Cloning of the Rhizomucor miehei 3-hydroxy-3-methylglutaryl-coenzyme A reductase gene and its heterologous expression in Mucor circinelloides.
In this study, the gene hmgR encoding the 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) was cloned and characterized in the zygomycete fungus Rhizomucor miehei. The hmgR gene comprises a total of 3,585 bp including the coding sequence of a 1,058 amino acids length putative protein and five introns (137, 83, 59, 60 and 69 bp in length) dispersed in the whole coding region. Southern hybridization analysis revealed that the gene is present only in one copy in the R. miehei genome. The isolated Rhizomucor gene was expressed in the related fungus, Mucor circinelloides. Transformants harbouring the Rhizomucor hmgR gene in an autoreplicative plasmid proved to be more tolerant to statins (e.g. lovastatin, simvastatin, and fluvastatin), the competitive inhibitors of the HMG-CoA reductase, than the original M. circinelloides strain. At the same time, heterologous expression of the Rhizomucor hmgR did not affect the carotenoid production of M. circinelloides.
Lukács
Gyöngyi
G
Department of Microbiology, Faculty of Sciences and Informatics, University of Szeged, Közép fasor 52, 6726, Szeged, Hungary.
Papp
Tamás
T
Somogyvári
Ferenc
F
Csernetics
Arpád
A
Nyilasi
Ildikó
I
Vágvölgyi
Csaba
C
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005658:Fungi / Q000145:classification / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D025782:Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009089:Mucor / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D020103:Rhizomucor / Q000201:enzymology*; D016415:Sequence Alignment
D005656:Fungal Proteins; D025782:Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent