Reaction #2146729

ord-4b893fa5b05d4628aae3bddcb32aaf81

Reaction equation

O=C(O)CCCC[C@@H]1SC[C@@H]2NC(=O)N[C@@H]21
Biotin
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
O=C(O)c1ccc(O)c(O)c1
protocatechuic acid
[Na+].[OH-]
NaOH
O=P([O-])(O)O.[K+]
KH2PO4
O=P([O-])([O-])O.[K+].[K+]
K2HPO4
O=S(=O)(O)CCCN1CCOCC1
MOPS
O=S(=O)([O-])[O-].[NH4+].[NH4+]
ammonium sulfate
[Ca+2].[Cl-].[Cl-]
CaCl2
→
N[C@@H](CCC(=O)O)C(=O)O
L-Glutamic Acid

Reactants

O=C(O)CCCC[C@@H]1SC[C@@H]2NC(=O)N[C@@H]21
Biotin
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
O=C(O)c1ccc(O)c(O)c1
protocatechuic acid
[Na+].[OH-]
NaOH
O=P([O-])(O)O.[K+]
KH2PO4
O=P([O-])([O-])O.[K+].[K+]
K2HPO4
—
FeSO4.7H2O
—
MgSO4.7H2O
O=S(=O)(O)CCCN1CCOCC1
MOPS
O=S(=O)([O-])[O-].[NH4+].[NH4+]
ammonium sulfate
[Ca+2].[Cl-].[Cl-]
CaCl2
—
MnSO4.7H2O

Reagents

O=S(=O)([O-])[O-].[Cu+2]
CuSO4

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Workup

  1. 1
    OtherOn the CM2B agar medium, the 2A-1R strain could form colonies at substantially the same rate as that of the wild-type strain ATCC13869
  2. 2
    Othera considerably decreased colony-forming rate
  3. 3
    OtherAccordingly, a gene that can recover the growth of the 2A-1R strain in the minimum medium

Procedure

On the CM2B agar medium, the 2A-1R strain could form colonies at substantially the same rate as that of the wild-type strain ATCC13869. However, on the minimum plate medium (20 g/l glucose, 2.64 g/l ammonium sulfate, 0.5 g/l KH2PO4, 0.5 g/l K2HPO4, 0.25 g/l MgSO4.7H2O, 0.01 g/l FeSO4.7H2O, 0.01 g/l MnSO4.7H2O, 0.01 g/l CaCl2, 0.02 mg/l CuSO4, 40 g/l MOPS, 30 mg/l protocatechuic acid, 200 μg/l VB1.Hu, 300 μg/l Biotin, 20 g/l agar, adjusted to pH 6.7 with NaOH), the 2A-1R strain showed a considerably decreased colony-forming rate as compared to the wild-type ATCC13869 strain. Accordingly, a gene that can recover the growth of the 2A-1R strain in the minimum medium was found.

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US08278074B2uspto-grants-2012_10