BRENDA - Enzyme Database show
show all sequences of 5.4.99.53

The photosynthetic bacteria Rhodobacter capsulatus and Synechocystis sp. PCC 6803 as new hosts for cyclic plant triterpene biosynthesis

Loeschcke, A.; Dienst, D.; Wewer, V.; Hage-Huelsmann, J.; Dietsch, M.; Kranz-Finger, S.; Hueren, V.; Metzger, S.; Urlacher, V.B.; Gigolashvili, T.; Kopriva, S.; Axmann, I.M.; Drepper, T.; Jaeger, K.E.; PLoS ONE 12, e0189816 (2017)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene MRN1, recombinant expression in Rhodobacter capsulatus strain SB1003. and Synechocystis sp. PCC 6803 using promoters Pnif and PcoaT, coexpression with CAS1, LUP1, and THAS1. Subcloning in Saccharomyces cerevisiae strain GIL77, both marnerol and the putative hydroxymarnerol are detected in the reference yeast strain expressing MRN1. Isolation of triterpenes, while cycloartenol and lupeol represent typical plant triterpenes with tetracyclic plant sterol and pentacyclic plant triterpene scaffolds, respectively, thalianol and marneral exhibit more unusual tri- and monocyclic structures
Arabidopsis thaliana
Engineering
Amino acid exchange
Commentary
Organism
additional information
2,3-oxidosqualene production is 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, in the two alternative hosts for biosynthesis of cyclic plant triterpenes, the metabolically versatile photosynthetic alpha-proteobacterium Rhodobacter capsulatus strain SB1003 and cyanobacterium Synechocystis sp. PCC 6803, triterpene pathway design, overview. While successful accumulation of 2,3-oxidosqualene can be detected by LC-MS analysis in both hosts, cyclase expression results in differential production profiles. CAS1 catalyzes conversion to only cycloartenol, but expression of LUP1 yields lupeol and a triterpenoid matching an oxidation product of lupeol, in both hosts. In contrast, THAS1 expression does not lead to cyclic product formation in either host, whereas MRN1-dependent production of marnerol and hydroxymarnerol is observed in Synechocystis but not in Rhodobacter capsulatus. 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
Arabidopsis thaliana
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(3S)-2,3-epoxy-2,3-dihydrosqualene
Arabidopsis thaliana
-
marneral
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Arabidopsis thaliana
Q9FJV8
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(3S)-2,3-epoxy-2,3-dihydrosqualene
-
749117
Arabidopsis thaliana
marneral
-
-
-
?
Cloned(Commentary) (protein specific)
Commentary
Organism
gene MRN1, recombinant expression in Rhodobacter capsulatus strain SB1003. and Synechocystis sp. PCC 6803 using promoters Pnif and PcoaT, coexpression with CAS1, LUP1, and THAS1. Subcloning in Saccharomyces cerevisiae strain GIL77, both marnerol and the putative hydroxymarnerol are detected in the reference yeast strain expressing MRN1. Isolation of triterpenes, while cycloartenol and lupeol represent typical plant triterpenes with tetracyclic plant sterol and pentacyclic plant triterpene scaffolds, respectively, thalianol and marneral exhibit more unusual tri- and monocyclic structures
Arabidopsis thaliana
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
2,3-oxidosqualene production is 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, in the two alternative hosts for biosynthesis of cyclic plant triterpenes, the metabolically versatile photosynthetic alpha-proteobacterium Rhodobacter capsulatus strain SB1003 and cyanobacterium Synechocystis sp. PCC 6803, triterpene pathway design, overview. While successful accumulation of 2,3-oxidosqualene can be detected by LC-MS analysis in both hosts, cyclase expression results in differential production profiles. CAS1 catalyzes conversion to only cycloartenol, but expression of LUP1 yields lupeol and a triterpenoid matching an oxidation product of lupeol, in both hosts. In contrast, THAS1 expression does not lead to cyclic product formation in either host, whereas MRN1-dependent production of marnerol and hydroxymarnerol is observed in Synechocystis but not in Rhodobacter capsulatus. 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
Arabidopsis thaliana
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(3S)-2,3-epoxy-2,3-dihydrosqualene
Arabidopsis thaliana
-
marneral
-
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(3S)-2,3-epoxy-2,3-dihydrosqualene
-
749117
Arabidopsis thaliana
marneral
-
-
-
?
General Information
General Information
Commentary
Organism
physiological function
the OSC MRN1 from Arabidopsis thaliana catalyzes the formation of the seco-triterpene marneral via an unusual chair-boat substrate conformation, resulting in a further atypical, monocyclic triterpenoid structure, LC-MS analysis
Arabidopsis thaliana
General Information (protein specific)
General Information
Commentary
Organism
physiological function
the OSC MRN1 from Arabidopsis thaliana catalyzes the formation of the seco-triterpene marneral via an unusual chair-boat substrate conformation, resulting in a further atypical, monocyclic triterpenoid structure, LC-MS analysis
Arabidopsis thaliana
Other publictions for EC 5.4.99.53
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
749117
Loeschcke
The photosynthetic bacteria R ...
Arabidopsis thaliana
PLoS ONE
12
e0189816
2017
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730614
Go
Identification of marneral syn ...
Arabidopsis thaliana
Plant J.
72
791-804
2012
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683590
Shibuya
Origin of structural diversity ...
Arabidopsis thaliana
J. Am. Chem. Soc.
129
1450-1455
2007
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713761
Xiong
An Arabidopsis oxidosqualene c ...
Arabidopsis thaliana
Angew. Chem.
45
1285-1288
2006
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