Cloning of the gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase from terpenoid antibiotic-producing Streptomyces strains.
We have isolated a mutant lacking 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase activity from a terpenoid antibiotic (terpentecin) producer, Streptomyces griseolosporeus MF730-N6, which uses both the mevalonate and nonmevalonate pathways for the formation of isopentenyl diphosphate, by screening terpentecin non-producing mutants. Terpentecin is known to be synthesized via the mevalonate pathway. The gene encoding HMG-CoA reductase (hmgg) was cloned and identified by complementation of the mutant, using a self-cloning system developed in this study for strain MF730-N6. The corresponding hmgs gene for HMG-CoA reductase was also cloned from Streptomyces sp. KO-3988, which produces the terpenoid antibiotic furaquinocin. Sequence analysis of hmgg and hmgs showed that both genes encode polypeptides of 353 amino acids which are 84% identical to each other. A search of protein sequence databases revealed that both gene products were also similar to HMG-CoA reductases from a variety of other organisms, including Streptomyces sp. CL190 (hmgg is 89% and hmgs 85% identical to its CL190 homolog), sea urchin (40.3 and 40.5%), German cockroach (37.6 and 38.4%), and Camptotheca acuminata (39.7 and 40.8%).
Dairi
T
T
Biotechnology Research Center, Toyama Prefectural University, Japan.
Motohira
Y
Y
Kuzuyama
T
T
Takahashi
S
S
Itoh
N
N
Seto
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D003001:Cloning, Molecular; D004224:Diterpenes / Q000378:metabolism; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D006801:Humans; D006903:Hydroxymethylglutaryl CoA Reductases / Q000235:genetics*; D008798:Mevalonic Acid / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009285:Naphthoquinones / Q000378:metabolism; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D004224:Diterpenes; D009285:Naphthoquinones; C047785:terpentecin; D006903:Hydroxymethylglutaryl CoA Reductases; D008798:Mevalonic Acid