Glycerol Utilization Research Articles

Photocatalytic glycerol reforming on Pt, Au and Cu supported by reduced TiO2 under visible light irradiation

The commercial feasibility of biodiesel production highly depends on the effective utilization of byproduct glycerol through an environmentally friendly process like photocatalytic reforming. However, the process should be possible under visible light if solar energy is used. In this work, this process was studied over Pt, Au, Au–Cu, and Cu on reduced TiO2 (by NaBH4), which is more effective under visible light than pristine TiO2, for the first time in literature. Au, Au–Cu, and Cu were loaded by deposition precipitation with urea, while Pt was loaded by impregnation. Although the performance under UV–vis and white TiO2 is still higher, the visible light performance of Pt, Au–Cu, and Cu is much better over reduced TiO2 (91, 36, and 27 μmol/h. gcat respectively) than those over pristine TiO2 (9, 14 and 18 μmol/h. gcat respectively). This indicates that reduced TiO2 has the potential for the visible light reforming of glycerol.

Transcriptomic and metabolomic analyses revealed regulation mechanism of mixotrophic Cylindrotheca sp. glycerol utilization and biomass promotion

BackgroundDiatoms have been viewed as ideal cell factories for production of some high-value bioactive metabolites, such as fucoxanthin, but their applications are restrained by limited biomass yield. Mixotrophy, by using both CO2 and organic carbon source, is believed effective to crack the bottleneck of biomass accumulation and achieve a sustainable bioproduct supply.ResultsGlycerol, among tested carbon sources, was proved as the sole that could significantly promote growth of Cylindrotheca sp. with illumination, a so-called growth pattern, mixotrophy. Biomass and fucoxanthin yields of Cylindrotheca sp., grown in medium with glycerol (2 g L−1), was increased by 52% and 29%, respectively, as compared to the autotrophic culture (control) without compromise in photosynthetic performance. As Cylindrotheca sp. was unable to use glycerol without light, a time-series transcriptomic analysis was carried out to elucidate the light regulation on glycerol utilization. Among the genes participating in glycerol utilization, GPDH1, TIM1 and GAPDH1, showed the highest dependence on light. Their expressions decreased dramatically when the alga was transferred from light into darkness. Despite the reduced glycerol uptake in the dark, expressions of genes associating with pyrimidine metabolism and DNA replication were upregulated when Cylindrotheca sp. was cultured mixotrophically. Comparative transcriptomic and metabolomic analyses revealed amino acids and aminoacyl-tRNA metabolisms were enhanced at different timepoints of diurnal cycles in mixotrophic Cylindrotheca sp., as compared to the control.ConclusionsConclusively, this study not only provides an alternative for large-scale cultivation of Cylindrotheca, but also pinpoints the limiting enzymes subject to further metabolic manipulation. Most importantly, the novel insights in this study should aid to understand the mechanism of biomass promotion in mixotrophic Cylindrotheca sp.

Vacuum-assisted adsorption of Candida rugosa lipase with nanotubes for monooleoglycerol synthesis

A surplus of crude glycerol has been produced as a by-product due to the escalating production of biofuels. Utilization of crude glycerol for monoolein production which offers a different approach using the abandoned crude glycerol from the biodiesel sector is the main aim of this study. Immobilized lipase was used in this investigation to catalyse the direct esterification of oleic acid and glycerol, which resulted in monoolein. The immobilized Candida rugosa lipase onto the halloysite nanotubes (HNTs) has shown remarkably promising attainment of lipase activity and loading capacity. The immobilised lipase was optimised for crucial parameters (HNT to lipase ratio, pH, temperature, and pressure) via response surface methodology (RSM). The recommended optimum conditions obtained are pH 6, 38.6 °C, and 0.33 atm (81.92% lipase activity). To analyse influences of reaction period, temperature, mixing rate, and crude glycerol to oleic acid molar ratio on the esterification reaction was carried out using the optimized conditions of immobilization process. The optimum conditions for physically immobilised lipase were to yield 88.97% monoolein produced at 42 °C in a substrate ratio of 1:4 (oleic acid: glycerol) stirred at 200 rotations per minute (rpm) with 16 h’ incubation time. Based on the research's findings, it was confirmed that HNT has good application potential for enzyme immobilization carriers.

Novel evolved Yarrowia lipolytica strains for enhanced growth and lipid content under high concentrations of crude glycerol

BackgroundYarrowia lipolytica is a well-studied oleaginous yeast known for its ability to accumulate and store intracellular lipids, while growing on diverse, non-conventional substrates. Amongst them, crude glycerol, a low-cost by-product of the biodiesel industry, appears to be an interesting option for scaling up a sustainable single-cell oil production process. Adaptive laboratory evolution (ALE) is a powerful tool to force metabolic adaptations endowing tolerance to stressful environmental conditions, generating superior phenotypes with industrial relevance.ResultsY. lipolytica MUCL 28849 underwent ALE in a synthetic medium with increasing concentration of pure or crude glycerol as a stressing factor (9–20% v/v) for 520 generations. In one case of pure glycerol, chemical mutagenesis with ethyl methanesulfonate (EMS) was applied prior to ALE. Growth profile, biomass production and lipid content of 660 evolved strains (EVS), revealed 5 superior isolates; exhibiting from 1.9 to 3.6-fold increase of dry biomass and from 1.1 to 1.6-fold increase of lipid concentration compared to the parental strain, when grown in 15% v/v crude glycerol. NGS for differential gene expression analysis, showed induced expression in all EVS affecting nucleosomal structure and regulation of transcription. As strains differentiated, further changes accumulated in membrane transport and protein transport processes. Genes involved in glycerol catabolism and triacylglycerol biosynthesis were overexpressed in two EVS. Mismatches and gaps in the expressed sequences identified altered splicing and mutations in the EVS, with most of them, affecting different components of septin ring formation in the budding process. The selected YLE155 EVS, used for scale-up cultivation in a 3L benchtop bioreactor with 20% v/v crude glycerol, achieved extended exponential phase, twofold increase of dry biomass and lipid yields at 48 h, while citric acid secretion and glycerol consumption rates were 40% and 50% lower, respectively, compared to the parental strain, after 24 h of cultivation.ConclusionALE and EMS-ALE under increasing concentrations of pure or crude glycerol generated novel Y. lipolytica strains with enhanced biomass and lipid content. Differential gene expression analysis and scale-up of YLE155, illustrated the potential of the evolved strains to serve as suitable “chassis” for rational engineering approaches towards both increased lipid accumulation, and production of high-added value compounds, through efficient utilization of crude glycerol.

Transcriptomic profiling reveals differences in the adaptation of two Tetragenococcus halophilus strains to a lupine moromi model medium

BackgroundTetragenococcus (T.) halophilus is a common member of the microbial consortia of food fermented under high salt conditions. These comprises salty condiments based on soy or lupine beans, fish sauce, shrimp paste and brined anchovies. Within these fermentations this lactic acid bacterium (LAB) is responsible for the formation of lactic and other short chain acids that contribute to the flavor and lower the pH of the product. In this study, we investigated the transcriptomic profile of the two T. halophilus strains TMW 2.2254 and TMW 2.2256 in a lupine moromi model medium supplied with galactose. To get further insights into which genomic trait is important, we used a setup with two strains. That way we can determine if strain dependent pathways contribute to the overall fitness. These strains differ in the ability to utilize L-arginine, L-aspartate, L-arabinose, D-sorbitol, glycerol, D-lactose or D-melibiose. The lupine moromi model medium is an adapted version of the regular MRS medium supplied with lupine peptone instead of casein peptone and meat extract, to simulate the amino acid availabilities in lupine moromi.ResultsThe transcriptomic profiles of the T. halophilus strains TMW 2.2254 and TMW 2.2256 in a lupine peptone-based model media supplied with galactose, used as simulation media for a lupine seasoning sauce fermentation, were compared to the determine potentially important traits. Both strains, have a great overlap in their response to the culture conditions but some strain specific features such as the utilization of glycerol, sorbitol and arginine contribute to the overall fitness of the strain TMW 2.2256. Interestingly, although both strains have two non-identical copies of the tagatose-6P pathway and the Leloir pathway increased under the same conditions, TMW 2.2256 prefers the degradation via the tagatose-6P pathway while TMW 2.2254 does not. Furthermore, TMW 2.2256 shows an increase in pathways required for balancing out the intracellular NADH/NADH+ ratios.ConclusionsOur study reveals for the first time, that both versions of tagatose-6P pathways encoded in both strains are simultaneously active together with the Leloir pathway and contribute to the degradation of galactose. These findings will help to understand the strain dependent features that might be required for a starter strain in lupine moromi.

Investigation of two metabolic engineering approaches for (R,R)-2,3-butanediol production from glycerol in Bacillus subtilis

BackgroundFlux Balance Analysis (FBA) is a well-known bioinformatics tool for metabolic engineering design. Previously, we have successfully used single-level FBA to design metabolic fluxes in Bacillus subtilis to enhance (R,R)-2,3-butanediol (2,3-BD) production from glycerol. OptKnock is another powerful technique for devising gene deletion strategies to maximize microbial growth coupling with improved biochemical production. It has never been used in B. subtilis. In this study, we aimed to compare the use of single-level FBA and OptKnock for designing enhanced 2,3-BD production from glycerol in B. subtilis.ResultsSingle-level FBA and OptKnock were used to design metabolic engineering approaches for B. subtilis to enhance 2,3-BD production from glycerol. Single-level FBA indicated that deletion of ackA, pta, lctE, and mmgA would improve the production of 2,3-BD from glycerol, while OptKnock simulation suggested the deletion of ackA, pta, mmgA, and zwf. Consequently, strains LM01 (single-level FBA-based) and MZ02 (OptKnock-based) were constructed, and their capacity to produce 2,3-BD from glycerol was investigated. The deletion of multiple genes did not negatively affect strain growth and glycerol utilization. The highest 2,3-BD production was detected in strain LM01. Strain MZ02 produced 2,3-BD at a similar level as the wild type, indicating that the OptKnock prediction was erroneous. Two-step FBA was performed to examine the reason for the erroneous OptKnock prediction. Interestingly, we newly found that zwf gene deletion in strain MZ02 improved lactate production, which has never been reported to date. The predictions of single-level FBA for strain MZ02 were in line with experimental findings.ConclusionsWe showed that single-level FBA is an effective approach for metabolic design and manipulation to enhance 2,3-BD production from glycerol in B. subtilis. Further, while this approach predicted the phenotypes of generated strains with high precision, OptKnock prediction was not accurate. We suggest that OptKnock modelling predictions be evaluated by using single-level FBA to ensure the accuracy of metabolic pathway design. Furthermore, the zwf gene knockout resulted in the change of metabolic fluxes to enhance the lactate productivity.

STUDY OF ACID CATALYSTS IN THE GLYCEROL ETHERIFICATION WITH BENZYL ALCOHOL

Glycerol (Gly) is the major by-product of the biodiesel production (approximately 10 wt.%). There are different research focusing on developing glycerol utilization to produce value-added chemicals. One of them is the etherification with alcohols in the presence of acidic catalysts. In the present work, the catalytic behaviour of SiO2-based system functionalized with heteropolyacids (HPA: H3O40PW12.xH2O) was studied in the etherification of Gly with benzyl alcohol (BA). The solids were impregnated with different solutions of a HPA (28, 35 and 40%) and characterized. Very good catalytic performance was achieved (94% conversion and 78% of selectivity to ME1: 3-benzyloxy-1, 2-propanediol) after 360 min of reaction, at 120°C and Gly: BA= 3:1, with 10 wt.% of SiO2-HPA35. This system was compared with functionalized carbon-based catalysts (Cst-SO3H and Ccs-SO3H) and it was observed that the results were similar, conversion over 90% and selectivity between 70 -80% to some of the desired products (ME1).

Process coupling of CO2 and glycerol comprehensive utilization based on anionic 2D nanomaterial

AbstractWe developed an approach to realize the comprehensive utilization of CO2 and glycerol by smartly design a nanocatalyst, which concerns the coupling of catalytic reactions and the separation or capture process. Specifically, MgFe‐CO32−‐LDHs with different thickness were fabricated by employing atmospheric CO2 as carbon source and an atomic‐level interface was constructed by introducing Pt single atoms to obtain series of Pt/MgFe‐LDHs nanocatalysts. During a catalytic assessment, the CO32− confined within interlayer was in situ converted when integrated with glycerol oxidation under irradiation. The optimal nanocatalyst exhibited a decent conversion efficiency of CO32−‐CO‐CH4 and near 100% selectivity of secondary hydroxy group in glycerol. DFT calculation, isotope‐labeled experiments and in situ spectrograph characterization revealed possible coupling reaction mechanism. More importantly, the nanocatalyst exhibited impressive reusability for at least six 6 runs. The developed multiple coupling process will be an alternative and promising procedure for the waste and low‐valued sources synergetic utilization.

Lignin depolymerization for aromatic compounds over Ni-Ce/biochar catalyst under aqueous-phase glycerol

Developing an advanced catalytic system for the purposeful depolymerization of lignin into aromatic compounds has presented a significant prospect for green manufacturing. In this paper, the catalytic process of microporous biochar (BC) derived from lignin supported Ni-Ce catalysts (xNi-Ce/BC) coupling with the aqueous-phase glycerol medium was reported for the depolymerization of Kraft lignin to guaiacol and 4-alkyl guaiacols. Under the optimal conditions, 3Ni-Ce/BC yielded 59.02 % of lignin oil and simultaneously realized the highest yields of guaiacol (243.94 mg/g lignin) and 4-alkyl guaiacols (265.65 mg/g lignin). The characterization results revealed BC promoted the formation of metallic Ni sites and the interaction of Ni with CeO2 drove the generation of Ni-CeO2-x interfaces and oxygen vacancies (OV). These could adsorb and activate the CC and CO bonds of lignin and its depolymerized fragments to form reactive intermediates. Then, the Ni sites activated the aqueous-phase glycerol to form adsorbed H atoms, which then spilled over to the adjacent OV to stabilize reactive intermediates. Subsequently, the optimal distribution between Brønsted acid sites (BAS) and Lewis acid sites (LAS) in 3Ni-Ce/BC enhanced the yields of guaiacol and 4-alkyl guaiacols. The kinetic analysis adopting 2-phenoxy-1-phenylethanol as the β-O-4 bond model demonstrated that 3Ni-Ce/BC significantly reduced both the bond dissociation energy of β-O-4 bond and the apparent activation energy. Finally, the possible reaction mechanism of lignin depolymerization catalyzed by 3Ni-Ce/BC catalyst was proposed. This work provides a feasible method for the simultaneous utilization of lignin wastes and crude glycerol (the by-products of biodiesel market).

High production of acetoin from glycerol by Bacillus subtilis 35.

Acetoin is a high-value volatile compound widely applied in the chemical, food, and pharmaceutical industries. Despite the promising use of waste glycerol as a substrate in several microbial syntheses, acetoin production by natural microorganisms from glycerol as a sole carbon source has never been reported. The present study investigates the innate ability of Bacillus subtilis 35 (DSM 113,620) to convert glycerol into acetoin and 2,3-butanediol. The fermentation was directed towards acetoin production by medium selection and process parameter optimization using response surface design methodology. Thus, the fed batch conducted under optimized conditions received 77.9g/L acetoin with a productivity of 0.85g/L h and a yield of 0.36g/g. The obtained acetoin concentration is the highest from glycerol reported to date, comparable to the highest values gained from glucose. Transcription analysis of the gene cluster glpPFKD showed that all four genes responsible for the utilization of glycerol were expressed. This natural ability of the strain, along with its non-pathogenic nature, defines B. subtilis 35 as a very promising candidate for acetoin production from glycerol on an industrial scale. KEY POINTS: • The highest microbial production of acetoin from glycerol. • Process parameter optimization directs glycerol conversion to acetoin production. • B. subtilis 35 is promising for industrial acetoin production from glycerol.

Molecular mechanism of GylR-mediated regulation of glycerol metabolism in Streptomyces clavuligerus NRRL 3585.

Glycerol is a readily available and low-cost simple polyol compound, which can be used as a carbon source for microorganisms to produce various value-added products. Understanding the underlying regulatory mechanism in glycerol metabolism is critical for making better use of glycerol for diverse applications. In a few reported Streptomyces strains, the glycerol utilization gene cluster (glp operon) was shown to be regulated by the IclR family transcriptional regulator GylR. However, the molecular regulatory mechanism mediated by GylR has not been fully elucidated. In this study, we first analyzed the available Actinobacteria genomes in the NCBI Genome database, and found that the glp operon-like gene clusters are conserved in Streptomyces and several other genera of Actinobacteria. By taking Streptomyces clavuligerus NRRL 3585 as a model system, we identified that GylR represses the expressions of glp operon and gylR by directly binding to their promoter regions. Both glycerol-3-phosphate and dihydroxyacetone phosphate can induce the dissociation of GylR from its binding sequences. Furthermore, we identified a minimal essential operator site (a palindromic 18-bp sequence) of GylR-like regulators in Streptomyces. Our study for the first time reported the binding sequences and effector molecules of GylR-like proteins in Streptomyces. The molecular regulatory mechanism mediated by GylR presumably exists widely in Streptomyces. Our findings would facilitate the design of glycerol utilization pathways for producing valuable products. Moreover, our study provided new basic elements for the development of glycerol-inducible regulatory tools for synthetic biology research in the future.

Enhancing erythritol production from crude glycerol in a wild-type Yarrowia lipolytica by metabolic engineering.

Background: Erythritol is a zero-calorie sweetener that is widely used in the food, pharmaceutical, and medical industries. Crude glycerol is the main by-product of biodiesel, and the effective utilization of crude glycerol will help to improve biodiesel viability. Previous studies on the production of erythritol from Y. lipolytica using crude glycerol as a carbon source have focused on optimizing the fermentation process of the mutant Y. lipolytica Wratislavia K1, while metabolic engineering has not been successfully applied. Results: To this end, we engineered the yeast Y. lipolytica to increase the productivity of this strain. Wild strains tolerant to high concentrations of crude glycerol were screened and identified. A series of rational metabolic approaches were employed to improve erythritol production. Among them, the engineered strain Y-04, obtained by tandem overexpression of GUT1 and GUT2, significantly increased glycerol assimilation by 33.3%, which was consistent with the results of RT-qPCR analysis. The effects of tandem overexpression of GUT1, GUT2, TKL1, and TAL1 on erythritol synthesis were also evaluated. The best results were obtained using a mutant that overexpressed GUT1, GUT2, and TKL1 and knocked out EYD1. The final Y-11 strain produced 150 g/l erythritol in a 5-L bioreactor with a yield and productivity of 0.62 g/g and 1.25 g/l/h, respectively. To the best of our knowledge, this is the highest erythritol yield and productivity from crude glycerol ever reported in Y. lipolytica. Conclusion: This work demonstrated that overexpression of GUT1, GUT2, and TKL1 and knockdown of EYD1 could be used to improve crude glycerol utilization and erythritol synthesis in Y. lipolytica. The process parameters such as erythritol yield and productivity were significantly elevated, which is valuable for industrial applications. Crude glycerol, as a carbon source, could efficiently restrict the synthesis of by-products while enhancing the generation of erythritol, compared to glucose. This indicates considerable potential for synthesizing value-added products from crude glycerol by Y. lipolytica.

Adaptations of endolithic communities to abrupt environmental changes in a hyper-arid desert

The adaptation mechanisms of microbial communities to natural perturbations remain unexplored, particularly in extreme environments. The extremophilic communities of halite (NaCl) nodules from the hyper-arid core of the Atacama Desert are self-sustained and represent a unique opportunity to study functional adaptations and community dynamics with changing environmental conditions. We transplanted halite nodules to different sites in the desert and investigated how their taxonomic, cellular, and biochemical changes correlated with water availability, using environmental data modeling and metagenomic analyses. Salt-in strategists, mainly represented by haloarchaea, significantly increased in relative abundance at sites characterized by extreme dryness, multiple wet/dry cycles, and colder conditions. The functional analysis of metagenome-assembled genomes (MAGs) revealed site-specific enrichments in archaeal MAGs encoding for the uptake of various compatible solutes and for glycerol utilization. These findings suggest that opportunistic salt-in strategists took over the halite communities at the driest sites. They most likely benefited from compounds newly released in the environment by the death of microorganisms least adapted to the new conditions. The observed changes were consistent with the need to maximize cellular bioenergetics when confronted with lower water availability and higher salinity, providing valuable information on microbial community adaptations and resilience to climate change.

Metabolic pathways and ΔpH regulation in Escherichia coli during the fermentation of glucose and glycerol in the presence of formate at pH 6.5: the role of FhlA transcriptional activator.

Escherichia coli is able to ferment mixed carbon sources and produce various fermentation end-products. In this study, the function of FhlA protein in the specific growth rate (µ), metabolism, regulation of ΔpH and proton ATPase activity was investigated. Reduced µ in fhlA mutant of ∼25% was shown, suggesting the role of FhlA in the growth process. The utilization rate of glycerol is decreased in fhlA ∼ 2 fold, depending on the oxidation-reduction potential values. Bacteria regulate the activity of hydrogenase enzymes during growth depending on the external pH, which manifests as a lack of hydrogen gas generation during glycerol utilization at pH values below 5.9. It is suggested that cells maintain ΔpH during the fermentative growth via formate-lactate-succinate exchange. The decrement of the value of pHin, but not of pHex in mutant cells, is regulating ΔpH and consequently proton motive force generation.

Polyhydroxyalkanoates (PHAs) Production from Residual Glycerol by Wild Type Cupriavidus necator

PurposeThe main objective of the present study is to assess and improve fermentation conditions to produce polyhydroxyalkanoates (PHAs), such as the homopolymer poly(3-hydroxybutyrate) (PHB) and the copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), from renewable and inexpensive industrial waste by wild type Cupriavidus necator.MethodsUntreated residual glycerol, a byproduct generated by biodiesel industry, was used as the main carbon source by wild type C. necator. The bacterial adaptation stage, the glycerol concentration and other medium components were adjusted to enhance the PHB production. Fed batch fermentation with a pulse feeding strategy of propionic acid solution was employed for PHBV production.ResultsC. necator, previously adapted to glycerol utilization, was able to accumulate PHB up to 4.1 g/l under optimal substrate concentration conditions. The bacterium could also synthesize up to 3.4 g/l PHBV, with a mole fraction of 3-hydroxyvalerate (3HV) up to 12% by fed-batch fermentation with a pulsed propionic acid feed strategy.ConclusionThis is the first report on the use of residual glycerol as ideal feedstock for PHA production by wild type C. necator strain.Graphical Abstract

Glycerol conversion by aerobic granular sludge

Glycerol is abundantly present in wastewater from industries such as biodiesel production facilities. Glycerol is also a potential carbon source for microbes that are involved in wastewater nutrient removal processes. The conversion of glycerol in biological phosphorus removal of aerobic granular sludge processes has not been explored to date. The current study describes glycerol utilization by aerobic granular sludge and enhanced biological phosphorus removal (EBPR). Robust granules with good phosphorus removal capabilities were formed in an aerobic granular sludge sequencing batch reactor fed with glycerol. The interaction between the fermentative conversion of glycerol and product uptake by polyphosphate accumulating organisms (PAO) was studied using stoichiometric and microbial community analysis. Metagenomic, metaproteomic and microscopic analysis identified a community dominated by Actinobacteria (Tessaracoccus and Micropruina) and a typical PAO known as Ca. Accumulibacter. Glycerol uptake facilitator (glpF) and glycerol kinase (glpK), two proteins involved in the transport of glycerol into the cellular metabolism, were only observed in the genome of the Actinobacteria. The anaerobic conversion appeared to be a combination of a substrate fermentation and product uptake-type reaction. Initially, glycerol fermentation led mainly to the production of 1,3-propanediol (1,3-PDO) which was not taken up under anaerobic conditions. Despite the aerobic conversion of 1,3-PDO stable granulation was observed. Over time, 1,3-PDO production decreased and complete anaerobic COD uptake was observed. The results demonstrate that glycerol-containing wastewater can effectively be treated by the aerobic granular sludge process and that fermentative and polyphosphate accumulating organisms can form a food chain in glycerol-based EBPR processes.

Improving glycerol utilization during high-temperature xylitol production with Kluyveromyces marxianus using a transient clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 system

Glycerol is an ideal co-substrate for xylitol production with Kluyveromyces marxianus. This study demonstrated that K. marxianus catabolizes glycerol through the Gut1-Gut2 pathway instead of the previously speculated NADPH-dependent Gcy1-Dak1 pathway using the transient clustered regularly interspaced short palindromic repeats/ CRISPR-associated protein 9 (CRISPR/Cas9) system. Additionally, Utr1p was demonstrated to mediate NADPH generation through NADH phosphorylation. YZB392, which was constructed by integrating Utr1 into the Ypr1 site in the strain overexpressing NcXyl1 and CiGxf1 and harboring disrupted Xyl2, exhibited enhanced glycerol utilization for xylitol production (from 2.50- to 3.30- g/L after consuming 1 g/L glycerol). Fed-batch fermentation at 42 °C with YZB392 yielded 322.07 g/L xylitol, which is the highest known xylitol titer obtained via biological method. Feeding crude glycerol, xylose mother liquor, and corn steep liquor powder into a bioreactor resulted in the production of 235.69 g/L xylitol. This study developed a platform for xylitol production from industrial by-products.

Engineering synthetic microbial consortium for cadaverine biosynthesis from glycerol.

1,5-pentanediamine (cadaverine) is a C5 platform chemical, also an important raw material for bio-polyamide PA5X. With increasing concerns about the depletion of fossil resources and global environmental protection, cadaverine bio-production has attracted more attentions. Here, a microbial consortium consisting of Corynebacterium glutamicum cgl-FDK and Escherichia coli BL-ABST-Spy was constructed to de novo synthesize cadaverine utilizing glycerol as the sole carbon resource. The glycerol utilization pathway was initially constructed in C. glutamicum cgl-FDK to produce lysine from glycerol. Then, the pyridoxal 5'-phosphate (PLP) biosynthesis pathway and SpyTag/SpyCatcher protein-ligation system for lysine decarboxylase (CadA) and cadaverine-lysine antiporter protein (CadB) were introduced into E. coli BL-ABST-Spy to synthesize cadaverine from lysine. Furthermore, the fermentation conditions of microbial consortium were optimized and the cadaverine production reached 9.3g/L with glycerol as the sole carbon source. This work provides a promising strategy for efficiently producing cadaverine from glycerol with an artificial microbial consortium.

A comprehensive review on catalytic etherification of glycerol to value-added products

Abstract The increase in biodiesel production has resulted in the oversupply of glycerol into the market. Purified and processed glycerol has found many direct applications in pharmaceuticals, food, etc. However, the cost of processing and market value of processed glycerol has driven the research of direct utilization of crude glycerol to industrially essential chemicals. Various methods and research have been devoted to using glycerol to produce value-added products separately. Glycerol can undergo several transformation reactions like hydrogenation, oxidation, alcoholysis, and etherification. Etherification of glycerol can be divided into three main reactions: self-etherification, using alcohol, and olefins and these products have vast applications such as fuel additives, plasticizer, etc. The current review presents a comprehensive summary of glycerol etherification to value-added products and their applications. The catalytic system developed along with reaction conditions and the factors responsible for the better activity is also discussed. Overall, the review presents a detailed discussion on the catalytic system developed, the utilization of different alcohols and olefins, and the application of products. Moreover, the environmental and economic aspects of the etherification of glycerol via various conversion routes while assessing the process parameters needs to be tackled to attain wider adoption of the process.

The influence of hydrogen production on the formation of metabolic pathways and regulation of ΔpH in Escherichia coli

Hydrogen (H2) metabolism in Escherichia coli occurs via reversible membrane-associated hydrogenase enzymes (Hyd). Hyd-3 and Hyd-4 with formate dehydrogenase H (FDH-H) form formate hydrogen lyase complexes. The changes of metabolic pathways and ΔpH (pHin-pHex) regulation during fermentation of glucose, glycerol and formate in non H2-producing hypF (lack of all Hyds) and fdhF (lack of FDH-H) mutants at pH 7.5 were investigated. It was shown that specific growth rate was higher by ∼23% in hypF and fdhF, compared to wild type (wt), suggesting the negative effect of H2 on bacterial growth. Moreover, it was shown that H2 generation did not have a vital role in glucose and glycerol utilization rate at 0–72 h. The utilization of external formate was detected in wt (∼2.6 mM) and hypF (∼0.68 mM), but not in fdhF, due to the absence of enzyme responsible for formate metabolism. Nevertheless, the changes in ΔpH were not evident at 3 h. The ratio of generated end-products and regulation of ΔpH at late log (6 h) and exponential phase (24, 72 h) were various in hypF and fdhF due to formate disproportionation in hypF and proton generation, therewith absence of H2 generation. Taken together it can be concluded that bacteria regulate generation of fermentation end-products via balancing the concentration of acids and ethanol to maintain ΔpH and redox potential values. The results obtained are important for development and regulation of H2 production technology when applying mixed carbon sources.