No Cover Image

Journal article 837 views

Investigating conservation of the albaflavenone biosynthetic pathway and CYP170 bifunctionality in streptomycetes.

SC Moody, B Zhao, L Lei, Jonathan Mullins Orcid Logo, MR Waterman, Steven Kelly Orcid Logo, DC Lamb

FEBS J., Volume: 279, Issue: 9, Pages: 1640 - 1649

Swansea University Authors: Jonathan Mullins Orcid Logo, Steven Kelly Orcid Logo

Full text not available from this repository: check for access using links below.

DOI (Published version): 10.1111/j.1742-4658.2011.08447.x.

Abstract

Albaflavenone, a tricyclic sesquiterpene antibiotic, is biosynthesized in Streptomyces coelicolor A3(2) by enzymes encoded in a two-gene operon. Initially, sesquiterpene cyclase catalyzes the cyclization of farnesyl diphosphate to the terpenoid epi-isozizaene, which is oxidized to the final albaflav...

Full description

Published in: FEBS J.
Published: FEBS 2012
URI: https://cronfa.swan.ac.uk/Record/cronfa10672
Tags: Add Tag
No Tags, Be the first to tag this record!
first_indexed 2013-07-23T12:04:02Z
last_indexed 2021-10-30T02:21:56Z
id cronfa10672
recordtype SURis
fullrecord <?xml version="1.0"?><rfc1807><datestamp>2021-10-29T09:39:26.4152715</datestamp><bib-version>v2</bib-version><id>10672</id><entry>2012-04-19</entry><title>Investigating conservation of the albaflavenone biosynthetic pathway and CYP170 bifunctionality in streptomycetes.</title><swanseaauthors><author><sid>4cf2dddedbe1dacb506ec925fdbd5b40</sid><ORCID>0000-0003-0144-2962</ORCID><firstname>Jonathan</firstname><surname>Mullins</surname><name>Jonathan Mullins</name><active>true</active><ethesisStudent>false</ethesisStudent></author><author><sid>b17cebaf09b4d737b9378a3581e3de93</sid><ORCID>0000-0001-7991-5040</ORCID><firstname>Steven</firstname><surname>Kelly</surname><name>Steven Kelly</name><active>true</active><ethesisStudent>false</ethesisStudent></author></swanseaauthors><date>2012-04-19</date><deptcode>BMS</deptcode><abstract>Albaflavenone, a tricyclic sesquiterpene antibiotic, is biosynthesized in Streptomyces&#x2003;coelicolor&#x2003;A3(2) by enzymes encoded in a two-gene operon. Initially, sesquiterpene cyclase catalyzes the cyclization of farnesyl diphosphate to the terpenoid epi-isozizaene, which is oxidized to the final albaflavenone by cytochrome&#x2003;P450 (CYP)170A1. Additionally, this CYP is a bifunctional enzyme, being able to also generate farnesene isomers from farnesyl diphosphate, owing to a terpene synthase active site moonlighting on the CYP molecule. To explore the functionality of this operon in other streptomycetes, we have examined culture extracts by GC/MS and established the presence of albaflavenone in five Streptomyces species. Bioinformatics examination of the predicted CYP170 primary amino acid sequences revealed substitutions in the CYP terpene synthase active site. To examine whether the terpene synthase site was catalytically active in another CYP170, we characterized the least related CYP170 orthologue from Streptomyces albus (CYP170B1). Following expression and purification, CYP170B1 showed a normal reduced CO difference spectrum at 450&#x2003;nm, in contrast to the unusual 440-nm peak observed for S.&#x2003;coelicolor A3(2) CYP170A1. CYP170B1 can catalyze the conversion of epi-isozizaene to albaflavenone, but was unable to catalyze the conversion of farnesyl diphosphate to farnesene. Molecular modeling with our crystal structure of CYP170A1 suggests that the absence of key amino acids for binding the essential terpene synthase cofactor Mg(2+) may be the explanation for the loss of CYP170B1 bifunctionality.</abstract><type>Journal Article</type><journal>FEBS J.</journal><volume>279</volume><journalNumber>9</journalNumber><paginationStart>1640</paginationStart><paginationEnd>1649</paginationEnd><publisher>FEBS</publisher><placeOfPublication/><isbnPrint/><isbnElectronic/><issnPrint/><issnElectronic/><keywords/><publishedDay>31</publishedDay><publishedMonth>1</publishedMonth><publishedYear>2012</publishedYear><publishedDate>2012-01-31</publishedDate><doi>10.1111/j.1742-4658.2011.08447.x.</doi><url/><notes/><college>COLLEGE NANME</college><department>Biomedical Sciences</department><CollegeCode>COLLEGE CODE</CollegeCode><DepartmentCode>BMS</DepartmentCode><institution>Swansea University</institution><apcterm/><lastEdited>2021-10-29T09:39:26.4152715</lastEdited><Created>2012-04-19T11:17:05.7740338</Created><path><level id="1">Faculty of Medicine, Health and Life Sciences</level><level id="2">Swansea University Medical School - Medicine</level></path><authors><author><firstname>SC</firstname><surname>Moody</surname><order>1</order></author><author><firstname>B</firstname><surname>Zhao</surname><order>2</order></author><author><firstname>L</firstname><surname>Lei</surname><order>3</order></author><author><firstname>Jonathan</firstname><surname>Mullins</surname><orcid>0000-0003-0144-2962</orcid><order>4</order></author><author><firstname>MR</firstname><surname>Waterman</surname><order>5</order></author><author><firstname>Steven</firstname><surname>Kelly</surname><orcid>0000-0001-7991-5040</orcid><order>6</order></author><author><firstname>DC</firstname><surname>Lamb</surname><order>7</order></author></authors><documents/><OutputDurs/></rfc1807>
spelling 2021-10-29T09:39:26.4152715 v2 10672 2012-04-19 Investigating conservation of the albaflavenone biosynthetic pathway and CYP170 bifunctionality in streptomycetes. 4cf2dddedbe1dacb506ec925fdbd5b40 0000-0003-0144-2962 Jonathan Mullins Jonathan Mullins true false b17cebaf09b4d737b9378a3581e3de93 0000-0001-7991-5040 Steven Kelly Steven Kelly true false 2012-04-19 BMS Albaflavenone, a tricyclic sesquiterpene antibiotic, is biosynthesized in Streptomyces coelicolor A3(2) by enzymes encoded in a two-gene operon. Initially, sesquiterpene cyclase catalyzes the cyclization of farnesyl diphosphate to the terpenoid epi-isozizaene, which is oxidized to the final albaflavenone by cytochrome P450 (CYP)170A1. Additionally, this CYP is a bifunctional enzyme, being able to also generate farnesene isomers from farnesyl diphosphate, owing to a terpene synthase active site moonlighting on the CYP molecule. To explore the functionality of this operon in other streptomycetes, we have examined culture extracts by GC/MS and established the presence of albaflavenone in five Streptomyces species. Bioinformatics examination of the predicted CYP170 primary amino acid sequences revealed substitutions in the CYP terpene synthase active site. To examine whether the terpene synthase site was catalytically active in another CYP170, we characterized the least related CYP170 orthologue from Streptomyces albus (CYP170B1). Following expression and purification, CYP170B1 showed a normal reduced CO difference spectrum at 450 nm, in contrast to the unusual 440-nm peak observed for S. coelicolor A3(2) CYP170A1. CYP170B1 can catalyze the conversion of epi-isozizaene to albaflavenone, but was unable to catalyze the conversion of farnesyl diphosphate to farnesene. Molecular modeling with our crystal structure of CYP170A1 suggests that the absence of key amino acids for binding the essential terpene synthase cofactor Mg(2+) may be the explanation for the loss of CYP170B1 bifunctionality. Journal Article FEBS J. 279 9 1640 1649 FEBS 31 1 2012 2012-01-31 10.1111/j.1742-4658.2011.08447.x. COLLEGE NANME Biomedical Sciences COLLEGE CODE BMS Swansea University 2021-10-29T09:39:26.4152715 2012-04-19T11:17:05.7740338 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine SC Moody 1 B Zhao 2 L Lei 3 Jonathan Mullins 0000-0003-0144-2962 4 MR Waterman 5 Steven Kelly 0000-0001-7991-5040 6 DC Lamb 7
title Investigating conservation of the albaflavenone biosynthetic pathway and CYP170 bifunctionality in streptomycetes.
spellingShingle Investigating conservation of the albaflavenone biosynthetic pathway and CYP170 bifunctionality in streptomycetes.
Jonathan Mullins
Steven Kelly
title_short Investigating conservation of the albaflavenone biosynthetic pathway and CYP170 bifunctionality in streptomycetes.
title_full Investigating conservation of the albaflavenone biosynthetic pathway and CYP170 bifunctionality in streptomycetes.
title_fullStr Investigating conservation of the albaflavenone biosynthetic pathway and CYP170 bifunctionality in streptomycetes.
title_full_unstemmed Investigating conservation of the albaflavenone biosynthetic pathway and CYP170 bifunctionality in streptomycetes.
title_sort Investigating conservation of the albaflavenone biosynthetic pathway and CYP170 bifunctionality in streptomycetes.
author_id_str_mv 4cf2dddedbe1dacb506ec925fdbd5b40
b17cebaf09b4d737b9378a3581e3de93
author_id_fullname_str_mv 4cf2dddedbe1dacb506ec925fdbd5b40_***_Jonathan Mullins
b17cebaf09b4d737b9378a3581e3de93_***_Steven Kelly
author Jonathan Mullins
Steven Kelly
author2 SC Moody
B Zhao
L Lei
Jonathan Mullins
MR Waterman
Steven Kelly
DC Lamb
format Journal article
container_title FEBS J.
container_volume 279
container_issue 9
container_start_page 1640
publishDate 2012
institution Swansea University
doi_str_mv 10.1111/j.1742-4658.2011.08447.x.
publisher FEBS
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
document_store_str 0
active_str 0
description Albaflavenone, a tricyclic sesquiterpene antibiotic, is biosynthesized in Streptomyces coelicolor A3(2) by enzymes encoded in a two-gene operon. Initially, sesquiterpene cyclase catalyzes the cyclization of farnesyl diphosphate to the terpenoid epi-isozizaene, which is oxidized to the final albaflavenone by cytochrome P450 (CYP)170A1. Additionally, this CYP is a bifunctional enzyme, being able to also generate farnesene isomers from farnesyl diphosphate, owing to a terpene synthase active site moonlighting on the CYP molecule. To explore the functionality of this operon in other streptomycetes, we have examined culture extracts by GC/MS and established the presence of albaflavenone in five Streptomyces species. Bioinformatics examination of the predicted CYP170 primary amino acid sequences revealed substitutions in the CYP terpene synthase active site. To examine whether the terpene synthase site was catalytically active in another CYP170, we characterized the least related CYP170 orthologue from Streptomyces albus (CYP170B1). Following expression and purification, CYP170B1 showed a normal reduced CO difference spectrum at 450 nm, in contrast to the unusual 440-nm peak observed for S. coelicolor A3(2) CYP170A1. CYP170B1 can catalyze the conversion of epi-isozizaene to albaflavenone, but was unable to catalyze the conversion of farnesyl diphosphate to farnesene. Molecular modeling with our crystal structure of CYP170A1 suggests that the absence of key amino acids for binding the essential terpene synthase cofactor Mg(2+) may be the explanation for the loss of CYP170B1 bifunctionality.
published_date 2012-01-31T03:12:08Z
_version_ 1763750055623262208
score 11.037056

Similar Items