title: Cloning, expression, characterization and application of atcA, atcB and atcC from Pseudomonas sp. for the production of L-cysteine.
abstract: An isolate of a Pseudomonas sp. uses the L-NCC (N-carbamoyl-L-cysteine) pathway to convert DL-2-amino-Δ(2)-thiazoline-4-carboxylic acid (DL-ATC) to L-cysteine. Genes encoding ATC racemase (AtcA), L-ATC hydrolase (AtcB) and L-NCC amidohydrolase (AtcC), involved in this pathway, were cloned from the Pseudomonas sp. and expressed in Escherichia coli BL21 via pET-28a(+). The resulting enzymes were purified, their functions identified, and their biochemical properties are described. In vitro catalysis experiments, using these enzymes, revealed that the bioconversion rate of L-cysteine from DL-ATC in the presence of AtcA was more efficient than in the absence of AtcA. This is the first report describing simultaneous cloning and expression of atcA, atcB and atcC and characterization of their enzymes for L-cysteine production from DL-ATC via the L-NCC pathway, enabling the complete L-NCC pathway to be elucidated.
authors.affiliation: College of Pharmacy, State Key Laboratory of Medicinal Chemical Biology and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, China. duankangjing@163.com
authors.forename: Jingjing
authors.initials: J
authors.lastname: Duan
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.forename: Hongzhi
authors.initials: H
authors.lastname: Zhao
authors.forename: Jun
authors.initials: J
authors.lastname: Du
authors.forename: Fang
authors.initials: F
authors.lastname: Bai
authors.forename: Gang
authors.initials: G
authors.lastname: Bai
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D001711:Biotransformation; D003001:Cloning, Molecular; D003545:Cysteine / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006867:Hydrolases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D053858:Metabolic Networks and Pathways / Q000235:genetics; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics* / Q000302:isolation & purification; D019998:Racemases and Epimerases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D013844:Thiazoles; C015674:2-amino-delta(2)-thiazoline-4-carboxylic acid; D006867:Hydrolases; D000581:Amidohydrolases; C045389:2-amino-delta(2)-thiazoline-4-carboxylate hydrolase; D019998:Racemases and Epimerases; D003545:Cysteine