Different strategies for multi-enzyme cascade reaction for chiral vic-1,2-diol production
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The stereoselective three-enzyme cascade for the one-pot synthesis of (1S,2S)-1-phenylpropane-1,2-diol ((1S,2S)-1-PPD) from inexpensive starting substrates, benzaldehyde and acetaldehyde, was explored. By coupling stereoselective carboligation catalyzed by benzoylformate decarboxylase (BFD), L-selective reduction of a carbonyl group with alcohol dehydrogenase from Lactobacillus brevis (ADHLb) as well as the coenzyme regeneration by formate dehydrogenase (FDH), enantiomerically pure diastereoselective 1,2-diol was produced. Two different multi-enzyme system approaches were applied: the sequential two-step one-pot and the simultaneous one-pot cascade. All enzymes were kinetically characterized. The impact of acetaldehyde on the BFD and ADHLb stability was investigated. To overcome the kinetic limitation of acetaldehyde in the carboligation reaction and to reduce its influence on the enzyme stability, experiments were performed in two different excesses of acetaldehyde (100 and 300%). Due to the ADHLb deactivation by acetaldehyde, the simultaneous one-pot cascade proved not to be the first choice for the investigated three-enzyme system. In the sequential cascade with 300% acetaldehyde excess a 100% yield of vic 1,2-diol was reached.
KeywordsMulti-enzyme cascade reaction Asymmetric reduction Stereoselectivity Alcohol dehydrogenase Benzoylformate decarboxylase
This work was supported by University of Zagreb short-term financial scientific research support under the title “Mathematical modeling of the biocatalytic synthesis of industrially interesting products”. The authors would like to thank Prof. Martina Pohl from the Institute of Bio- and Geosciences, IBG-1: Biotechnology, Research center Jülich, Germany for the gift of alcohol dehydrogenase from Lactobacillus brevis and Davor Valinger from Faculty of Food Technology and Biotechnology, University of Zagreb for the isolation of enzyme benzoylformate decarboxylase.
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Conflict of interest
The authors declare that they have no conflict of interest.
- 4.Burton SG, le Roes-Hill M (2008) In: Garcia-Junceda E (ed) Multi-step enzyme catalysis—biotransformations and chemoenzymatic synthesis. Wiley, WeinheimGoogle Scholar
- 8.Rios-Solisa L, Morrisa P, Granta C, Odeleyea AO, Hailesb HC, Warda JM, Dalbya PA, Baganza F, Lyea GJ (2015) Modelling and optimisation of the one-pot, multi-enzymatic synthesis of chiral amino-alcohols based on microscale kinetic parameter determination. Chem Eng Sci 122:360–372CrossRefGoogle Scholar