Part:BBa_K5059001

TDH3-AAS-TPS1

This is a composite part that integrates a strong constitutive promoter (BBa_K124002), Kozak sequence (BBa_K165002), Team Cornell's BBA_K5059000 BioBrick that encodes for alpha-amyrin synthase (AAS), and a strong terminator (BBa_K2926005). Implementing these BioBricks together allows for increased expression of AAS, outputting more alpha-amyrin for ursolic acid production.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Figure 1: Metabolic Pathway for Producing Ursolic Acid in Yeast

Within this pathway, TDH3-AAS-TPS1 is the first enzyme required to catalyze ursolic acid production. Specifically, it catalyzes the conversion of 2,3-oxidosqualene to alpha-amyrin, which is the direct precursor for ursolic acid through the action of Cytochrome P450, which is activated via Cytochrome P450 reductase donating an electron from NADPH to it.

Functionality

Catalytic Efficiency

Substrate	Product	Km (µM)	kcat (min^-1)	kcat/Km (min^-1/µM)
1. 2,3-oxidosqualene	alpha-amyrin	50.07	43.4	0.87
2. alpha-amyrin	Ursolic Acid	24.5	35	1.43

This data comes from Dr. Yu et al.[3].