Diol Products Research Articles

Metabolic Engineering and Regulation of Diol Biosynthesis from Renewable Biomass in Escherichia coli.

As bulk chemicals, diols have wide applications in many fields, such as clothing, biofuels, food, surfactant and cosmetics. The traditional chemical synthesis of diols consumes numerous non-renewable energy resources and leads to environmental pollution. Green biosynthesis has emerged as an alternative method to produce diols. Escherichia coli as an ideal microbial factory has been engineered to biosynthesize diols from carbon sources. Here, we comprehensively summarized the biosynthetic pathways of diols from renewable biomass in E. coli and discussed the metabolic-engineering strategies that could enhance the production of diols, including the optimization of biosynthetic pathways, improvement of cofactor supplementation, and reprogramming of the metabolic network. We then investigated the dynamic regulation by multiple control modules to balance the growth and production, so as to direct carbon sources for diol production. Finally, we proposed the challenges in the diol-biosynthesis process and suggested some potential methods to improve the diol-producing ability of the host.

Allosteric Regulation of the Soluble Epoxide Hydrolase by Nitro Fatty Acids: a Combined Experimental and Computational Approach

The human soluble epoxide hydrolase (hsEH) is a key regulator of epoxy fatty acid (EpFA) metabolism. Inhibition of sEH can maintain endogenous levels of beneficial EpFAs and reduce the levels of their corresponding diol products, thus ameliorating a variety of pathological conditions including cardiovascular, central nervous system and metabolic diseases. The quest for orthosteric drugs that bind directly to the catalytic crevice of hsEH has been prolonged and sustained over the past decades, but the disappointing outcome of clinical trials to date warrants alternative pharmacological approaches. Previously, we have shown that hsEH can be allosterically inhibited by the endogenous electrophilic lipid 15-deoxy-Δ12,14-Prostaglandin-J2, via covalent adduction to two cysteines, C423 and C522. In this study, we explore the properties and behaviour of three electrophilic lipids belonging to the class of the nitro fatty acids, namely 9- and 10-nitrooleate and 10-nitrolinoleate. Biochemical and biophysical investigations revealed that, in addition to C423 and C522, nitro fatty acids can covalently bind to additional nucleophilic residues in hsEH C-terminal domain (CTD), two of which predicted in this study to be latent allosteric sites. Systematic mapping of the protein mutational space and evaluation of possible propagation pathways delineated selected residues, both in the allosteric patches and in other regions of the enzyme, envisaged to play a role in allosteric signalling. The responses elicited by the ligands on the covalent adduction sites supports future fragment-based design studies of new allosteric effectors for hsEH with increased efficacy and selectivity.

Biosynthesizing structurally diverse diols via a general route combining oxidative and reductive formations of OH-groups

Diols encompass important bulk and fine chemicals for the chemical, pharmaceutical and cosmetic industries. During the past decades, biological production of C3-C5 diols from renewable feedstocks has received great interest. Here, we elaborate a general principle for effectively synthesizing structurally diverse diols by expanding amino acid metabolism. Specifically, we propose to combine oxidative and reductive formations of hydroxyl groups from amino acids in a thermodynamically favorable order of four reactions catalyzed by amino acid hydroxylase, L-amino acid deaminase, α-keto acid decarboxylase and aldehyde reductase consecutively. The oxidative formation of hydroxyl group from an alkyl group is energetically more attractive than the reductive pathway, which is exclusively used in the synthetic pathways of diols reported so far. We demonstrate this general route for microbial production of branched-chain diols in E. coli. Ten C3-C5 diols are synthesized. Six of them, namely isopentyldiol (IPDO), 2-methyl-1,3-butanediol (2-M-1,3-BDO), 2-methyl-1,4-butanediol (2-M-1,4-BDO), 2-methyl-1,3-propanediol (MPO), 2-ethyl-1,3-propanediol (2-E-1,3-PDO), 1,4-pentanediol (1,4-PTD), have not been biologically synthesized before. This work opens up opportunities for synthesizing structurally diverse diols and triols, especially by genome mining, rational design or directed evolution of proper enzymes.

Process Design and Comprehensive 3e Analysis of Low Carbon Diol Production with and Without Heat Integration

Process Design and Comprehensive 3e Analysis of Low Carbon Diol Production with and Without Heat Integration

Spongy titanosilicate promotes the catalytic performance and reusability of WO3 in oxidative cleavage of methyl oleate.

A tungsten containing catalyst catalyzed oxidative cleavage of methyl oleate (MO) by employing H2O2 as an oxidant and is known as an efficient approach for preparing high value-added chemicals, however, the tungsten leaching problem remains unresolved. In this work, a binary catalyst consisting of tungsten oxide (WO3) and spongy titanosilicate (STS) zeolite is proposed for MO oxidative cleavage. The function of STS in this catalyst is investigated. On the one hand, STS converts MO to 9,10-epoxystearate (MES), which further forms nonyl aldehyde (NA) and methyl azelaaldehydate (MAA) with the catalysis of WO3. In this way, MO oxidation and hydrolysis that generates unwanted diol product 9,10-dihydroxystearate (MDS) decreases obviously. On the other hand, STS decomposes peroxide and promotes the conversion of soluble peroxotungstate to insoluble polytungstate. Meanwhile, these tungsten species are allowed to precipitate on its surface instead of remaining in the liquid phase owing to its relative large specific area. Therefore, tungsten leaching can be reduced from 37.0% to 1.2%. Due to the cooperation of WO3 and STS, 94.4% MO conversion and oxidative cleavage product selectivity of 63.1% are achieved, and the WO3–STS binary catalyst maintains excellent catalytic performance for 8 recycling reactions.

When metabolic prowess is too much of a good thing: how carbon catabolite repression and metabolic versatility impede production of esterified \u03b1,\u03c9-diols in Pseudomonas putida KT2440

BackgroundMedium-chain-length α,ω-diols (mcl-diols) are important building blocks in polymer production. Recently, microbial mcl-diol production from alkanes was achieved in E. coli (albeit at low rates) using the alkane monooxygenase system AlkBGTL and esterification module Atf1. Owing to its remarkable versatility and conversion capabilities and hence potential for enabling an economically viable process, we assessed whether the industrially robust P. putida can be a suitable production organism of mcl-diols.ResultsAlkBGTL and Atf1 were successfully expressed as was shown by oxidation of alkanes to alkanols, and esterification to alkyl acetates. However, the conversion rate was lower than that by E. coli, and not fully to diols. The conversion was improved by using citrate instead of glucose as energy source, indicating that carbon catabolite repression plays a role. By overexpressing the activator of AlkBGTL-Atf1, AlkS and deleting Crc or CyoB, key genes in carbon catabolite repression of P. putida increased diacetoxyhexane production by 76% and 65%, respectively. Removing Crc/Hfq attachment sites of mRNAs resulted in the highest diacetoxyhexane production. When the intermediate hexyl acetate was used as substrate, hexanol was detected. This indicated that P. putida expressed esterases, hampering accumulation of the corresponding esters and diesters. Sixteen putative esterase genes present in P. putida were screened and tested. Among them, Est12/K was proven to be the dominant one. Deletion of Est12/K halted hydrolysis of hexyl acetate and diacetoxyhexane. As a result of relieving catabolite repression and preventing the hydrolysis of ester, the optimal strain produced 3.7 mM hexyl acetate from hexane and 6.9 mM 6-hydroxy hexyl acetate and diacetoxyhexane from hexyl acetate, increased by 12.7- and 4.2-fold, respectively, as compared to the starting strain.ConclusionsThis study shows that the metabolic versatility of P. putida, and the associated carbon catabolite repression, can hinder production of diols and related esters. Growth on mcl-alcohol and diol esters could be prevented by deleting the dominant esterase. Carbon catabolite repression could be relieved by removing the Crc/Hfq attachment sites. This strategy can be used for efficient expression of other genes regulated by Crc/Hfq in Pseudomonas and related species to steer bioconversion processes.

Renewable synthesis of branched diols as polymer precursors from biomass-derived lactones and 2-pyrones

Hydric alcohols are important synthetic intermediates used to manufacture polymer, fine chemicals, and commodity products. Recent environmental problems due to fossil fuel-based non-degradable products have call for the advancement in research towards developing an alternate source that can sustainably substitute petroleum. In this direction, we have conceptualized a combined fermentation and catalytic process to obtain biorenewable branched diols from biomass. The significant yield of the desired diol products was obtained from different lignocellulose-based cyclic esters under optimized conditions. In order to know the universality of our process, saturated, partially saturated and unsaturated lactones were tested, which gave up to 99% conversion and (∼68–91%) diol’s yield. Finally, a plausible mechanism for 6-amyl-α-pyrone hydrogenation is proposed which is more likely to follow a pathway involving ring-opening reaction via partial hydrogenation of the double bond.

Species-Specific Differences in C-5 Sterol Desaturase Function Influence the Outcome of Azole Antifungal Exposure.

The azole antifungals inhibit sterol 14α-demethylase (S14DM), leading to depletion of cellular ergosterol and the synthesis of an aberrant sterol diol that disrupts membrane function. In Candida albicans, sterol diol production is catalyzed by the C-5 sterol desaturase enzyme encoded by ERG3. Accordingly, mutations that inactivate ERG3 enable the fungus to grow in the presence of the azoles. The purpose of this study was to compare the propensities of C-5 sterol desaturases from different fungal pathogens to produce the toxic diol upon S14DM inhibition and thus contribute to antifungal efficacy. The coding sequences of ERG3 homologs from C. albicans (CaERG3), Candida glabrata (CgERG3), Candida auris (CaurERG3), Cryptococcus neoformans (CnERG3), Aspergillus fumigatus (AfERG3A-C) and Rhizopus delemar (RdERG3A/B) were expressed in a C. albicans erg3Δ/Δ mutant to facilitate comparative analysis. All but one of the Erg3p-like proteins (AfErg3C) at least partially restored C-5 sterol desaturase activity and to corresponding degrees rescued the stress and hyphal growth defects of the C. albicans erg3Δ/Δ mutant, confirming functional equivalence. Each C-5 desaturase enzyme conferred markedly different responses to fluconazole exposure in terms of the MIC and residual growth observed at supra-MICs. Upon fluconazole-mediated inhibition of S14DM, the strains expressing each homolog also produced various levels of 14α-methylergosta-8,24(28)-dien-3β,6α-diol. The RdErg3A and AfErg3A proteins are notable for low levels of sterol diol production and failing to confer appreciable azole sensitivity upon the C. albicans erg3Δ/Δ mutant. These findings suggest that species-specific properties of C-5 sterol desaturase may be an important determinant of intrinsic azole sensitivity.

Facile pinacol coupling of aliphatic ketones by Brook rearrangement in the presence of samarium species

Herein we report a practical pinacol coupling reaction, in which ketones (aldehydes) react smoothly with Sm and TMSBr to afford the diol products with Sm(II) or (III) siliyl species generated in situ. This reported method affords poor yields for aromatic ketone substrates and good yields for aliphatic ketones. Therefore, it distinguishes from most reductive coupling approaches that are more effective for aromatic carbonyl compounds and provides a facile and robust approach for the pinacol coupling of aliphatic ketones. Mechanistic studies also indicated the pinacolization probably proceeded via an anionic instead of radical coupling pathway involving the Brook rearrangement in the presence of samarium (II or III) silyl species.

Recent Advancements in the Development of Osmium Catalysts for Various Oxidation Reactions: A New Era?

ABSTRACT Symmetric and asymmetric dihydroxylation reactions catalyzed by osmium have emerged as the most efficient route in the one-step synthesis of diols. This is because on an industrial scale 1,2-diols are prepared via a two-step procedure. Still, it is a challenge to prepare and apply osmium catalysts in dihydroxylation catalysis because of their safety and environmental implications; e.g., volatile, highly toxic, product contamination, etc. To date, few efforts were reported to comprehensively overcome these drawbacks with great success by heterogenizing to an insoluble matrix, including diminishment in catalytic performance. Hence, the need for efficacious solid-supported osmium catalysts to subdue these difficulties is a topic of great importance. This mini review provides a concise overview of some of the most encouraging solid-supported osmium catalysts reported over the last few years, which proved to be more efficient under mild conditions as opposed to other immobilized osmium catalytic systems. The summary discusses the latest developments in the synthetic heterogenizing and homogenization strategies, structure analysis and investigation toward symmetric and asymmetric dihydroxylation for diol production. Specific emphasis is focused on their mechanistic considerations, their reusability and improvement in their safety, and environmental implications.

A simple route for the synthesis of novel norcantharimide derivatives via acidolysis with hydrochloric acid(gas)

AbstractIn this work, seven new norcantharimide derivatives were synthesized by an acidolysis method. The compounds were prepared by acidolyzing trans‐1,4‐diacetate and trans‐1,2‐chloroacetate structures, which were obtained by stereospecific cleavage of the internal etheric bond of the tricyclic imides. The HCl(gas) was produced from the reaction of H2SO4 with NaCl. The resulting gas was bubbled into the reaction mixture. Trans‐1,4‐diacetate and trans‐1,2‐chloroacetate were thus acidolyzed, and the corresponding diol and halohydrin products were obtained respectively in moderate overall yields from low‐cost starting materials, using simple and easily scalable chemistry. The products were characterized by means of spectroscopic techniques. The synthesized compounds have high potential as anticancer agents and can be valuable for studies in this area.

Photo‐Induced Dihydroxylation of Alkenes with Diacetyl, Oxygen, and Water

AbstractHerein reported is a photo‐induced production of vicinal diols from alkenes under mild reaction conditions. The present dihydroxylation method using diacetyl (= butane‐2,3‐dione), oxygen, and water dispenses with toxic reagents and intractable waste generation.

Synthesis of a Divinyl-functionalized Diamantane-Analogue from naturally occurring myo-Inositol and its application to polymer synthesis via the Thiol-ene reaction

Myo-inositol, a naturally occurring cyclic compound obtained from rice bran, was transformed into a divinyl derivative bearing a rigid hexaoxadiamantane (HODAM) core composed of two orthoester moieties. The divinyl compound underwent radical polyaddition with dithiols to afford the corresponding semi-crystalline polythioethers, the melting temperatures (Tm) of which were above 240 °C. Furthermore, the divinyl compound was transformed into a HODAM-bearing diol via the radical addition of 2-mercaptoethanol to the vinyl groups. The diol product was used as a rigid monomer in polyurethane synthesis. The obtained polyurethanes exhibited glass transition temperatures (Tg) ranging from 55 to 110 °C.

Biocatalytic synthesis of non-vicinal aliphatic diols

Biocatalysts are receiving increased attention in the field of selective oxyfunctionalization of C-H bonds, with cytochrome P450 monooxygenases (CYP450s), and the related peroxygenases, leading the field. Here we report on the substrate promiscuity of CYP505A30, previously characterized as a fatty acid hydroxylase. In addition to its regioselective oxyfunctionalization of saturated fatty acids (ω-1 - ω-3 hydroxylation), primary fatty alcohols are also accepted with similar regioselectivities. Moreover, alkanes such as n-octane and n-decane are also readily accepted, allowing for the production of non-vicinal diols through sequential oxygenation.

Structure of the fungal hydroxylase, CYP505A30, and rational transfer of mutation data from CYP102A1 to alter regioselectivity

Regioselective oxyfunctionalisation ofn-alkanes and production of non-vicinal diols by evolved CYP505A30 through rational transfer of knowledge between protein scaffolds.

Recent advances in microbial biosynthesis of C3 – C5 diols: Genetics and process engineering approaches

Diols derived from renewable feedstocks have significant commercial interest in polymer, pharmaceutical, cosmetics, flavors and fragrances, food and feed industries. In C3-C5 diols biological processes of 1,3-propanediol, 1,2-propanediol and 2,3-butanediol have been commercialized as other isomers are non-natural metabolites and lack natural biosynthetic pathways. However, the developments in the field of systems and synthetic biology paved a new path to learn, build, construct, and test for efficient chassis strains. The current review addresses the recent advancements in metabolic engineering, construction of novel pathways, process developments aimed at enhancing in production of C3-C5 diols. The requisites on developing an efficient and sustainable commercial bioprocess for C3-C5 diols were also discussed.

A silicododecamolybdate/pyridinium-tetrazole hybrid molecular salt as a catalyst for the epoxidation of bio-derived olefins

The hybrid polyoxometalate (POM) salt (Hptz)4[SiMo12O40]∙nH2O (1) (ptz = 5-(2-pyridyl)tetrazole) has been prepared, characterized by X-ray crystallography, and examined as a catalyst for the epoxidation of cis-cyclooctene (Cy) and bio-derived olefins, namely dl-limonene (Lim; a naturally occurring monoterpene found in the rinds of citrus fruits), methyl oleate and methyl linoleate (fatty acid methyl esters (FAMEs) obtained by transesterification of vegetable oils). The crystal structure of 1 consists of α-Keggin-type heteropolyanions, [SiMo12O40]4-, surrounded by space-filling and charge-balancing 2-(tetrazol-5-yl)pyridinium (Hptz+) cations, as well as by a large number of water molecules of crystallization (n = 9). The water molecules mediate an extensive three-dimensional (3D) hydrogen-bonding network involving the inorganic anions and organic cations. For the epoxidation of the model substrate Cy in a nonaqueous system (tert-butylhydroperoxide as oxidant), the catalytic performance of 1 (100% epoxide yield at 24 h, 70 °C) was superior to that of the tetrabutylammonium salt (Bu4N)4[SiMo12O40] (2) (63% epoxide yield at 24 h), illustrating the role of the counterion Hptz+ in enhancing catalytic activity. The hybrid salt 1 was effective for the epoxidation of Lim (69%/85% conversion at 6 h/24 h) and the FAMEs (87–88%/100% conversion at 6 h/24 h), leading to useful bio-based products (epoxides, diepoxides and diol products).

Microbial upgrading of acetate into 2,3-butanediol and acetoin by E. coli W

BackgroundAcetate is an abundant carbon source and its use as an alternative feedstock has great potential for the production of fuel and platform chemicals. Acetoin and 2,3-butanediol represent two of these potential platform chemicals.ResultsThe aim of this study was to produce 2,3-butanediol and acetoin from acetate in Escherichia coli W. The key strategies to achieve this goal were: strain engineering, in detail the deletion of mixed-acid fermentation pathways E. coli W ΔldhA ΔadhE Δpta ΔfrdA 445_Ediss and the development of a new defined medium containing five amino acids and seven vitamins. Stepwise reduction of the media additives further revealed that diol production from acetate is mediated by the availability of aspartate. Other amino acids or TCA cycle intermediates did not enable growth on acetate. Cultivation under controlled conditions in batch and pulsed fed-batch experiments showed that aspartate was consumed before acetate, indicating that co-utilization is not a prerequisite for diol production. The addition of aspartate gave cultures a start-kick and was not required for feeding. Pulsed fed-batches resulted in the production of 1.43 g l−1 from aspartate and acetate and 1.16 g l−1 diols (2,3-butanediol and acetoin) from acetate alone. The yield reached 0.09 g diols per g acetate, which accounts for 26% of the theoretical maximum.ConclusionThis study for the first time showed acetoin and 2,3-butanediol production from acetate as well as the use of chemically defined medium for product formation from acetate in E. coli. Hereby, we provide a solid base for process intensification and the investigation of other potential products.

Co2(CO)8-Catalyzed Reactions of Acetals or Lactones with Hydrosilanes and Carbon Monoxide. A New Access to the Preparation of 1,2-Diol Derivatives through Siloxymethylation

Abstract The Co2(CO)8-catalyzed reaction of acetals with hydrosilanes and CO under mild reaction conditions (an ambient temperature under an ambient CO pressure), leading to the production of vicinal diols is reported. A siloxymethyl group can be introduced via the cleavage of one of two alkoxy groups in the acetal. The effects of the types of hydrosilanes, acetals, solvents, and reaction temperatures on the yield of siloxymethylation products were examined in detail. The reactivity for hydrosilanes is as follows; HSiMe3 > HSiEtMe2 > HSiEt2Me > HSiEt3. Hemiacetal esters are more reactive than dimethyl acetals. The polarity of the solvent used also has a significant effect on both the course of the reaction as well as the reaction rate. The site-selective siloxymethylation can be achieved in the case of cyclic acetals such as tetrahydrofuran (THF) and tetrahydropyrane (THP) derivatives, depending on the nature of the oxygen substituent attached adjacent to the oxygen atom in the ring. When 2-alkoxy THF or THP derivatives are used as substrates, the siloxymethylation takes place with cleavage of the ring C-O bond. In contrast, the reaction of 2-acetoxy THF or THP derivatives results in siloxymethylation with the cleavage of C-OAc bond. The ring-opening siloxymethylation of lactones was also examined.

3-25 高密度炭素担持Cu微粒子触媒を用いたエリスリトールの水素化脱酸素によるC4ジオール合成

3-25 高密度炭素担持Cu微粒子触媒を用いたエリスリトールの水素化脱酸素によるC4ジオール合成