Genetic analysis of the phosphinothricin-tripeptide biosynthetic pathway of Streptomyces viridochromogenes Tü494.
Streptomyces viridochromogenes Tü494 produces the antibiotic phosphinothricyl-alanyl-alanine (Ptt). Ptt-non-producing mutants were isolated following N-methyl-N'-nitro-N-nitrosoguanidine (NTG) or UV light treatment of spore suspensions. In co-synthesis and bioconversion experiments the mutational blocks in the biosynthetic pathway were localized. The mutant NTG1 was analysed in detail. This mutant acts as a secretor for all other mutants. From bioconversion experiments with presumptive precursors circumstantial evidence was obtained that NTG1 is mutated in a gene involved in the alanylation of N-acetyl-demethyl-phosphinothricin. Using a cosmid gene library the DNA region complementing the defective gene of mutant NTG1 was isolated on a 4-kb BamHI fragment. Subcloning experiments showed that a 3-kb BglII/BamHI fragment is sufficient for complementation of mutant NTG1.
Alijah
R
R
Universität Bielefeld, Lehrstuhl für Genetik, Federal Republic of Germany.
Dorendorf
J
J
Talay
S
S
Pühler
A
A
Wohlleben
W
W
D000900:Anti-Bacterial Agents / Q000378:metabolism*; D003360:Cosmids; D004269:DNA, Bacterial / Q000302:isolation & purification; D004352:Drug Resistance, Microbial / Q000235:genetics; D005816:Genetic Complementation Test; D015698:Genomic Library; D009154:Mutation; D009943:Organophosphorus Compounds / Q000378:metabolism*; D015183:Restriction Mapping; D013171:Spores, Bacterial / Q000528:radiation effects; D013302:Streptomyces / Q000235:genetics*; D014466:Ultraviolet Rays
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D009943:Organophosphorus Compounds; C003122:bialaphos