Simultaneous binding of bFGF to both FGFR and integrin maintains properties of primed human induced pluripotent stem cells.

Introduction: Basic fibroblast growth factor (bFGF, FGF2) and integrin α6ß1 are important for maintaining the pluripotency of human pluripotent stem cells (hPSCs). Although bFGF-integrin binding contributes to biofunctions in cancer cells, the relationship in hPSCs remains unclear. Methods: To investigate the relationship between bFGF and integrin in human induced pluripotent stem cells (hiPSCs), we generated recombinant human bFGF wild-type and mutant proteins, that do not bind to integrin, FGFR, or both. We then cultured hiPSCs with these recombinant bFGF proteins. To evaluate the abilities of recombinant bFGF proteins in maintaining hPSC properties, pluripotent markers, ERK activity, and focal adhesion structure were analyzed through flow cytometry, immunofluorescence (IF), and immunoblotting (IB). Result: We identified an interaction between bFGF and integrin α6ß1 in vitro and in hiPSCs. The integrin non-binding mutant was incapable of inducing the hPSC properties, such as proliferation, ERK activity, and large focal adhesions at the edges of hiPSC colonies. Signaling induced by bFGF-FGFR binding was essential during the first 24 h after cell seeding for maintaining the properties of hPSCs, followed by a shift towards intracellular signaling via the bFGF-integrin interaction. The mixture of the two bFGF mutants also failed to maintain hPSC properties, indicating that bFGF binds to both FGFR and integrin. Conclusion: Our study demonstrates that the integrin-bFGF-FGFR ternary complex maintains the properties of hPSCs via intracellular signaling, providing insights into the functional crosstalk between bFGF and integrins in hiPSCs.

A comprehensive review on nanocatalysts and nanobiocatalysts for biodiesel production in Indonesia, Malaysia, Brazil and USA.

Biodiesel is an alternative to fossil-derived diesel with similar properties and several environmental benefits. Biodiesel production using conventional catalysts such as homogeneous, heterogeneous, or enzymatic catalysts faces a problem regarding catalysts deactivation after repeated reaction cycles. Heterogeneous nanocatalysts and nanobiocatalysts (enzymes) have shown better advantages due to higher activity, recyclability, larger surface area, and improved active sites. Despite a large number of studies on this subject, there are still challenges regarding its stability, recyclability, and scale-up processes for biodiesel production. Therefore, the purpose of this study is to review current modifications and role of nanocatalysts and nanobiocatalysts and also to observe effect of various parameters on biodiesel production. Nanocatalysts and nanobiocatalysts demonstrate long-term stability due to strong Brønsted-Lewis acidity, larger active spots and better accessibility leading to enhancethe biodiesel production. Incorporation of metal supporting positively contributes to shorten the reaction time and enhance the longer reusability. Furthermore, proper operating parameters play a vital role to optimize the biodiesel productivity in the commercial scale process due to higher conversion, yield and selectivity with the lower process cost. This article also analyses the relationship between different types of feedstocks towards the quality and quantity of biodiesel production. Crude palm oil is convinced as the most prospective and promising feedstock due to massive production, low cost, and easily available. It also evaluates key factors and technologies for biodiesel production in Indonesia, Malaysia, Brazil, and the USA as the biggest biodiesel production supply.

Effects of biodegradation, biotoxicity and microbial community on biostimulation of sulfolane.

To evaluate the effectiveness of biostimulation in remediating soil-free groundwater and groundwater with soil, experiments were conducted using soil and groundwater samples that were contaminated with sulfolane. The main objective was to characterize the differences in sulfolane removal efficiency and biotoxicity between in situ soil-free groundwater and groundwater with soil and different concentrations of dissolved oxygen (1 mg/L and 5 mg/L) and various nutrient salts (in situ and spiked). Optimizing the nutrient salt conditions improved the removal efficiency of sulfolane by 1.8-6.5 that under in situ nutrient salt conditions. Controlling the dissolved oxygen concentration enhanced the efficiency of removal of sulfolane by 1.5-4.5 times over that at the simulated in situ dissolved oxygen concentration, suggesting that the degradation of sulfolane by indigenous microorganisms requires nutrient salts more than it requires dissolved oxygen. Biotoxicity data showed that the luminescence inhibition of Aliivibrio fischeri by sulfolane was lower in the biostimulated samples than in the pre-treated samples. Biostimulation reduced the biotoxicity of the treated samples by 42-51%, revealing that it was effective in removing sulfolane and reducing biotoxicity. Microbial community analysis showed that the biostimulation did not change the dominant species in the original in situ community, and increased the proportion of sulfolane-degraders. The outcome of this study can be used to set parameters for the remediation of groundwater that is contaminated by sulfolane in oil refineries.

Insect biorefinery: A circular economy concept for biowaste conversion to value-added products.

With rapid growing world population and increasing demand for natural resources, the production of sufficient food, feed for protein and fat sources and sustainable energy presents a food insecurity challenge globally. Insect biorefinery is a concept of using insect as a tool to convert biomass waste into energy and other beneficial products with concomitant remediation of the organic components. The exploitation of insects and its bioproducts have becoming more popular in recent years. This review article presents a summary of the current trend of insect-based industry and the potential organic wastes for insect bioconversion and biorefinery. Numerous biotechnological products obtained from insect biorefinery such as biofertilizer, animal feeds, edible foods, biopolymer, bioenzymes and biodiesel are discussed in the subsequent sections. Insect biorefinery serves as a promising sustainable approach for waste management while producing valuable bioproducts feasible to achieve circular bioeconomy.

Progress on microalgae cultivation in wastewater for bioremediation and circular bioeconomy.

Water usage increased alongside its competitiveness due to its finite amount. Yet, many industries still rely on this finite resource thus recalling the need to recirculate their water for production. Circular bioeconomy is presently the new approach emphasizing on the 'end-of-life' concept with reusing, recycling, and recovering materials. Microalgae are the ideal source contributing to circular bioeconomy as it exhibits fast growth and adaptability supported by biological rigidity which in turn consumes nutrients, making it an ideal and capable bioremediating agent, therefore allowing water re-use as well as its biomass potential in biorefineries. Nevertheless, there are challenges that still need to be addressed with consideration of recent advances in cultivating microalgae in wastewater. This review aimed to investigate the potential of microalgae biomass cultivated in wastewater. More importantly, how it'll play a role in the circular bioeconomy. This includes an in-depth look at the production of goods coming from wastes tattered by emerging pollutants. These emerging pollutants include microplastics, antibiotics, ever-increasingly sewage water, and heavy metals which have not been comprehensively compared and explored. Therefore, this review is aiming to bring new insights to researchers and industrial stakeholders with interest in green alternatives to eventually contribute towards environmental sustainability.

Safety Assessment of 3S, 3'S Astaxanthin Derived from Metabolically Engineered K. marxianus.

Previous reviews have already explored the safety and bioavailability of astaxanthin, as well as its beneficial effects on human body. The great commercial potential in a variety of industries, such as the pharmaceutical and health supplement industries, has led to a skyrocketing demand for natural astaxanthin. In this study, we have successfully optimized the astaxanthin yield up to 12.8 mg/g DCW in a probiotic yeast and purity to 97%. We also verified that it is the desired free-form 3S, 3'S configurational stereoisomer by NMR and FITR that can significantly increase the bioavailability of astaxanthin. In addition, we have proven that our extracted astaxanthin crystals have higher antioxidant capabilities compared with natural esterified astaxanthin from H. pluvialis. We also screened for potential adverse effects of the pure astaxanthin crystals extracted from the engineered probiotic yeast by dosing SD rats with 6, 12, and 24 mg/kg/day of astaxanthin crystals via oral gavages for a 13-week period and have found no significant biological differences between the control and treatment groups in rats of both genders, further confirming the safety of astaxanthin crystals. This study demonstrates that developing metabolically engineered microorganisms provides a safe and feasible approach for the bio-based production of many beneficial compounds, including astaxanthin.

Improvement of Enzymatic Saccharification and Ethanol Production from Rice Straw Using Recycled Ionic Liquid: The Effect of Anti-Solvent Mixture.

One of the major concerns for utilizing ionic liquid on an industrial scale is the cost involved in the production. Despite its proven pretreatment efficiency, expenses involved in its usage hinder its utilization. A better way to tackle this limitation could be overcome by studying the recyclability of ionic liquid. The current study has applied the Box-Behnken design (BBD) to optimize the pretreatment condition of rice straw through the usage of 1-ethyl-3-methylimidazolium acetate (EMIM-Ac) as an ionic liquid. The model predicted the operation condition with 5% solid loading at 128.4 °C for 71.83 min as an optimum pretreatment condition. Under the optimized pretreatment condition, the necessity of the best anti-solvent was evaluated among water, acetone methanol, and their combinations. The study revealed that pure methanol is the suitable choice of anti-solvent, enhancing the highest sugar yield. Recyclability of EMIM-Ac coupled with anti-solvent was conducted up to five recycles following the predicted pretreatment condition. Fermentation studies evaluated the efficacy of recycled EMIM-Ac for ethanol production with 89% more ethanol production than the untreated rice straw even after five recycles. This study demonstrates the potential of recycled ionic liquid in ethanol production, thereby reducing the production cost at the industrial level.

Adipose-Derived Stem Cell-Incubated HA-Rich Sponge Matrix Implant Modulates Oxidative Stress to Enhance VEGF and TGF-ß Secretions for Extracellular Matrix Reconstruction In Vivo.

This study demonstrated both adipose-derived stem cells (ASCs) in vitro and in vivo combined with three-dimensional (3D) porous sponge matrices on implant wound healing. Sponge matrices were created from hyaluronic acid (HA), collagen (Col), and gelatin (Gel), constructing two types: HA-L (low content) and HA-H (high content), to be cross-linked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC). Fourier transform infrared spectroscopy method verified carboxyl groups of HA and amino groups of Col and Gel reacting between the raw materials and scaffolds to identify the successive cross-linking. The swelling ratios of two types of sponge matrices were analyzed by water absorption capabilities, and the results displayed both over 30-fold dry scaffold weight enhancements. In biodegradation tests, matrices were hydrolyzed over time by three cutaneous enzymes, hyaluronidase, lysozyme, and collagenase I. ASCs from rats were cultured within the HA-H scaffold, demonstrating higher antioxidative abilities and secretions on related genes and proteins compared to the other two groups. The ASC HA-H matrix promoted cell proliferation to stimulate capillary angiogenesis inducer secretions, including vascular endothelial growth factor (VEGF) and transforming growth factor-ß (TGF-ß). In vivo histological examinations showed ASCs from implanted HA-H implant transported into the subcutis, and rat skin cells also infiltrated into the original matrix zone to increase the extracellular matrix (ECM) reconstructions. Our experimental data revealed that the ASC HA-H sponge implant was effective in improving wound repair.

MYCL promotes iPSC-like colony formation via MYC Box 0 and 2 domains.

Human induced pluripotent stem cells (hiPSCs) can differentiate into cells of the three germ layers and are promising cell sources for regenerative medicine therapies. However, current protocols generate hiPSCs with low efficiency, and the generated iPSCs have variable differentiation capacity among different clones. Our previous study reported that MYC proteins (c-MYC and MYCL) are essential for reprogramming and germline transmission but that MYCL can generate hiPSC colonies more efficiently than c-MYC. The molecular underpinnings for the different reprogramming efficiencies between c-MYC and MYCL, however, are unknown. In this study, we found that MYC Box 0 (MB0) and MB2, two functional domains conserved in the MYC protein family, contribute to the phenotypic differences and promote hiPSC generation in MYCL-induced reprogramming. Proteome analyses suggested that in MYCL-induced reprogramming, cell adhesion-related cytoskeletal proteins are regulated by the MB0 domain, while the MB2 domain regulates RNA processes. These findings provide a molecular explanation for why MYCL has higher reprogramming efficiency than c-MYC.

Interferences of Waxes on Enzymatic Saccharification and Ethanol Production from Lignocellulose Biomass.

Wax is an organic compound found on the surface of lignocellulose biomass to protect plants from physical and biological stresses in nature. With its small mass fraction in biomass, wax has been neglected from inclusion in the design of the biorefinery process. This study investigated the interfering effect of wax in three types of lignocellulosic biomass, including rice straw (RS), Napier grass (NG), and sugarcane bagasse (SB). In this study, although small fractions of wax were extracted from RS, NG, and SB at 0.57%, 0.61%, and 1.69%, respectively, dewaxing causes changes in the plant compositions and their functional groups and promotes dissociations of lignocellulose fibrils. Additionally, dewaxing of biomass samples increased reducing sugar by 1.17-, 1.04-, and 1.35-fold in RS, NG, and SB, respectively. The ethanol yield increased by 1.11-, 1.05-, and 1.23-fold after wax removal from RS, NG, and SB, respectively. The chemical composition profiles of the waxes obtained from RS, NG, and SB showed FAME, alcohol, and alkane as the major groups. According to the conversion rate of the dewaxing process and ethanol fermentation, the wax outputs of RS, NG, and SB are 5.64, 17.00, and 6.00 kg/ton, respectively. The current gasoline price is around USD 0.903 per liter, making ethanol more expensive than gasoline. Therefore, in order to reduce the cost of ethanol in the biorefinery industry, other valuable products (such as wax) should be considered for commercialization. The cost of natural wax ranges from USD 2 to 22 per kilogram, depending on the source of the extracted wax. The wax yields obtained from RS, SB, and NG have the potential to increase profits in the biorefining process and could provide an opportunity for application in a wider range of downstream industries than just biofuels.

Microencapsulation of Curcumin in Crosslinked Jelly Fig Pectin Using Vacuum Spray Drying Technique for Effective Drug Delivery.

Microencapsulation of curcumin in jelly fig pectin was performed by the vacuum spray drying (VSD) technique. The VSD was advanced with a low inlet temperature of 80-90 °C and low pressure of 0.01 mPa. By the in situ cross-linking with multivalent calcium ions, jelly fig pectin produced stable curcumin encapsulated microparticles. The physiochemical characteristics of microparticles were thoroughly investigated. The results revealed that 0.75 w/w% of jelly fig pectin and inlet temperature of 90 °C could be feasible for obtaining curcumin microparticles. The VSD technique showed the best encapsulation efficiency and yield and loading efficiency was up to 91.56 ± 0.80%, 70.02 ± 1.96%, and 5.45 ± 0.14%, respectively. The curcumin was readily released into simulated gastrointestinal fluid with 95.34 ± 0.78% cumulative release in 24 h. The antioxidant activity was stable after being stored for six months and stored as a solution for seven days at room temperature before analysis. Hence, the VSD technique could be applicable for the microencapsulation of bioactive compounds such as curcumin to protect and use in the food/pharmaceutical industry.

Effect of dewaxing on saccharification and ethanol production from different lignocellulosic biomass.

Dewaxing effects on the pretreatment, saccharification and fermentation are rarely reported due to the low abundance of wax in lignocellulose. This study aimed to investigate the effect of wax removal on saccharification and ethanol yield from lignocellulose by using Rice straw (RS), Napier grass (NG), and sugarcane bagasse (SB). The wax contents of 0.56%, 1.7%, and 0.6% were obtained from RS, NG and SB after the wax extraction, respectively. The alkaline pretreatment was applied in combination with dewaxing to decipher the synergistic effect of these treatments. Dewaxing and alkaline pretreatment of lignocellulosic biomass showed changes in the plant compositions. Removal of wax from RS, NG and SB showed significant changes in the surface morphology and functional groups. A higher yield of sugars and ethanol was observed in dewaxed and alkaline pretreated samples. The ethanol yields of 75.4%, 89.85%, and 74% from RS, NG, and SB were obtained after fermentation, respectively.

Differential effects of inorganic salts on cellulase kinetics in enzymatic saccharification of cellulose and lignocellulosic biomass.

Inorganic salt pretreatment of lignocellulosic biomass has proven to be an efficient way to increase the efficiency of enzymatic saccharification. However, it is not clear that this improvement is the result of modification of the lignocellulosic substrate after pretreatment, or removal of inhibitor, or enhancement of cellulase or a combination of these events. Therefore, this study aimed to analyze the effects of inorganic salts on kinetics of cellulase enzymes (celluclast 1.5L and accellerase 1500). Two substrates rich in cellulose content [carboxymethylcellulose (CMC), avicel (AV)] and lignocellulose substrate [sugarcane bagasse (SB)] were considered. The enzymatic saccharification was carried with and without the addition of inorganic salts (NaCl and KCl) at 0.5 M and 1.0 M concentration. The kinetic parameters, Km and Vm, were determined to mechanically understand the pattern of inhibition and enhancement of inorganic salts on enzymatic saccharification. The kinetics parameters of celluclast 1.5L and accellerase 1500 for hydrolysis of CMC and AV with NaCl showed uncompetitive inhibition. Whereas, influences of KCl on both cellulase were differentiated to function in inhibition or enhancement modes when challenged with different substrates. On the other hand, enzymatic hydrolysis efficiencies of SB using both cellulases were enhanced under addition of NaCl and KCl, by increasing Vm of celluclast 1.5L from 0.303 to 0.635 mg/mL min (0.5 M KCl) and accellerase 1500 from 0.383 to 0.719 mg/mL min (1.0 M NaCl). The details of kinetic analysis in this work revealed the mechanism of inorganic salts on cellulase kinetics to be involved in substrate modification and removal of inhibitor.

Encapsulation and Characterization of Nanoemulsions Based on an Anti-oxidative Polymeric Amphiphile for Topical Apigenin Delivery.

Apigenin (Apig) is used as a model drug due to its many beneficial bio-activities and therapeutic potentials. Nevertheless, its poor water solubility and low storage stability have limited its application feasibility on the pharmaceutical field. To address this issue, this study developed nanoemulsions (NEs) using an anti-oxidative polymeric amphiphile, d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS), hydrogenated soy lecithin (HL), black soldier fly larvae (BSFL) oil, and avocado (AV) oil through pre-homogenization and ultrasonication method. Addition of TPGS (weight ratios 100 and 50% as compared to HL) into NEs effectively reduced particle size and phase transition region area of NEs with pure HL. Incorporation of Apig into NEs made particle size increase and provided a disorder effect on intraparticle molecular packing. Nevertheless, the encapsulation efficiency of NEs for Apig approached to about 99%. The chemical stability of Apig was significantly improved and its antioxidant ability was elevated by incorporation with BSFL oil and AV oil NEs, especially for NEs with single TPGS. NEs with single TPGS also exhibited the best Apig skin deposition. For future application of topical Apig delivery, NEs-gel was formed by the addition of hyaluronic acid (HA) into NEs. Their rheological characteristics were dominated by the surfactant ratios of HL to TPGS.

Fast Dissolving Electrospun Nanofibers Fabricated from Jelly Fig Polysaccharide/Pullulan for Drug Delivery Applications.

The fast-dissolving drug delivery systems (FDDDSs) are developed as nanofibers using food-grade water-soluble hydrophilic biopolymers that can disintegrate fast in the oral cavity and deliver drugs. Jelly fig polysaccharide (JFP) and pullulan were blended to prepare fast-dissolving nanofiber by electrospinning. The continuous and uniform nanofibers were produced from the solution of 1% (w/w) JFP, 12% (w/w) pullulan, and 1 wt% Triton X-305. The SEM images confirmed that the prepared nanofibers exhibited uniform morphology with an average diameter of 144 ± 19 nm. The inclusion of JFP in pullulan was confirmed by TGA and FTIR studies. XRD analysis revealed that the increased crystallinity of JFP/pullulan nanofiber was observed due to the formation of intermolecular hydrogen bonds. The tensile strength and water vapor permeability of the JFP/pullulan nanofiber membrane were also enhanced considerably compared to pullulan nanofiber. The JFP/pullulan nanofibers loaded with hydrophobic model drugs like ampicillin and dexamethasone were rapidly dissolved in water within 60 s and release the encapsulants dispersive into the surrounding. The antibacterial activity, fast disintegration properties of the JFP/pullulan nanofiber were also confirmed by the zone of inhibition and UV spectrum studies. Hence, JFP/pullulan nanofibers could be a promising carrier to encapsulate hydrophobic drugs for fast-dissolving/disintegrating delivery applications.

Core-Shell Encapsulation of Lipophilic Substance in Jelly Fig (Ficus awkeotsang Makino) Polysaccharides Using an Inexpensive Acrylic-Based Millifluidic Device.

The polysaccharides extracted from the achenes of jelly fig, Ficus awkeotsang Makino, were mainly composed of low methyl pectin and used as a novel shell material for encapsulating lipophilic bioactives in the core of microcapsule. The polysaccharide microcapsules with oil core were prepared using a novel acrylic-based millifluidic device developed in this study. To investigate the physiochemical properties of and find the suitable formula of polysaccharide shells, the films casted with jelly fig polysaccharide were thoroughly characterized. For the preparation of microcapsules, the millifluidic device was optimized by controlling the flow rate to obtain uniform spherical shape with a core diameter of 1.4-1.9 mm and the outer diameter of 2.1-2.8 mm. The encapsulation efficiency was around 90%, and the microcapsules displayed a clear boundary between the polysaccharide shell and oil core. Encapsulation of curcumin in the microcapsules was prepared to test the applicability of the device and processes developed in this study, and the results showed that the microencapsulation could enhance the stability of curcumin against external environment. Overall, the results suggested that the jelly fig polysaccharides and the developed millifluidic device can be useful for the preparation of core-shell microcapsules for encapsulation of lipophilic bioactives.

Development and Characterization of Nano-emulsions Based on Oil Extracted from Black Soldier Fly Larvae.

Insect-based biorefinery is seen as a potential alternative approach to manufacturing foods, feeds, and fuel because of the increasing demand for renewable and sustainable products. Insect oil and protein are the two major components that can be quantitatively obtained from insect farming. However, very few attempts have been conducted to utilize insect oil for the production of value-added products. In this study, the oil extracted from the black soldier fly (Hermetia illucens) larvae (BSFL) was used as a novel feedstock for preparing nano-emulsions. The nano-emulsions were prepared with BSFL oil, hydrogenated lecithin (HL), and d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) in water using pre-homogenization followed by ultrasonication. The morphology and the particle size of nano-emulsions were affected by ratios of HL to TPGS. Moreover, the nano-emulsions showed a nearly Newtonian liquid behavior and the presence of TPGS was able to improve the storage stability of HL nano-emulsions. The addition of TPGS could eliminate the phase transition region of HL nano-emulsions but did not provide a significant change for the molecular mobility in the HL nano-emulsions. In summary, the BSFL oil could be used as a renewable feedstock for formulating nano-emulsions from the aspect of high value-added applications and physicochemical characteristics of the nano-emulsions could be adjusted by the mixed surfactant ratio, surfactant to oil ratio, and oil content. Graphical Abstract The physicochemical characteristics and optimization of nano-emulsions based on black soldier fly larvae oil were investigated.

Evaluation of Macaranga tanarius as a biomass feedstock for fermentable sugars production.

Macaranga tanarius is a fast-growing tree species that could be potentially utilized as a biomass feedstock for biorefinery. The average productivity of M. tanarius biomass was estimated to be ~19.2 ton/ha if the above-ground biomass is harvested bi-annually. Different pretreatment approaches were investigated to increase the enzymatic digestibility of foliage and woody biomass. The results indicated that no pretreatment was required for the foliage biomass while sequential acid/alkali pretreatment was necessary for the woody biomass before enzymatic hydrolysis. For the woody biomass, the delignification was 34.5% after sequential dilute acid/alkali pretreatment. The reducing sugar yields from enzymatic hydrolysis of foliage and pretreated woody biomass were 0.31 and 0.42 g/g dry biomass, respectively. The results also showed that both hydrolysates were fermentable by lactic acid bacteria. Overall, the results suggested that M. tanarius could be a potential feedstock for biorefinery based on the findings and processes derived from this study.

Integration Process for Protein Extraction from Microalgae Using Liquid Biphasic Electric Flotation (LBEF) System.

Microalgae are the most promising sources of protein, which have high potential due to their high-value protein content. Conventional methods of protein harnessing required multiple steps, and they are generally complex, time consuming, and expensive. Currently, the study of integration methods for microalgae cell disruption and protein recovery process as a single-step process is attracting considerable interest. This study aims to investigate the novel approach of integration method of electrolysis and liquid biphasic flotation for protein extraction from wet biomass of Chlorella sorokiniana CY-1 and obtaining the optimal operating conditions for the protein extraction. The optimized conditions were found at 60% (v/v) of 1-propanol as top phase, 250 g/L of dipotassium hydrogen phosphate as bottom phase, crude microalgae loading of 0.1 g, air flowrate of 150 cc/min, flotation time of 10 min, voltage of 20 V and electrode's tip touching the top phase of LBEF. The protein recovery and separation efficiency after optimization were 23.4106 ± 1.2514% and 173.0870 ± 4.4752%, respectively. Comparison for LBEF with and without the aid of electric supply was also conducted, and it was found that with the aid of electrolysis, the protein recovery and separation efficiency increased compared to the LBEF without electrolysis. This novel approach minimizes the steps for overall protein recovery from microalgae, time consumption, and cost of operation, which is beneficial in bioprocessing industry.

Optimization of High Solids Dilute Acid Hydrolysis of Spent Coffee Ground at Mild Temperature for Enzymatic Saccharification and Microbial Oil Fermentation.

Soluble coffee, being one of the world's most popular consuming drinks, produces a considerable amount of spent coffee ground (SCG) along with its production. The SCG could function as a potential lignocellulosic feedstock for production of bioproducts. The objective of this study is to investigate the possible optimal condition of dilute acid hydrolysis (DAH) at high solids and mild temperature condition to release the reducing sugars from SCG. The optimal condition was found to be 5.3 % (w/w) sulfuric acid concentration and 118 min reaction time. Under the optimal condition, the mean yield of reducing sugars from enzymatic saccharification of defatted SCG acid hydrolysate was 563 mg/g. The SCG hydrolysate was then successfully applied to culture Lipomyces starkeyi for microbial oil fermentation without showing any inhibition. The results suggested that dilute acid hydrolysis followed by enzymatic saccharification has the great potential to convert SCG carbohydrates to reducing sugars. This study is useful for the further developing of biorefinery using SCG as feedstock at a large scale.