Managing plastic waste remains an urgent environmental concern and switching to biodegradable plastics can reduce the dependence on depleting fossil fuels. This study emphasises the efficacy of macroalgae wastes, Eucheuma denticulatum residues (EDRs), as potential alternate feedstock to produce l-lactic acid (l-LA), the monomer of polylactic acid, through fermentation. An innovative environmental friendly strategy was explored in this study to develop a glucose platform from EDRs: pretreatment with microwave-assisted autohydrolysis (MAA) applied to enhance enzymatic hydrolysis of EDRs. The results indicate that MAA pretreatment significantly increased the digestibility of EDRs during the enzymatic hydrolysis process. The optimum pretreatment conditions were 120 °C and 50 min, resulting in 96.5% of enzymatic digestibility after 48 h. The high l-LA yield of 98.6% was obtained using pretreated EDRs and supplemented with yeast extract. The energy analysis implies that MAA pretreatment could further improve the overall energy efficiency of the process.
Poplar trees rapidly yield wood and are therefore suitable as a biofuel feedstock; however, the quality of poplar is modest, and the profitability of poplar cultivation depends on the efficiency of the harvesting process. This study offers a simple and sustainable technique to harvest lignocellulosic resources from poplar for bioethanol production. The proposed two-step pretreatment method increased the surface lignin content and decreased the surface polysaccharide content. The cellulose content increased to 54.9% and the xylan content decreased to 6.7% at 5% AC. The cellulose yield of poplar residues (Populus L.) reached 65.5% by this two-step acetic acid (AC) and sodium sulphite (SS) treatment method. Two-step pretreatment using 5% AC and 4% SS obtained a recovery of nearly 80% of the total available fermentable sugar. The surface characterization showed a higher porosity in treated samples, which improved their hydrolysability. This method decreased the amount of lignin in plant biomass, making it applicable for further wood resource recovery or waste recycling for biorefinery purposes at very low costs.
Pectinases are outstanding multienzymes, which have the potential to produce new emerging pectic-oligosaccharides (POS) via enzymatic hydrolysis of pectin. However, free pectinase is unable to undergo repeated reaction for the production of POS. This study proposed a sustainable biocatalyst of pectinases known as cross-linked pectinase aggregates (CLPA). Pectinase from Aspergillus aculeatus was successfully precipitated using 2 mg/mL pectinase and 60 % acetone for 20 min at 20 °C, which remained 36.3 % of its initial activity. The prepared CLPA showed the highest activity recovery (85.0 %), under the optimised conditions (0.3 % (v/v) starch and glutaraldehyde mixture (St/Ga), 1.5: 1 of St/Ga, 25 °C, 1.5 h). Furthermore, pectin-degrading enzymes from various sources were used to produce different CLPA. The alteration of pectinase secondary structure gave high stability in acidic condition (pH 4), thermostability, deactivation energy and half-life, and improved storage stability at 4 °C for 30 days. Similarly to their free counterpart, the CLPA exhibited comparable enzymatic reaction kinetics and could be reused eight times with approximately 20 % of its initial activity. The developed CLPA does not only efficaciously produced POS from pectin as their free form, but also exhibited better operational stability and reusability, making it more suitable for POS production.
The effects of steam explosion (SE) pretreatment on the structural properties of lignin isolated from coconut husk (CH) biomass via soda pulping were investigated in this work. The isolated SE lignin was classified as dilute acid impregnation SE lignin (ASEL), water impregnation SE lignin (WSEL), and 2-naphthol impregnation SE lignin (NSEL). The various types of functional groups isolated from SE lignin were characterized and compared using a variety of complementary analyses: FTIR spectroscopy, NMR spectroscopy, GPC chromatography, HPAEC-PAD chromatography and thermal analyses. It was revealed that ASEL has the highest solid recovery with 55.89 % yield as well as the highest sugars content compared to WSEL (45.66 % yield) and NSEL (49.37 % yield). Besides, all isolated SE lignin contain a significant quantity of non-condensed G-type and S-type units but less amount of H-type units as supported by previous research. The SE lignin produced lignin with higher molecular weight (Mw ASEL: 72725 g mol-1 > Mw WSEL: 13112 g mol-1 > Mw NSEL: 6891 g mol-1) seems to influence the success of the synthesis reaction of phenolic resins. Because of the large variances in the physicochemical properties of SE lignin polymers, their structural properties were increased toward numerous alternative techniques in lignin-based applications.
This study investigated an innovative strategy of incorporating surfactants into alkaline-catalyzed glycerol pretreatment and enzymatic hydrolysis to improve lignocellulosic biomass (LCB) conversion efficiency. Results revealed that adding 40 mg/g PEG 4000 to the pretreatment at 195 °C obtained the highest glucose yield (84.6%). This yield was comparable to that achieved without surfactants at a higher temperature (240 °C), indicating a reduction of 18.8% in the required heat input. Subsequently, Triton X-100 addition during enzymatic hydrolysis of PEG 4000-assisted pretreated substrate increased glucose yields to 92.1% at 6 FPU/g enzyme loading. High-solid fed-batch semi-simultaneous saccharification and co-fermentation using this dual surfactant strategy gave 56.4 g/L ethanol and a positive net energy gain of 1.4 MJ/kg. Significantly, dual assistance with surfactants rendered 56.3% enzyme cost savings compared to controls without surfactants. Therefore, the proposed surfactant dual-assisted promising approach opens the gateway to economically viable enzyme-mediated LCB biorefinery.
To express and determine the hydrolytic activity of a cellobiohydrolase (TTCBH6B) from the thermophilic fungus Thielavia terrestris in Pichia pastoris.
Pretreatment with pure, mixed, and diluted deep eutectic solvents (DESs) was evaluated for its effect on Napier grass through compositional and characterization studies. The morphological changes of biomass caused by pretreatment were analyzed by FTIR and XRD. The cellulose and hemicellulose content after pretreatment using mixed DES increased and decreased 1.29- and 4.25-fold, respectively, when compared to untreated Napier grass. The crystallinity index (CrI. %) of mixed DES sample increased due to the maximum removal of hemicellulose (76 %) and delignification of 62 %. The material costs of ChCl/FA and ChCl/LA for a single run are ≈2.16 USD and ≈1.65 USD, respectively. Pure DES showed that ChCl/LA pretreatment enhanced delignification efficiency and that ChCl/FA increased hemicellulose removal. It was estimated that a single run using ChCl/LA:ChCl/FA to achieve maximum hemicellulose and lignin removal would cost approximately ≈1.89 USD. Future work will evaluate the effect of DES mixture on enzyme digestibility and ethanol production from Napier grass. HYPOTHESES: Deep eutectic solvent (DES) pretreatment studies on the fractionation of lignocellulosic biomass have grown exponentially. The use of pure and diluted DES has been reported to improve saccharification efficiency, delignification, and cellulose retention (Gundupalli et al., 2022). These studies have reported maximum lignin removal but also a lower effect on hemicellulose removal from lignocellulosic biomass. It was hypothesized that mixing two pure DESs could result in maximum removal of hemicellulose and lignin after pretreatment. To our knowledge, no studies have been performed to investigate the efficiency of pretreatment using a DES mixture and compared the outcome with pure and diluted DESs. Furthermore, it was hypothesized that using two pure DESs in a mixed form could lower the material cost for each experimental run. Process efficiency was determined by compositional, XRD, and FTIR analysis. Avenues for future research include determining glucose and ethanol yields during the enzymatic saccharification and fermentation processes.
A new cutting-edge lignocellulose fractionation technology for the co-production of glucose, native-like lignin, and furfural was introduced using mannitol (MT)-assisted p-toluenesulfonic acid/pentanol pretreatment, as an eco-friendly process. The addition of optimized 5% MT in pretreatment enhanced the delignification rate by 29% and enlarged the surface area and biomass porosity by 1.07-1.80 folds. This increased the glucose yield by 45% (from 65.34 to 94.54%) after enzymatic hydrolysis relative to those without MT. The extracted lignin in the organic phase of pretreatment exhibited β-O-4 bonds (61.54/100 Ar) properties of native cellulosic enzyme lignin. Lignin characterization and molecular docking analyses revealed that the hydroxyl tails of MT were incorporated with lignin and formed etherified lignin, which preserved high lignin integrity. The solubilized hemicellulose (96%) in the liquid phase of pretreatment was converted into furfural with a yield of 83.99%. The MT-assisted pretreatment could contribute to a waste-free biorefinery pathway toward a circular bioeconomy.
In recent years, with increasing emphasis on healthy, green, and sustainable consumption concepts, plant-based foods have gained popularity among consumers. As widely sourced plant-based raw materials, legume proteins are considered sustainable and renewable alternatives to animal proteins. However, legume proteins have limited functional properties, which hinder their application in food products. LAB fermentation is a relatively natural processing method that is safer than chemical/physical modification methods and can enrich the functional properties of legume proteins through biodegradation and modification. Therefore, changes in legume protein composition, structure, and functional properties and their related mechanisms during LAB fermentation are described. In addition, the specific enzymatic hydrolysis mechanisms of different LAB proteolytic systems on legume proteins are also focused in this review. The unique proteolytic systems of different LAB induce specific enzymatic hydrolysis of legume proteins, resulting in the production of hydrolysates with diverse functional properties, including solubility, emulsibility, gelability, and foamability, which are determined by the composition (peptide/amino acid) and structure (secondary/tertiary) of legume proteins after LAB fermentation. The correlation between LAB-specific enzymatic hydrolysis, protein composition and structure, and protein functional properties will assist in selecting legume protein raw materials and LAB strains for legume plant-based food products and expand the application of legume proteins in the food industry.
Palm pressed fibre (PPF) is a lignocellulose biomass generated from palm oil mill that is rich in cellulose. The present work aimed to combine acid hydrolysis followed by high-pressure homogenisation (HPH) to produce nanocrystal cellulose (CNC) with enhanced physicochemical properties from PPF. PPF was alkaline treated, bleached, acid hydrolysed and homogenised under high pressure condition to prepare CNC. The effects of homogenisation pressure (10, 30, 50, 70 MPa) and cycles (1, 3, 5, 7) on the particle size, zeta potential and rheological properties of CNC produced were investigated. HPH was capable of producing CNC with better stability. Results revealed that utilizing 1 cycle of homogenisation at a pressure of 50 MPa resulted in CNC with the smallest dimension, highest aspect ratio, moderate viscosity and exceptionally high zeta potential. Subsequently, 0.15% (CNC 0.15 -PE) and 0.30% (CNC 0.30 -PE) of CNC was used to stabilise oil-in-water emulsions and their stability was evaluated against different pH, temperature and ionic strength. All the CNC-stabilised emulsions demonstrated good thermal stability. CNC 0.30 -PE exhibited larger droplets but higher stability than CNC 0.15 -PE. In short, CNC with gel like structure has a promising potential to serve as a natural Pickering emulsifier to stabilise oil-in-water emulsion in various food applications.
Herein, choline chloride/oxalic acid (ChCl/OA) and choline chloride/oxalic acid/ethylene glycol (ChCl/OA/EG) pretreatments of oil palm empty fruit bunches (EFB) and mesocarp fibers (MSF) were conducted to achieve protection of the lignin structure, while improving the enzymatic efficiency of the solid residues. Under the operating conditions of 90 °C and 6 h, ChCl/OA/EG demonstrated a higher lignin extraction selectivity and obtained solid residues with higher hemicellulose content compared to ChCl/OA. The digestibility of glucan and xylan in solid residues obtained using ChCl/OA/EG achieved 98.56 % and 95.63 %, respectively, for EFB and 75.95 % and 88.60 %, for MSF. Uncondensed lignin enriched with 71.79-81.61 % of β-O-4 bonds was obtained from EFB and MSF using ChCl/OA/EG. 2D HSQC NMR and the density functional theory calculation confirmed that substituting the lignin Cα position by ethylene glycol changed the local potentials of the β-O-4 bonds, impeding the attack of protons (H+). The higher β-O-4 linkage content in ChCl/OA/EG-Ls led to the formation of several oxygenated alkyl methoxy phenols and alkyl methoxy phenols were promoted during the pyrolysis. Moreover, molecular dynamics simulations showed that the main factor affecting lignin extraction and dissolution in this study was the diffusion coefficient of lignin in DESs.
Microalgae are a sustainable source of bioactive proteins for personal care and cosmetic applications. Conventional alkaline extraction can achieve high protein recovery but relies on harsh chemical conditions that often compromise protein integrity, reduce bioactivity, and raise environmental concerns. Moreover, the extracted proteins often require additional enzymatic hydrolysis to enhance their functionality, thus increasing process complexity. This study proposes a dual-function Savinase-assisted in situ extraction strategy for the simultaneous extraction and functionalization of protein from Nannochloropsis oculata (N. oculata). Process optimization using improvement in protein extraction yield (IPEY) as the performance indicator identified optimal conditions of pH 7, 50 °C, 0.5 wt% Savinase and 2 h hydrolysis, under which a maximum IPEY of 85.1% was achieved. Fluorescence spectroscopy and zeta potential measurements confirmed that Savinase remained structurally stable under the optimized conditions while promoting protein solubilization. The resulting protein hydrolysates exhibited enhanced functional properties, with ABTS•+ radical scavenging activity reaching 83.3% at 5 mg/mL, representing a 56.9% increase compared with the non-enzymatic control. In addition, UV absorption analysis revealed a critical wavelength (λc) of 383.6 nm, indicating broad-spectrum photoprotective potential. Overall, the proposed one-step Savinase-assisted process provides a mild, sustainable, and effective strategy for producing bioactive microalgal protein hydrolysates with strong potential as natural ingredients in skin-protective cosmetic formulations.
This study aims to produce biochar and sugars from a macroalga Eucheuma denticulatum using dilute sulfuric acid hydrolysis along with microwave-assisted heating. The reactions were operated at sulfuric acid concentrations of 0.1 and 0.2M, reaction temperatures of 150-170°C and a heating time of 10min. Compared to the raw macroalga, biochar qualities were improved with increased carbon content and lower ash and moisture contents. The calorific value of the biochar could be intensified up to 45%, and 39% of energy yield was recovered. Apart from producing biochar, the highest total reducing sugars were 51.47g/L (74.84% yield) along with a low by-product 5-HMF of 0.20g/L, when the biomass was treated under the optimum conditions at 160°C with 0.1M H2SO4. Thus, this study demonstrated that macroalgae could be potentially used as biomass feedstock under microwave-assisted acid hydrolysis for the production of biofuel and value-added products.
For the first time, a highly efficient Cr(NO3)3 catalysis system was proposed for optimization the yield and crystallinity of nanocellulose end product. A five-level three-factor central composite design coupled with response surface methodology was employed to elucidate parameters interactions between three design factors, namely reaction temperature (x1), reaction time (x2) and concentration of Cr(NO3)3 (x3) over a broad range of process conditions and determine the effect on crystallinity index and product yield. The developed models predicted the maximum nanocellulose yield of 87% at optimum process conditions of 70.6°C, 1.48h, and 0.48M Cr(NO3)3. At these conditions, the obtained nanocellulose presented high crystallinity index (75.3%), spider-web-like interconnected network morphology with the average width of 31.2±14.3nm. In addition, the yielded nanocellulose rendered a higher thermal stability than that of original cellulosic source and expected to be widely used as reinforcement agent in bio-nanocomposites materials.
This study involves extraction of sulfated polysaccahride (SP) from brown seaweed (Turbinaria turbinata). Eight processing conditions affecting enzyme aided extraction (EAE) were screened using Plackett-Burman design. Three significant factors (hydrolysis time, enzyme concentration and extraction stage) were optimized using Faced Centred Central Composite Design in Random Surface Methods. Micrograph obtained using Field Emission Scanning Electron Microscopy revealed that cellulase degradation ruptured the seaweed cell matrix thus caused increase in the release of SP. The optimum conditions for extraction of SP from T. turbinata are: extraction stage of 2, hydrolysis time of 19.5 h and enzyme concentration of 1.5 μl/ml to produce 25.13% yield. The SP obtained from cellulase treated T. turbinata is a suitable anti-inflammatory agent for pharmaceutical applications.
Among the chemical pretreatments available for pretreating biomass, the inorganic salt is considered to be a relatively new but simple reagent that offers comparable pentose (C5) sugar recoveries as the conventional dilute acid hydrolysis. This study investigated the effects of different concentrations (1.5-6.0% (v/v)) of H2O2 or Na2S2O8 in facilitating CuSO4·5H2O pretreatment for improving pentose sugar recovery from oil palm fronds. The best result was observed when 0.2 mol/L of CuSO4·5H2O was integrated with 4.5% (v/v) of Na2S2O8 to recover 8.2 and 0.9 g/L of monomeric xylose and arabinose, respectively in the liquid fraction. On the other hand, an addition of 1.5% (v/v) of H2O2 yielded approximately 74% lesser total pentose sugars as compared to using 4.5% (v/v) Na2S2O8. By using CuSO4·5H2O alone (control), only 0.8 and 1.0 g/L xylose and arabinose, respectively could be achieved. The results mirrored the importance of using chemical additives together with the inorganic salt pretreatment of oil palm fronds. Thus, an addition of 4.5% (v/v) of Na2S2O8 during CuSO4·5H2O pretreatment of oil palm fronds at 120 °C and 30 min was able to attain a total pentose sugar yield up to ∼40%.
A study was carried out to determine the process parameters and optimization for the hydrolysis of protein precipitate from cockle (Anadara granosa) meat wash water. Precipitation of the protein in the wash water was done using pH manipulation (pH3-8). The precipitate was hydrolyzed using hydrochloric acid (HCl) and optimized for HCl volume, HCl concentration and hydrolysis time using response surface methodology (RSM) based on a central composite rotatable design. Based on the results, hydrolysis of cockle meat wash water precipitate was carried out by precipitation of the wash water at pH4. Optimum condition for the hydrolysis of 2.0 g of cockle meat wash water precipitate was 25 mL of 1 N HCl for 10 h which resulted in nitrogen content (NC) of 0.7% and degree of hydrolysis (DH) of 55%. NC and DH were significantly influenced only by the hydrolysis time.
Cellulose nanomaterial with rod-like structure and highly crystalline order, usually formed by elimination of the amorphous region from cellulose during acid hydrolysis. Cellulose nanomaterial with the property of biocompatibility and nontoxicity can be used for enzyme immobilization. In this work, urease enzyme was used as a model enzyme to study the surface modification of cellulose nanomaterial and its potential for biosensor application. The cellulose nanocrystal (CNC) surface was modified using 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-mediated oxidation to introduce the carboxyl group at C6 primary alcohol. The success of enzyme immobilization and surface modification was confirmed using chemical tests and measured using UV-Visible spectrophotometer. The immobilization strategy was then applied for biosensor application for urea detection. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) techniques were used for electroanalytical characterization of the urea biosensor.
: Hydrogen (H2) is a clean energy carrier which can help to solve environmental issues with the depletion of fossil fuels. Sodium borohydride (NaBH4) is a promising candidate material for solid state hydrogen storage due to its huge hydrogen storage capacity and nontoxicity. However, the hydrolysis of NaBH4 usually requires expensive noble metal catalysts for a high H2 generation rate (HGR). Here, we synthesized high-aspect ratio copper nanowires (CuNWs) using a hydrothermal method and used them as the catalyst for the hydrolysis of NaBH4 to produce H2. The catalytic H2 generation demonstrated that 0.1 ng of CuNWs could achieve the highest volume of H2 gas in 240 min. The as-prepared CuNWs exhibited remarkable catalytic performance: the HGR of this study (2.7 × 1010 mL min-1 g-1) is ~3.27 × 107 times higher than a previous study on a Cu-based catalyst. Furthermore, a low activation energy (Ea) of 42.48 kJ mol-1 was calculated. Next, the retreated CuNWs showed an outstanding and stable performance for five consecutive cycles. Moreover, consistent catalytic activity was observed when the same CuNWs strip was used for four consecutive weeks. Based on the results obtained, we have shown that CuNWs can be a plausible candidate for the replacement of a costly catalyst for H2 generation.
An innovative approach was developed by incorporating high-pressure CO2 into the separate hydrolysis-fermentation of aspen leftover branches, aiming to enhance the bioethanol production efficiency. The high-pressure CO2 significantly increased the 72-h enzymatic hydrolysis yield of converting aspen into glucose from 53.8% to 82.9%. The hydrolysis process was performed with low enzyme loading (10 FPU g-1 glucan) with the aim of reducing the cost of fuel bioethanol production. The ethanol yield from fermentation of the hydrolyzed glucose using yeast (Saccharomyces cerevisiae) was 8.7 g L-1, showing increment of 10% compared with the glucose control. Techno-economic analysis indicated that the energy consumption of fuel bioethanol production from aspen branch chips was reduced by 35% and the production cost was cut 44% to 0.615 USD L-1, when 68 atm CO2 was introduced into the process. These results furtherly emphasized the low carbon footprint of this sustainable energy production approach.