Universität zu Köln

Loeschcke, Anita, Dienst, Dennis, Wewer, Vera ORCID: 0000-0002-0199-5228, Hage-Huelsmann, Jennifer, Dietsch, Maximilian, Kranz-Finger, Sarah, Hueren, Vanessa, Metzger, Sabine, Urlacher, Vlada B., Gigolashvili, Tamara ORCID: 0000-0002-0416-4796, Kopriva, Stanislav ORCID: 0000-0002-7416-6551, Axmann, Ilka M., Drepper, Thomas ORCID: 0000-0002-0096-8084 and Jaeger, Karl-Erich (2017). The photosynthetic bacteria Rhodobacter capsulatus and Synechocystis sp PCC 6803 as new hosts for cyclic plant triterpene biosynthesis. PLoS One, 12 (12). SAN FRANCISCO: PUBLIC LIBRARY SCIENCE. ISSN 1932-6203

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Abstract

Cyclic triterpenes constitute one of the most diverse groups of plant natural products. Besides the intriguing biochemistry of their biosynthetic pathways, plant triterpenes exhibit versatile bioactivities, including antimicrobial effects against plant and human pathogens. While prokaryotes have been extensively used for the heterologous production of other classes of terpenes, the synthesis of cyclic triterpenes, which inherently includes the two-step catalytic formation of the universal linear precursor 2,3-oxidosqualene, is still a major challenge. We thus explored the suitability of the metabolically versatile photosynthetic alpha-proteobacterium Rhodobacter capsulatus SB1003 and cyanobacterium Synechocystis sp. PCC 6803 as alternative hosts for biosynthesis of cyclic plant triterpenes. Therefore, 2,3-oxidosqualene production was implemented and subsequently combined with different cyclization reactions catalyzed by the representative oxidosqualene cyclases CAS1 (cycloartenol synthase), LUP1 (lupeol synthase), THAS1 (thalianol synthase) and MRN1 (marneral synthase) derived from model plant Arabidopsis thaliana. While successful accumulation of 2,3-oxidosqualene could be detected by LC-MS analysis in both hosts, cyclase expression resulted in differential production profiles. CAS1 catalyzed conversion to only cycloartenol, but expression of LUP1 yielded lupeol and a triterpenoid matching an oxidation product of lupeol, in both hosts. In contrast, THAS1 expression did not lead to cyclic product formation in either host, whereas MRN1-dependent production of marnerol and hydroxymarnerol was observed in Synechocystis but not in R. capsulatus. Our findings thus indicate that 2,3-oxidosqualene cyclization in heterologous phototrophic bacteria is basically feasible but efficient conversion depends on both the respective cyclase enzyme and individual host properties. Therefore, photosynthetic alpha-proteo- and cyanobacteria are promising alternative candidates for providing new bacterial access to the broad class of triterpenes for biotechnological applications.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Loeschcke, Anita UNSPECIFIED UNSPECIFIED UNSPECIFIED Dienst, Dennis UNSPECIFIED UNSPECIFIED UNSPECIFIED Wewer, Vera UNSPECIFIED orcid.org/0000-0002-0199-5228 UNSPECIFIED Hage-Huelsmann, Jennifer UNSPECIFIED UNSPECIFIED UNSPECIFIED Dietsch, Maximilian UNSPECIFIED UNSPECIFIED UNSPECIFIED Kranz-Finger, Sarah UNSPECIFIED UNSPECIFIED UNSPECIFIED Hueren, Vanessa UNSPECIFIED UNSPECIFIED UNSPECIFIED Metzger, Sabine UNSPECIFIED UNSPECIFIED UNSPECIFIED Urlacher, Vlada B. UNSPECIFIED UNSPECIFIED UNSPECIFIED Gigolashvili, Tamara UNSPECIFIED orcid.org/0000-0002-0416-4796 UNSPECIFIED Kopriva, Stanislav UNSPECIFIED orcid.org/0000-0002-7416-6551 UNSPECIFIED Axmann, Ilka M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Drepper, Thomas UNSPECIFIED orcid.org/0000-0002-0096-8084 UNSPECIFIED Jaeger, Karl-Erich UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-207657
DOI:	10.1371/journal.pone.0189816
Journal or Publication Title:	PLoS One
Volume:	12
Number:	12
Date:	2017
Publisher:	PUBLIC LIBRARY SCIENCE
Place of Publication:	SAN FRANCISCO
ISSN:	1932-6203
Language:	English
Faculty:	Faculty of Mathematics and Natural Sciences
Divisions:	Faculty of Mathematics and Natural Sciences > Department of Biology > Botanical Institute
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language ARABIDOPSIS-THALIANA; HETEROLOGOUS EXPRESSION; OXIDOSQUALENE CYCLASE; LOTUS-JAPONICUS; BIOTECHNOLOGICAL PRODUCTION; CYANOBACTERIAL INOCULATION; HYDROCARBON PRODUCTION; CYCLOARTENOL SYNTHASE; SQUALENE SYNTHASE; MOLECULAR-CLONING Multiple languages Multidisciplinary Sciences Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/20765