title: Characterization of a Unique Pathway for 4-Cresol Catabolism Initiated by Phosphorylation in Corynebacterium glutamicum.
abstract: 4-Cresol is not only a significant synthetic intermediate for production of many aromatic chemicals, but also a priority environmental pollutant because of its toxicity to higher organisms. In our previous studies, a gene cluster implicated to be involved in 4-cresol catabolism, creCDEFGHIR, was identified in Corynebacterium glutamicum and partially characterized in vivo. In this work, we report on the discovery of a novel 4-cresol biodegradation pathway that employs phosphorylated intermediates. This unique pathway initiates with the phosphorylation of the hydroxyl group of 4-cresol, which is catalyzed by a novel 4-methylbenzyl phosphate synthase, CreHI. Next, a unique class I P450 system, CreJEF, specifically recognizes phosphorylated intermediates and successively oxidizes the aromatic methyl group into carboxylic acid functionality via alcohol and aldehyde intermediates. Moreover, CreD (phosphohydrolase), CreC (alcohol dehydrogenase), and CreG (aldehyde dehydrogenase) were also found to be required for efficient oxidative transformations in this pathway. Steady-state kinetic parameters (Km and kcat) for each catabolic step were determined, and these results suggest that kinetic controls serve a key role in directing the metabolic flux to the most energy effective route.
authors.affiliation: From the CAS Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology and the University of the Chinese Academy of Sciences, Beijing 100049, China, and.
authors.forename: Lei
authors.initials: L
authors.lastname: Du
authors.affiliation: From the CAS Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology and.
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.affiliation: From the CAS Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology and.
authors.forename: Feifei
authors.initials: F
authors.lastname: Qi
authors.affiliation: From the CAS Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology and the University of the Chinese Academy of Sciences, Beijing 100049, China, and.
authors.forename: Xianliang
authors.initials: X
authors.lastname: Zheng
authors.affiliation: the State Key Laboratory of Microbial Resources, and Environmental Microbiology and Biotechnology Research Center at Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Chengying
authors.initials: C
authors.lastname: Jiang
authors.affiliation: the CAS Key Laboratory of Bio-based Materials at Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, Shandong 266101, China.
authors.forename: Ailei
authors.initials: A
authors.lastname: Li
authors.affiliation: the CAS Key Laboratory of Bio-based Materials at Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, Shandong 266101, China.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Wan
authors.affiliation: the State Key Laboratory of Microbial Resources, and Environmental Microbiology and Biotechnology Research Center at Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China liusj@im.ac.cn.
authors.forename: Shuang-Jiang
authors.initials: SJ
authors.lastname: Liu
authors.affiliation: From the CAS Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology and lishengying@qibebt.ac.cn.
authors.forename: Shengying
authors.initials: S
authors.lastname: Li
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D055162:Biocatalysis; D048230:Corynebacterium glutamicum / Q000201:enzymology / Q000235:genetics*; D003408:Cresols / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics; D007700:Kinetics; D053858:Metabolic Networks and Pathways; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry / Q000235:genetics; D010766:Phosphorylation; D010770:Phosphotransferases / Q000737:chemistry / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D003408:Cresols; C032538:4-cresol; D000255:Adenosine Triphosphate; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D010770:Phosphotransferases; D010744:Phosphoric Monoester Hydrolases
keywords: 4-cresol; Corynebacterium glutamicum; biodegradation; cytochrome P450; gram-positive bacteria; microbiology; phosphorylation
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