Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant fusion proteins in Saccharomyces cerevisiae strain W303-1a. coexpression of wild-type enzyme with dammarenediol-II synthase from Panax ginseng and a Arabidopsis thaliana NADPH-cytochrome P450 reductase (ATR1). The low metabolic flux through PPDS results in a low productivity of protopanaxadiol, but is increased with the fusion enzymes | Panax ginseng |
Protein Variants | Comment | Organism |
---|---|---|
additional information | to increase the low metabolic flux through PPDS and increase the productivity of protopanaxadiol production, enzyme PPDS is modified through transmembrane domain truncation and construction of self-sufficient PPDS-ATR1 fusion proteins. The fusion enzymes exhibit approximately 4.5fold increase in catalytic activity and 71.1% increase in protopanaxadiol production compared with PPDS and ATR1 co-expression. The engineered yeast carrying fusion protein effectively converts 96.8% of dammarenediol-II into protopanaxadiol, bioreactor in fed-batch fermentation. Construction of four fusion proteins named as PPDS-linker1-46tATR1, PPDSlinker2-46tATR1, PPDS-linker3-46tATR1, and PPDS-nolinker-46tATR1, respectively. In addition, 31tPPDS heme domain and 46tATR1 reductase domain are linked by polypeptide GSTSSGSG. Method optimization, overview. Evaluation of the oxidative stress in yeast cells induced by co-expression of PPDS and ATR1 | Panax ginseng |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
endoplasmic reticulum membrane | the enzyme contains a transmembrane domain | Panax ginseng | 5789 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
dammarenediol-II + [reduced NADPH-hemoprotein reductase] + O2 | Panax ginseng | - |
protopanaxadiol + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Panax ginseng | H2DH16 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dammarenediol-II + [reduced NADPH-hemoprotein reductase] + O2 | - |
Panax ginseng | protopanaxadiol + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
cytochrome P450-type protopanaxadiol synthase | - |
Panax ginseng |
PPDS | - |
Panax ginseng |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Panax ginseng |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Panax ginseng |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
cytochrome P450 | - |
Panax ginseng | |
NADPH-hemoprotein reductase | A flavoprotein containing both FMN and FAD. This enzyme catalyses the transfer of electrons from NADPH, an obligatory two-electron donor, to microsomal P-450 monooxygenases, EC 1.14.14._ | Panax ginseng |