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KMID : 0545120140240040475
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Journal of Microbiology and Biotechnology
2014 Volume.24 No. 4 p.475 ~ p.482
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Biotransformation of Eugenol via Protocatechuic Acid by Thermophilic Geobacillus sp. AY 946034 Strain
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Grazina Giedraityte
Lilija Kalediene
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Abstract
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The metabolic pathway of eugenol degradation by thermophilic Geobacillus sp. AY 946034 strain was analyzed based on the lack of data about eugenol degradation by thermophiles. TLC, GC-MS, and biotransformation with resting cells showed that eugenol was oxidized through coniferyl alcohol, and ferulic and vanillic acids to protocatechuic acid before the aromatic ring was cleaved. The cell-free extract of Geobacillus sp. AY 946034 strain grown on eugenol showed a high activity of eugenol hydroxylase, feruloyl-CoA synthetase, vanillate-Odemethylase, and protocatechuate 3,4-dioxygenase. The key enzyme, protocatechuate 3,4- dioxygenase, which plays a crucial role in the degradation of various aromatic compounds, was purified 135-fold to homogeneity with a 34% overall recovery from Geobacillus sp. AY 946034. The relative molecular mass of the native enzyme was about 450 ± 10 kDa and was composed of the non-identical subunits. The pH and temperature optima for enzyme activity were 8 and 60oC, respectively. The half-life of protocatechuate 3,4-dioxygenase at the optimum temperature was 50 min.
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KEYWORD
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thermophilic bacteria, eugenol, protocatechuate 3, 4-dioxygenase, metabolism
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