Cloned (Comment) | Organism |
---|---|
recombinant expression of GST-tagged S-limonene synthase mutant in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha | Mentha spicata |
Protein Variants | Comment | Organism |
---|---|---|
H579A | site-directed mutagenesis, the mutant produces only 56.3% S-limonene | Mentha spicata |
L423A/S454A | site-directed mutagenesis, mutation M2, the mutant shows altered substrate specificity and product profile compared to the wild-type enzyme | Mentha spicata |
M458A | site-directed mutagenesis, the mutant produces only 3.2% S-limonene | Mentha spicata |
additional information | S-limonene synthase is converted to pinene or phellandrene synthases by site-directed mutagenesis, product profiles of mutant enzymes, overview | Mentha spicata |
N345A | site-directed mutagenesis, the mutant produces only 20.9%imonene | Mentha spicata |
N345A | site-directed mutagenesis, the mutation enlarges the active site, the mutant shows altered substrate specificity and product profilecompared to the wild-type enzyme | Mentha spicata |
N345A/L423A/S454A | site-directed mutagenesis, mutation M3 enlarges the active site, the mutant shows altered substrate specificity and product profile compared to the wild-type enzyme, the mutant acts as a pinene synthase and produces about 70% pinenes and has about 2fold increase in the yield of overall terpene products | Mentha spicata |
N345A/L423A/S454G | site-directed mutagenesis, mutation M5, the mutant shows altered substrate specificity and product profile compared to the wild-type enzyme, products of M5 are composed of 9.8% alpha-pinene, 52.7% beta-pinene and 13.4% limonene, it has about 2fold increase in the yield of overall terpene products | Mentha spicata |
N345G/L423A/S454A | site-directed mutagenesis, mutation M4, the mutant shows altered substrate specificity and product profile compared to the wild-type enzyme, products of M4 are composed of 18.81% alpha-pinene, 44.00% beta-pinene, and 16.2% limonene | Mentha spicata |
S454A | site-directed mutagenesis, the mutant produces only 73.7% S-limonene | Mentha spicata |
S454A | site-directed mutagenesis, the mutation enlarges the active site, the mutant shows altered substrate specificity and product profile compared to the wild-type enzyme | Mentha spicata |
S454G | site-directed mutagenesis, the mutation enlarges the active site, the mutant shows altered substrate specificity and product profile compared to the wild-type enzyme, the mutant enzyme produces about 46% alpha- and beta-pinenes and only about 52% limonene | Mentha spicata |
T349A | site-directed mutagenesis, the mutant produces only 60.3% limonene | Mentha spicata |
W324A | site-directed mutagenesis, the mutant produces only 18.1% S-limonene | Mentha spicata |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | - |
Mentha spicata | 9507 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Mentha spicata |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mentha spicata | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant GST-tagged S-limonene synthase mutant from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography | Mentha spicata |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
leaf | - |
Mentha spicata | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
geranyl diphosphate | stereochemistry is not specified in the publication, reaction of recombinantly expressed S-limonene sythase mutant N345A/L423A/S454A enzyme lacking the N-terminal transit peptide | Mentha spicata | beta-pinene + diphosphate | - |
? | |
additional information | product profiles of mutant enzymes, overview | Mentha spicata | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
More | cf. EC 4.2.3.120 | Mentha spicata |
pinene synthase | - |
Mentha spicata |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Mentha spicata |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Mentha spicata |
General Information | Comment | Organism |
---|---|---|
additional information | plasticity of the active site | Mentha spicata |
physiological function | mutationally converting S-limonene synthase from Mentha spicata (UniProt ID Q9SW76, EC 4.2.3.16) to beta-pinene synthase (EC 4.2.3.122 and 4.2.3.120) or phellandrene synthase (EC 4.2.3.51) reveals the plasticity of the active site | Mentha spicata |