Biotransformation of Eugenol via Protocatechuic Acid by Thermophilic Geobacillus sp. AY 946034 Strain
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초록
The metabolic pathway of eugenol degradation by thermophilic Geobacillus sp. AY 946034strain 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 oxidizedthrough coniferyl alcohol, and ferulic and vanillic acids to protocatechuic acid before thearomatic ring was cleaved. The cell-free extract of Geobacillus sp. AY 946034 strain grown oneugenol 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. AY946034. The relative molecular mass of the native enzyme was about 450 ± 10 kDa and wascomposed of the non-identical subunits. The pH and temperature optima for enzyme activitywere 8 and 60oC, respectively. The half-life of protocatechuate 3,4-dioxygenase at the optimumtemperature was 50 min.
Abstract
The metabolic pathway of eugenol degradation by thermophilic Geobacillus sp. AY 946034strain 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 oxidizedthrough coniferyl alcohol, and ferulic and vanillic acids to protocatechuic acid before thearomatic ring was cleaved. The cell-free extract of Geobacillus sp. AY 946034 strain grown oneugenol 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. AY946034. The relative molecular mass of the native enzyme was about 450 ± 10 kDa and wascomposed of the non-identical subunits. The pH and temperature optima for enzyme activitywere 8 and 60oC, respectively. The half-life of protocatechuate 3,4-dioxygenase at the optimumtemperature was 50 min.
참고문헌 (13)
- Aoki K, "Purification and characterization of catechol 1,2-dioxygenase from anilineassimilating Rhodococcus erythropolis An-13", Agric. Biol. Chem, 48, pp.2087~2095 (1984)
- Bradford MM., "A rapid and sensitive method for quantitation of protein utilizing the principle of proteindye binding", Anal. Biochem, 72, pp.248~254 (1976)
- Brandt K, "Characterization of eugenol hydroxylase genes (ehyA", ehyB) from the new eugenol degrading Pseudomonas sp. strain OPS1, 2001, pp.56, 724~730
- Bubinas A, "Protocatechuate 3, 4-dioxygenase from thermophilic Geobacillus sp. bacterium", Biologija, 1, pp.31~34 (2007)
- Bubinas A, "Degradation of naphthalene by thermophilic bacteria via a pathway, through protocatechuic acid", Cent. Eur. J. Biol, 1, pp.61~68 (2008)
- Bustard MT, "Biodegradation of high-concentration isopropanol by a solvent-tolerant thermophile, Bacillus pallidus", Extremophiles, 6, pp.319~323 (2002)
- Chen CL, "Aromatic acids produced during degradation of lignin in spruce wood by Phanerochaete chrysosporium", Holzforschung, 36, pp.3~9 (1982)
- Civolani C, "Bioconversion of ferulic acid into vanillic acid by means of a vanillate-negative mutant of Pseudomonas fluorescens strain BF13", Appl. Environ. Microbiol, 66, pp.2311~2317 (2000)
- Furukava H, "Novel double bondtransferring hydroxylation reactions in microbial metabolism of eugenol", Ann. NY Acad. Sci, 864, pp.81~86 (1998)
- Furukawa H, "Purification and characterization of eugenol dehydrogenase from Pseudomonas fluorescens E118", Arch. Microbiol, 171, pp.37~43 (1998)
- Dongfei Han, "Bacterial Biotransformation of Phenylpropanoid Compounds for Producing Flavor and Fragrance Compounds", Applied Biological Chemistry, 56(2), pp.125~133 (2013), 한국응용생명화학회
- Iwagami SG, "Characterization of the protocatechuic acid catabolic gene cluster from Streptomyces sp. strain 2065", Appl. Environ. Microbiol, 66, pp.1499~1508 (2000)
- De Jang E, "Purification and characterization of vanillyl-alcohol oxidase from Penicillium simplicissimum: a novel aromatic alcohol oxidase containing covalently bound FAD", Eur. J. Biochem, 208, pp.651~657 (1992)