Significant Enhancement of 5-Hydroxymethylfural Productivity from D-Fructose with SG(SiO2) in Betaine:Glycerol-Water for Efficient Synthesis of Biobased 5-(Hydroxymethyl)furfurylamine.

5-Hydroxymethyl-2-furfurylamine (5-HMFA) as an important 5-HMF derivative has been widely utilized in the manufacture of diuretics, antihypertensive drugs, preservatives and curing agents. In this work, an efficient chemoenzymatic route was constructed for producing 5-(hydroxymethyl)furfurylamine (5-HMFA) from biobased D-fructose in deep eutectic solvent Betaine:Glycerol-water. The introduction of Betaine:Glycerol could greatly promote the dehydration of D-fructose to 5-HMF and inhibit the secondary decomposition reactions of 5-HMF, compared with a single aqueous phase. D-Fructose (200 mM) could be catalyzed to 5-HMF (183.4 mM) at 91.7% yield by SG(SiO2) (3 wt%) after 90 min in Betaine:Glycerol (20 wt%), and at 150 °C. E. coli AT exhibited excellent bio-transamination activity to aminate 5-HMF into 5-HMFA at 35 °C and pH 7.5. After 24 h, D-fructose-derived 5-HMF (165.4 mM) was converted to 5-HMFA (155.7 mM) in 94.1% yield with D-Ala (D-Ala-to-5-HMF molar ratio 15:1) in Betaine:Glycerol (20 wt%) without removal of SG(SiO2), achieving a productivity of 0.61 g 5-HMFA/(g substrate D-fructose). Chemoenzymatic valorization of D-fructose with SG(SiO2) and E. coli AT was established for sustainable production of 5-HMFA, which has potential application.

Biodegradation of alkali lignin by a newly isolated Rhodococcus pyridinivorans CCZU-B16.

Based on the Prussian blue spectrophotometric method, one high-throughput screening strategy for screening lignin-degrading microorganisms was built on 24-well plate at room temperature. One high activity of alkali lignin-degrading strain Rhodococcus pyridinivorans CCZU-B16 was isolated from soil. After the optimization of biodegradation, 30.2% of alkali lignin (4 g/L) was degraded under the nitrogen-limited condition (30/1 of C/N ratio; g/g) at 30 °C for 72 h. It was found that syringyl (S) units and guaiacyl (G) in lignin decreased after biodegradation. Moreover, the accumulated lipid in cells had a fatty acid profile rich in C16 and C18 with four major constituent fatty acids including palmitic acid (C16:0; 22.4%), palmitoleic acid (C16:1; 21.1%), stearic acid (C18:0; 16.2%), and oleic acid (C18:1; 23.1%). In conclusion, Rhodococcus pyridinivorans CCZU-B16 showed high potential application in future.

Effective pretreatment of dilute NaOH-soaked chestnut shell with glycerol-HClO4-water media: structural characterization, enzymatic saccharification, and ethanol fermentation.

In this study, an effective pretreatment of dilute NaOH-soaked chestnut shell (CNS) with glycerol-HClO4-water (88.8:1.2:10, w/w/w) media at 130 °C for 30 min was successfully demonstrated. Results revealed that the combination pretreatment removed 66.0 % of lignin and 73.7 % of hemicellulose in untreated CNS. The changes in the structural features (crystallinity, morphology, and porosity) of the solid residue of CNS were characterized with Fourier transform infrared spectroscopy, fluorescent microscope, scanning electron microscopy, and X-ray diffraction. Biotransformation of glycerol-HClO4-water pretreated-NaOH-soaked CNS (50 g/L) with a cocktail of enzymes for 72 h, the reducing sugars and glucose were 39.7 and 33.4 g/L, respectively. Moreover, the recovered hydrolyzates containing 20 g/L glucose had no inhibitory effects on the ethanol-fermenting microorganism, and the ethanol production was 0.45 g/g glucose within 48 h. In conclusion, this combination pretreatment shows promise as pretreatment solvent for wheat straw, although the in-depth exploration of this subject is needed.

Biotransformation of 1,3-propanediol cyclic sulfate and its derivatives to diols by Rhodococcus sp.

Rhodococcus sp. CGMCC 4911 transformed 1,3-propanediol cyclic sulfate (1,3-PDS) and its derivatives into corresponding diols. Ethylene sulfate, glycol sulfide, 1,3-PDS, and 1,2-propanediol cyclic sulfate were effectively hydrolyzed with growing cells. (R)-1,2-Propanediol (>99 % e.e.) was obtained at 44 % yield with growing cells. Glycol sulfide, ethylene sulfate, and 1,3-PDS were converted into the corresponding diols at 94.6, 96.3, and 98.3 %, respectively. Optimal reaction conditions with lyophilized resting cells were 30 °C, pH 7.5, and cell dosage 17.9 mg cell dry wt/ml. 1,3-Propanediol was obtained from 50 mM 1,3-PDS at 97.2 % yield by lyophilized cells after 16 h. Lyophilized cells were entrapped in calcium alginate with a half-life of 263 h at 30 °C, and the total operational time of the immobilized biocatalysts could reach over 192 h with a high conversion rate.

Biosynthesis of terephthalic acid, isophthalic acid and their derivatives from the corresponding dinitriles by tetrachloroterephthalonitrile-induced Rhodococcus sp.

The nitrilase from Rhodococcus sp. CCZU10-1 catalyses the hydrolysis of dinitriles to acids without the formation of amides and cyanocarboxylic acids. It was induced by benzonitrile and its analogues (tetrachloroterephthalonitrile > Îµ-caprolactam > benzonitrile > phenylacetonitrile), and had activity towards aromatic nitriles (terephthalonitrile > tetrachloroterephthalonitrile > isophthalonitrile > tetrachloroisophthalonitrile > tetrafluoroterephthalonitrile > benzonitrile). After the optimization, the highest nitrilase induction [311 U/(g DCW)] was achieved with tetrachloroterephthalonitrile (1 mM) in the medium after 24 h at 30 °C after optimum enzyme activity was at pH 6.8 and at 30 °C. Efficient biocatalyst recycling was achieved by cell immobilization in calcium alginate, with a product-to-biocatalyst ratios of 776 g terephthalic acid/g DCW and 630 g isophthalic acid/g DCW.

Antibacterial, antioxidant and fruit packaging ability of biochar-based silver nanoparticles-polyvinyl alcohol-chitosan composite film.

Silver nanoparticles were prepared by loading Ag+ into biochar of waste barley distillers' grains shell by reduction with trisodium citrate, and this silver-loaded biochar was introduced into polyvinyl alcohol-chitosan. Various analysis with Fourier Transform Infrared spectroscopy, X-ray diffraction, Thermogravimetric analysis, and water contact angle revealed that biochar-based silver nanoparticle was incorporated into the polyvinyl alcohol-chitosan film, the biochar-based silver nanoparticles-polyvinyl alcohol-chitosan (C-Ag-loaded PVA/CS) composite film had good thermostability and hydrophobicity. Through the analysis via disk diffusion method, the composite containing 3 % of biochar-based silver nanoparticles-polyvinyl alcohol-chitosan had high antibacterial activity (inhibition zone: 18 mm against E. coli and 15 mm against S. aureus), and the bacterial membrane permeability was measured, indicating that C-Ag-loaded PVA/CS composite film could destroy the cell membrane, release intracellular substances, and have high antioxidant activity. During the storage, the weight loss rate of the biochar-based silver nanoparticles-polyvinyl alcohol-chitosan plastic wrap group was 0.14 %, and the titratable acid content only decreased by 0.061 %, which had a good effect on extending the shelf life of blueberries. The C-Ag-loaded PVA/CS composite film could also delay deterioration of blueberries and prolong storage time. Overall, this composite film had potential in food packaging and extending food shelf-life aspects.

Efficient synthesis of vanillylamine through bioamination of lignin-derived vanillin by recombinant E. coli containing ω-transaminase from Caulobacter sp. D5 in dimethyl sulfoxide-water.

Lignin is a plentiful and readily accessible renewable resource. Vanillylamine is a crucial raw material used to synthesize pharmaceuticals and high-value furan compounds that can be acquired by aminating lignin-derived vanillin (Van). However, effectually achieving the biocatalytic synthesis of vanillylamine has remained challenging. In this study, a dimethyl sulfoxide (DMSO)-H2O (1:9, vol/vol) bioreaction medium was constructed, and a recombinant E. coli ATA1012 carrying ω-transaminase from Caulobacter sp. D5 was used as the ω-transaminase biocatalyst to acquire the effectual biocatalytic synthesis of vanillylamine. Under optimized bioreaction conditions (37 ℃ and pH 7.5) by supplementary of isopropylamine (IPA) (Van/IPA = 1:5, mol/mol), 80-100 mM Van could be effectually converted by ATA1012 whole cells in DMSO-H2O (1:9, vol/vol) within 12 h, yielding 91.2 %-95.4 % vanillylamine, with >99 % selectivity. An efficient amination process was developed using ATA1012 with superior transaminase catalytic activity and substrate tolerance to effectively convert Van to vanillylamine in a DMSO-H2O medium.

Highly enantioselective oxidation of racemic phenyl-1,2-ethanediol to optically pure (R)-(-)-mandelic acid by a newly isolated Brevibacterium lutescens CCZU12-1.

Enantioselective oxidation of racemic phenyl-1,2-ethanediol into (R)-(-)-mandelic acid by a newly isolated Brevibacterium lutescens CCZU12-1 was demonstrated. It was found that optically active (R)-(-)-mandelic acid (e.e.p > 99.9 %) is produced leaving the other enantiomer (S)-(+)-phenyl-1,2-ethanediol intact. Using fed-batch method, a total of 172.9 mM (R)-(-)-mandelic acid accumulated in the reaction mixture after the seventh feed. Moreover, oxidation of phenyl-1,2-ethanediol using calcium alginate-entrapped resting cells was carried out in the aqueous system, and efficient biocatalyst recycling was achieved as a result of cell immobilization in calcium alginate, with a product-to-biocatalyst ratio of 27.94 g (R)-(-)-mandelic acid g⁻¹ dry cell weight cell after 16 cycles of repeated use.

Highly enantioselective oxidation of phenyl methyl sulfide and its derivatives into optically pure (S)-sulfoxides with Rhodococcus sp. CCZU10-1 in an n-octane-water biphasic system.

Enantiopure sulfoxides can be prepared via the asymmetric oxidation of sulfides using sulfide monooxygenases. The n-octane-water biphasic system was chosen for the bio-oxidation of a water-insoluble phenyl methyl sulfide (PMS) by Rhodococcus sp. CCZU10-1. In this n-octane-water system, the optimum reaction conditions were obtained. (S)-phenyl methyl sulfoxide ((S)-PMSO) with >99.9 % enantiomeric excess formed at 55.3 mM in the n-octane-water biphasic system. Using fed-batch method, a total of 118 mM (S)-PMSO accumulated in 1-L reaction mixture after the 7th feed, and no (R)-PMSO and sulfone were detected. Moreover, Rhodococcus sp. CCZU10-1 displayed fairly good activity and enantioselectivity toward other sulfides. In conclusion, Rhodococcus sp. CCZU10-1 is a promising biocatalyst for synthesizing highly optically active sulfoxides.

Efficient co-production of reducing sugars and xylooligosaccharides via clean hydrothermal pretreatment of rape straw.

Hydrothermal treatment was applied to pretreat rape straw for the efficient co-production of reducing sugars and xylooligosaccharides. It was observed that hydrothermal treatment using water as solvent and catalyst destructed the compact structure of rape straw and increased its enzymatic digestion efficiency from 24.6% to 92.0%. Xylooligosaccharide (3.3 g/L) was acquired after the treatment under 200 °C for 60 min (severity factor Log Ro = 4.7). With increasing pretreatment intensity from 3.1 to 5.4, the hemicellulose removal increased from 14.4% to 100%, and the delignification was raised from 12% to 44%. Various characterization proved that the surface morphology of treated material showed a porous shape, while the cellulose accessibility, lignin surface area and lignin hydrophobicity were greatly improved. Consequently, hydrothermal pretreatment played a vital role in the sustainable transformation of biomass to valuable biobased compounds, and had a wide range of application prospects in lignocellulosic biorefining.

Comprehensive understanding of enzymatic saccharification of Betaine:Lactic acid-pretreated sugarcane bagasse.

Green solvents, especially deep eutectic solvents (DESs), are widely applied to pretreat biomass for enhancing its enzymatic hydrolysis. In this work, lactic acid was selected as the hydrogen-bond-donor to prepare Betaine-base DES (Betaine:LA), The DES was utilized to pretreat sugarcane bagasse (SCB) at 160 ℃ for 80 min (severity factor LogR0 = 3.67). The influences of Betaine:LA treatment on the chemical composition, crystal and microstructure structure of cellulose, and cellulase digestion were investigated. The results showed that the lignin (47.1%) and xylan (44.6%) were removed, the cellulase digestibility of Betaine:LA-treated SCB was 4.2 times that of the raw material. This improved efficiency was attributed to the enhanced accessibility of cellulose, the weakened surface area of lignin, the declined hydrophobicity, and the decreased crystallinity of cellulose. Several compelling linear correlations were fitted between enzymatic hydrolysis and these alterations of physicochemical features, comprehensively understanding enzymatic saccharification of Betaine:LA-pretreated SCB.

Biological valorization of lignin-derived vanillin to vanillylamine by recombinant E. coli expressing ω-transaminase and alanine dehydrogenase in a petroleum ether-water system.

Vanillylamine, as an important drug precursor and fine chemical intermediate, has great economic value. By constructing a strategy of double enzyme co-expression, one newly constructed recombinant E. coli HNIQLE-AlaDH expressing ω-transaminase from Aspergillus terreus and alanine dehydrogenase from Bacillus subtilis was firstly used aminate lignin-derived vanillin to vanillylamine by using a relatively low dosage of amine donors (vanillin:L-alanine:isopropylamine = 1:1:1, mol/mol/mol). In addition, in a two-phase system (water:petroleum ether = 80:20 v/v), the bioconversion of vanillin to vanillylamine was catalyzed by HNIQLE-AlaDH cell under the ambient condition, and the vanillylamine yield was 71.5%, respectively. This double-enzyme HNIQLE-AlaDH catalytic strategy was applied to catalyze the bioamination of furfural and 5-hydroxymethylfurfural with high amination efficiency. It showed that the double-enzyme catalytic strategy in this study promoted L-alanine to replace D-alanine to participate in bioamination of vanillin and its derivatives, showing a great prospect in the green biosynthesis of biobased chemicals from biomass.

An efficient and sustainable furfurylamine production from biomass-derived furfural by a robust mutant ω-transaminase biocatalyst.

Furfurylamine is a key furan-based compound for manufacturing perfumes, fibers, additives, medicines and agrochemicals. It can be obtained by amination of furfural by ω-transaminase (AtAT) from Aspergillus terreus. In this work, site-directed mutant of amino acid residues [Threonine (T) at AT130 was mutated to Methionine (M) and Glutamic acid (E) at AT133 was mutated to Phenylalanine (F)] was used to change in the flexible region of AtAT. The transamination activity and thermostability were significantly improved. In ChCl:MA (30 wt%), furfural (500 mM) was efficiently transformed into furfurylamine (92% yield) with TMEF after 12 h. 101.3 mM of biomass-derived furfural and 129.7 mM of D-xylose-derived furfural were wholly converted into furfurylamine within 5 h, achieving the productivity of 0.465 g furfurylamine/(g xylan in corncob) and 0.302 g furfurylamine/(g D-xylose). This established chemoenzymatic conversion strategy by bridging chemocatalysis and biocatalysis could be utilized in the valorisation of renewable biomass to valuable furans.

Effective one-pot chemoenzymatic cascade catalysis of biobased feedstock for synthesizing 2,5-diformylfuran in a sustainable reaction system.

2,5-Diformylfuran, which can be prepared via the oxidation of biobased HMF, has received considerable attention because of its potential applications in producing furan-based chemicals and functional materials, such as biofuels, polymers, fluorescent material, vitrimers, surfactants, antifungal agents and medicines. This work aimed to develop an efficient one-pot process for chemoenzymatic transformation of biobased substrate to 2,5-diformylfuran with deep eutectic solvent (DES) Betaine:Lactic acid ([BA][LA]) catalyst and oxidase biocatalyst in [BA][LA]-H2O. Using waste bread (50 g/L) and D-fructose (18.0 g/L) as feedstocks in [BA][LA]-H2O (15:85, vol/vol), the yields of HMF were 32.8% (15 min) and 91.6% (90 min) at 150 °C, respectively. These prepared HMF could be biologically oxidized to 2,5-diformylfuran by Escherichia coli pRSFDuet-GOase, achieving a productivity of 0.631 g 2,5-diformylfuran/(g fructose) and 0.323 g 2,5-diformylfuran/(g bread) after 6 h under the mild performance condition. This bioresourced intermediate 2,5-diformylfuran was effectively synthesized from biobased feedstock in an environmentally-friendly system.

Efficient co-production of xylooligosaccharides, furfural and reducing sugars from yellow bamboo via the pretreatment with biochar-based catalyst.

The research on the efficient use of biomass to produce chemical products has received extensive attention. In this work, a novel heterogeneous biocarbon-based heterogeneous catalyst AT-Sn-YB was prepared using yellow bamboo (YB) as a carrier, and its physical properties were proved to be good by various characterization and stability experiments. In the γ-valerolactone/water (3:1, v/v) medium containing 100 mM CuCl2, the use of AT-Sn-YB (3.6 wt%) under 170 °C for 20 min was applied to catalyze YB into furfural (80.3% yield), accompanied with 2.8 g/L xylooligosaccharides. The YB solid residue obtained from treatment was efficiently saccharified to reducing sugars (17.2 g/L). Accordingly, comprehensive understanding of efficiently co-producing xylooligosaccharides, furfural and reducing sugars from YB was demonstrated via the pretreatment with biochar-based catalyst. This study innovatively used a new type of solid acid to complete the efficient co-production of chemical products, and realized the value-added utilization of yellow bamboo.

Chemobiocatalytic transfromation of biomass into furfurylamine with mixed amine donor in an eco-friendly medium.

Biobased furfurylamine (FAM) is a versatile platform molecule for producing additives, pharmaceuticals, and pesticides. Recombinant E. coli HNND-AlaDH was created by co-expressing L-alanine dehydrogenase (AlaDH) and mutated Aspergillus terreus ω-transaminase (HNND), aiming to convert furfural (FUR) into FAM using inexpensive L-alanine and isopropylamine as mixed amine donors. In ChCl:FA:OA (10 wt%), pineapple peel, bagasse, barley shell, peanut shell, and corn stalk could be efficiently transformed into FUR under 170 °C for 10 min. Pineapple peel produced a high titer of FUR (183.3 mM). Additionally, the viscosity, surface tension and polarity of ChCl:FA:OA were explored. The biomass-derived FUR was fully transformed to FAM by HNND-AlaDH with amine donor (1:1:1 of L-Ala/isopropylamine/FUR mol/mol/mol) within 300 min. Accordingly, the FAM productivity was 0.58 g/(g xylan in pineapple peel). This chemobiocatalytic strategy established through the combination of chemocatalysis and biocatalysis could be applied to convert renewable biomass into valuable organic amines.

Combined sulfuric acid and choline chloride/glycerol pretreatment for efficiently enhancing enzymatic saccharification of reed stalk.

In this study, an efficient combination of pretreatment solvents involving Choline chloride/Glycerol (ChCl/Gly) and H2SO4 was firstly developed to assess the pretreatment performance and determine optimal pretreatment conditions. The results illustrated that the H2SO4-[ChCl/Gly] combination efficiently removed lignin (52.6%) and xylan (80.5%) from the pretreated reed stalk, and subsequent enzymatic hydrolysis yielded 91.1% of glucose. Furthermore, several characterizations were conducted to examine the structural and morphological changes of the reed stalk, revealing apparently enhanced accessibility (128.4 to 522.6 mg/g), reduced lignin surface area (357.9 to 229.5 m2/g), and substantial changes on biomass surface. Based on the aforementioned study, possible mechanisms for the H2SO4-[ChCl/Gly] pretreatment of reed stalks were proposed. The comprehensive understanding of combined H2SO4-[ChCl/Gly] pretreatment system for enhancing the saccharification of the reed stalk was interpreted in this work. Overall, this novel approach could be efficiently applied to pretreat and saccharify reed stalks, empowering the biomass refining industry.

Efficient production of hydroxymethyl-2-furfurylamine by chemoenzymatic cascade catalysis of bread waste in a sustainable approach.

In this study, efficient and sustainable conversion of waste bread (WB) to 5-hydroxymethyl-2-furoamine (HMFA) was achieved in a cascade reaction in betaine:malonic acid (B:MA) - water. 5-HMF (30.3 wt% yield) was synthesized from WB (40.0 g/L) in B:MA - water (B:MA, 18 wt%) in 45 min at 190 °C. By using the newly created recombinant E. coli HNILGD-AlaDH cells expressing L-alanine dehydrogenase (AlaDH) and ω-transaminase mutant HNILGD as biocatalyst, the WB-valorized 5-HMF was biologically aminated into HMFA in a high yield (92.1%) at 35 °C for 12 h through in situ removal of the amino transfer by-products of the amine donor, greatly reducing amine donor dosage (from D-Ala/5-HMF = 16/1 to D-Ala/5-HMF = 2/1, mol/mol) and improving the productivity of HMFA (0.282 g HMFA per g WB). This two-step chemical-enzymatic cascade reaction strategy with B:MA and HNILGD-AlaDH whole-cell provides a new idea for the chemoenzymatic synthesis of valuable furan chemicals from waste biomass.

Transformation of bread waste into 2,5-furandimethanol via an efficient chemoenzymatic approach in a benign reaction system.

Via combination catalysis with deep eutectic solvent lactic acid:betaine (chemocatalyst) and HMFOMUT cell (biocatalyst: E. coli HMFOMUT whole-cell), one-pot manufacture of 2,5-furandimethanol from waste bioresource was constructed in a chemoenzymatic approach. With bread waste (50 g/L) as substrate, the 5-hydroxymethylfuran yield reached 44.2 Cmol% (based on bread waste) by lactic acid:betaine (15 wt%) at 180 °C for 15 min. With glucose as co-substrate, HMFOMUT could transform 5-hydroxymethylfurfural (150 mM) to 2,5-furandimethanol (84.5 % yield) after 1 day at 37 °C and pH 7.0. In lactic acid:betaine-H2O, HMFOMUT effectively converted bread-derived 5-hydroxymethylfurfural into 2,5-furandimethanol in a productivity of 700 kg 2,5-furandimethanol per kg 5-hydroxymethylfurfural (230 kg 2,5-furandimethanol per kg bread). In an eco-friendly lactic acid:betaine system, an effective one-pot chemoenzymatic strategy was firstly developed to convert bread waste into 2,5-furandimethanol, which would reduce the operation cost and has potential application value for valorizing waste food bioresource into value-added furan.

An efficient chemoenzymatic cascade strategy for transforming biomass into furfurylamine with lobster shell-based chemocatalyst and mutated ω-transaminase biocatalyst in methyl isobutyl ketone-water.

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.