P450 Cytochromes and antibiotic biosynthesis
We study the structure of microbial P450 cytochromes to decipher substrate recognition detrminants and the structural transitions involved in the antibiotic tailoring steps, crucial for obtaining the final active compound or to engineer the production of new ones.
We demonstrated the structural selection mechanism involved in the activity of the EryK P450, leading to Erythromycin A production and we engineered a variant of the enzyme that could eliminate the presence of the shunt product Erythromycin B.
We are currently analysing the substrate specificity and enzyme dynamics of the OleP P450, endowed with substrate promiscuity and carrying out a rare and potentially very interesting epoxidation within the antibiotic oleandomycin.
Substrate-induced conformational change in cytochrome P450 OleP.
Parisi G, Montemiglio LC, Giuffrè A, Macone A, Scaglione A, Cerutti G, Exertier C, Savino C, Vallone B.
FASEB J. 2019 Feb;33(2):1787-1800. doi: 10.1096/fj.201800450RR. Epub 2018 Sep 12.
Functional analysis and crystallographic structure of clotrimazole bound OleP, a cytochrome P450 epoxidase from Streptomyces antibioticus involved in oleandomycin biosynthesis.
Montemiglio LC, Parisi G, Scaglione A, Sciara G, Savino C, Vallone B.
Biochim Biophys Acta. 2016 Mar;1860(3):465-75. doi: 10.1016/j.bbagen.2015.10.009. Epub 2015 Oct 22.
Redirecting P450 EryK specificity by rational site-directed mutagenesis.
Montemiglio LC, Macone A, Ardiccioni C, Avella G, Vallone B, Savino C.
Biochemistry. 2013 May 28;52(21):3678-87. doi: 10.1021/bi400223j. Epub 2013 May 16.
Azole drugs trap cytochrome P450 EryK in alternative conformational states.
Montemiglio LC, Gianni S, Vallone B, Savino C.
Biochemistry. 2010 Nov 2;49(43):9199-206. doi: 10.1021/bi101062v.
Investigating the structural plasticity of a cytochrome P450: three-dimensional structures of P450 EryK and binding to its physiological substrate.
Savino C, Montemiglio LC, Sciara G, Miele AE, Kendrew SG, Jemth P, Gianni S, Vallone B.
J Biol Chem. 2009 Oct 16;284(42):29170-9. doi: 10.1074/jbc.M109.003590. Epub 2009 Jul 22.
Free PMC Article
Cloning, expression, purification, crystallization and preliminary X-ray crystallographic analysis of C-12 hydroxylase EryK from Saccharopolyspora erythraea.
Savino C, Sciara G, Miele AE, Kendrew SG, Vallone B.
Protein Pept Lett. 2008;15(10):1138-41.