Method for isolating and subsequently purifying carbamate-hydrolase, method for producing BrCN-degraded peptide of carbamate-hydrolase, synthetic oligonucleotide, method for isolating carbamate-hydrolase-gene, and novel strain of Arthrobacter oxydans

摘要

The present invention relates to a process for the isolation and characterisation of a gene-enzyme system for the inactivation of the herbicide phenmedipham. The enzyme is a carbamate hydrolase from Arthrobacter oxidans which is responsible for the hydrolytic cleavage of the -OOC-N linkage between the two phenyl nuclei of phenmedipham. The process includes the detection of carbamate hydrolase, the identification of the amino-acid sequence of two BrCN-cleaved peptides of carbamate hydrolase, the synthesis of oligonucleotides for the specific detection of the carbamate hydrolase sequence by hybridisation, and the identification of the coding region, the cloning and the determination of the nucleotide sequence of the carbamate hydrolase gene from Arthrobacter oxidans.