Inability of muconate cycloisomerases to cause dehalogenation during conversion of 2-chloro-cis,cis-muconate.
The conversion of 2-chloro-cis,cis-muconate by muconate cycloisomerase from Pseudomonas putida PRS2000 yielded two products which by nuclear magnetic resonance spectroscopy were identified as 2-chloro- and 5-chloromuconolactone. High-pressure liquid chromatography analyses showed the same compounds to be formed also by muconate cycloisomerases from Acinetobacter calcoaceticus ADP1 and Pseudomonas sp. strain B13. During 2-chloro-cis,cis-muconate turnover by the enzyme from P. putida, 2-chloromuconolactone initially was the major product. After prolonged incubation, however, 5-chloromuconolactone dominated in the resulting equilibrium. In contrast to previous assumptions, both chloromuconolactones were found to be stable at physiological pH. Since the chloromuconate cycloisomerases of Pseudomonas sp. strain B13 and Alcaligenes eutrophus JMP134 have been shown previously to produce the trans-dienelactone (trans-4-carboxymethylene-but-2-en-4-olide) from 2-chloro-cis,cis-muconate, they must have evolved the capability to cleave the carbon-chlorine bond during their divergence from normal muconate cycloisomerases.
Institut für Mikrobiologie, Universität Stuttgart, Germany.
M D
MD
Vollmer
P
P
Fischer
H J
HJ
Knackmuss
M
M
Schlömann
D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000302:isolation & purification / Q000378:metabolism; D016954:Acinetobacter calcoaceticus / Q000201:enzymology; D000272:Adipates / Q000378:metabolism*; D019753:Intramolecular Lyases; D007535:Isomerases / Q000302:isolation & purification / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D011549:Pseudomonas / Q000201:enzymology; D016958:Pseudomonas putida / Q000201:enzymology*; D013011:Sorbic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D013379:Substrate Specificity
D000272:Adipates; C088142:5-chloromuconolactone; C088140:2-chloromuconic acid; C088141:2-chloromuconolactone; D007535:Isomerases; D019753:Intramolecular Lyases; C021811:muconate cycloisomerase; D015107:4-Butyrolactone; D013011:Sorbic Acid