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6.1 水解反应 6.1.1 机理和动力学Attack of another alcohol R2-OH leads to another ester R1-CO-OR2.This is an interesterification reaction,called enzymatic“acyl transfer”.An incoming amine R3-NH2 results in the formation of an amide R1-CO-NH-R3,yielding an enzymatic aminolysis of esterPeracids of type R1-CO-OOH are formed when hydrogen peroxides is acting as the nucleophile.Hydrazinolysis provides access to hydrazides,and the action of hydroxylamine results in the formation of hydroxamic acid derivatives.Enantioface Differentiation:(achiral substrate)Enantiotopos Differentiation:(Prochiral substrate)Single step process Double step process Asymmetrization of meso-substrates Single step process Double step process Single-step kineticsDouble-step kinetics Asymmetrization of a meso-diacetate Enantiomer Differentiation Kinetic resolution with in-situ racemization kR/kS=enzymatic hydrolysisi of enantiomers R/Skracsub/kracprod=racemization of substrate/productkspont=spontaneous hydrolysisTo overcome the occurrence of the undesired“wrong”enantiomer:Repeated resolutionIn-situ racemizationIn-situ inversionkinetics resolution with in situ inversion Irreversible reaction:Enzymatc kinetic resolution(irreversible reaction)For the product:For the substrate:c=conversione.e.=enantiomeric excess of substrate(s)or product(p)E=Enantiomeric RatioThe dependence of the selectivity and the conversion of the reaction isDependence of optical purities of the conversion Two-step-enzymatic resolution For the product:For the substrate:c=conversione.e.=enantiomeric excess of substrate(s)or product(p)E=Enantiomeric RatioK=equilibrium constant Reversible reaction Enzymatic kinetic resolution(reversible reaction)sequential enzymatic kinetic resolutionconditions:aqueous buffer,Absidia glauca cells Sequential enzymatic resolution by hydrolysis and esterification R=C5H11;conditions:I-octane,hexanoic acid,Pseudomonas sp.Lipase Mechanism of sequential enzymatic kinetic resolution via hydrolysis-esterification.Sequential enzymatic kinetic resolution via hydrolysis-esterification 6.1.2 Hydrolysis of the Amide BondApplication :Amino acids:optically pure starting materialsfor pharma-.and agro chemicals.Or artifical sweeteners:non natural amino acidsD-phenglglycine and p-hydroxy derivatineD-valineWorld production of amino acid using Enzymatic processes(1980-1992)Amino acid Amountt/a L-Ala 250 L-Asp 5000 L-2,4-dihydroxyphenylalanine 200 L-Met 240 L-Phe 5000 L-Trp 200 L-Val 150 D-Phenylglycine 1000 D-p-hydroxyphenylglycine 1000_Important enzymatic routes to enantiomerically pure-amino acidsEsterase-MethodEnzymatic resolution of Amino acid esters Via the esterase-MethodResolution of N-acetyl amino acid esters by-chymotrypsinDynamic resolution of N-acetyl aminoacid estersR1R2productYield(%)e.e.(%)Ph-CH2-Ph-CH2-L-Phe9298Ph-CH2-n-Bu-L-Phe92984-Hydroxyphenyl-CH2-Ph-CH2-L-Tyr95974-Hydroxyphenyl-CH2-n-PrL-Tyr9597(CH3)2CH-CH2-Ph-CH2-L-Leu8793n-Bu-Ph-CH2-L-NorLeu8790EtPh-CH2-L-NorLeu8791Amidase-MethodEnzymatic resolution of Amino Acid Amides Via the Amidase-MethodAcylase-MethodEnzymatic resolution of N-Acyl Amino acids via the Acylase-MethodEnzymatic resolution of N-Acyl Amino acids for natural product synthesisHydantoinase-MethodEnzymatic resolution of N-Acyl Amino Acids Via the D-hydantoinase-MethodEnzymatic resolution of N-Acyl Amino Acids Via the L-hydantoinase-MethodLactamase-MethodEnzymatic resolution of bicyclic-and-Lactams via the Lactamase-MethodEsterases and ProteasesTypes of substrates for esterase and protease6.1.3 Ester HydrolysisPig Liver Esterase and-chymotrypsin Mild hydrolysis by PLERegioselective ester hydrolysis by PLERegioselective hydrolysis of E/Z-diastereotopic diesters by PLEAsymmetrization of prochiral malonates by PLE and-chymotrypsin_ Enzyme R Configuration e.e.%_ PLE*Ph-S 86 PLE C2H5-S 73 PLE n-C3H7-S 52 PLE n-C4H9-S 58_ PLE n-C5H11-R 46 PLE n-C6H13-R 87 PLE n-C7H15-R 88 PLE p-MeO-C6H4-CH2-R 82 PLE t-Bu-O-CH2-R 96_-chymotrypsin Ph-CH2-R 100_*The ethyl ester was usedAsymmetrization of prochiral diestersChemo-enzymatic synthesis of-methyl-L-amino acidsAsymmetrization of prochiral glutarates_Hydrolase R1 R2 Product e.e%_-chymotrypsin*AcNH-H B 79-chymotrypsin*OH-H B 85-chymotrypsin Ph-CH2-O-H B 84-chymotrypsin Ph-CO-O-H B 92-chymotrypsin CH3OCH2O-H B 93_ PLE CH3-H B 90 PLE AcNH-H B 93 PLE OH-H A 12 PLE t-Bu-CO-NH-H A 93 PLE Ph-CH2-O-CONH-H A 93 PLE Ph-CH2-CH=CH-CH2-H A 88 PLE OH-CH3 A 99_Acinetobacter sp.*OH-H B 95Arthrobacter sp.*OH-H A 95*The corresponding ethyl esters were used.Asymmetrization of acyclic meso-dicarboxylates by-chymotrypsin and PLEAcyctic meso-dicarboxylic acid ester.With succinic and glutaric acid backbonewere also good substrates for PLE,-chymotrypsin.Asymmetrization of polycyclic meso-1,2-dicarboxylates by PLEAsymmetrization of cyclic meso-diacetates by PLE and ACEAsymmetrization of N-containing cyclic meso-diesters by PLEResolution of acyclic carboxylic esters by PLEResolution of racemic estersResolution of cyclic carboxylic esters by PLEResolution of cyclic trans-1,2-diacetates by PLEMicrobial resolution of masked hydroxyaldehyde by Bacillus sp.Microbial EsterasesMicrobial resolution of secondary acetates by Bakers yeastSynthetic potential of chiral 1-alkyn-3-olsResolution of-substituted propionates by carboxylesterase Resolution of-nitro-methyl carboxylates by-chymotrypsinRegioselective ester-hydrolyses catalyzed by subtilisinChemo-and enantioselective ester-hydrolyses catalyzed by penicillin acylaseproteasee.e.%e.e.%subtilisin2525Aspergillus aryzase prot8888Resolution of bulky esters by subtilisin and AspergillusResolution with in situ racemization by protease from streptomyces griseusOptimization of SelectivityOptimization of PLE-catalyzed hydrolysis by substrate modificationXconfiguratione.e.%HCH3-CO-CH2=CH-CO-C2H5-CO-RRRR419386N-C4H9-CO-(CH3)2CH-CO-C6H11-CO-(CH3)3C-CO-Ph-CH2-O-CO-(E)-CH3-CH=CH-CO-SSSSSS25479939397Medium:e.e%H2O55H2O/DMSO72H2O/DMF84H2O/t-BuOH96Medium:e.e.%H2O17H2O/MeOH 9:197H2O/DMSO 1:197Medium:e.e.%H2O25H2O/DMSO 1:193Bn=-CH2-PhR=3,4-dimethoxyphenylSelectivity enhancement of PLE by addition of organic cosolventsLipasesEsterase-and lipase-kineticsSubstrate-types for lipase Preferrde enantiomer:”Kazlauskas-rule”Sequence rule order of largemedium assumedSteric requirements of lipasesRegio-and enantioselective hydrolysis of dimethyl-methylsuccinateRegioselective hydrolysis of carbohydrate esters by PLLXe.e.%40728886867899Asymmetric hydrolysis cyclic meso-diacetates by PPLXe.e.%5064946866908896RCosoventConfiguratione.e.%n-C7H15-(E)-n-C5H11-CH=CH-(E)-n-C5H11-CH=CH-i-Pr2ONonei-Pr2OSSS708495(CH3)2CH-(CH2)2-(E)-(CH3)2CH-CH=CH-(E)-(CH3)2CH-CH=CH-i-Pr2ONonei-Pr2OSSS729097(Z)-n-C5H11-CH=CH-(Z)-(CH3)2CH-CH=CH-i-Pr2Oi-Pr2ORR5315Ph-Ph-P-CH3-C6H4-2-naphtyl-NoneTolueneNonenoneSSSS85-92999696Ph-CH2-O-Ph-CH2-O-i-Pr2Oi-Pr2ORR88-9194Asymmetrization of prochiral 1,3-propanediol diesters by PPLR1Re.e.%Selectivity(E)HCH3535HC2H58828Hn-C3H79245Hn-C4H99695Resolution of epoxy-esters by PPLEnzymeReaction RateSelectivity(E)Crude PPLgood300Pure PPLNo reaction-chymotrypsinNo reaction-Cholesterol esterasefast17Novel hydrolasegood210Resolution of bicyclic acetate by hydrolases present in crude PPLEnantioselective hydrolysis of lactones by PPLLipase catalyzed resolution of oxazolin-5-onesEnzymatic resolution of a cyclic enol ester by CRLEnzymatic resolution of 1,3-dioxolane-4-carboxylates by CRL.Enzymatic resolution of cyclohexane-1,2,3-triol esters by CRL and PSL.Structural Variations:A=endo-,exo-configurationR=Me,n-C3H7,n-C7H15B=-O-,-CH2-,-CH2-CH2-Sa,Ss=H,-OMe,-CO2Me,-CO2-t-Bu,-O-CH2-PhSx=H,bond,-O-,-O-C(Me)2-O-,-OH Sn=H,-OH,-O-C(Me)2-O-Substrate-model for CRL(Candida rugosa lipase)RegionRequirementsASite of reactionmust be endo,R may be n-alkyl,preferably n-C3H7BBridgeMay contain hetero atoms,but must be smallSa,Ssanti-syn SubstituentsMethylene bridges may carry small ester,ether or acetal groupsSxexo-Substituentsallowed region,substituents may be largeSnendo-Substituentsforbidden region,substituents(if any)must be very small-Site-electrons in this area enhance the selectivityc After treatment with deoxycholate and ethanol/ether.Selectivity enhancement of CRL by non-covalent enzyme modification.Lipasen/Distanceae.e.%CRLCRLCRL1/32/43/5786285PSLPSLPSL1/32/43/5489879a Number of bonds between the prochiral center and the site of reaction.Asymmetrization of esters having remote prochiral centers by lipases.Re.e.Acid%e.e.Ester%p-NO2-C6H4-p-Cl-C6H4-Ph-C6H11-9791929898989898Resolution of sulfoxide esters by PSLResolution of dithioacetal esters by PSL.R1R2Selectivity(E)Me-Me-Me-Me-Me-MeCl-CH2-n-C3H7-Me-O-CH2-Me-S-CH2-7621429Ph-CH2-CH2-Ph-CH=CH-Ph-Ph-S-CH2-Ph-S-CH2-Ph-S-CH2-365574Resolution of-acyloxynitriles by PSL.LipaseXe.e.Alcohol%e.e.Ester%Selectivity(E)CRLPSLPSLHH-O-(CH2)2-O-92999974989953200200Resolution of bicyclo3.3.0octane systems by CRL and PSL.Asymmetrization of bis(acyloxymethyl)tetrahydrofurans by lipases.LipaseRSolvent systeme.e.%CRLPSLMSLMSLHHHMebufferbufferbufferBuffer12819999PPLPPLMeMebufferbuffer/hexane2085R1R2Selectivity(E)Me-Et-Me-S-Me-S-Me-Me-Me-Me-S-CH2-802030180Resolution of-substituted-acyloxy esters by Aspergillus sp.Lipase.Lipasee.e.Ester%ConfigurationSelectivity(E)CRL70R12PSLAlcaligenes sp.Chromobacterium sp.Arthrobacter sp.93939699SSSS8888160200Hydrolysis of cyanohydrin esters using microbial lipases.Optimization of selectivityTypes of Bichiral esters.Optimization of selectivity Bichiral estersEnhanced alcohol resolution by CRL using bichiral esters.Enantioselective inhibition of LipaseSelectivity enhancement of CRL by enantioselective inhibition.Selectivity enhancement of PSL by enantioselective inhibition.Selectivity enhancement of CRL by covalent enzyme modification.Chemical modification of lipases6.1.4 Hydrolysis and Formation of Phosphate Esters*Hydrolysis of Phosphate EstersMild enzymatic hydrolysis of phosphate esters.Resolution of threonine O-phosphate using acid phosphatase.Resolution of carbocyclic nucleoside analogues.*Formation of Phosphate EstersEnzymatic recycling of ATP from ADP.Enzymatic recycling of ATP from adenosine and/or AMP.Enzymatic synthesis of carbocyclic ATP-mimics.Phosphorylation of hexose derivatives by hexokinase.Phosphorylation of dihydroxyacetone by glycerol kinase.Phosphorylation of D-ribose and enzymatic synthesis of UMP.Enantioselective phosphorylation of glycerol derivatives.Substrate model for glycerol kinase.6.1.5 Hydrolysis of EpoxidesBiodegradation of aromatics.Mechanism of microsomal epoxide hydrolase Substrate types for MEHAsymmerization of cyclic cis-meso-epoxides by EHn Optical Purity of Diol%microsomal EH cytosolic EH 1 90 60 2 94 22 3 40 30 4 70 no reactionR1 R2 Conversion%e.e.Diol%selectivity(E)t-Bu H 47 8837H t-Bu 537417Resolution of substituted cyclohexene oxides by MEHR Conversion%e.e.Diol%Selectivity(E)Ph-1890(CH3)2CH-CH2-Torlopsis candida 95 Ph-Alcaligenes faecalis 100 Re.e.Amide%e.e.Acid%e.e.Nitrile%CH3-CH2-CH(CH3)-99 8773 Cl76 99 OCH376 99 6.2 Addition and Elimination Reactions*Cyanohydrin Fornation R1R2e.e.%Ph-H 94Ph-CH2-H 40p-MeO-Ph-H 932-furyl-H 98 n-C3H7-H 92-96 n-C5H11-H 67t-Bu-H 73(E)-CH3-CH=CH-H 69C2H5-Me 76n-C4H9-Me 98(CH3)2CH-(CH2)2-Me 98CH2=CH(CH3)-Me 94Cl-(CH2)3-Me 84 Table Synthesis(R)-cyanohydrins from aldehydes and ketones R1R2e.e.%Ph-H 96-97m-HO-Ph-H 91-98p-HO-Ph-H 94-99m-MeO-Ph-H 89 m-PhO-Ph-H 96m-Br-Ph-H 92p-Cl-Ph-H 54n-C5H11-H 84n-C8H17-H85CH2=CH-H 84Table Sythesis of(S)-cyanohydrines from aldehydes XYield%H90Cl-60CH3-61C2H5-60(CH3)2CH-54 n-C3H7-49 n-C4H9-0
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