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Literature summary extracted from

  • Moser, S.; Strohmeier, G.A.; Leitner, E.; Plocek, T.J.; Vanhessche, K.; Pichler, H.
    Whole-cell (+)-ambrein production in the yeast Pichia pastoris (2018), Metab. Eng. Commun., 7, e00077 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.B20 gene BmTC, recombinant coexpression of enzyme BmTC with AaSHC (EC 5.4.99.17) mutant D377C in a squalene epoxidase Erg1-deficient Pichia pastoris strain leading to increased production of (+)-ambrein, recombinant expression of enzyme BmTC mutant D373C Priestia megaterium

Protein Variants

EC Number Protein Variants Comment Organism
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 Priestia megaterium
4.2.1.B20 additional information 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 Priestia megaterium

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.B20 3-deoxyachilleol A Priestia megaterium
-
 (+)-ambrein
-
 ?
4.2.1.B20 8alpha-hydroxypolypoda-13,17,21-triene Priestia megaterium
-
 onoceranoxide + 14beta-hydroxyonocera-8(26)-ene
-
 ?
4.2.1.B20 squalene Priestia megaterium
-
 8alpha-hydroxypolypoda-13,17,21-triene
-
 ?
4.2.1.B20 tetraprenyl-beta-curcumene + H2O Priestia megaterium
-
 baciterpenol A
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.B20 Priestia megaterium
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.B20 3-deoxyachilleol A
-
 Priestia megaterium (+)-ambrein
-
 ?
4.2.1.B20 3-deoxyachilleol A substrate is produced by AaSHC mutant D377C Priestia megaterium (+)-ambrein
-
 ?
4.2.1.B20 8alpha-hydroxypolypoda-13,17,21-triene
-
 Priestia megaterium onoceranoxide + 14beta-hydroxyonocera-8(26)-ene
-
 ?
4.2.1.B20 additional information for analysis, the GC-MS method used cannot be used to separate the highly similar compounds such as squalene and 3-deoxyachilleol, or 8alpha-hydroxypolypoda-13,17,21-triene and (+)-ambrein, product analysis by NMR spectrometry Priestia megaterium ?
-
 ?
4.2.1.B20 squalene
-
 Priestia megaterium 8alpha-hydroxypolypoda-13,17,21-triene
-
 ?
4.2.1.B20 squalene via 3-deoxyachilleol A, by mutant BmTC D373C Priestia megaterium (+)-ambrein
-
 ?
4.2.1.B20 tetraprenyl-beta-curcumene + H2O
-
 Priestia megaterium baciterpenol A
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.2.1.B20 BmeTC
-
 Priestia megaterium
4.2.1.B20 tetraprenyl-beta-curcumene cyclase
-
 Priestia megaterium