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Literature summary for 4.2.3.57 extracted from
- Functional characterization and transient expression manipulation of a new sesquiterpene synthase involved in beta-caryophyllene accumulation in Ocimum (2016), Biochem. Biophys. Res. Commun., 473, 265-271 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene OkBCS, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic tree, quantitative RT-PCR expression analysis, recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli TOP10 cells, transient overexpression in Ocimum kilimandscharicum via Agrobacterium tumefaciens strain GV3101 mediated transfection | Ocimum kilimandscharicum |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | gene OkBCS silencing in Ocimum kilimandscharicum plants via Agrobacterium tumefaciens strain GV3101 mediated transfection. beta-Caryophyllene and alpha-humulene levels decrease by about 20% in silenced plants compared to controls. The silencing effect is more prominent in systemic leaves with 95% reduction in OkBCS transcripts against 51% decrease in local leaves | Ocimum kilimandscharicum |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | steady state kinetics | Ocimum kilimandscharicum | |
| 0.1255 | - |
(2E,6E)-farnesyl diphosphate | pH not specified in the publication, 30°C, recombinant His-tagged enzyme | Ocimum kilimandscharicum |  |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required | Ocimum kilimandscharicum | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (2E,6E)-farnesyl diphosphate | Ocimum kilimandscharicum | - |
beta-caryophyllene + diphosphate | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Ocimum kilimandscharicum | A0A0E3KJK7 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant C-terminally His6-tagged enzyme from Escherichia coli TOP10 cells by nickel affinity chromatography to homogeneity | Ocimum kilimandscharicum |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| flower | high enzyme expression level | Ocimum kilimandscharicum | - |
| leaf | high enzyme expression level | Ocimum kilimandscharicum | - |
| additional information | expression variation of OkBCS well corroborate with beta-caryophyllene levels in different tissues from five Ocimum species | Ocimum kilimandscharicum | - |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (2E,6E)-farnesyl diphosphate | - |
Ocimum kilimandscharicum | beta-caryophyllene + diphosphate | - |
? | |
| (2E,6E)-farnesyl diphosphate | the enzyme stereochemistry is not determined in the publication. Recombinant enzyme OkBCS converts farnesyl diphosphate to beta-caryophyllene as a major product (94%) and 6% alpha-humulene | Ocimum kilimandscharicum | beta-caryophyllene + diphosphate | - |
? | |
| additional information | enzyme OkBCS is not active with geranyl diphosphate (GPP) as substrate, besides a weak hydrolysis activity converting GPP to geraniol, identification of substrates and products by GC-MS analysis | Ocimum kilimandscharicum | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 63600, about, sequence calculation | Ocimum kilimandscharicum |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| BCS | - |
Ocimum kilimandscharicum |
| More | cf. EC 4.2.3.89 | Ocimum kilimandscharicum |
| OkBCS | - |
Ocimum kilimandscharicum |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Ocimum kilimandscharicum |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 4.06 | - |
(2E,6E)-farnesyl diphosphate | pH not specified in the publication, 30°C, recombinant His-tagged enzyme | Ocimum kilimandscharicum | |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Ocimum kilimandscharicum | sequence calculation | - |
5.66 |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to typical class of sesquiterpene synthases from angiosperms, clade TPS-a. Across five Ocimum species, OkBCS shows the highest expression in Ocimum kilimandscharicum followed by Ocimum americanum, Ocimum tenuiflorum, and Ocimum gratissimum while Ocimum basilicum hadve trace levels | Ocimum kilimandscharicum |
| physiological function | agro-infiltration based transient expression manipulation with OkBCS overexpression and silencing confirms its role in beta-caryophyllene biosynthesis | Ocimum kilimandscharicum |
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