Cytochrome P-450terp. Isolation and purification of the protein and cloning and sequencing of its operon - PubMed (original) (raw)
Comparative Study
. 1992 Jul 15;267(20):14193-203.
Affiliations
- PMID: 1629218
Free article
Comparative Study
Cytochrome P-450terp. Isolation and purification of the protein and cloning and sequencing of its operon
J A Peterson et al. J Biol Chem. 1992.
Free article
Abstract
Cytochromes P-450 are extremely important in the oxidative metabolism of a variety of endogenous and exogenous compounds in pro- and eukaryotic organisms. Progress in understanding the structure and mechanism of action of this superfamily of enzymes has been hampered by the properties of the eukaryotic enzymes and the availability of only one well-characterized prokaryotic enzyme as a model. We report here the isolation of a Pseudomonas species which will utilize a monoterpene natural product, alpha-terpineol, as its sole source of carbon and energy. Approximately 1% of the soluble protein in the cell-free extract is a novel cytochrome P-450 (P-450terp). This enzyme and its associated iron sulfur protein electron carrier (terpredoxin) have been purified to homogeneity and their NH2-terminal amino acid sequences determined. The amino acid sequences of six tryptic peptide fragments of cytochrome P-450terp have also been determined. This sequence information was used to clone the gene encoding cytochrome P-450terp. Three clones representing approximately 8 kilobase pairs of unique sequences were selected and sequenced. Five non-overlapping open reading frames (ORFs) were found in the sequences, and the translated sequences were used to search the Protein Identification Resource for comparable proteins. The ORFs were identified as: 1) an alcohol dehydrogenase, 2) an aldehyde dehydrogenase, 3) cytochrome P-450terp, 4) terpredoxin reductase, and 5) terpredoxin. The identification of both the cytochrome P-450terp and terpredoxin DNA sequence was confirmed by the presence of each of the corresponding amino acid sequences found in the purified proteins. The five ORFs were bounded on both the 5' and 3' ends by consensus factor-independent terminator sequences. A consensus promoter sequence was found immediately 5' to the first ORF. These results indicate that we have sequenced the complete terp operon. Comparison of the amino acid sequence of cytochrome P-450terp to that of all other cytochromes P-450 has shown that it is the first member of the gene family CYP108. Preliminary characterization of the chemical and physical properties and the preparation of crystals of this new cytochrome P-450, suitable for x-ray diffraction analysis, indicate that it will be useful in comparison studies with other members of this class of proteins.
Similar articles
- Cytochrome P450(cin) (CYP176A), isolation, expression, and characterization.
Hawkes DB, Adams GW, Burlingame AL, Ortiz de Montellano PR, De Voss JJ. Hawkes DB, et al. J Biol Chem. 2002 Aug 2;277(31):27725-32. doi: 10.1074/jbc.M203382200. Epub 2002 May 16. J Biol Chem. 2002. PMID: 12016226 - Cloning and sequencing of a phenol hydroxylase gene of Pseudomonas pseudoalcaligenes strain MH1: a bacterium able to mineralize various aromatic compounds.
Zouari H, Moukha S, Labat M, Sayadi S. Zouari H, et al. Appl Biochem Biotechnol. 2002 Jul-Dec;102-103(1-6):261-76. doi: 10.1385/abab:102-103:1-6:261. Appl Biochem Biotechnol. 2002. PMID: 12396129 - The Pseudomonas oleovorans alkBAC operon encodes two structurally related rubredoxins and an aldehyde dehydrogenase.
Kok M, Oldenhuis R, van der Linden MP, Meulenberg CH, Kingma J, Witholt B. Kok M, et al. J Biol Chem. 1989 Apr 5;264(10):5442-51. J Biol Chem. 1989. PMID: 2647719 - Identification and molecular characterization of the eugenol hydroxylase genes (ehyA/ehyB) of Pseudomonas sp. strain HR199.
Priefert H, Overhage J, Steinbüchel A. Priefert H, et al. Arch Microbiol. 1999 Dec;172(6):354-63. doi: 10.1007/s002030050772. Arch Microbiol. 1999. PMID: 10591845
Cited by
- Cooperative Substrate Binding Controls Catalysis in Bacterial Cytochrome P450terp (CYP108A1).
Gable JA, Poulos TL, Follmer AH. Gable JA, et al. J Am Chem Soc. 2023 Feb 13:10.1021/jacs.2c12388. doi: 10.1021/jacs.2c12388. Online ahead of print. J Am Chem Soc. 2023. PMID: 36779970 Free PMC article. - Current Advances in the Bacterial Toolbox for the Biotechnological Production of Monoterpene-Based Aroma Compounds.
Soares-Castro P, Soares F, Santos PM. Soares-Castro P, et al. Molecules. 2020 Dec 28;26(1):91. doi: 10.3390/molecules26010091. Molecules. 2020. PMID: 33379215 Free PMC article. Review. - A new approach to understanding structure-function relationships in cytochromes P450 by targeting terpene metabolism in the wild.
Wong NR, Liu X, Lloyd H, Colthart AM, Ferrazzoli AE, Cooper DL, Zhuang Y, Esquea P, Futcher J, Pochapsky TM, Matthews JM, Pochapsky TC. Wong NR, et al. J Inorg Biochem. 2018 Nov;188:96-101. doi: 10.1016/j.jinorgbio.2018.08.006. Epub 2018 Aug 6. J Inorg Biochem. 2018. PMID: 30170307 Free PMC article. - CYP101J2, CYP101J3, and CYP101J4, 1,8-Cineole-Hydroxylating Cytochrome P450 Monooxygenases from Sphingobium yanoikuyae Strain B2.
Unterweger B, Bulach DM, Scoble J, Midgley DJ, Greenfield P, Lyras D, Johanesen P, Dumsday GJ. Unterweger B, et al. Appl Environ Microbiol. 2016 Oct 27;82(22):6507-6517. doi: 10.1128/AEM.02067-16. Print 2016 Nov 15. Appl Environ Microbiol. 2016. PMID: 27590809 Free PMC article. - Identification, characterization and molecular adaptation of class I redox systems for the production of hydroxylated diterpenoids.
Görner C, Schrepfer P, Redai V, Wallrapp F, Loll B, Eisenreich W, Haslbeck M, Brück T. Görner C, et al. Microb Cell Fact. 2016 May 23;15:86. doi: 10.1186/s12934-016-0487-6. Microb Cell Fact. 2016. PMID: 27216162 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources