While remarkable progress has been made in the development of peptide medicines, many problems related to peptide synthesis remain unresolved. Previously, we reported electrochemical peptide synthesis using a phosphine as a potentially recyclable coupling reagent. However, there was room for improvement from the point of view of reaction efficiency, especially in the carboxylic acid activation step and the peptide bond formation step. To overcome these challenges, we searched for the optimal phosphine. Among phosphines with various electronic properties, we found that electron-rich triaryl phosphines improved the reaction efficiency. Consequently, we successfully performed electrochemical peptide synthesis on sterically hindered and valuable amino acids. We also synthesized oligopeptides that were challenging with our previous method. Finally, we examined the effect of substituents on the phosphine cations, and gained some insights into reactivity, which will aid researchers designing reactions involving phosphine cations.

A facile biphasic system composed of choline chloride (ChCl)-based deep eutectic solvent (DES) and methyl isobutyl ketone (MIBK) was developed to realize the furfural production, lignin separation and preparation of fermentable glucose from Eucalyptus in one-pot. Results showed that the ChCl/1,2-propanediol/MIBK system owned the best property to convert hemicelluloses into furfural. Under the optimal conditions (MRChCl:1,2-propanediol = 1:2, raw materials:DES:MIBK ratio = 1:4:8 g/g/mL, 0.075 mol/L AlCl3·6H2O, 140 °C, and 90 min), the furfural yield and glucose yield reached 65.0 and 92.2 %, respectively. Meanwhile, the lignin with low molecular weight (1250-1930 g/mol), low polydispersity (DM = 1.25-1.53) and high purity (only 0.08-2.59 % carbohydrate content) was regenerated from the biphasic system. With the increase of pretreatment temperature, the β-O-4, β-β and β-5 linkages in the regenerated lignin were gradually broken, and the content of phenolic hydroxyl groups increased, but the content of aliphatic hydroxyl groups decreased. This research provides a new strategy for the comprehensive utilization of lignocellulose in biorefinery process.

New routes for biomass valorization have been developing by the scientific community. The aim of this work was developing a novel OrganoCat-based protocol and deeply understand the structure of the obtained lignins. Microwave-assisted OrganoCat-based process was performed using a biphasic system (ethyl acetate and oxalic acid or HCl) at mild conditions. OrganoCat-based lignins (OCLs) were characterized by compositional analysis, FTIR, 1H, 13C, 1H13C HSQC, 31P NMR, TGA and GPC. The solubility of OCLs in different organic solvents and their antioxidant capacity against DPPH were investigated. The spectroscopic analyses showed that OCLs have high residual extractives and the lignin motifs were preserved. OCLs have presented lower thermal stability than MWL, but showed great antioxidant activities and high solubility in a wide range of organic solvents. A novel biorefinery protocol yielded coconut shell lignins with peculiar structural and compositional features and several technological applications through an eco-friendly, sustainable and relatively low-cost biphasic pulping process.

Developing an environmentally friendly and efficient pretreatment to utilize wheat straw is essential to a sustainable future. An acid biphasic system with 2-methyltetrahydrofuran (2-MeTHF) organic solvent and dilute p-toluenesulfonic acid (p-TsOH) were employed for the simultaneous fractionation of three components. Results showed that the biphasic system had excellent cellulose protection and high removal of hemicellulose and lignin. In detail, Under the optimal conditions (0.1 M p-TsOH, 2-MeTHF: H2O = 1:1 (v:v), 140 °C, 3 h), mostly cellulose retained in the residues (95.69%), 57.18% of lignin was removed and high yield of hemicellulose-based C5 sugars was achieved (77.49%). In the further process of dehydration of pre-hydrolysate dichloromethane (DCM) as an organic phase, the yield of furfural was 80.07% (170 °C-80 min). The saccharification of residue reached 95.82%. p-TsOH/2-MeTHF/H2O pretreatment was desirable for high selectivity fractionation. Important chemicals for bioenergy including furfural, monosaccharides and lignin are obtained.

Biobased furfural is a sustainable alternative to petrochemical intermediates for bulk chemicals and fuel production. However, existing methods for the conversion of xylose or lignocelluloses in mono-/bi-phasic systems to furfural involve non-selective sugar isolation or lignin condensation, limiting the valorisation of lignocelluloses. Herein, we used diformylxylose (DFX), a xylose derivative that is formed during the lignocellulosic fractionation process with formaldehyde protection, as a substitute for xylose to produce furfural in biphasic systems. Under kinetically optimized conditions, over 76 mol% of DFX could be converted to furfural in water-methyl isobutyl ketone system at a high reaction temperature with a short reaction time. Finally, isolation of xylan in eucalyptus wood as DFX with formaldehyde protection followed by converting DFX in a biphasic system gave a final furfural yield of 52 mol% (on the basis of xylan in wood), which was more than two times of that without formaldehyde. Combined with the value-added utilization of formaldehyde-protected lignin, this study would enable the full and efficient utilization of lignocellulosic biomass components and further improve the economics of the formaldehyde protection fractionation process.

To solve the insufficient availability of mogrol, an 11α-hydroxy aglycone of mogrosides in Siraitia grosvenorii, snailase was employed as the enzyme to completely deglycosylate LHG extract containing 50% mogroside V. Other commonly used glycosidases performed less efficiently. Response surface methodology was conducted to optimize the productivity of mogrol, which peaked at 74.7% in an aqueous reaction. In view of the differences in water-solubility between mogrol and LHG extract, we employed an aqueous-organic system for the snailase-catalyzed reaction. Of five tested organic solvents, toluene performed best and was relatively well tolerated by snailase. After optimization, biphasic medium containing 30% toluene (v/v) could produce a high-quality mogrol (98.1% purity) at a 0.5 L scale with a production rate of 93.2% within 20 h. This toluene-aqueous biphasic system would not only provide sufficient mogrol to construct future synthetic biology systems for the preparation of mogrosides, but also facilitate the development of mogrol-based medicines.

Sustainable biodesulfurization (BDS) processes require the use of microbial biocatalysts that display high activity against the recalcitrant heterocyclic sulfur compounds and can simultaneously withstand the harsh conditions of contact with petroleum products, inherent to any industrial biphasic BDS system. In this framework, the functional microbial BDS-related diversity in a naturally oil-exposed ecosystem, was examined through a 4,6-dimethyl-dibenzothiophene based enrichment process. Two new Rhodococcus sp. strains were isolated, which during a medium optimization process revealed a significantly enhanced BDS activity profile when compared to the model strain R. qingshengii IGTS8. In biocatalyst stability studies conducted in biphasic mode using partially hydrodesulfurized diesel under various process conditions, the new strains also presented an enhanced stability phenotype. In these studies, it was also demonstrated for all strains, that the BDS activity losses were decoupled from the overall cells\' viability, in addition to the fact that the use of whole-broth biocatalyst positively affected BDS performance.

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.

To date, an efficient process for manufacturing valuable furan compounds from available renewable resources has gained great attention via a chemoenzymatic route. In this study, a sulfonated tin-loaded heterogeneous catalyst CLUST-Sn-LS using lobster shell as biobased carrier was prepared to convert corncob (75.0 g/L) into furfural (122.5 mM) at 170 °C for 30 min in methyl isobutyl ketone (MIBK)-H2O biphasic system (2:1, v/v). To improve furfurylamine yield, a novel recombinant E. coli TFTS harboring robust mutant Aspergillus terreus ω-transaminase [hydrophilic threonine (T) at position 130 was site-directed mutated to hydrophobic phenylalanine (F)] was constructed to transform 300-500 mM furfural into furfurylamine (90.1-93.6 % yield) at 30 °C and pH 7.5 in MIBK-H2O. Corncob was converted to furfurylamine in MIBK-H2O with a high productivity of 0.461 g furfurylamine/(g xylan). This constructed chemoenzymatic method coupling bio-based chemocatalyst CLUST-Sn-LS and mutant ω-transaminase biocatalyst in a biphasic system could efficiently convert lignocellulose into furfurylamine.

The developing of pretreatment method to break the biomass barrier of lignocellulosic is a challenging task for achieve high value utilization. A fast microwave-assisted choline chloride/1,2-propanediol/methyl isobutyl ketone biphasic system was constructed for pretreating Eucalyptus to the production of furfural and cellulose-rich residues and the extraction of lignin. Results showed that the combination of AlCl3·6H2O and HCl had the best catalytic ability for furfural production among the examined catalysts. Under the optimal conditions (140 °C, 15 min, 0.075 M AlCl3·6H2O, 0.05 M HCl), the furfural yield of 55.4 %, the glucose yield of 90.3 % and the delignification rate of 92.4 % could be achieved. Moreover, the extracted lignin samples with a low polydispersity (1.55-1.73) and molecular weight (1380-2040 g/mol) are promising to act as precursor for the value-add products processing. These findings demonstrated an ultrafast pretreatment process with excellent results in biomass fractionation and comprehensive utilization of biomass components.