期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 67, 期 31, 页码 8548-8558出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.9b02655
关键词
Gordonia polyisoprenivorans; esterase; bis(2-ethylhexyl) phthalate; phthalate esters; biodegradation
资金
- National Key Research and Development of China [2016YFA0204300]
- National Natural Science Foundation of China [21506055, 21505044]
- Natural Science Foundation of Shanghai [18ZR1409900, 18DZ1112703]
- Fundamental Research Funds for the Central Universities [22221818014]
Herein, we report a double enzyme system to degrade 12 phthalate esters (PAEs), particularly bulky PAEs, such as the widely used bis(2-ethylhexyl) phthalate (DEHP), in a one-pot cascade process. A PAE-degrading bacterium, Gordonia sp. strain 5F, was isolated from soil polluted with plastic waste. From this strain, a novel esterase (GoEst15) and a mono(2-ethylhexyl) phthalate hydrolase (GoEstM1) were identified by homology-based cloning. GoEst15 showed broad substrate specificity, hydrolyzing DEHP and 10 other PAEs to monoalkyl phthalates, which were further degraded by GoEstMl to phthalic acid. GoEst15 and GoEstM1 were heterologously coexpressed in Escherichia coli BL21 (DE3), which could then completely degrade 12 PAEs (5 mM), within 1 and 24 h for small and bulky substrates, respectively. To our knowledge, GoEst15 is the first DEHP hydrolase with a known protein sequence, which will enable protein engineering to enhance its catalytic performance in the future.
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