Thermostable enzymes in organic synthesis.  2.  Asymmetric reduction of ketones with alcohol dehydrogenase from Thermoanaerobium brockii

Keinan, Ehud; Hafeli, Eva K.; Seth, K. K.; Lamed, Raphael

doi:10.1021/ja00261a026

Cited by 293 publications

(146 citation statements)

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“…The stereoselectivity for the reduction of acyclic ketones has been investigated using alcohol dehydrogenase from Thermoanaerobium brockii [37] and alcohol dehydrogenase from T. ethanolicus [6]. In both cases, the stereoselectivity was found to depend on the ketone structure.…”

Section: Stereoselectivity Of Zn (Ii) Ladh-catalyzed Ketone Reductionsmentioning

confidence: 99%

Substrate specificity and stereoselectivity of horse liver alcohol dehydrogenase

Adolph

Maurer

Schneider-Bernlöhr

et al. 1991

European Journal of Biochemistry

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1. The steady-state parameters k,,, and K , and the rate constants of hydride transfer for the substrates isopropanol/acetone; (9-2-butano1, (R)-2-butanol/2-butanone; (9-2-pentano1, (R)-2-pentanol/2-pentanone; 3-pentanol/3-pentanone; (S)-2-octanol and (R)-2-octanol have been determined for the native Zn(I1)-containing horse-liver alcohol dehydrogenase (LADH) and the specific active-site-substituted Co(1I)LADH.2. A combined evaluation of steady-state kinetic data and rate constants obtained from stopped-flow measurements, allowed the determination of all rate constants of the following ordered bi-bi mechanism: E $ E . NAD 3. On the basis of the different substrate specificities of LADH and yeast alcohol dehydrogenase (YADH), a procedure has been developed to evaluate the enantiomeric product composition of ketone reductions. 2-Butanone and 2-pentanone reductions revealed (S)-Z-butanol(86%) and (S)-2-pentanol(95%) as the major products.4. The observed enantioselectivity implies the existence of two productive ternary complexes; E . NADH .(pro-S) 2-butdnone and E . NADH . (pro-R) 2-butanone. All rate constants describing the kinetic pathways of the system (S)-2-butanol, (R)-2-butanol/2-butanone have been determined. These data have been used to estimate the expected enantiomer product composition of 2-butanone reductions using apparent k,,,/K, values for the two different ternary-complex configurations of 2-butanone. Additionally, these data have been used for computer simulations of the corresponding reaction cycles. Calculated, simulated and experimental data were found to be in good agreement. Thus, the system (5')-2-butanol, (R)-2-butanol/2-butanone is the first example of a LADHcatalyzed reaction for which the stereochemical course could be described in terms of rate constants of the underlying mechanism. 5. The effects of Co(I1) substitution on the different steps of the kinetic pathway have been investigated. The free energy of activation is higher for alcohol oxidation and lower for ketone reduction when catalyzed by Co(1I)LADH in comparison to Zn(1I)LADH. However, the free energies of binding are affected by metal substitution in such a way that the enantioselectivity of ketone reduction is not significantly changed by the substitution of Co(I1) for Zn(I1).6. Evaluation of the data shows that substrate specificity and stereoselectivity result from combination of the free energies of binding and activation, with differences in binding energies as the dominating factors. In this regard, the interactions of substrate molecules with the protein moiety are dominant over the interactions with the catalytic metal ion.

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Section: Stereoselectivity Of Zn (Ii) Ladh-catalyzed Ketone Reductionsmentioning

confidence: 99%

Substrate specificity and stereoselectivity of horse liver alcohol dehydrogenase

Adolph

Maurer

Schneider-Bernlöhr

et al. 1991

European Journal of Biochemistry

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“…Also their catalytic mechanism is identical: the catalytic machinery consists of an amino acid triad (serine, histidine and aspartate or glutamate) with the serine acting as a nucleophile, attacking the carbonyl carbon of the substrate ester, while the histidine acts as an amphiphile catalyzing the withdrawal of the alcohol moiety by transferring a proton on the ester oxygen. [4] Most [5] solvent variation, [6] immobilization, [7] the modification of parameters like temperature, [8] pH, [9] and pressure, [10] or by modification of the enzyme's structure. Several attempts to vary enantioselectivity of esterases and lipases by directed evolution, [11][12][13] and by rational protein engineering were successful.…”

Section: Introductionmentioning

confidence: 99%

A Molecular Mechanism of Enantiorecognition of Tertiary Alcohols by Carboxylesterases

et al. 2003

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“…Ketones possessing a Cs chain (such as 2-pentanone) or larger give the ($)-alcohol with excellent optical yields [86]. This reversal of the enzyme stereospecificity has also been observed for other enzymes [87, 881 and can be explained by the structure of the active site in the enzyme.…”

Section: Ohmentioning

confidence: 58%

“…Meanwhile, several applications have been described using this enzyme for synthetic purposes, e. g. the gram-scale conversion of NADP to NADPH applying 0.3% 2-propanol as the POI. small site large site NAD(P) H reductant with a coenzyme recycling number of 20000 [85] or the reduction of the 2-ketone to the chiral (S)-(+)-2-pentanol in the presence of 2-propanol [85], erroneously assigned as (R)-(+)-2-pentanol [86]. Ketones possessing a Cs chain (such as 2-pentanone) or larger give the ($)-alcohol with excellent optical yields [86].…”

Section: Ohmentioning

confidence: 99%

“…Diketones are further interesting substrates for alcohol dehydrogenases. Ketoesters with 1 $located carbonyl function can be reduced by the TbrADH enzyme, optically pure (9-lactones are obtained spontaneously under reaction conditions out of the 5-hydroxyester [89]. Chiral aliphatic chloroalcohols are valuable building blocks that can be prepared from the corresponding chloroketones with TbrADH [89].…”

Section: Ohmentioning

confidence: 99%

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Dehydrogenases for the synthesis of chiral compounds

Hummel

Kula

1989

European Journal of Biochemistry

329

126

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Thermostable enzymes in organic synthesis. 2. Asymmetric reduction of ketones with alcohol dehydrogenase from Thermoanaerobium brockii

Cited by 293 publications

References 0 publications

Substrate specificity and stereoselectivity of horse liver alcohol dehydrogenase

Substrate specificity and stereoselectivity of horse liver alcohol dehydrogenase

A Molecular Mechanism of Enantiorecognition of Tertiary Alcohols by Carboxylesterases

Dehydrogenases for the synthesis of chiral compounds

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