Enzyme Cocktail Research Articles

Bioconversion of the invasive seaweed Rugulopteryx okamurae into enzymes and polyhydroxyalkanoates

The invasive macroalgae Rugulopteryx okamurae has been used as raw material for the production of hydrolytic enzymes and polyhydroxyalkanoates (PHA), bioplastic precursors. The fungus Aspergillus oryzae, grown in solid state fermentation (SSF) on the macroalga biomass, produced several hydrolytic enzymes, mainly xylanase and alginate lyase. Also, SSF was used as biological pretreatment of the macroalga to enhance the release of fermentable sugars after enzyme hydrolysis for subsequent PHA production. To increase sugar concentration and reduce the total cost of the process, the enzyme hydrolysis was performed at a high solid load and stirring rate, reducing the enzyme cocktail dose used by 63.5 %. At these conditions, the hydrolysate contained 18.52 g/L of reducing sugars, which were fermented with Cupriavidus necator, reaching a maximum of 288.0 mg PHA/L at 24 h.

Conversion of industrially relevant lignocellulosic biomass into monomers: By customization of cellulolytic enzyme cocktail using simplex-lattice mixture designing (SLMD)

The use of differently pre-treated lignocellulosic biomass (LCB), which contains varying levels of cellulose, hemicellulose and lignin content, is profitable during biofuel production but also require unique enzymatic preparations to convert these differently pre-treated LCB into their monomeric constituents. To address this question, this study employed simplex-lattice mixture design strategy to customize lignocellulolytic enzymatic cocktails for hydrolysis of different pre-treated substrates, obtained from PRAJ and IOCL Industries. In-house recombinant fungal auxiliary/accessory enzymes were evaluated to work in synergy with Cellic®CTec3. The mixtures of recombinant proteins derived from Scytalidium thermophilum CM-4T with Cellic®CTec3 were found to optimally hydrolyse diluted acid pre-treated rice straw slurries obtained from PRAJ and IOCL Industry using Simplex-lattice mixture designing (SLMD) and showed 70.39 % and 84.46 % saccharification efficiency, respectively.

Guiding the selectivity of commercial glycosidase preparation towards the production of rutinose

Aromase™ H2 is an enzymatic cocktail marketed to intensify the aroma, promote clarity and overall flavor of tea by releasing aromatic compounds and antioxidants that are naturally glycosylated. The hydrolysis of a wide range of glycoconjugates is the key to its action, which can arise from the substrate promiscuity of the main diglycosidase or other enzymes that are in the mixture. The characterization focusing on the activity toward flavonoids at different pH values allowed to increase the cocktail ratio of diglycosidase to α-rhamnosidase activity. In that way, the free disaccharide rutinose was selectively obtained in a laboratory scale reactor with 90 % conversion after 3.5 h reaction at pH 8. Purification was carried out by liquid-liquid extraction, recovering the aglycone hesperetin. Subsequent extraction with activated carbon allowed a recovery of 84 % of the rutinose. This work demonstrates the practical application of a commercial cocktail for the selective hydrolysis of rutinosylated compounds.

Light Exposure of Lignin Affects the Saccharification Efficiency of LPMO-Containing Cellulolytic Enzyme Cocktails

Light Exposure of Lignin Affects the Saccharification Efficiency of LPMO-Containing Cellulolytic Enzyme Cocktails

Optimizing the Extraction of Bioactive Compounds from Porphyra linearis (Rhodophyta): Evaluating Alkaline and Enzymatic Hydrolysis for Nutraceutical Applications.

Porphyra sensu lato is one of the most economically significant and widely cultured and consumed algae in the world. Porphyra species present excellent nutraceutic properties due to their bioactive compounds (BACs). This research aimed to find the most efficient aqueous extraction method for BACs by examining alkaline and enzymatic hydrolysis. Alkaline hydrolysis with 2.5% sodium carbonate (SC) and at 80 °C proved optimal for extracting all BACs (phycobiliproteins, soluble proteins, polyphenols, and carbohydrates) except mycosporine-like amino acids (MAAs), which were best extracted with water only, and at 80 °C. Enzymatic hydrolysis, particularly with the 'Miura' enzymatic cocktail (cellulase, xylanase, glycoside hydrolase, and β-glucanase), showed superior results in extracting phycoerythrin (PE), phycocyanin (PC), soluble proteins, and carbohydrates, with increases of approximately 195%, 510%, 890%, and 65%, respectively, compared to the best alkaline hydrolysis extraction (2.5% SC and 80 °C). Phenolic content analysis showed no significant difference between the 'Miura' cocktail and 2.5% SC treatments. Antioxidant activity was higher in samples from alkaline hydrolysis, while extraction of MAAs showed no significant difference between water-only and 'Miura' treatments. The study concludes that enzymatic hydrolysis improves the efficiency of BACs extraction in P. linearis, highlighting its potential for the nutraceutical industry, and especially with respect to MAAs for topical and oral UV-photoprotectors.

Multi-objective optimization of lignocellulolytic enzyme cocktail production from Pseudolagarobasidium acaciicola TDW-48 by artificial neural network-genetic algorithm (ANN-GA) strategy and its application in lignocellulose waste bioconversion

Multi-objective optimization of lignocellulolytic enzyme cocktail production from Pseudolagarobasidium acaciicola TDW-48 by artificial neural network-genetic algorithm (ANN-GA) strategy and its application in lignocellulose waste bioconversion

Mimicking the enzymatic plant cell wall hydrolysis mechanism for the degradation of polyethylene terephthalate

Plastic pollution presents a global challenge, impacting ecosystems, wildlife, and economies. Polyethylene terephthalate (PET), widely used in products like bottles, significantly contributes to this issue due to poor waste collection. In recent years, there has been increasing interest in plant biomass-degrading enzymes for plastic breakdown, due to the structural and physicochemical similarities between natural and synthetic polymers. Filamentous fungi involved in hemicellulose degradation have developed a complex mode of action that includes not only enzymes but also biosurfactants; surface-active molecules that facilitate enzyme-substrate interactions. For this reason, this study aimed to mimic the mechanism of biomass degradation by repurposing plant cell wall degrading enzymes including a cutinase and three esterases to cooperatively contribute to PET degradation. Surfactants of different charge were also introduced in the reactions, as their role is similar to biosurfactants, altering the surface tension of the polymers and thus improving enzymes’ accessibility. Notably, Fusarium oxysporum cutinase combined with anionic surfactant exhibited a 2.3- and 1.6-fold higher efficacy in hydrolyzing amorphous and semi-crystalline PET, respectively. When cutinase was combined with either of two ferulic acid esterases, it resulted in complete conversion of PET intermediate products to TPA, increasing the overall product release up to 1.9– fold in presence of surfactant. The combination of cutinase with a glucuronoyl esterase demonstrated significant potential in plastic depolymerization, increasing degradation yields in semi-crystalline PET by up to 1.4-fold. The approach of incorporating enzyme cocktails and surfactants emerge as an efficient solution for PET degradation in mild reaction conditions, with potential applications in eco-friendly plastic waste management.

Performance of broiler chickens fed palm kernel cake with enzyme supplementation

Abstract The purpose of this study was examined the performance of broiler chickens fed palm kernel cake (PKC) with enzyme supplementation. This research used 1,260 Indian River strain male broiler chickens divided into seven treatments and six replications, with 30 chickens in each replication. This research used two types of cocktail enzymes, enzyme one and enzyme two. Enzyme one consists of mannanase 182 g/ton (182×103 U), NSPase 200 g/ton (α – amylase 54×103 U, selulase 500 ×103 U, xylanase 375×103 U, and protease 180×103 U) and protease 130 g/ton (240.5 U), while enzyme two consists of mannanase 182 g/ton (182×103 U), NSPase 400 g/ton (α – amylase 108×103 U, selulase 106 U, xylanase 750×103 U, and protease 360×103 U), and protease 260 g/ton (481 Unit enzyme). This study’s treatment was: P0 - basal feed (control), P1 – P0 with 10% PKC, P2 – P0 with 20% PKC, P3 – P0 with 10% PKC and enzyme one, P4 – P0 with 10% PKC and enzyme one, P5 – P0 with PKC 20% and enzyme two, and P6 – P0 with PKC 20% and enzyme two. Chickens were kept for 35 days. The variables observed included feed intake (FI), average daily gain (ADG), Feed conversion Ratio (FCR), Performance Index (PI), and carcass percentage. The data were analyzed using Analysis of Variance, followed by Duncan’s Multiple Range Test (DMRT). Enzyme addition did not affect feed consumption. The P3 had the largest ADG (2,219.75 g), the lowest FCR (1.59), and had the best FCR compared to the other treatments. The P4 and P6 could increase ADG compared P0, but did not increase compared to P3. The addition of 10% PKC could reduce FCR compared to the control, while the addition of 20% PKC did not reduce FCR. The addition of 10% PKC could increase PI by 19.07% (P1) and 28.58% (P3) compared to the control. The P4 and P6 did not increase PI. The P3 could increase carcass percentage by 6.15% and 6.19% (P5) compared to the control. Carcass percentage in P3 and P5 was highest than the control which was 71.53% and 71.56%. This research concluded that the P3 treatment consisting of basal feed with 10% PKC and the addition of enzymes mannanase 182 g/ton, NSPase 200 g/ton, and protease 130 g/ton could increase ADG, PI, and carcass percentage, and reduced FCR.

Recycling of printed Xerographic paper using Aspergillus assiutensis enzyme cocktail: an integrated approach to sustainable development.

To overcome the human and animal survivability risk, sustainable development is the only option on earth that can be achieved through the maximum use of renewable environmental resources. Recycling of waste paper is an emerging waste management approach to conserve natural resources. Herein, we studied enzyme-mediated process to recycle the xerographic paper by using the crude fungal extract from indigenously isolated fungi identified asAspergillus assiutensis. The fungal enzyme cocktail has been characterized for the production of multiple enzymes namely cellulase, amylase, xylanase, pectinase, and protease. All these enzymes have pH optima in the acidic range and except cellulase and all the enzymes are stable from 10 to 80C. In the zymogram analysis, pectinase, xylanase, amylase, and cellulase were detected at 68kDa, ~ 54kDa, 38kDa, and 30kDa, respectively. Also, the presence of protease was confirmed by the clear zone at 68, 31, and 16kDa. A 26% decrease in the kappa number and reduction in Hex A of the pulp was observed on the treatment of the pulp with enzyme as compared to the control pulp without any treatment. The physical and chemical properties of the pulp were also improved by enzyme-mediated pulping as compared to the control The physiochemical parameter of the effluent like TDS was reduced (397ppm) significantly in comparison to chemical deinking process and it was within the permissible limit. BOD and alkalinity were reduced when the enzymes and chemical dosage were used in combination. These results indicate that chemi-enzymatic deinking is most promising to reduce or remove the pollution parameters including ink and this approach can be used in the paper and pulp industry for sustainable development.

Smart Contact Lens for Colorimetric Visualization of Glucose Levels in the Body Fluid.

Frequent blood glucose monitoring is a crucial routine for diabetic patients. Traditional invasive methods can cause discomfort and pain and even pose a risk of infection. As a result, researchers have been exploring noninvasive techniques. However, a limited number of products have been developed for the market due to their high cost. In this study, we developed a low-cost, highly accessible, and noninvasive contact lens-based glucose monitoring system. We functionalized the surface of the contact lens with boronic acid, which has a strong but reversible binding affinity to glucose. To achieve facile conjugation of boronic acid, we utilized a functional coating layer called poly(tannic acid). The functionalized contact lens binds to glucose in body fluids (e.g., tear) and releases it when soaked in an enzymatic cocktail, allowing for the glucose level to be quantified through a colorimetric assay. Importantly, the transparency and oxygen permeability of the contact lens, which are crucial for practical use, were maintained after functionalization, and the lenses showed high biocompatibility. Based on the analysis of colorimetric data generated by the smartphone application and ultraviolet-visible (UV-vis) spectra, we believe that this contact lens has a high potential to be used as a smart diagnostic tool for monitoring and managing blood glucose levels.

Efficient enzyme-free method to assess the development and maturation of the innate and adaptive immune systems in the mouse colon

Researchers who aim to globally analyze the gastrointestinal immune system via flow cytometry have many protocol options to choose from, with specifics generally tied to gut wall layers of interest. To get a clearer idea of the approach we should use on full-thickness colon samples from mice, we first undertook a systematic comparison of three tissue dissociation techniques: two based on enzymatic cocktails and the other one based on manual crushing. Using flow cytometry panels of general markers of lymphoid and myeloid cells, we found that the presence of cell-surface markers and relative cell population frequencies were more stable with the mechanical method. Both enzymatic approaches were associated with a marked decrease of several cell-surface markers. Using mechanical dissociation, we then developed two minimally overlapping panels, consisting of a total of 26 antibodies, for serial profiling of lymphoid and myeloid lineages from the mouse colon in greater detail. Here, we highlight how we accurately delineate these populations by manual gating, as well as the reproducibility of our panels on mouse spleen and whole blood. As a proof-of-principle of the usefulness of our general approach, we also report segment- and life stage-specific patterns of immune cell profiles in the colon. Overall, our data indicate that mechanical dissociation is more suitable and efficient than enzymatic methods for recovering immune cells from all colon layers at once. Additionally, our panels will provide researchers with a relatively simple tool for detailed immune cell profiling in the murine gastrointestinal tract, regardless of life stage or experimental conditions.

Discovery of Potent Glycosidases Enables Quantification of Smoke-Derived Phenolic Glycosides through Enzymatic Hydrolysis.

When grapes are exposed to wildfire smoke, certain smoke-related volatile phenols (VPs) can be absorbed into the fruit, where they can be then converted into volatile-phenol (VP) glycosides through glycosylation. These volatile-phenol glycosides can be particularly problematic from a winemaking standpoint as they can be hydrolyzed, releasing volatile phenols, which can contribute to smoke-related off-flavors. Current methods for quantitating these volatile-phenol glycosides present several challenges, including the requirement of expensive capital equipment, limited accuracy due to the molecular complexity of the glycosides, and the utilization of harsh reagents. To address these challenges, we proposed an enzymatic hydrolysis method enabled by a tailored enzyme cocktail of novel glycosidases discovered through genome mining, and the generated VPs from VP glycosides can be quantitated by gas chromatography-mass spectrometry (GC-MS). The enzyme cocktails displayed high activities and a broad substrate scope when using commercially available VP glycosides as the substrates for testing. When evaluated in an industrially relevant matrix of Cabernet Sauvignon wine and grapes, this enzymatic cocktail consistently achieved a comparable efficacy of acid hydrolysis. The proposed method offers a simple, safe, and affordable option for smoke taint analysis.

265 Effect of L-glutamine and enzyme supplementation of liquid creep feed from d 8 to 28 of age on pre-weaning growth and health of pigs

Abstract The provision of liquid creep feed has been shown to increase dry matter intake (DMI) compared with dry creep feeding. The greater DMI associated with liquid feeding means that it could be used as an effective route for feed additive supplementation of suckling pigs to optimize growth and health at weaning. The objective of this study was to assess the effect of L-glutamine or enzyme supplementation of liquid creep feed on growth and medication usage in suckling pigs. Sows (n = 60) were blocked on parity, previous number of piglets weaned and body weight (BW) at d 107 of gestation, and randomly assigned to one of 3 dietary treatments from d 8 of age to weaning at d 28: 1) liquid starter (control); 2) control supplemented with 10 kg of L-glutamine per tonne of starter diet (glutamine); and 3) control supplemented with a cocktail of enzymes (lipase, protease and α-amylase included at 200 g, 50 g and 112.5 g, respectively per tonne of starter diet; enzymes). Individual piglets were weighed on d 8, 14, 21 and 27 of age. Litter dry matter intake, the volume of antibiotics and anti-inflammatories administered per pig and per sow were recorded for the pre-weaning period. Data were analyzed using the mixed models procedure in SAS (version 9.4; SAS Institute Inc.) with treatment as a fixed effect, and day post-partum as the repeated measure. Block was included as a random effect and the number of pigs/litter at d 8 and the average pig BW at d 8 were included as co-variates when significant in the model. Means separation was by Tukey’s adjustment for multiple comparisons. The piglet nested within sow was the experimental unit. The DMI of creep feed did not differ between treatments (P > 0.05). From d 14 to 21, glutamine decreased average daily gain (ADG) by 7% compared with the control (P < 0.05). However, ADG did not differ between treatment groups from d 8 to 14 or from d 21 to 27. Body weight was not affected by treatment at d 8 or 14 (P > 0.05). However, at d 21, glutamine tended to reduce BW by 2% compared with the control (P = 0.09) and at d 27, it tended to reduce BW by 2% compared with the control and enzymes (P = 0.08). The volume of antibiotics or anti-inflammatories administered to piglets or sows was not affected by treatment. However, glutamine tended to increase diarrhea incidence by 12% between d 8 and 27 compared with the control (P = 0.09). In conclusion, supplementing liquid creep feed with 10 kg of glutamine/tonne of feed reduced pre-weaning growth and tended to increase diarrhea incidence in piglets. In addition, supplementing liquid creep feed with enzymes had no effect on pre-weaning growth and medication usage in piglets.

Community dynamics and metagenomic analyses reveal Bacteroidota's role in widespread enzymatic Fucus vesiculosus cell wall degradation

Enzymatic degradation of algae cell wall carbohydrates by microorganisms is under increasing investigation as marine organic matter gains more value as a sustainable resource. The fate of carbon in the marine ecosystem is in part driven by these degradation processes. In this study, we observe the microbiome dynamics of the macroalga Fucus vesiculosus in 25-day-enrichment cultures resulting in partial degradation of the brown algae. Microbial community analyses revealed the phylum Pseudomonadota as the main bacterial fraction dominated by the genera Marinomonas and Vibrio. More importantly, a metagenome-based Hidden Markov model for specific glycosyl hydrolyses and sulphatases identified Bacteroidota as the phylum with the highest potential for cell wall degradation, contrary to their low abundance. For experimental verification, we cloned, expressed, and biochemically characterised two α-L-fucosidases, FUJM18 and FUJM20. While protein structure predictions suggest the highest similarity to a Bacillota origin, protein–protein blasts solely showed weak similarities to defined Bacteroidota proteins. Both enzymes were remarkably active at elevated temperatures and are the basis for a potential synthetic enzyme cocktail for large-scale algal destruction.

Enzyme-mediated ultrasound-assisted extraction of rutin, quercetin, and other antioxidant and antidiabetic compounds from Holarrhena antidysenterica seeds and optimization as per RSM

Rutin and quercetin are valuable bioactive natural products. The current study successfully discovered an efficient extraction protocol for these and other bioactive compounds from Holarrhena antidysenterica (HA) seeds based on an enzyme-mediated ultrasound-assisted extraction (EM-UAE). Extraction process optimization was done based on response surface methodology with seven response factors including total extraction yield (%), amount of rutin (AOR, mg/g), amount of quercetin (AOQ, mg/g), total phenolic content (TPC, mg gallic acid equivalents/g dry weight (mg GAE/g DW), total flavonoids content (TFC, mg rutin equivalents/g DW (mg RE/g DW) anti-radical activity (ARA, %), and α-amylase inhibitory activity (AIA, %). High-performance liquid chromatography (HPLC) was used for quantification of rutin and quercetin. The optimal extraction yield, AOR, AOQ, TPC, TFC, ARA, and AIA were 32.39 %, 71.43 mg/g, 23.68 mg/g, 178.63 mg GAE/g DW, 100.22 mg RE/g DW, 76.41 %, and 77.48 %, respectively. The optimal conditions were 91 min time, 6.1 mL enzyme cocktail, pH 4.8, and 48 ℃ temperature. For ARA, EC50 (µg/mL) of HA was 9.75 ± 0.27 compared to 6.10 ± 0.38 of ascorbic acid standard. For AIA, IC50 (µg/mL) of HA and acarbose standard were 51.97 ± 0.21 and 47.89 ± 0.61 μg/mL, respectively. The validation study strongly supported the predicted model with a very low relative standard deviation (0.45–3.37 %). The model was statistically significant and efficient for obtaining rutin, quercetin, and other antioxidant and antidiabetic compounds from Holarrhena antidysenterica seeds on the industrial level.

Production of Mannooligosaccharides from Açaí Seed by Immobilized β-Mannanase

In this work, an enzyme cocktail with β-mannanase as the main activity was immobilized on epoxy resin foams filled with fibers from annatto capsules. The catalytic system was characterized by SEM, FTIR, and a mechanical crush resistance test. The behavior of the pH and temperature for the hydrolysis of the locust bean gum were also studied. With the same substrate and with respect to the free enzyme, the immobilized enzyme showed an activity retention of 79.61%. Its operational stability in ten reuse cycles did not show any statistically significant loss of activity. This catalytic system was used to study the preferential release of MOS of two to five degrees of polymerization from mannan present in dried and ground açaí seeds, which were not subjected to any other pretreatment. Using an experimental response surface design, the predicted quadratic models for the M2–M5 MOS content were obtained and they fit well with the experimental data, predicting a production range between 0.435 and 20 g/L of MOS (M2–M5). In addition, the production reached about 12 g/L under the optimized conditions. These results indicate that the used foamed epoxy resin supports and immobilization methodology are suitable for catalyzing the hydrolysis of mannan from açaí seeds.

Fungal systems for lignocellulose deconstruction: From enzymatic mechanisms to hydrolysis optimization

AbstractLignocellulosic biomass is an abundant renewable feedstock, but its complex structure of lignocellulose poses barriers to its enzymatic hydrolysis and fermentation. Fungi possess diverse lignocellulolytic enzyme systems that synergistically deconstruct lignocellulose into soluble sugars for fermentation. This review elucidates recent advances in understanding the molecular mechanisms underpinning fungal degradation of lignocellulose. We analyze major enzyme classes tailored by fungi to depolymerize cellulose, hemicellulose, and lignin. Highlighted are the concerted actions and intimate partnerships between these biomass‐degrading enzymes. Current challenges impeding large‐scale implementation of enzymatic hydrolysis are discussed, along with emerging biotechnological opportunities. Advanced pretreatments, high‐throughput enzyme engineering platforms, and machine learning or artificial intelligence‐guided lignocellulolytic enzyme cocktail optimization represent promising ways to improve hydrolytic efficiencies. Elucidating the coordinated interplay and regulation of fungal lignocellulolytic machinery can facilitate optimization of fungal biotechnology platforms. Harnessing the efficiency of fungal biomass deconstruction promises to enhance the development of biorefinery processes for sustainable bioenergy.

The rotamer of the second-sphere histidine in AA9 lytic polysaccharide monooxygenase is pH dependent

Lytic polysaccharide monooxygenases (LPMOs) catalyze a reaction that is crucial for the biological decomposition of various biopolymers and for the industrial conversion of plant biomass. Despite the importance of LPMOs, the exact molecular-level nature of the reaction mechanism is still debated today. Here, we investigated the pH-dependent conformation of a second-sphere histidine (His) that we call the stacking histidine, which is conserved in fungal AA9 LPMOs and is speculated to assist catalysis in several of the LPMO reaction pathways.Using constant-pH and accelerated molecular dynamics simulations, we monitored the dynamics of the stacking His in different protonation states for both the resting Cu(II) and active Cu(I) forms of two fungal LPMOs. Consistent with experimental crystallographic and neutron diffraction data, our calculations suggest that the side chain of the protonated and positively charged form is rotated out of the active site toward the solvent. Importantly, only one of the possible neutral states of histidine (HIE state) is observed in the stacking orientation at neutral pH or when bound to cellulose. Our data predict that, in solution, the stacking His may act as a stabilizer (via hydrogen bonding) of the Cu(II)-superoxo complex after the LPMO-Cu(I) has reacted with O2 in solution, which, in fine, leads to H2O2 formation. Also, our data indicate that the HIE-stacking His is a poor acid/base catalyst when bound to the substrate and, in agreement with the literature, may play an important stabilizing role (via hydrogen bonding) during the peroxygenase catalysis.Our study reveals the pH titration midpoint values of the pH-dependent orientation of the stacking His should be considered when modeling and interpreting LPMO reactions, whether it be for classical LPMO kinetics or in industry-oriented enzymatic cocktails, and for understanding LPMO behavior in slightly acidic natural processes such as fungal wood decay.

Trichoderma koningiopsis fermentation in airlift bioreactor for bioherbicide production.

During scaling of fermentations, choosing a bioreactor is fundamental to ensure the product's quality. This study aims to produce bioherbicides using Trichoderma koningiopsis fermentation, evaluating process parameters in an Airlift bioreactor. As a response, we quantified the production of enzymes involved in the bioherbicide activity (amylase, cellulase, laccase, lipase, and peroxidase). In addition, it evaluated the agronomic efficiency of the fermented extract optimized through tests that promoted soybean growth and nodulation, soybean seed germination, and in vitro phytopathogen control. As a result of optimizing the scaling bioprocess, it was possible to obtain an adequate fermentation condition, which, when applied to soybean seeds, had beneficial effects on their growth. It allowed the production of an enzyme cocktail. These results add a crucial biotechnological potential factor for the success of the optimized formulation in the Airlift bioreactor, in addition to presenting relevant results for the scientific community.

Optimization and characterization of aqueous enzyme-assisted solvent extraction of apricot kernel oil.

Aqueous enzyme-assisted solvent extraction (AE-SE) of oil from apricot (Prunus armeniaca L.) kernel was investigated in this study and the operational parameters were optimized. After preliminary screening, a cocktail of enzymes (Celluclast 1.5 L and Alcalase Pure 2.4 L [60:40, v/v)] of 1% (v/w) was chosen. The extraction process parameters: temperature (40-60 °C), time (1-5 h), and pH (4-9) were optimized using Box-Behnken design to achieve the highest oil yield (%) and antioxidant activity (DPPH, %). Under optimized conditions, i.e., temperature 40 °C, time 2.5 h, and pH 8.28, the highest oil yield and DPPH were 47.93% and 67.31%, respectively. The gas chromatography analysis disclosed that apricot kernel oil extracted by solvent extraction and AE-SE have similar fatty acid compositions, and the oil is rich in unsaturated fatty acids. The physicochemical analysis showed AE-SE method produces high-quality oil and can be substituted as green technology for industrial oil extraction purposes.