Hydrolysis Time Research Articles

Amino acid requirements of the infant: the amino acid composition of human breast milk.

The recommended amino acid requirements of the infant are based on the amino acid composition of mature human breast milk. The amino acid composition of breast milk is usually determined following either acid or alkaline (for tryptophan) hydrolysis. For accuracy, however, the known effect of hydrolysis time on amino acid composition should be accounted for. Also, ideally the amino acid composition of breast milk should be given in units of digested (assumed to be absorbed) amino acids. A review of the literature is presented which gives mean total amino acid concentrations in mature human milk (n = 26 studies), mean hydrolysis correction factors (n = 3 studies) and mean true ileal amino acid digestibility coefficients (n = 3 studies, suckling piglet). There were differences between the estimates of amino acid concentration corrected for hydrolysis time and digestibility, and current FAO (2013) recommendations that were not corrected for these factors. The values based on the published literature up until 2023 (mg/g true protein) corrected for hydrolysis time and digestibility gave higher values (more than 16% higher) for leucine, lysine and threonine, and considerably higher values (greater than 30%) for histidine and tryptophan. Current recommendations may need revision.

Enzymatic hydrolysis and biological activities of Konjac glucomannan hydrolysate in different degree of polymerisation

SummaryThe study explores the enhanced functional and bioactive properties of Konjac Glucomannan Hydrolysate (KGMH) by partially degrading Konjac Glucomannan (KGM) using β‐mannanase over 60 min. KGM concentrations (40% and 50% w/w) were treated with 200–300 U g−1 of the enzyme over 60 min. The hydrolysis of KGM was monitored by colorimetry, with DP values ranging from 4.20 to 6.16 for 40% KGM and 4.10 to 4.60 for 50% KGM. MALDI‐TOF‐MS analysis confirmed typical oligosaccharides with DP values from 2 to 9 and some acetyl substitutions. The optimal conditions of 40% KGM with 250 U g−1 enzyme and varying hydrolysis times produced KGMHs with a wide range of DPs, demonstrating in vitro antioxidant and anti‐glycation activities. The results showed significant bioactivities (P < 0.05) positively correlated with lower DP values. This study emphasises the potential of KGMH as a novel functional food ingredient, highlighting its bioactive properties and the significant impact of DPs on the biological functionality of saccharides.

Optimization, isolation, identification and molecular mechanisms in B16F10 melanoma cells of a novel tyrosinase inhibitory peptide derived from split gill mushrooms

Hyperpigmentation often arises from an imbalance in melanogenesis, primarily due to the overexpression of tyrosinase (TYR). While the inhibition of TYR presents a common approach to skin whitening, it can lead to undesirable side effects. Thus, there is growing interest in safe and natural alternatives for TYR inhibition. Bioactive compounds and peptides sourced from split gill mushrooms hold promise in this regard. This study aims to optimize the conditions for papain-mediated hydrolysis of split gill mushroom protein to inhibit TYR activity, utilizing response surface methodology (RSM) and central composite design (CCD). Optimal conditions were determined at a temperature of 46.70 °C, a hydrolysis time of 217.09 min, and an enzyme-to-substrate ratio (E/S) of 1.1%. Under these conditions, the resulting hydrolysates exhibited significant TYR inhibition, with an IC50 value of 117.86 μg/mL and a degree of hydrolysis (DH) of 87.97%. Further purification via ultrafiltration and RP-HPLC yielded a peptide, Tyr-Ala-Ser-Ile-Leu-Leu (YASILL or YL-6), identified through LC-Q-TOF-MS/MS, which competitively inhibited TYR. YL-6 demonstrated an IC50 value of 3.97 mM for mono-phenolase activity and 6.75 mM for di-phenolase activity. Molecular docking analysis revealed hydrogen bonds and hydrophobic interactions between TYR and YL-6. Treatment of B16F10 cells with YL-6 across concentrations ranging from 10-3000 μM showed no cytotoxic effects.The inhibition of melanin synthesis was investigated via qRT-PCR along with Western blot in MITF, TYR, TRP-1, and TRP-2. The results obtained in this research may prove significant in guiding the development of commercially viable cosmetic products to whiten the skin.

Optimization of the pumpkin pectin paste production process

Taking into account the mentioned advantages, the use of vegetable and fruit pastes with increased pectin content has great potential in therapeutic and preventive nutrition. They not only contribute to the removal of radionuclides, heavy metals and toxins from the body but also provide additional enrichment of the body with a complex of natural vitamins, tannins and sugars. These natural substances help to improve the work of the gastrointestinal tract, increase the general tone of the body and help strengthen the immune system. Optimized technological production processes, in particular the hydromodule of hydrolysis of raw materials, correctly selected temperature regimes and evaporation methods that ensure maximum efficiency of pectin extraction from raw materials. This makes it possible to achieve high quality of the finished product with full preservation of the beneficial properties of pectin substances. Modern technological methods guarantee the preservation of the maximum number of active components, which makes the product even more valuable for consumers. For the production of pectin pastes, Gleisdorfer Olkurbis, Hamlet, Pivdenny, Miranda and Dana Styrian Bare-seeded pumpkins were selected. The hydrolysis process was carried out using lactic acid and whey, which contributed to a faster process and acted as preservatives. According to the results of experimental studies, rational hydrolysis conditions were established: hydro modulus (1:14–1:17), concentration of hydrolyzers, temperature regime, hydrolysis time, and evaporation method. The evaluation of the product included the analysis of dry matter content, colour, pectin content and gel-forming properties. According to the results of the experiments, it was determined that the rational conditions for the hydrolysis of pumpkin raw materials are a temperature within 80°C, a duration of the process of 65 minutes and a hydromodule of 15–16. Rational conditions for evaporation in a vacuum are established, which allow obtaining high-quality pectin paste with a pectin content of 1.5% and a dry matter content within 70%. A specialized technology for the production of pectin pastes from pumpkin and other vegetable raw materials has been developed, which allows you to effectively preserve the natural properties and nutrients characteristic of local fruits and vegetables. Technological and hardware-technological production schemes have been developed, which will contribute to the organization of production in the areas where raw materials are grown, reducing transportation costs and contributing to regions' sustainable development. Localization of production contributes to the creation of new jobs and support of the local economy Thus, the use of vegetable and fruit pastes with increased pectin content has a great potential for strengthening the health of the population. They are a rich source of natural vitamins and microelements, which contribute to the improvement of the general condition of the body, increase immunity and protect against harmful effects of the environment. The growing demand for natural and healthy food products makes pectin pastes a promising direction for investment and development of new technologies in the food industry.

Acid Hydrolysis of Quinoa Starch to Stabilize High Internal Phase Emulsion Gels.

Starch nanocrystals (SNCs) to stabilize high internal phase emulsions (HIPEs) always suffer low production efficiency from acid hydrolysis. Due to its small granule size, Quinoa starch (QS) was selected to produce SNCs as a function of acid hydrolysis time (0-4 days), and their structural changes and potential application as HIPEs' stabilizers were further explored. With increasing the acid hydrolysis time from 1 day to 4 days, the yield of QS nanocrystals decreased from 30.4% to 10.8%, with the corresponding degree of hydrolysis increasing from 51.2% to 87.8%. The occurrence of QS nanocrystals was evidenced from the Tyndall effect and scanning electron microscopy with particle size distribution. The relative crystallinity of QS subjected to different hydrolysis times (0-4 days) increased from 22.27% to 26.18%. When the acid hydrolysis time of QS was 3 and 4 days, their HIPEs showed self-standing after inversion, known as high internal phase emulsion gels (HIPE gels), closely related to their densely packed interfacial architecture around oil droplets, seen on an optical microscope, and relatively high apparent viscosity. This study could provide a theoretical guidance for the efficient production and novel emulsification of SNCs from QS to HIPE gels.

Formation of self-assembled fibril aggregates of trypsin-controllably hydrolyzed soy protein and its regulation on stability of high internal phase Pickering emulsions

The mechanisms of trypsin hydrolysis time on the structure of soy protein hydrolysate fibril aggregates (SPHFAs) and the stability of SPHFAs-high internal phase Pickering emulsions (HIPPEs) were investigated. SPHFAs were prepared using soy protein hydrolysate (SPH) with different trypsin hydrolysis time (0 min–120 min) to stabilize SPHFAs-HIPPEs. The results showed that moderate trypsin hydrolysis (30 min, hydrolysis degree of 2.31 %) induced SPH unfolding and increased the surface hydrophobicity of SPH, thereby promoting the formation of flexible SPHFAs with maximal thioflavin T intensity and ζ-potential. Moreover, moderate trypsin hydrolysis improved the viscoelasticity of SPHFAs-HIPPEs, and SPHFAs-HIPPEs remained stable after storage at 25 °C for 80 d and heating at 100 °C for 1 h. Excessive trypsin hydrolysis (> 30 min) decreased the stability of SPHFAs-HIPPEs. In conclusion, moderate trypsin hydrolysis promoted the formation of flexible SPHFAs with high surface charge by inducing SPH unfolding, thereby promoting the stability of SPHFAs-HIPPEs.

Direct analysis in real time mass spectrometry (DART-MS/MS) for rapid urine opioid detection in a clinical setting

Background and aimsCurrent laboratory methods for opioid detection involve an initial screening with immunoassays which offers efficient but non-specific results and a subsequent liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmation which offers accurate results but requires extensive sample preparation and turnaround time. Direct Analysis in Real Time (DART) tandem mass spectrometry is evaluated as an alternative approach for accurate opioid detection with efficient sample preparation and turnaround time. Materials and methodsDART-MS/MS was optimized by testing the method with varying temperatures, operation modes, extraction methods, hydrolysis times, and vortex times. The method was evaluated for 12 opioids by testing the analytical measurement range, percent carryover, precision studies, stability, and method-to-method comparison with LC-MS/MS. ResultsDART-MS/MS shows high sensitivity and specificity for the detection of 6-acetylmorphine, codeine, hydromorphone, oxymorphone, hydrocodone, naloxone, buprenorphine, norfentanyl, and fentanyl in urine samples. However, its performance was suboptimal for norbuprenorphine, morphine and oxycodone. ConclusionIn this proof-of-concept study, DART-MS/MS is evaluated for its rapid quantitative definitive testing of opioids drugs in urine. Further research is needed to expand its application to other areas of drug testing.

A novel ultrasound-assisted enzyme extraction method of total flavonoids from Viticis Fructus and processed Viticis Fructus: Comparison of in vitro antioxidant activity

In this study, it is the first that the Viticis Fructus (VF) was used as the raw material for extracting total flavonoids using the ultrasound-assisted enzyme extraction (UAE) method. Response surface methodology was employed to determine the optimal extraction parameters. The optimal conditions were as follows: 60 % ethanol solution as the extract solvent, material–liquid ratio of 1:25, pH value of 4, enzyme addition amount of 1.5 %, enzymatic hydrolysis time of 30 min, enzymatic hydrolysis temperature of 40 ℃, and ultrasonic time of 50 min. Comparing the total flavonoid yield of VF and processed VF (PVF) extracted using different methods, it was observed that UAE resulted in a higher total flavonoid yield compared to traditional ultrasound extraction and enzyme extraction. Additionally, the total flavonoid yield of PVF extracted by all three methods was generally higher than that of VF. The PVF solution extracted by UAE also demonstrated better in vitro antioxidant activity compared to VF. These results suggest that UAE is an effective method to enhance the activity of natural total flavonoids. The study of the physicochemical properties and in vitro antioxidant activity of VF and PVF showed that the total flavonoid yield and antioxidant activity significantly increased after VF stir-frying, indicating that their efficacy can also be enhanced.

Hierarchical porous structure design and water activation in hydrogels containing hyperbranched peach gum polysaccharide for efficient solar water evaporation

Solar-powered interfacial evaporation is a developing and sustainable technique increasingly utilized in desalination and wastewater purification. This technology involves the creation of cellulose nanofiber (CNF)/polylactic acid (PLA) composite aerogels through the Pickering emulsion approach. Self-floating aero-hydrogel (E-VGP) with a hierarchical porous structure was formed on a viscous mixture containing polyvinyl alcohol (PVA), peach gum polysaccharide (PGP), and polypyrrole (PPy) via an in-situ polymerization process. Furthermore, by modifying the hydrolysis time of PGP with a hyperbranched polyhydroxy structure, VGP hybrid hydrogels of varying microscopic molecular sizes were produced. Additionally, solar vapor generators (SVG) with diverse macroscopic structures were fabricated using molds. The V8G4-12hP0.2 hybrid hydrogel, synthesized using PGP hydrolyzed for 12 h, exhibited an evaporation enthalpy of water at 1204 J g−1. This capacity effectively activates water and enables low enthalpy evaporation. Conversely, the macrostructural design allows the cylindrical rod raised sundial-shaped structure of SVG3 to possess an expanded evaporation area, minimize energy loss, and even harness additional energy from its nonradiative side. Consequently, this micro-macrostructural design enables SVG3 to attain an exceptionally high evaporation rate of 3.13 kg m−2 h−1 under 1 Sun exposure. Moreover, SVG3 demonstrates robust water purification abilities, suggesting significant potential for application in both desalination and industrial wastewater treatment.

Ultrasound-assisted hydrolysis of food waste using glucoamylase: Statistical optimization and mechanistic analysis with molecular simulations

This paper reports investigations in food waste hydrolysis using ternary approach that combines statistical optimization, ultrasound-assisted enhancement of hydrolysis kinetics, and molecular simulations that provide physical insight into the process. Initial optimization of hydrolysis parameters (Box–Behnken design) resulted in total reducing sugar yield of 263.4 mg/g biomass in 42 h. Sonication of hydrolysis mixture at 35 kHz at 20 % duty cycle yielded 4× reduction in hydrolysis time with 22 % enhancement in TRS yield (320 mg/g biomass). Analysis of GLCM's secondary structure through FTIR spectra deconvolution revealed significant changes induced by sonication. Sonication led to reduction in α-helix, and increase in random coil content. Molecular dynamics simulations unveiled majority of amino acid residues associated with GLCM binding pocket in α-helix and random coil regions. Consequently, sonication widened the binding pockets, facilitating easier transport of substrate and product. This effect translated into faster kinetics of enzymatic food waste hydrolysis.

Effects and optimization of hydrolysis conditions on the diameter distance ratio for metatitanic acid

Controlling and predicting the narrow particle size distribution of metatitanic acid could help to improve pigment and application performances of TiO2. To prepare metatitanic acid with narrow particle size distribution, the hydrolysis conditions of industrial TiOSO4 solution were optimized by using the response surface methodology of Box-Behnken design method. The effects of hydrolysis conditions such as TiO2 concentration, aging time and hydrolysis time on the diameter distance ratio and structure of metatitanic acid were investigated. The prediction model including the hydrolysis variables with good fitting result was established to predict the diameter distance ratio or optimize the hydrolysis variable values. The regression equation model was significant and reliable with actual correlation coefficient R2 of 0.9989. The hydrolysis conditions greatly influenced the quality and quantity of hydrolysis induced crystal nucleus, hydrolysis rate, completeness degree of hydrolysis, crystallization growth and aggregation rate, and eventually affected the particle size distribution of metatitanic acid. The synergistic interaction for TiO2 concentration and hydrolysis time was very significant. The verification experiments confirmed that the diameter distance ratio could reach the average value of 1.100 under the optimal conditions. The prepared metatitanic acid had anatase TiO2 phase structure, with average particle size of 1.621–1.819 µm. And this helped to improve the high quality and green development of the titanium dioxide industry.

Optimization of Hydrolytic Enzyme Production from Tempeh Starter to Increase In Vitro Pepsin Digestibility of Black Soldier Fly Maggot

This research aims to evaluate the optimal level and incubation time for the production of hydrolytic enzymes from tempeh starter (TS), in order to increase the in vitro pepsin digestibility of Black Soldier Fly (BSF), Hermetia illucens, maggot flour. The materials used were commercial tempeh starter, pepsin, and 15 days of age dried BSF maggot. In this study, two experiments were conducted. In a factorial completely randomized design, the analysis of variance (ANOVA) was performed to assess the significance of treatment effects, and orthogonal polynomial contrasts were employed to identify specific linear and quadratic trends among the group means. In experiment 1, research was carried out to determine the optimal starter level and incubation time to produce hydrolytic enzymes from tempeh starter. In experiment 2, crude enzyme from the best treatment in the first stage was used to increase the pepsin digestibility of BSF maggot flour. The optimal protein content and enzyme activities of tempeh starter was 0.10-0.13% starter with an incubation time of 2.00-3.47 days. Meanwhile, the optimal pepsin digestibility of BSF maggot flour was 1.51% enzymes with a hydrolysis time of 23.87 hours. This analysis measurement of protein content and enzymatic activity of tempeh starter and in vitro digestibility of BSF flour represent the preliminary methodology used to initially select hydrolyzed BSF flour to be a source of protein in animal feed.

Improving stereoselectivity of phosphotriesterase (PTE) for kinetic resolution of chiral phosphates.

Specific stereoisomer is paramount as it is vital for optimizing drug efficacy and safety. The quest for the isolation of desired stereoisomer of active pharmaceutical ingredients or key intermediates drives innovation in drug synthetic and biocatalytic methods. Chiral phosphoramidate is an important building block for the synthesis of antiviral drugs such as remdesivir and sofosbuvir. Given the clinical potency of the (Sp)-diastereomer of the drugs, an enzyme capable of completely hydrolyzing the (Rp)-diastereomer is needed to achieve the purified diastereomers via biocatalytic reaction. In this study, protein engineering of phosphotriesterase (PTE) was aimed to improve the specificity. Employing rational design and site-directed mutagenesis, we generated a small library comprising 24 variants for activity screening. Notably, W131M and I106A/W131M variants demonstrated successful preparation of pure (Sp)-diastereomer of remdesivir and sofosbuvir precursors within a remarkably short hydrolysis time (<20min). Our work unveils a promising methodology for producing pure stereoisomeric compounds, utilizing novel biocatalysts to enable the chemoenzymatic synthesis of phosphoramidate nucleoside prodrugs.

Technological and Bioactive Properties of Collagen Extracts Obtained from Bullfrog (Lithobates catesbeianus) Skin

ABSTRACT The objective was to extract type I collagen and determine its ideal extraction conditions, technological parameters, and antioxidant properties. A 2 × 3factorial design was used, with variations in the type of enzyme and hydrolysis time. The treatments that presented the highest yield based on protein content (70.46% and 64.57%) and collagen content (34.65 and 49.05 g/100 g) were the samples subjected to acid hydrolysis-enzymatic for 12 h (CP12 and CB12). CB12 demonstrated greater water absorption capacity (WA), while CP12 demonstrated greater water solubility (WS) and antioxidant activity. CP12 is the most promising collagen extract for use as a bioactive ingredient in foods.

Effect of the addition of protein hydrolysates from grape seed meal residue to red wines in warm regions in the stabilization stage

The effect of adding protein hydrolysates from defatted grape seed meals, a residue of the grape pomace industry, on the improvement of color stabilization of Syrah wines in warm climates was evaluated. Factors such as the type of raw material, the hydrolysis time, and the doses of protein hydrolysate were assessed. Hydrolysates were added to the wines at the stabilization stage and scrutinized over five months focusing on the polyphenolic composition, copigmentation and polymerization, as well as color by differential tristimulus colorimetry. The addition of 3 g/L protein hydrolysates from 1-h hydrolysis depicted a color stabilization effect, leading to higher values of chroma (C*ab), phenolic compounds (benzoic acids and flavan-3-ols), and copigmentation, although assuming a slightly more aged-related tonality. These data obtained provide an interesting insight, not reported so far, on the use of this oenological alternative to avoid common color losses of red wines elaborated in warm climate.

Effects of silane hydrolysis time on the physicochemical properties of bioplastics based on starch and epoxidized soybean oil

This work investigated the effects of 3-aminopropyl triethoxy silane (APTES) hydrolysis time on the physicochemical properties of the resulting starch/epoxidized soybean oil (ESO) bioplastics comprehensively. FTIR analysis confirmed that APTES hydrolyzed for 4 h had the best modification effect on starch. The results of XRD and TGA demonstrated the successful silylation of starch by APTES despite hydrolysis time. Silylation treatment reduced the thermal stability of starch slightly, but enhanced the thermal stability of the resultant bioplastics, revealing better interaction between silylated starch and ESO. The interfacial adhesion of starch and ESO in the bioplastics was obviously enhanced when APTES was hydrolyzed for 2–24 h. The bioplastics with APTES hydrolyzed for 2–4 h showed more desirable tensile properties as the silane hydrolysis was complete and self-condensation of hydrolyzed silanes was avoided. The bioplastics containing silylated starch still showed superior opacity and biodegradability.

Enhancement of terephthalic acid recovered from PET waste using a combination of citric acid and dimethyl sulfoxide extraction

This study aimed to develop an eco-friendly, cost-efficient, and practically viable method for extracting terephthalic acid (H2BDC) from polyethylene terephthalic (PET) waste. Dimethyl sulfoxide (DMSO) was combined with either citric acid (C6H8O7) or H2SO4 to enhance the particle size of H2BDC, and the optimum conditions during the acidification step were determined. Additionally, response surface methodology was employed to examine the influence and interaction of extractant (NaOH) concentration, hydrolysis temperature, and time on the optimal H2BDC yield and recovery ratio. Experimental results demonstrated that NaOH concentration significantly impacted both H2BDC yield and recovery ratio, surpassing the effects of hydrolysis temperature and time. Under optimal conditions involving a temperature of 200 °C and a 12 h reaction time with 5% NaOH, the model predicted a 100% yield and recovery ratio, which closely matched the experimental results of 99% and 100% for yield and recovery ratio, respectively. To enhance particle size, a combination of DMSO and C6H8O7 was more effective than H2SO4. The maximum particle size achieved was 57.4 µm under the following optimum conditions: premixing 5 M C6H8O7 with DMSO at a 35:75 mL ratio and maintaining a reaction temperature of 75 °C for 40 min. The study demonstrated the stability and consistency of the method. The H2BDC yield remained between 96 and 98% with high purity over eight consecutive cycles of using the DMSO and C6H8O7 mixture. The findings highlight the importance of integrating C6H8O7 and DMSO to enhance H2BDC quality, meeting commercial product criteria with evidence of high purity and large particle size. This method presents a promising solution for extracting H2BDC from PET waste, with potential implications for the recycling industry and a positive environmental impact.

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

In the context of biorefinery, researchers have been looking for lignocellulosic biomasses and ideal treatments to produce economically viable biofuels. In this scenario, the bamboo culm appears as a plant matrix of great potential, given the high cellulose content of low crystallinity. Thus, the objective and differential of this work was to determine the best conditions for enzymatic hydrolysis of cellulose extracted from bamboo culm and to evaluate its potential application in the production of bioethanol through Separate Hydrolysis and Fermentation (SHF) and Saccharification and Simultaneous Fermentation (SSF) by Saccharomyces cerevisiae modified via CRISPR/Cas9. The average cellulose extraction yield was 41.87 % with an extraction efficiency of 86.76 %. In general, as the hydrolysis time increased, an increase in glucose production was observed in almost all assays, with higher hydrolysis efficiency values at 72 h. The results ranged from 2.09 to 19.8 g/L of glucose obtained with efficiency values of 10.47 to 99 %. The best conditions were found in test 5 (temperature of 36 °C and pH 5.0, with only 10 FPU/g of substrate Cellic Ctec2 Novozymes ® cocktail). It is observed that for all hydrolysis times the independent variables pH and temperature were significant under the hydrolysis efficiency, showing a negative effect, indicating that higher values of the same promote lower values of the response variable. For bioethanol production, a maximum concentration of 7.84 g/L was observed for the SSH process after 4 h of fermentation, while for the SSF process it was 12.6 g/L after 24 h of fermentation, indicating the large potential of the simultaneous process together with the application of bamboo culm biomass for high production of biofuel.

Enzymatic hydrolysis of chicken viscera and bones: Rest raw material characterization and evaluation of industrially relevant process parameters on product yields

Enzymatic hydrolysis is an efficient processing method for valorizing chicken rest raw material by generating new food- or feed ingredients. This paper characterizes two types of chicken rest raw material: viscera and a mixture of bones, skin and remaining meat, and evaluates how industrially relevant hydrolysis process designs affect overall yield of all products, including hydrolysate, sediment, lipid, and emulsion. Eleven hydrolyses were performed using viscera and bone materials, endogenous and commercial enzymes, pre-treatments, variations in water addition and hydrolysis times. Compared to bone materials, viscera had higher proteolytic activity, more readily water-soluble components, and lower effect of adding commercial enzymes on product yields. The process of heating the raw material to hydrolysis temperature greatly impacted product yields, representing about 50 % of the overall hydrolysate production from viscera during hydrolysis. Pre-inactivation of endogenous enzymes reduced initial, but not final, hydrolysate yields. Adding commercial enzymes to pre-inactivated viscera had no effect on yields compared to not adding enzymes. Reducing water addition lowered initial hydrolysate yield from bone material. Lipid yield reflected lipid content in the raw material, and thermal pre-separation of lipids did not increase the total lipid recovery. In general, hydrolysis of viscera generated more emulsion than the bone material.

Production of cellulose nano/microfibers through simultaneous milling and enzymatic hydrolysis with an optimized cocktail of cellulase/xylanase/LPMO

Cellulose nanofibers (CNF) and microfibrillated cellulose (MFC) are biodegradable micro and nanomaterials derived from plant cellulose. The properties of CNF, influenced by fibril length, impact the strength of cellulose-based nanomaterials. This study uses enzymatic milling to explore pretreatments, enzymatic optimization, and CNF/MFC production. The study found that hammer milling and lamination are effective pretreatments for preserving fibers. The optimal enzymatic mixture contained 75 % cellulase and 25 % xylanase, yielding 68.7 % and 2.86 %/h productivity, respectively. The addition of the LPMO enzyme increased the yield by 10–13 %, depending on its concentration. Enzymatic milling was introduced as an innovative approach to CNF production, reducing hydrolysis time and creating milder processing conditions. The combined process of ball milling and enzymatic hydrolysis on pre-ground pulp in the hammer mill achieved a maximum CNF yield of 45.83 % and productivity of 13.10 %/h. X-ray diffraction showed that the lamination-pre-treated pulp had a high crystallinity index of 84.34 %. Scanning electron microscopy revealed fibers with diameters ranging from 52 nm to 296 nm. These findings support the search for sustainable and eco-friendly alternatives to non-renewable resources, enabling sustainable CNF production at room temperature with reduced processing time.