Biodegradation of Phthalate Esters by a Newly Isolated Acinetobacter sp. Strain LMB-5 and Characteristics of Its Esterase

Phthalate esters(PAEs) are extensively applied in industry, and they migrate to environment during the process of production,employ, and treatment and are difficult to be degraded in nature. However, some microorganisms could use them as the carbon source to growth. In this study, an Acinetobacter sp. strain LMB-5, capable of utilizing PAEs, was isolated from a vegetable greenhouse soil.The degradation capability of strain LMB-5 was also investigated by incubation in mineral salt medium containing different PAEs,dimethyl phthalate(DMP), diethyl phthalate(DEP), di-n-butyl phthalate(DBP), and di-(2-ethylhexyl) phthalate(DEHP). The strain could grow well with DMP, DEP, DBP, and DEHP. When the concentration of DBP increased from 100 to 400 mg L^(-1), the half-life extended from 9.5 to 15.5 h. In the concentration range of DBP, the degradation ability of strain LMB-5 could be described by first-order kinetics. During the biodegradation of DBP, three intermediates, 1,2-benzenedicarboxylic acid,butyl methyl ester, DMP,and phthalic acid(PA) were detected, and the proposed pathway of DBP was identified. By analysis of bioinformatics, one esterase was cloned from the genome of LMB-5 and expressed in Escherichia coli. It displayed an ability to break the ester bonds of DBP. The enzyme exhibited maximal activity at pH 7.0 and 40℃ with DBP as the substrate. It was activated by Cu^(2+) and Fe^(3+) and had a high activity in the presence of low concentrations of methanol or dimethylsulfoxide(each 10%, volume:volume). The Acinetobacter sp. strain LMB-5 may make a contribution to the remediation of soils polluted by PAEs in the future.

Phthalate esters （PAEs） are extensively applied in industry, and they migrate to environment during the process of production, employ, and treatment and axe difficult to be degraded in nature. However, some microorganisms could use them as the carbon source to growth. In this study, an Acinetobacter sp. strain LMB-5, capable of utilizing PAEs, was isolated from a vegetable greenhouse soil. The degradation capability of strain LMB-5 was also investigated by incubation in mineral salt medium containing different PAEs, dimethyl phthalate （DMP）, diethyl phthalate （DEP）, di-n-butyl phthalate （DBP）, and di-（2-ethylhexyl） phthalate （DEHP）. The strain could grow well with DMP, DEP, DBP, and DEHP. When the concentration of DBP increased from 100 to 400 mg L-1, the half-life extended from 9.5 to 15.5 h. In the concentration range of DBP, the degradation ability of strain LMB-5 could be described by first-order kinetics. During the biodegradation of DBP, three intermediates, 1,2-benzenedicaxboxylic acid,butyl methyl ester, DMP, and phthalic acid （PA） were detected, and the proposed pathway of DBP was identified. By analysis of bioinformatics, one esterase was cloned from the genome of LMB-5 and expressed in Escherichia coll. It displayed an ability to break the ester bonds of DBP. The enzyme exhibited maximal activity at pH 7.0 and 40 ℃ with DBP as the substrate. It was activated by Cu2＋ and Fe3＋ and had a high activity in the presence of low concentrations of methanol or dimethylsulfoxide （each 10%, volume：volume）. The Acinetobacter sp. strain LMB-5 may make a contribution to the remediation of soils polluted by PAEs in the future.