EC Number   |
Protein Variants |
Reference |
|---|
  4.2.1.B20 | D373C |
site-directed mutagenesis, the mutant enzyme BmeTC (D373C) leads to an improved enzyme that is able to catalyze the conversion starting from squalene to 3-deoxyachilleol and further to (+)-ambrein far more efficiently than the described two-enzyme cascade |
748565 |
| 4.2.1.B20 | more |
establishing of an (+)-ambrein production system by coexpressing the onoceroid synthase BmeTC from Bacillus megaterium with Saccharomyces cerevisiae squalene synthase (gene ScERG9) and Alicyclobacillus acidocaldarius squalene-hopene synthase mutant D377C (mutated gene shc) in Escherichia coli strain MEP43 (with deleted the beta-carotene synthesis module). The squalene synthase produces squalene, SHC enzyme mutant D377C produces 3-deoxyachilleol A from squalene, and BmTC forms ambrein from it, method optimization. Optimal temperature for ambrein production is 30-34°C |
747395 |
| 4.2.1.B20 | more |
for the biosynthesis of hydrophobic compounds such as terpenoids in Pichia pastoris, i.e.whole-cell production of (+)-ambrein, a central enzyme in the sterol biosynthesis pathway, squalene epoxidase Erg1, is targeted so that intracellular squalene levels in Pichia pastoris are strongly enhanced. Heterologous expression of AaSHC (EC 5.4.99.17) mutant D377C and enzyme BmeTC, and development of suitable methods to analyze all products of the engineered strain provide conclusive evidence of whole-cell (+)-ambrein production. Engineering of BmeTC leads to a remarkable one-enzyme system that is by far superior to the cascade, thereby increasing (+)-ambrein levels approximately 7fold in shake flask cultivation. Upscaling to 5 l bioreactor yields more than 100 mg/l of (+)-ambrein, demonstrating that metabolically engineered yeast Pichia pastoris represents a valuable, whole-cell system for high-level production of (+)-ambrein. Method evaluation and optimization, detailed overview |
748565 |
| 4.2.1.B20 | Y167A |
site-directed mutagenesis, the enzyme mutant produces unnatural tricyclic triterpenol, deoxyarabidiol ((5E)-6,10-dimethyl-2-[(3R,3aR,5aS,9aS,9bR)-3a,6,6,9a-tetramethyldodecahydro-1H-cyclopenta[a]naphthalen-3-yl]undeca-5,9-dien-2-ol), product ratios compared to wild-type, overview |
747544 |
| 4.2.1.B20 | Y167A/L596F |
site-directed mutagenesis, the mutant catalyzes only the synthesis of 8alpha-hydroxypolypoda-13,17,21-triene and 3-deoxyarabidiol ((5E)-6,10-dimethyl-2-[(3R,3aR,5aS,9aS,9bR)-3a,6,6,9a-tetramethyldodecahydro-1H-cyclopenta[a]naphthalen-3-yl]undeca-5,9-dien-2-ol), no formation of baciterpenol from tetraprenyl-beta-curcumene or onoceranoxide and 14beta-hydroxyonocera-8(26)-ene from 8alpha-hydroxypolypoda-13,17,21-triene, product ratios compared to wild-type, overview |
747544 |
| 4.2.1.B20 | Y167F |
site-directed mutagenesis, the enzyme mutant forms small quantity of tricyclic compounds, product ratios compared to wild-type, overview |
747544 |
| 4.2.1.B20 | Y167L |
site-directed mutagenesis, the enzyme mutant forms small quantity of tricyclic compounds, product ratios compared to wild-type, overview |
747544 |
| 4.2.1.B20 | Y167W |
site-directed mutagenesis, the enzyme mutant forms small quantity of tricyclic compounds, product ratios compared to wild-type, overview |
747544 |
