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Literature summary for 2.5.1.148 extracted from

  • Thapa, H.R.; Naik, M.T.; Okada, S.; Takada, K.; Molnar, I.; Xu, Y.; Devarenne, T.P.
    A squalene synthase-like enzyme initiates production of tetraterpenoid hydrocarbons in Botryococcus braunii Race L (2016), Nat. Commun., 7, 11198 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.5.1.148

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Botryococcus braunii
recombinant expression of the enzyme in Escherichia coli Botryococcus braunii
recombinant expression of the enzyme in Escherichia coli, LOS coexpression in Saccharomyces cerevisiae with Arabidopsis thaliana GGPP synthase-11 (AtGGPPS11) resulting in lycopaoctaene production, which is undetectable when AtGGPPS11 is expressed without LOS. Recombinantly expressed in a Saccharomyces cerevisiae SS knockout strain, LOS restores ergosterol prototrophy, indicating its ability to produce squalene in vivo Botryococcus braunii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.07
-
 geranylgeranyl diphosphate pH 6.8, 37°C Botryococcus braunii
0.13
-
 (2E,6E)-farnesyl diphosphate pH and temperature not specified in the publication Botryococcus braunii

Localization

Localization Comment Organism GeneOntology No. Textmining
endoplasmic reticulum
-
 Botryococcus braunii 5783
-
endoplasmic reticulum lycopaoctaene synthase (LOS) activity is localized to a membrane system, possibly the endoplasmic reticulum as is seen for squalene synthase Botryococcus braunii 5783
-
membrane lycopaoctaene synthase (LOS) activity is localized to a membrane system, possibly the endoplasmic reticulum as is seen for squalene synthase Botryococcus braunii 16020
-
microsome
-
 Botryococcus braunii
-
-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 (2E,6E)-farnesyl diphosphate + NADPH + H+ Botryococcus braunii
-
 squalene + 2 diphosphate + NADP+
-
 ?
2 geranylgeranyl diphosphate + NADPH + H+ Botryococcus braunii LOS catalyzes the condensation of two GGPP units in the first half reaction to form the cyclopropylcarbinyl diphosphate intermediate PLPP, with concomitant release of one molecule of inorganic diphosphate. In the second half reaction, the PLPP cyclopropyl ring is cleaved and rearranged to form a 1-1' linkage and further reduction by NADPH forms lycopaoctaene. Without NADPH only the reaction intermediate PLPP accumulates lycopaoctaene + 2 diphosphate + NADP+
-
 ?
2 geranylgeranyl diphosphate + NADPH + H+ Botryococcus braunii the enzyme is involved in biosynthesis of (14E,18E)-lycopadiene. Lycopaoctaene i.e. 15,15'-dihydrophytoene = (6E,10E,14E,18E,22E,26E)-2,6,10,14,19,23,27,31-octamethyldotriaconta-2,6,10,14,18,22,26,30-octaene lycopaoctaene + 2 diphosphate + NADP+
-
 ?
additional information Botryococcus braunii squalene synthase enzyme diversification results in the production of specialized tetraterpenoid oils in race L of Botryococcus braunii ?
-
 ?
additional information Botryococcus braunii the enzyme also acts as lycopaoctaene synthase and uses alternative C15 and C20 prenyl diphosphate substrates to produce combinatorial hybrid hydrocarbons, but almost exclusively uses GGPP in vivo. Squalene synthase enzyme diversification results in the production of specialized tetraterpenoid oils in race L of Botryococcus braunii. Race L produces the C40 tetraterpenoid hydrocarbon lycopadiene. Ozonolysis experiments suggest lycopatriene and lycopapentaene share an identical reduced C20 moiety with lycopadiene. NMR spectroscopy confirms identity and structure ?
-
 ?

Organism

Organism UniProt Comment Textmining
Botryococcus braunii A0A142ZC57 race L
-
Botryococcus braunii A0A142ZC57 race L, Songkla Nakarin strain
-
Botryococcus braunii A0A144YEA5 race L
-

Purification (Commentary)

Purification (Comment) Organism
-
 Botryococcus braunii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(2E,6E)-farnesyl diphosphate + geranylgeranyl diphosphate + NADPH + H+
-
 Botryococcus braunii (6E,10E,14E,18E,22E)-2,6,10,14,19,23,27-heptamethyloctacosa-2,6,10,14,18,22,26-heptaene + NADP+ + 2 diphosphate
-
 ?
(2E,6E)-farnesyl diphosphate + phytyl diphosphate + NADPH + H+ when LOS is supplied with (2E,6E)-farnesyl diphosphate and phythyl diphoshate, squalene production predominates with small amounts of (6E,10E,14E)-2,6,10,15,19,23,27-heptamethyloctacosa-2,6,10,14,tetraene and lycopadiene Botryococcus braunii squalene + (6E,10E,14E)-2,6,10,15,19,23,27-heptamethyloctacosa-2,6,10,14,tetraene + lycopadiene + NADP+
-
 ?
2 (2E,6E)-farnesyl diphosphate + NADPH + H+
-
 Botryococcus braunii squalene + 2 diphosphate + NADP+
-
 ?
2 geranylgeranyl diphosphate
-
 Botryococcus braunii diphosphate + prephytoene diphosphate
-
 ?
2 geranylgeranyl diphosphate + NADH + H+ overall reaction. Lycopaoctaene i.e. 15,15'-dihydrophytoene = (6E,10E,14E,18E,22E,26E)-2,6,10,14,19,23,27,31-octamethyldotriaconta-2,6,10,14,18,22,26,30-octaene. The enzyme uses both NADH and NADPH as reducing agents for lycopaoctaene production, with preference for NADPH Botryococcus braunii lycopaoctaene + 2 diphosphate + NAD+
-
 ?
2 geranylgeranyl diphosphate + NADPH + H+ LOS catalyzes the condensation of two GGPP units in the first half reaction to form the cyclopropylcarbinyl diphosphate intermediate PLPP, with concomitant release of one molecule of inorganic diphosphate. In the second half reaction, the PLPP cyclopropyl ring is cleaved and rearranged to form a 1-1' linkage and further reduction by NADPH forms lycopaoctaene. Without NADPH only the reaction intermediate PLPP accumulates Botryococcus braunii lycopaoctaene + 2 diphosphate + NADP+
-
 ?
2 geranylgeranyl diphosphate + NADPH + H+ the LOS reaction uses a cyclopropyl intermediate Botryococcus braunii lycopaoctaene + 2 diphosphate + NADP+
-
 ?
2 geranylgeranyl diphosphate + NADPH + H+ the enzyme is involved in biosynthesis of (14E,18E)-lycopadiene. Lycopaoctaene i.e. 15,15'-dihydrophytoene = (6E,10E,14E,18E,22E,26E)-2,6,10,14,19,23,27,31-octamethyldotriaconta-2,6,10,14,18,22,26,30-octaene Botryococcus braunii lycopaoctaene + 2 diphosphate + NADP+
-
 ?
2 geranylgeranyl diphosphate + NADPH + H+ overall reaction. Lycopaoctaene i.e. 15,15'-dihydrophytoene = (6E,10E,14E,18E,22E,26E)-2,6,10,14,19,23,27,31-octamethyldotriaconta-2,6,10,14,18,22,26,30-octaene. The enzyme uses both NADH and NADPH as reducing agents for lycopaoctaene production, with preference for NADPH Botryococcus braunii lycopaoctaene + 2 diphosphate + NADP+
-
 ?
geranylgeranyl diphosphate + phytyl diphosphate + NADPH + H+ LOS incubation with GGPP and phythyl diphoshate produces lycopadiene and lycopapentaene as minor products and lycopaoctaene as the major product Botryococcus braunii lycopaoctaene + lycopadiene + lycopapentaene + NADP+
-
 ?
additional information squalene synthase enzyme diversification results in the production of specialized tetraterpenoid oils in race L of Botryococcus braunii Botryococcus braunii ?
-
 ?
additional information the enzyme also acts as lycopaoctaene synthase and uses alternative C15 and C20 prenyl diphosphate substrates to produce combinatorial hybrid hydrocarbons, but almost exclusively uses GGPP in vivo. Squalene synthase enzyme diversification results in the production of specialized tetraterpenoid oils in race L of Botryococcus braunii. Race L produces the C40 tetraterpenoid hydrocarbon lycopadiene. Ozonolysis experiments suggest lycopatriene and lycopapentaene share an identical reduced C20 moiety with lycopadiene. NMR spectroscopy confirms identity and structure Botryococcus braunii ?
-
 ?
additional information detection of lycopaoctaene synthase activity from an algal homogenate in an assay similar to that of squalene synthase supports the notion that an lycopaoctaene synthase enzyme may be similar to a typical squalene synthase enzyme Botryococcus braunii ?
-
 ?
additional information the enzyme can accept (2E,6E)-farnesyl diphosphate and phytyl diphosphate as substrates, and is also able to catalyse the condensation of two different substrate molecules, forming chimeric products. However, the use of these alternative substrates is not significant in vivo Botryococcus braunii ?
-
 ?
prephytoene diphosphate + NADPH + H+ lycopaoctaene i.e. 15,15'-dihydrophytoene i.e. (6E,10E,14E,18E,22E,26E)-2,6,10,14,19,23,27,31-octamethyldotriaconta-2,6,10,14,18,22,26,30-octaene Botryococcus braunii lycopaoctaene + diphosphate + NADP+
-
 ?

Synonyms

Synonyms Comment Organism
LOS
-
 Botryococcus braunii
lycopaoctaene synthase
-
 Botryococcus braunii
squalene synthase-like enzyme
-
 Botryococcus braunii
SSL
-
 Botryococcus braunii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay Botryococcus braunii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0114
-
 geranylgeranyl diphosphate pH 6.8, 37°C Botryococcus braunii
0.0205
-
 (2E,6E)-farnesyl diphosphate pH and temperature not specified in the publication Botryococcus braunii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.8
-
 assay at Botryococcus braunii

Cofactor

Cofactor Comment Organism Structure
NADH the enzyme uses both NADH and NADPH as reducing agents for lycopaoctaene production, with preference for NADPH Botryococcus braunii
NADPH
-
 Botryococcus braunii
NADPH the enzyme uses both NADH and NADPH as reducing agents for lycopaoctaene production, with preference for NADPH Botryococcus braunii

General Information

General Information Comment Organism
evolution two SSL cDNAs are identified and named based on the function of their encoded proteins as detailed below: Squalene synthase from race L (LSS) and synthase (LOS). Both the LSS and LOS proteins contain all five conserved activity domains, the transmembrane domain and the NADPH-binding residues found in typical squalene synthase enzymes. As LOS may have arisen from an SS paralogue that evolved to accept GGPP as substrate for lycopaoctaene production, LOS may have retained the ability to use (2E,6E)-farnesyl diphosphate to produce squalene. LSS is a true squalene synthase enzyme, whereas LOS appears to be a promiscuous squalene synthase-like (SSL) enzyme with broader substrate chain length and saturation specificity Botryococcus braunii
evolution two SSL cDNAs are identified and named based on the function of their encoded proteins as detailed below: Squalene synthase from race L (LSS) and synthase (LOS). Both the LSS and LOS proteins contain all five conserved activity domains, the transmembrane domain and the NADPH-binding residues found in typical squalene synthase enzymes. LSS is a true squalene synthase enzyme, whereas LOS appears to be a promiscuous squalene synthase-like (SSL) enzyme with broader substrate chain length and saturation specificity Botryococcus braunii
metabolism lycopadiene biosynthesis from C20 prenyl diphosphate intermediates can proceed via two possible biosynthetic routes: the first entails C20 geranylgeranyl diphosphate (GGPP) reduction by GGPP reductase to produce C20 phytyl diphosphate. Two molecules of phythyl diphoshate then undergo head-to-head condensation (1-1' linkage) to produce lycopadiene. The second possibility is the head-to-head condensation of two GGPP molecules to produce lycopaoctaene, followed by stepwise enzymatic reduction to produce lycopadiene Botryococcus braunii
metabolism the enzyme is involved in biosynthesis of (14E,18E)-lycopadiene. Lycopaoctaene i.e. 15,15'-dihydrophytoene = (6E,10E,14E,18E,22E,26E)-2,6,10,14,19,23,27,31-octamethyldotriaconta-2,6,10,14,18,22,26,30-octaene Botryococcus braunii
additional information there are three different races of Botryococcus braunii based on the hydrocarbons synthesized. Race A produces fatty acid-derived C23-C33 alkadienes and alkatrienes. Races B and L produce isoprenoid-derived hydrocarbons: methylsqualenes and C30-C37 botryococcene triterpenoids in race B and the C40 tetraterpenoid lycopadiene in race L Botryococcus braunii
additional information there are three different races of Botryococcus braunii based on the hydrocarbons synthesized. Race A produces fatty acid-derived C23-C33 alkadienes and lkatrienes. Races B and L produce isoprenoid-derived hydrocarbons: methylsqualenes and C30-C37 botryococcene triterpenoids in race B and the C40 tetraterpenoid lycopadiene in race L Botryococcus braunii
physiological function squalene synthase enzyme diversification results in the production of specialized tetraterpenoid oils in race L of Botryococcus braunii. Race L produces the C40 tetraterpenoid hydrocarbon lycopadiene. trans,trans-Lycopadiene is the predominant hydrocarbon (98% of total hydrocarbons) produced by race L, with a small amount of lycopatriene Botryococcus braunii
physiological function the enzyme also acts as lycopaoctaene synthase and uses alternative C15 and C20 prenyl diphosphate substrates to produce combinatorial hybrid hydrocarbons, but almost exclusively uses GGPP in vivo, detailed overview. Squalene synthase enzyme diversification results in the production of specialized tetraterpenoid oils in race L of Botryococcus braunii. Race L produces the C40 tetraterpenoid hydrocarbon lycopadiene. trans,trans-Lycopadiene is the predominant hydrocarbon (98% of total hydrocarbons) produced by race L, with a small amount of lycopatriene Botryococcus braunii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.000165
-
 geranylgeranyl diphosphate pH 6.8, 37°C Botryococcus braunii
0.162
-
 (2E,6E)-farnesyl diphosphate pH and temperature not specified in the publication Botryococcus braunii