Genetic characterization of the resorcinol catabolic pathway in Corynebacterium glutamicum.
Corynebacterium glutamicum grew on resorcinol as a sole source of carbon and energy. By genome-wide data mining, two gene clusters, designated NCgl1110-NCgl1113 and NCgl2950-NCgl2953, were proposed to encode putative proteins involved in resorcinol catabolism. Deletion of the NCgl2950-NCgl2953 gene cluster did not result in any observable phenotype changes. Disruption and complementation of each gene at NCgl1110-NCgl1113, NCgl2951, and NCgl2952 indicated that these genes were involved in resorcinol degradation. Expression of NCgl1112, NCgl1113, and NCgl2951 in Escherichia coli revealed that NCgl1113 and NCgl2951 both coded for hydroxyquinol 1,2-dioxygenases and NCgl1112 coded for maleylacetate reductases. NCgl1111 encoded a putative monooxygenase, but this putative hydroxylase was very different from previously functionally identified hydroxylases. Cloning and expression of NCgl1111 in E. coli revealed that NCgl1111 encoded a resorcinol hydroxylase that needs NADPH as a cofactor. E. coli cells containing Ncgl1111 and Ncgl1113 sequentially converted resorcinol into maleylacetate. NCgl1110 and NCgl2950 both encoded putative TetR family repressors, but only NCgl1110 was transcribed and functional. NCgl2953 encoded a putative transporter, but disruption of this gene did not affect resorcinol degradation by C. glutamicum. The function of NCgl2953 remains unclear.
Huang
Yan
Y
Institute of Microbiology, Chinese Academy of Sciences, ZhongGuanCun, Haidian, Beijing 100080, People's Republic of China.
Zhao
Ke-Xin
KX
Shen
Xi-Hui
XH
Chaudhry
Muhammad Tausif
MT
Jiang
Cheng-Ying
CY
Liu
Shuang-Jiang
SJ
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D048230:Corynebacterium glutamicum / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D049308:Dioxygenases / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism; D005810:Multigene Family; D044925:Oxidoreductases Acting on CH-CH Group Donors / Q000235:genetics / Q000378:metabolism; D012118:Resorcinols / Q000378:metabolism*
D001426:Bacterial Proteins; D012118:Resorcinols; D006899:Mixed Function Oxygenases; D049308:Dioxygenases; C041894:hydroxyquinol 1,2-dioxygenase; C037083:resorcinol hydroxylase; D044925:Oxidoreductases Acting on CH-CH Group Donors; C021530:maleylacetate reductase; C031389:resorcinol