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  • [Characterization of salicylate 5-hydroxylase for phenanthrene degradation using moderately halophilic Martelella sp. AD-3].

[Characterization of salicylate 5-hydroxylase for phenanthrene degradation using moderately halophilic Martelella sp. AD-3].

Wei sheng wu xue bao = Acta microbiologica Sinica (2012-11-24)
Fei Dong, Changzheng Cui, Tiancai Feng, Yaoyu Feng, Yongdi Liu
ABSTRACT

To identify the relationship between salicylate 5-hydroxylase activity and the degradation rate of phenanthrene by moderately halophilic Martelella sp. AD-3, and to investigate their enzymatic characteristics. The products resulted from degradation of salicylic acid using the crude enzyme produced by AD-3 was analyzed using HPLC analysis, and salicylate 5-hydroxylase activities at different conditions was calculated according to the change of NADH absorbance at 340 nm. The salicylate 5-hydroxylase activity was higher during log-phase and early stationary phase of AD-3, and the enzyme activity was in consistent with phenanthrene degradation rate. The enzyme production was induced by phenanthrene and salicylic acid. The highest enzyme activity, 132.8 nmol/(min x mg), from strain AD-3 was obtained under the phenanthrene concentration of 200 mg/L, 3% salinity and a pH of 9.0. The optimum conditions for degradation of salicylic acid using this enzyme were 30 degrees C , pH 7.5, and salinity 3%. The V(max) and K(m) for the enzyme were 200 nmol/(min x mg) and 8.7 micromol/L, respectively. The salicylate 5-hydroxylase was produced in the degradation of phenanthrene in AD-3. The enzyme activity was related to phenanthrene degradation rate.

MATERIALS
Product Number
Brand
Product Description

Supelco
Phenanthrene solution, certified reference material, 5000 μg/mL in methanol
Sigma-Aldrich
Phenanthrene, 98%
Sigma-Aldrich
Phenanthrene, sublimed grade, ≥99.5%
Supelco
Phenanthrene, analytical standard, for environmental analysis
Supelco
Phenanthrene, certified reference material, TraceCERT®