, associated with an intensification of the peaks of magnesium and potassium. Acid hydrolysis on the pretreated pod was optimized under low combined severities factors (CSF) conditions between 1.11 and 2.45, using response surface methodology. A three-variable Box-Behnken design (autoclaving time (X1), biomass to solvent ratio (X2), and sulfuric acid (H2SO4) concentration (X3)) was used. The quadratic model with an R2 of 0.98 proved to be the most suitable, giving the following optimal hydrolysis conditions: 26.95 min of autoclave, biomass to solvent ratio of 9.53% (w/v) and 4.92% (w/v) H2SO4.18.77 g/L of reducing sugars was obtained for a CSF of 2.06. In addition, a marked decrease in the content of phenolic compounds was observed, shifting from 19.45 g/L in the raw delignified sample to 3.11 g/L in the delignified sample after reduction the extractable compounds. Thus, it is proved in this study that optimization of the acid hydrolysis process of CPH, after delignification with KOH, can improve the acid hydrolysis of polysaccharides, under conditions of low combined severity factors.

Açai Seeds (Euterpe oleracea Mart) Are Agroindustrial Waste with High Potential to Produce Low-Cost Substrates after Acid Hydrolysis.

Açai seeds have been discarded improperly around the Amazonia region, but they can be seen as promising low-cost substrates for fermentation processes. The structural carbohydrates and physicochemical characterization of açai seeds from the Amazonia were assessed followed by the determination of the optimal hydrolysis conditions using H3PO4 (phosphoric acid) and H2SO4 (sulfuric acid) to obtain a liquor with high contents of simple carbohydrates and low levels of potential microbial inhibitors usually generated during acid hydrolysis of carbohydrates. A central composite rotational design was carried out varying the concentrations of diluted acid (0-5%, w/v), solids (0.1-25%, w/v), and hydrolysis time (9.5-110 min). Acid hydrolysis with H2SO4 was more effective in producing reducing sugars (15.9-103.1 g/L) than H3PO4 (2.9-33.9 g/L) during optimization. The optimal hydrolysis conditions with H2SO4 were 3.5% of acid (w/v), 25% of solids during 70 min at 121 °C, which provided a liquor with 55 g/L of reducing sugars and low levels of microbial inhibitors: acetic acid (1.8 g/L), hydroxymethyl furfural (338 mg/L), and furfural (10 mg/L). Thus, açai seeds were characterized as promising agroindustrial waste with high potential to be used as a low-cost substrate in biotechnological processes, comprising relevant environmental and bioeconomic aspects for the development of the Amazonia.

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.

Optimizing the First Step of the Biocatalytic Process for Green Leaf Volatiles Production: Lipase-Catalyzed Hydrolysis of Three Vegetable Oils.

Green leaf volatiles (GLVs), including short chain volatile aldehydes, are widely used in the flavor and food industries because of their fresh aroma. To meet the growing demand for natural GLVs with high added value, the use of biocatalytic processes appears as a relevant application. In such processes, vegetable oils are bioconverted into GLVs. First, the triacylglycerols of the oils are hydrolyzed by a lipase. Then, the free polyunsaturated fatty acids are converted by a lipoxygenase. Finally, volatile C6 or C9 aldehydes and 9- or 12-oxoacids are produced with a hydroperoxide lyase. Optimization of each biocatalytic step must be achieved to consider a scale-up. In this study, three oils (sunflower, hempseed, and linseed oils) and three lipases (Candida rugosa, Pseudomonas fluorescens, and Rhizomucor miehei lipases) have been tested to optimize the first step of the process. The experimental design and response surface methodology (RSM) were used to determine the optimal hydrolysis conditions for each oil. Five factors were considered, i.e., pH, temperature, reaction duration, enzyme load, and oil/aqueous ratio of the reaction mixture. Candida rugosa lipase was selected as the most efficient enzyme to achieve conversion of 96 ± 1.7%, 97.2 ± 3.8%, and 91.8 ± 3.2%, respectively, for sunflower, hempseed, and linseed oils under the defined optimized reaction conditions.

Investigation into Optimal Conditions for Enzymatic Hydrolysis of Cassava Starch to Glucose by Amylase from Rice

The challenge of finding locally available materials in abundance to meet up with the increase in demand for glucose syrup necessitated this study. Enzymatic hydrolysis of cassava starch to glucose using glucose amylase sourced from rice was conducted. A-3 factor with 6 levels factorial viz. substrate concentration (0.5, 1.0, 1.5, 2.0, 2.5 and 3.0% w/v), pH (4, 5, 6, 7, 8 and 9) and temperature (30, 40, 50, 60, 70 and 80 °C) experiment were employed. Rice malt was prepared and enzyme activated. Starch (substrate), buffer solutions, standard glucose solution and its calibration curve were also prepared. Starch was hydrolyzed by α-amylase and tested for presence of reducing sugar using Benedict solution. Time course of the reaction was studied and enzyme activity determined. It was observed that as reaction time increased (t), amount of glucose produced [P] initially increased but soon recorded infinitesimal increase and later assumed constant. The effects of substrate concentration, pH and temperature were found to be essential on glucose production. Statistical analysis on the effect of substrate concentration [S], reaction time and their interactions showed significant impact at probability level of (p) = 0.05.

Protein preparations from rapse processing waste: A review of the current status and development prospects of existing technologies

The demand for protein products is increasing due to the demographic growth of the world’s population. As an alternative to traditional sources of protein, waste from plant raw material processing is becoming increasingly popular. An important place in the global economy is occupied by oilseeds, in particular rapeseed, which production volumes are increasing in the Russian Federation every year. Rapeseed (Brassicaceae napus) is of great interest due to its high oil content (39.80–46.00%) and rich fatty acid composition, while cake and meal formed in the process of oil production are characterized by a significant content of crude protein (35.00–45.00%) and crude fiber (8.20–17.50%); however, they are used mainly as a feed additive. Recent studies on the processing of rapeseed waste indicate the value of this raw material as a source of dietary protein, which has a balanced amino acid profile and a high degree of digestibility (up to 85%). To obtain protein, rapeseed processing is envisaged: cleaning, grinding, cold pressing at a temperature of ≤ 40 °C, fat extraction with a solvent. At the next stages, the protein is extracted with 0.1–0.5 M NaCl at pH 5.3–12.0 and a temperature of 5–30 °C for 1 hour. The extracted protein is precipitated at the isoelectric point (pH 4.0) with HCl, separated from the mixture and neutralized. The result is a protein isolate with a protein content of 90.0–98.7%. It is possible to increase the quality and yield of the protein product due to the additional stage of processing the defatted cake with cellulolytic enzyme preparations. In this case, additional studies are required to determine the substrate specificity of commercial cellulase enzyme preparations and the optimal hydrolysis conditions. The parameters of extraction and precipitation of the protein in the case of using the stage of enzymatic lysis should also be specified.

Effect of Peanut Protein Treated with Alkaline Protease and Flavorzyme on BALB/c Mice.

This article aims to analyze the effects of enzyme treatment concentration, temperature, and time on peanut protein so as to obtain an optimal enzymatic hydrolysis condition for flavorzyme (Fla) and alkaline protease (Alk). The results were as follows: enzymatic hydrolysis temperature 60 °C and 55 °C, enzyme concentration 10% and 4%, enzymatic hydrolysis time 80 min and 60 min, and double enzyme hydrolysis ratio 2% Fla + 5% Alk, respectively. The BALB/c mice were sensitized with gavage of peanut protein before and after enzyme treatment to evaluate the effects of different enzyme treatments on peanut allergenicity. Compared with the mice sensitized with raw peanuts, the weight growth rate of the mice sensitized with enzyme treatment peanut increased but not as much as the control, the degranulation degree of mast cell and basophils decreased, the inflammatory infiltration and congestion in jejunum and lung tissue decreased, the expression of proinflammatory factors and thymic stromal lymphopoietin (TSLP) gene decreased, and the secretion of specific antibodies (IgE, and IgG) decreased, and the binding ability of peanut protein with peanut-specific IgE antibodies decreased as well. The results above indicate that the allergenicity of peanut protein decreases after enzyme treatment and the dual enzyme (Fla + Alk) treatment can be much more efficient.

Preparation of Hypoallergenic Whey Protein Hydrolysate by a Mixture of Alcalase and Prozyme and Evaluation of Its Digestibility and Immunoregulatory Properties.

Whey protein (WP) has nutritional value, but the presence of β-lactoglobulin (β-LG) and α-lactalbumin (α-LA) cause allergic reactions. In this study, hypoallergenic whey protein hydrolyate (HWPH) was prepared by decomposing β-LG and α-LA of WP using exo- and endo-type proteases. The enzyme mixing ratio and reaction conditions were optimized using response surface methodology (RSM). Degradation of α-LA and β-LG was confirmed through gel electrophoresis, and digestion, and absorption rate, and immunostimulatory response were measured using in vitro and in vivo systems. Through RSM analysis, the optimal hydrolysis conditions for degradation of α-LA and β-LG included a 1:1 mixture of Alcalase and Prozyme reacted for 10 h at a 1.0% enzyme concentration relative to substrate. The molecular weight of HWPH was <5 kDa, and leucine was the prominent free amino acid. Both in vitro and in vivo tests showed that digestibility and intestinal permeability were higher in HWPH than in WP. In BALB/c mice, as compared to WP, HWPH reduced allergic reactions by inducing elevated Type 1/Type 2 helper T cell ratio in the blood, splenocytes, and small intestine. Thus, HWPH may be utilized in a variety of low allergenicity products intended for infants, adults, and the elderly.

Antioxidant activity of bioactive peptides from peanuts protein hydrolysate

Peanut protein hydrolysate was produced from peanut protein isolate hydrolyzed using Alcalase® 2.4 L. Hydrolysis conditions were optimized using a response surface methodology. The two independent variables were the substrate/enzyme ratio and the hydrolysis time with lower and upper limits of (20:1-50:1 v/v) and (1-3 hours), respectively. Both were designed at three levels of central composite design from the response surface methodology and the degree of hydrolysis as the dependent variable. This study aimed to obtain the optimum conditions for enzymatic hydrolysis of peanut protein isolate using Alcalase to produce protein hydrolysate with the maximum degree of hydrolysis. The optimum hydrolysis conditions obtained were the substrate/enzyme ratio of 20:1 (v/v) and the hydrolysis time of 3 hours with a predicted degree of hydrolysis of 40.54%. The actual experimental results obtained a degree of hydrolysis of 40.35% (p&gt;0.05), which showed no difference between the predicted and experimental results. The highest antioxidant activity was obtained from the combination of these treatments, indicated by IC50 of 5.61 ppm.

Producing bacterial cellulose from industrial recycling paper waste sludge

This study aimed to produce bacterial cellulose from paper waste sludge (PWS) as a method of utilizing the cellulose source from the remaining pulp in the material. Initially, PWS was hydrolyzed by sulfuric acid to create an enriched-reducing sugar hydrolysate. One-factor experiments were conducted with a fixed amount of PWS (5 g) to investigate the influence of hydrolysis conditions, including water, sulfuric acid addition, temperature, and retention time, on the production yield of reducing sugars. Based on these results, the Box-Behnken model was designed to optimize the hydrolysis reaction. The optimal hydrolysis conditions were 10 ml/g of the sulfuric acid solution (30.9%) at 105.5 °C for 90 min of retention time 0.81 (gGE/g PWS), corresponding to a conversion yield of 40.5%). Subsequently, 100 ml of the filtered and neutralized PWS hydrolysate was used as the culture to produce the bacterial cellulose (BC) using Acetobacter xylinum, which produced 12 g/L of bacterial cellulose. The conversion yield of bacterial cellulose calculated as the ratio of the weight of produced bacterial cellulose to that of cellulose in PWS reached 33.3%. The structure of the obtained BC was analyzed using scanning electron microscopy (SEM) and X-ray diffraction (XRD) to indicate the formation of nano-cellulose fiber networks. This research proposed a combined method to convert paper waste sludge into bacterial cellulose, demonstrating the potential for waste utilization and sustainable production of paper industries for added-value products.

Enhanced Enzymatic Hydrolysis of High-Solids Content Corncobs by a Lytic Polysaccharide Monooxygenase from Podospora anserina S Mat+ for Valuable Monosaccharides

Lytic polysaccharide monooxygenase (LPMO) has been found to enhance cellulase activity and promote efficient utilization of corncob cellulose. To this end, a novel LPMO from Podospora anserina S mat+ (PaLPMO) through in silico screening was heterologously expressed in Escherichia coli BL21(DE3) and its biochemical properties were investigated. To reduce production costs, chaperones and site-directed mutagenesis were employed to enhance the protein’s solubility, yielding an 8% increase. Through single-factor experiments, the optimal hydrolysis condition for the highest glucose yield of 129.8 g/L was identified as 25% high-solids content of corncob at 60 °C and pH 5.5 using a PaLPMO-to-cellulase ratio of 1:3 (total protein concentration of 12 mg/g) and with the addition of 0.5 mmol/L Mn2+. This result represented a 24% improvement compared to cellulase alone and a 33% reduction in commercial cellulase usage with the same degree of hydrolysis. The hydrolysis products of high-solids content corncob degradation were further used to produce valuable d-allulose through glucose isomerase and d-allulose 3-epimerase catalysis in a ratio of 2:1, with a yield of 56.22 g/L fructose and 20.6 g/L d-allulose. The PaLPMO and cellulase complex significantly improved the degradation efficiency of corncob residue and reduced cellulase usage in lowering the production cost. These findings contribute to developing comprehensive corncob utilization and pave the way for sustainable and efficient energy and resource utilization.

Optimization of microfibrillated cellulose isolation from cocoa pod husk via mild oxalic acid hydrolysis: A response surface methodology approach

Theobroma cacao L. species, cultivated worldwide for its valuable beans, generates up to 72% weight of the fruit as waste. The lack of reutilization technologies in the cocoa agroindustry has hindered the exploitation of valuable bio-components applicable to the generation of high value added bioproducts. One such bioproduct is microfibrillated cellulose (MFC), a biopolymer that stands out for its desirable mechanical properties and biocompatibility in biomedical, packing, 3D printing, and construction applications. In this study, we isolated microfibrillated cellulose (MFC) from cocoa pod husk (CPH) via oxalic acid hydrolysis combined with a steam explosion. MFC isolation started with the Solid/Liquid extraction via Soxhlet, followed by mild citric acid hydrolysis, diluted alkaline hydrolysis, and bleaching pre-treatments. A Response Surface Methodology (RSM) was used to optimize the hydrolysis reaction at levels between 110 and 125 °C, 30–90 min at 5–10% (w/v) oxalic acid concentration. The cellulose-rich fraction was characterized by Fourier-Transform Infrared Spectroscopy (FTIR), Thermogravimetric Analysis (TGA), Differential Scanning Calorimetry (DSC), X-Ray Diffraction (XRD), and Scanning Electron Microscopy (SEM) analyses. Characterization analyses revealed a cellulose-rich polymer with fibers ranging from 6 to 10 μm, a maximum thermal degradation temperature of 350 °C, and a crystallinity index of 63.4% (peak height method) and 29.0% (amorphous subtraction method). The optimized hydrolysis conditions were 125 °C, 30 min, at 5% w/v oxalic acid: with a 75.7% yield. These results compare with MFC obtained through highly concentrated inorganic acid hydrolysis from different biomass sources. Thus, we show a reliable and greener alternative chemical treatment for the obtention of MFC.

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.