Application of L-2-Haloacid Dehalogenase from Thermophilic Archaea Sulfolobus Tokodaii in the Production of D-Lactic Acid†

doi:10.7503/cjcu20140739

Abstract

Abstract:

Optical pure lactic acid is a useful chiral intermediate in the synthesis of chiral drugs and materials engineering. For the purpose of environmental-friendly and high stereoselectivity, enzymes were used to resolve racemic mixtures. In this study, we cloned and expressed L-2-haloacid dehalogenase(ST2570) from thermophilic Archaea Sulfolobus tokodaii in Escherichia coli BL21(DE3). The recombinant enzyme was purified to a single band using heat treatment and Ni²⁺-NTA affinity chromatography. The optimum temperature and pH value of ST2570 were 70 ℃ and 9.5, respectively. The specific activity of purified ST2570 for the hydrolysis of 2-chloropropionic acid was 2963.33 U/mg. For the high enatioselectivity of ST2570, only L-2-chloropropionic acid can be hydrolyzed to produce D-lactic acid during the reaction. With the optimization, the best conditions to produce D-lactic acid were pH of 9.5, temperature of 60 ℃, 3 mol/L of buffer, 0.5 mol/L of 2-chloropropionic acid and 3×10⁴ U/L of ST2570.

Key words: L-2-Haloacid dehalogenase, 2-Chloropropionic acid, Lactic acid

CLC Number:

O621.25

TrendMD:

XIE Guiqiu, PAN Dong, He Wenlong, GAO Gui, GAO Renjun. Application of L-2-Haloacid Dehalogenase from Thermophilic Archaea Sulfolobus Tokodaii in the Production of D-Lactic Acid^†[J]. Chem. J. Chinese Universities, 2015, 36(4): 698.

Figures/Tables 4

Scheme 1 Hydrolysis reaction catalyzed by L-2-dehalogenase

Fig.1 Effects of the concentration of substrate on the reaction(A) and conversion(B) Concentration/(mol·L-1): a. 0.05; b. 0.1; c. 0.25; d. 0.5; e. 0.75; f. 1.0.

Fig.2 Effect of the initial concentration of substrate on yield of lactic acid

Fig.3 Effects of the amount of enzyme on the reaction time(A) and conversion(B)

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