ABSTRACT
Aliphatic polyesters are the most common type of biodegradable synthetic polymer used in many pharmaceutical applications nowadays. This report describes the ring-opening polymerization (ROP) of l-lactide (L-LA), ε-caprolactone (CL) and glycolide (Gly) in the presence of a simple, inexpensive and convenient PEG200-BiOct3 catalytic system. The chemical structures of the obtained copolymers were characterized by 1H- or 13C-NMR. GPC was used to estimate the average molecular weight of the resulting polyesters, whereas TGA and DSC were employed to determine the thermal properties of polymeric products. The effects of temperature, reaction time, and catalyst content on the polymerization process were investigated. Importantly, the obtained polyesters were not cyto- or genotoxic, which is significant in terms of the potential for medical applications (e.g., for drug delivery systems). As a result of transesterification, the copolymers obtained had a random distribution of comonomer units along the polymer chain. The thermal analysis indicated an amorphous nature of poly(l-lactide-co-ε-caprolactone) (PLACL) and a low degree of crystallinity of poly(ε-caprolactone-co-glycolide) (PCLGA, Xc = 15.1%), in accordance with the microstructures with random distributions and short sequences of comonomer units (l = 1.02-2.82). Significant differences in reactivity were observed among comonomers, confirming preferential ring opening of L-LA during the copolymerization process.
Subject(s)
Bismuth/chemistry , Caproates/chemistry , Dioxanes/chemistry , Lactones/chemistry , Polyglycolic Acid/chemistry , Polymerization , Caproates/chemical synthesis , Catalysis , Dioxanes/chemical synthesis , Lactones/chemical synthesis , Polyesters/chemical synthesis , Polyesters/chemistry , Polyglycolic Acid/chemical synthesis , TemperatureABSTRACT
Due to a unique mechanism that limits the possibility of hypoglycemia, the free fatty acid receptor (FFA1) is an attractive target for the treatment of type 2 diabetes. So far, however, none of the promising agonists have been able to enter the market. The most advanced clinical candidate, TAK-875, was withdrawn from phase III clinical trials due to liver safety issues. In this article, we describe the key aspects leading to the discovery of CPL207280 (13), the design of which focused on long-term safety. The introduction of small, nature-inspired acyclic structural fragments resulted in compounds with retained high potency and a satisfactory pharmacokinetic profile. Optimized synthesis and upscaling provided a stable, solid form of CPL207280-51 (45) with the properties required for the toxicology studies and ongoing clinical trials.
Subject(s)
Caproates/pharmacology , Drug Development , Receptors, G-Protein-Coupled/agonists , Animals , Caproates/chemical synthesis , Caproates/chemistry , Dose-Response Relationship, Drug , Humans , Molecular Structure , Rats , Rats, Wistar , Structure-Activity RelationshipABSTRACT
Aldo-keto reductase KmAKR-catalyzed asymmetric reduction offers a green approach to produce dichiral diol tert-butyl 6-substituted-(3R,5R/S)-dihydroxyhexanoates, which are important building blocks of statins. In our previous work, we cloned a novel gene of NADPH-specific aldo-keto reductase KmAKR (WT) from a thermotolerant yeast Kluyveromyces marxianus ZJB14056 and a mutant KmAKR-W297H/Y296W/K29H (Variant III) has been constructed and displayed strict diastereoselectivity towards tert-butyl 6-cyano-(5R)-hydroxy-3-oxohexanoate ((5R)-1) but moderate activity and stability. Herein, to further co-evolve its activity and thermostability, we performed semi-rational engineering of Variant III by using a combinational screening strategy, consisting of tertiary structure analysis, loop engineering, and alanine scanning. As results, the "best" variant KmAKR-W297H/Y296W/K29H/Y28A/T63M (Variant VI) was acquired, whose Km, kcat/Km towards (5R)-1 was 0.66â¯mM and 210.77â¯s-1 mM-1, respectively, with improved thermostability (half-life of 14.13â¯h at 40⯰C). Combined with 1.5â¯g dry cell weight (DCW) L-1Exiguobacterium sibiricum glucose dehydrogenase (EsGDH) for NADPH regeneration, 4.5â¯g DCW L-1Variant VI completely reduced (5R)-1 of up to 450â¯g L-1 within 7.0â¯h at 40⯰C, yielding the corresponding optically pure tert-butyl 6-cyano-(3R,5R)-dihydroxyhexanoate ((3R,5R)-3, >99.5% d.e.p) with a space-time yield (STY) of 1.24â¯kg L-1 day-1, and this was the highest level documented in literatures so far on substrate loading and STY of producing (3R,5R)-3. Besides (5R)-1, Variant VI displayed strong activity on tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate ((5S)-2). 4.5â¯g DCW L-1Variant VI completely reduced 400â¯g L-1 (5S)-2, within 5.0â¯h at 40⯰C, yielding optically pure tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate ((3R,5S)-4, >99.5% d.e.p) with a STY of 1.34â¯kg L-1 day-1. In summary, Variant VI displayed industrial application potential in statins biomanufacturing.
Subject(s)
Aldo-Keto Reductases/chemistry , Caproates/chemical synthesis , Fungal Proteins/chemistry , Aldo-Keto Reductases/genetics , Enzyme Stability , Exiguobacterium/enzymology , Fungal Proteins/genetics , Glucose 1-Dehydrogenase/chemistry , Kluyveromyces/enzymology , Molecular Docking Simulation , Molecular Dynamics Simulation , Mutation , Protein Engineering , TemperatureABSTRACT
Catalytic processes in protecting-group-free syntheses of natural products are fast emerging towards achieving the goal of efficiency and economy in total synthesis. Present day sustainable development in synthesis of natural products does not permit the luxury of using stoichiometric reagents and protecting groups. Catalysis and step-economy can contribute significantly toward economy and efficiency of synthesis. This feature article details the ingenious efforts by many researchers in the last couple of years toward concise total syntheses, based on catalytic steps and protecting-group-free-strategies. These would again serve as guidelines in future development of reagents and catalysts aimed at achieving higher efficiency and chemoselectivity to the point that catalysis and protecting-group-free synthesis will be an accepted common practice.
Subject(s)
Biological Products/chemistry , Metals/chemistry , Alkylation , Berberine Alkaloids/chemical synthesis , Berberine Alkaloids/chemistry , Biological Products/chemical synthesis , Caproates/chemical synthesis , Caproates/chemistry , Catalysis , Diterpenes/chemical synthesis , Diterpenes/chemistry , Imidazolidines/chemical synthesis , Imidazolidines/chemistry , Oxidation-Reduction , Quinones/chemical synthesis , Quinones/chemistry , StereoisomerismABSTRACT
Here, we describe innovative synthesis of well-defined biocompatible N-(2-hydroxypropyl) methacrylamide (HPMA)-based polymer carriers and their drug conjugates with pirarubicin intended for controlled drug delivery and pH-triggered drug activation in tumor tissue. Polymer carrier synthesis was optimized to obtain well-defined linear HPMA-based polymer precursor with dispersity close to 1 and molar mass close to renal threshold with minimal synthesis steps. The developed synthesis enables preparation of tailored polymer nanomedicines with highly enhanced biological behavior in vivo, especially the biodistribution, urine elimination, tumor accumulation and anticancer activity. STATEMENT OF SIGNIFICANCE: The manuscript reports on novel synthesis and detailed physicochemical characterization and in vivo evaluation of well-defined biocompatible hydrophilic copolymers based on N-(2-hydroxypropyl)methacrylamide (HPMA) and their drug conjugates with pirarubicin enabling controlled drug delivery and pH-triggered drug activation in tumor tissue. Polymer carrier synthesis was optimized to obtain well-defined linear HPMA-based polymer precursor with minimal synthesis steps using controlled polymerization. Compared to previously published HPMA-based polymer drug conjugates whose polymer carriers were prepared by classical route via free radical polymerization, the newly prepared polymer drug conjugates exhibited enhanced biological behavior in vivo, especially the prolonged blood circulation, urine elimination, tumor accumulation and excellent anticancer activity. We believe that the newly prepared well-defined polymer conjugates could significantly enhance tumor therapy in humans.
Subject(s)
Acrylamides/therapeutic use , Antineoplastic Agents/therapeutic use , Doxorubicin/analogs & derivatives , Sarcoma, Experimental/drug therapy , Acrylamides/chemical synthesis , Acrylamides/pharmacokinetics , Animals , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/pharmacokinetics , Caproates/chemical synthesis , Caproates/pharmacokinetics , Caproates/therapeutic use , Cell Line, Tumor , Doxorubicin/chemical synthesis , Doxorubicin/pharmacokinetics , Doxorubicin/therapeutic use , Drug Delivery Systems , Mice , Nanomedicine/methods , PolymerizationABSTRACT
tert-Butyl (3R,5S)-6-chloro-3,5-dihydroxyhexanoate [(3R,5S)-CDHH] is the key chiral intermediate to synthesize the side chain of the lipid-lowering drug rosuvastatin. Carbonyl reductases showed excellent activity for the biosynthesis of (3R,5S)-CDHH. The requirement of cofactor NADH/NADPH leads to high cost for the industrial application of carbonyl reductases. In this study, a self-sufficient biocatalyst based on carbonyl reductase and NADP+ co-immobilization strategy was developed on an amino resin carrier LX-1000HAA (SCR-NADP+@LX-1000HAA). The self-sufficient biocatalyst achieved in situ cofactor regeneration and showed the activity recovery of 77.93% and the specific activity of 70.45 U/g. Asymmetric synthesis of (3R,5S)-CDHH using SCR-NADP+@LX-1000HAA showed high enantioselectivity (> 99% e.e.) and yield (98.54%). Batch reactions were performed for ten cycles without extra addition of NADP+, and the total yield of (3R,5S)-CDHH achieved at 10.56 g/g biocatalyst. The present work demonstrated the potential of the self-sufficient biocatalyst for the asymmetric biosynthesis of rosuvastatin intermediate.
Subject(s)
Alcohol Oxidoreductases/chemistry , Biocatalysis , Candida/enzymology , Caproates/chemical synthesis , Enzymes, Immobilized/chemistry , Fungal Proteins/chemistry , NADP/chemistry , Alcohol Oxidoreductases/genetics , Candida/genetics , Caproates/chemistry , Enzymes, Immobilized/genetics , Fungal Proteins/genetics , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Rosuvastatin Calcium/chemical synthesis , Rosuvastatin Calcium/chemistryABSTRACT
A novel and environmentally-friendly procedure for the preparation of polymer inclusion membranes (PIMs) containing an ionic liquid is presented for the first time. Traditionally, PIMs are prepared by a solvent casting method with the use of harmful organic solvents. Here we report a new solvent-free procedure based on a thermal-compression technique which involve the melting of the components of the PIM and the application of a high pressure to the melted specimen to form a flat-sheet film. In our study, we have tested different polymers, such as two cellulose derivatives as well as two thermoplastic polymers, polyurethane (TPU) and poli ε-caprolactone (PCL). The ionic liquid (IL) trioctylmethylammonium chloride (Aliquat 336) has been used to produce PIMs with a fixed composition of 70% polymer-30% IL (w/w). Both TPU and PCL polymers provide successful membranes, which have been thoroughly characterized. PIMs based on the polymer PCL showed a high stability. To test whether the properties of the IL were affected by the preparation conditions, the extraction ability of Aliquat 336 was investigated for both PCL and TPU membranes in terms of Cr(VI) extraction. Satisfactory values (90% extraction) were obtained for both membranes tested, showing this novel procedure as a green alternative for the preparation of PIMs with ILs.
Subject(s)
Ionic Liquids/chemistry , Polymers/chemical synthesis , Caproates/chemical synthesis , Caproates/chemistry , Green Chemistry Technology , Lactones/chemical synthesis , Lactones/chemistry , Membranes, Artificial , Polymers/chemistry , Polyurethanes/chemical synthesis , Polyurethanes/chemistry , Quaternary Ammonium Compounds/chemistryABSTRACT
Multi-enzyme cascade reactions capture the essence of nature's efficiency by increasing the productivity of a process. Here we describe one such three-enzyme cascade for the synthesis of 6-hydroxyhexanoic acid. Whole cells of Escherichia coli co-expressing an alcohol dehydrogenase and a Baeyer-Villiger monooxygenase (CHMO) for internal cofactor regeneration were used without the supply of external NADPH or NADP+. The product inhibition caused by the ε-caprolactone formed by the CHMO was overcome by the use of lipase CAL-B for in situ conversion into 6-hydroxyhexanoic acid. A stirred tank reactor under fed-batch mode was chosen for efficient catalysis. By using this setup, a product titre of >20 g L-1 was achieved in a 500 mL scale with an isolated yield of 81% 6-hydroxyhexanoic acid.
Subject(s)
Alcohol Dehydrogenase/genetics , Caproates/chemical synthesis , Escherichia coli Proteins/genetics , Escherichia coli/enzymology , Fungal Proteins/chemistry , Hydroxy Acids/chemical synthesis , Lipase/chemistry , Mixed Function Oxygenases/genetics , Alcohol Dehydrogenase/metabolism , Batch Cell Culture Techniques , Biocatalysis , Bioreactors , Caproates/chemistry , Caproates/metabolism , Coenzymes/biosynthesis , Coenzymes/chemistry , Escherichia coli/genetics , Escherichia coli Proteins/metabolism , Fungal Proteins/metabolism , Gene Expression , Hydroxy Acids/metabolism , Kinetics , Lactones/chemistry , Lactones/metabolism , Lipase/metabolism , Mixed Function Oxygenases/metabolism , NADP/biosynthesis , NADP/chemistryABSTRACT
The synthesis, absolute stereochemical configuration, complete biological characterization, mechanism of action and resistance, and pharmacokinetic properties of ( S)-(-)-acidomycin are described. Acidomycin possesses promising antitubercular activity against a series of contemporary drug susceptible and drug-resistant M. tuberculosis strains (minimum inhibitory concentrations (MICs) = 0.096-6.2 µM) but is inactive against nontuberculosis mycobacteria and Gram-positive and Gram-negative pathogens (MICs > 1000 µM). Complementation studies with biotin biosynthetic pathway intermediates and subsequent biochemical studies confirmed acidomycin inhibits biotin synthesis with a Ki of approximately 1 µM through the competitive inhibition of biotin synthase (BioB) and also stimulates unproductive cleavage of S-adenosyl-l-methionine (SAM) to generate the toxic metabolite 5'-deoxyadenosine. Cell studies demonstrate acidomycin selectively accumulates in M. tuberculosis providing a mechanistic basis for the observed antibacterial activity. The development of spontaneous resistance by M. tuberculosis to acidomycin was difficult, and only low-level resistance to acidomycin was observed by overexpression of BioB. Collectively, the results provide a foundation to advance acidomycin and highlight BioB as a promising target.
Subject(s)
Antitubercular Agents/pharmacology , Bacterial Proteins/antagonists & inhibitors , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/enzymology , Sulfurtransferases/antagonists & inhibitors , Thiazolidines/pharmacology , Tuberculosis/microbiology , Animals , Antitubercular Agents/chemical synthesis , Antitubercular Agents/chemistry , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Biological Products/chemical synthesis , Biological Products/chemistry , Biological Products/pharmacology , Biotin/biosynthesis , Caproates/chemical synthesis , Caproates/chemistry , Caproates/pharmacology , Drug Resistance, Bacterial , Humans , Kinetics , Mice , Microbial Sensitivity Tests , Mycobacterium tuberculosis/chemistry , Mycobacterium tuberculosis/genetics , Sulfurtransferases/chemistry , Sulfurtransferases/genetics , Sulfurtransferases/metabolism , Thiazolidines/chemical synthesis , Thiazolidines/chemistry , Tuberculosis/drug therapyABSTRACT
A mutant L206F/P207F/L259F of Talaromyces thermophilus lipase (TTL) exhibited high hydrolytic activity towards 2-carboxyethyl-3-cyano-5-methylhexanoic acid ethyl ester (CNDE) for synthesis of (S)-2-carboxyethyl-3-cyano-5-methylhexanoic acid (S-CCMA), a key chiral intermediate of pregabalin. However, low conversion at high CNDE concentration and unreusability of the free TTL mutant restricted its industrial applications. In this study, the TTL mutant was immobilized onto epoxy resin and its catalytic properties for kinetic resolution of CNDE were investigated. Under the optimized conditions, the immobilized lipase exhibited an increased catalytic efficiency even at a CNDE concentration of 3 M with 49.7% conversion and 95% ee p. The conversion retained higher than 46.3% even after 10 times repeated use of the immobilized lipase in n-heptane-water biphasic system. These results demonstrated great potential of the immobilized TTL mutant for industrial production of the chiral intermediate of pregabalin.
Subject(s)
Biocatalysis , Enzymes, Immobilized/chemistry , Lipase/chemistry , Pregabalin/chemistry , Caproates/chemical synthesis , Caproates/chemistry , Catalysis , Ethyl Ethers/chemistry , Kinetics , Pregabalin/chemical synthesis , Talaromyces/enzymology , Water/chemistryABSTRACT
Synthesized Pyridin-2-yl hexanoate was screened for its antibacterial and antibiofilm activities using the broth microdilution method and XTT assay respectively. The obtained results revealed that synthesized molecule inhibited the growth of pathogenic bacteria (MICs value ranged from 80 to 640⯵gâ¯ml-1). Moreover, it induced a strong antibiofilm effect against Gram positive cocci (BIC50 was 366, 378 and 412⯵gâ¯ml-1 for Streptococcus mutans ATCC 25175, Streptococcus salivarius ATCC 13419 and Streptococcus oralis ATCC 6249 respectively). Basis on these results, pyridin-2-yl hexanoate may be considered as effective compound with antibacterial and antibiofilm activities.
Subject(s)
Anti-Bacterial Agents/pharmacology , Biofilms/drug effects , Caproates/pharmacology , Escherichia coli/drug effects , Gram-Positive Bacteria/drug effects , Pyridines/pharmacology , Anti-Bacterial Agents/chemical synthesis , Biofilms/growth & development , Caproates/chemical synthesis , Escherichia coli/growth & development , Formazans/analysis , Gram-Positive Bacteria/growth & development , Microbial Sensitivity Tests , Microbial Viability/drug effects , Pyridines/chemical synthesis , Staining and LabelingABSTRACT
The extracellular crude Rhizomucor variabilis lipase was used for synthesis of flavor ester butyl caprylate and 1-butyl oleate often used as a diesel additive, a polyvinyl chloride plasticizer, a water-resisting agent, and an additive to hydraulic fluids. The influence of various reaction parameters such as the molar ratio, time, enzyme and substrate concentration, and effect of various fungal polysaccharides was estimated. The rate of catalyzed synthesis of esters largely depends on the solvent medium, and the maximum activity was found when n-hexane was used as a solvent. The maximum conversion yield of 58.2% and 59.3% was obtained for butyl caprylate and butyl oleate, respectively, under the following conditions: amount of free lipase 500â¯U; caprylic acid:butanol molar ratio 1:1; oleic acid:butanol molar ratio 2:1. The addition of naturally obtained fungal polysaccharides significantly enhanced the ester synthesis. The highest conversion rate of 95.2% was observed for butyl caprylate in the presence of AbEPS after 24â¯h with 500â¯U of free R. variabilis lipase. In the case of butyl oleate synthesis in the presence of LsPS, a maximum conversion yield of 91.2% was observed after the 24-h reaction.
Subject(s)
Caproates/chemical synthesis , Fungal Polysaccharides/chemistry , Fungal Proteins/chemistry , Lipase/chemistry , Oleic Acids/chemical synthesis , Rhizomucor/enzymology , Water/chemistry , Adsorption , Caproates/chemistry , Oleic Acids/chemistryABSTRACT
Protecting-group-free total synthesis of (+)-guadinomic acid is reported using δ-valerolactone as a readily available starting material. The protocol utilizes the recent hydroxyl-directed guanidylation of unactivated alkenes as an approach for direct stereoselective incorporation of the guanidine unit furnishing the natural product in 7 steps.
Subject(s)
Caproates/chemistry , Caproates/chemical synthesis , Guanidine/chemistry , Hydroxyl Radical/chemistry , Imidazolidines/chemistry , Imidazolidines/chemical synthesis , Catalysis , Chemistry Techniques, Synthetic , StereoisomerismABSTRACT
Structural modification of Cotton (Cot) fiber was carried out using ε-caprolactone (CL) as a monomer and oxydianiline (ODA) as a Schiff base forming agent in the presence of V2O5 catalyst in dimethylsulfoxide medium for 24h under air atmosphere (0.10MPa) at 80°C. The obtained products were analyzed for the characteristics and also for its adsorption of hexavalent chromium (Cr(VI)) and rhodamine 6G (R6G) dye molecules. The 1H NMR spectrum confirms the Schiff base formation at 8.7ppm. The adsorption study confirms the Pseudo second order kinetics with Langmuir adsorption model. The structurally modified Cot fiber based Schiff base exhibited the highest adsorption behavior through chemical interaction forces. The thermodynamic parameters were determined and confirmed the endothermic adsorption process.
Subject(s)
Aniline Compounds/chemistry , Caproates/chemistry , Cotton Fiber , Lactones/chemistry , Schiff Bases/chemistry , Adsorption , Aniline Compounds/chemical synthesis , Caproates/chemical synthesis , Chromium/chemistry , Dimethyl Sulfoxide/chemistry , Kinetics , Lactones/chemical synthesis , Schiff Bases/chemical synthesis , Thermodynamics , Water Pollutants, Chemical/chemistry , Water PurificationABSTRACT
Carbon dioxide assisted particle formation combined with electrospraying using supercritical CO2 (scCO2) as an aid (Carbon Dioxide Assisted Nebulization-Electrodeposition, CAN-ED) was used to produce Bortezomib loaded poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) P(3HB-co-3HHx) nanoparticles for sustained release. The morphology and structure of the prepared nanoparticles were investigated by SEM, TEM and FT-IR spectroscopy. Average diameter of particles obtained was 155nm and the average core sizes of P(3HB-co-3HHx) nanoparticles were between 6 and 13nm. The drug loading capacity, drug release and stability of Bortezomib loaded P(3HB-co-3HHx) nanoparticles were analyzed. The maximum loading capacity was achieved at pH=6.0 in phosphate buffer (K2HPO4/KH2PO4). It was found that temperature did not affect the stability of Bortezomib loaded nanoparticles and it was good both at 37°C and 4°C. This study pointed out that CAN-ED is a green method to produce P(3HB-co-3HHx) nanoparticles for pH responsive targeting of Bortezomib especially to parts of the body where size exclusion is not crucial.
Subject(s)
3-Hydroxybutyric Acid/chemical synthesis , Bortezomib/chemistry , Caproates/chemical synthesis , Drug Liberation , Nanoparticles/chemistry , 3-Hydroxybutyric Acid/chemistry , Bortezomib/therapeutic use , Caproates/chemistry , Carbon Dioxide/chemistry , Electroplating , Humans , Hydrogen-Ion Concentration , Microscopy, Electrochemical, Scanning , Nanoparticles/therapeutic use , Nanotechnology/methods , Spectroscopy, Fourier Transform Infrared , TemperatureABSTRACT
The synthesis and enzymatic reduction of several 6-substituted dioxohexanoates are presented. Two-step syntheses of tert-butyl 6-bromo-3,5-dioxohexanoate and the corresponding 6-hydroxy compound have been achieved in 89% and 59% yield, respectively. Regio- and enantioselective reduction of these diketones and of the 6-chloro derivative with alcohol dehydrogenase from Lactobacillus brevis (LBADH) gave the (5S)-5-hydroxy-3-oxo products with enantiomeric excesses of 91%, 98.4%, and >99.5%, respectively. Chain elongation of the reduction products by one carbon via cyanide addition, and by more than one carbon by Julia-Kocienski olefination, gave access to well-established statine side-chain building blocks. Application in the synthesis of the cholesterol-lowering natural compound solistatin is given.
Subject(s)
Amino Acids/chemistry , Hydroxymethylglutaryl-CoA Reductase Inhibitors/chemical synthesis , Lovastatin/analogs & derivatives , Alcohol Dehydrogenase/chemistry , Alcohol Dehydrogenase/metabolism , Amino Acids/chemical synthesis , Amino Acids/metabolism , Caproates/chemical synthesis , Caproates/chemistry , Caproates/metabolism , Hydroxymethylglutaryl-CoA Reductase Inhibitors/chemistry , Hydroxymethylglutaryl-CoA Reductase Inhibitors/metabolism , Levilactobacillus brevis/enzymology , Lovastatin/chemical synthesis , Lovastatin/chemistry , Lovastatin/metabolism , Models, Molecular , NADP/chemistry , NADP/metabolism , Oxidoreductases/metabolismABSTRACT
A computational approach for the simulation and prediction of a linear three-step enzymatic cascade for the synthesis of ϵ-caprolactone (ECL) coupling an alcohol dehydrogenase (ADH), a cyclohexanone monooxygenase (CHMO), and a lipase for the subsequent hydrolysis of ECL to 6-hydroxyhexanoic acid (6-HHA). A kinetic model was developed with an accuracy of prediction for a fed-batch mode of 37% for substrate cyclohexanol (CHL), 90% for ECL, and >99% for the final product 6-HHA. Due to a severe inhibition of the CHMO by CHL, a batch synthesis was shown to be less efficient than a fed-batch approach. In the fed-batch synthesis, full conversion of 100 mM CHL was 28% faster with an analytical yield of 98% compared to 49% in case of the batch synthesis. The lipase-catalyzed hydrolysis of ECL to 6-HHA circumvents the inhibition of the CHMO by ECL enabling a 24% higher product concentration of 6-HHA compared to ECL in case of the fed-batch synthesis without lipase. Biotechnol. Bioeng. 2017;114: 1215-1221. © 2017 Wiley Periodicals, Inc.
Subject(s)
Alcohol Dehydrogenase/chemistry , Caproates/chemical synthesis , Lactones/chemical synthesis , Lipase/chemistry , Oxygenases/chemistry , Enzyme Activation , Hydrolysis , Kinetics , Multienzyme Complexes/chemistry , Sorbic Acid/analogs & derivatives , Sorbic Acid/chemistry , Substrate SpecificityABSTRACT
To obtain materials useful for the biomedical field, toxic catalysts should be removed from the synthetic route of polymerization reactions and of their precursors. Lipase-catalyzed ring-opening polymerization and the synthesis of cyclic precursors can be performed with the same catalyst under different conditions. Here, we highlight the use of lipases as catalysts and optimization of their performance for both ring-closing and ring-opening polymerization, via varying parameters such as ring size, concentration, substrate molar ratio, temperature, and solvent. While the conditions for ring-closing reactions and ring-opening polymerizations of small molecules, such as ε-caprolactone, have been extensively explored using Candida antarctica lipase B (CALB), the optimization of macrocyclization, especially for more bulky substrates is surveyed here. Finally, recent methods and polymer architectures are summarized with an emphasis on new procedures for more sustainable chemistry, such as the use of ionic liquids as solvents and recycling of polyesters by enzymatic pathways.
Subject(s)
Candida/enzymology , Caproates/chemical synthesis , Fungal Proteins/chemistry , Lactones/chemical synthesis , Lipase/chemistry , Caproates/chemistry , Catalysis , Lactones/chemistryABSTRACT
BACKGROUND: Electro-spun scaffolds are utilized in a diverse spectrum of clinical targets, with an ever-increasing quantity of work progressing to clinical studies and commercialization. The limited number of conformations in which the scaffolds can be fabricated hampers their wide acceptance in clinical practice. MATERIALS & METHODS: Herein, we assessed a single-strep fabrication process for predesigned electro-spun scaffold preparation and the ramifications of the introduction of porosity (0, 30, 50, 70%) and pore shape (circle, rhomboid, square) on structural, mechanical (tensile and ball burst) and biological (dermal fibroblast and THP-1) properties. RESULTS: The collector design did not affect the fibrous nature of the scaffold. Modulation of the porosity and pore shape offered control over the mechanical properties of the scaffolds. Neither the porosity nor the pore shape affected cellular (dermal fibroblast and THP-1) response. CONCLUSION: Overall, herein we provide evidence that electro-spun scaffolds of controlled architecture can be fabricated with fibrous fidelity, adequate mechanical properties and acceptable cytocompatibility for a diverse range of clinical targets.
Subject(s)
Biocompatible Materials/chemistry , Caproates/chemistry , Lactones/chemistry , Tissue Engineering , Tissue Scaffolds/chemistry , Biocompatible Materials/chemical synthesis , Biocompatible Materials/therapeutic use , Caproates/chemical synthesis , Caproates/therapeutic use , Fibroblasts/drug effects , Humans , Lactones/chemical synthesis , Lactones/therapeutic use , PorosityABSTRACT
High activity of post-synthetically synthesized Sn-beta, producing novel caprolactone polymer building blocks, is demonstrated in Meerwein-Ponndorf-Verley (MPV), Oppenauer (OPO), Baeyer-Villiger (BV) and cascade reactions thereof with biomass-derived molecules.