Ammonia → L-Tryptophan

HRID 1955505Notes

Reaction equation

HRID 1955505 reaction equation
[NH:1]1[C:9]2[C:4](=[CH:5][CH:6]=[CH:7][CH:8]=2)[CH:3]=[CH:2]1.[NH2:10][CH2:11][C:12]([OH:14])=[O:13].[CH2:15]=O.[Na+].[Cl-].N>O.C1COCC1>[NH2:10][C@H:11]([C:12]([OH:14])=[O:13])[CH2:15][C:3]1[C:4]2[C:9](=[CH:8][CH:7]=[CH:6][CH:5]=2)[NH:1][CH:2]=1

Participants

8

Reactant 5

HCID 222AmmoniaHCID 712FormaldehydeHCID 750GlycineHCID 798Indole3×HCID 5234Sodium Chloride

Product 1

HCID 6305L-Tryptophan

Reagents, catalysts & solvents 2

HCID 962WaterSolventHCID 8028TetrahydrofuranSolvent

Conditions

Temperature
30 °C

Procedure

A 100 g of the frozen cells of Escherichia coli K-12 SH1001 prepared in Preparation Example 2 and 100 g of the frozen cells of Escherichia coli K-12 YK2009 (FERM BP-3244) prepared in Preparation Example 3 were suspended in 500 ml of the reaction solution (adjusted to pH 9.5 with 25% ammonia) containing 0.1 g of indole, 7.5 g of glycine, 1.6 g of formalin (formaldehyde content: 37%), 100 mg of PLP, 1.0 g of THF and 2.3 g of NaCl, and then water was added to make the total amount of the suspension 1,000 ml. This suspension was poured into a 3-liter jar fermentor (mfd. by Able Inc.) and reacted with stirring at 30° C. The pH of the reaction solution was adjusted to maintain at 9.5 with 25% ammonia throughout the reaction. Also, indole was added continuously or intermittently so that the indole concentration wouldn't exceed 2 mmol/l while monitoring the indole concentration in the reaction solution by a high-performance liquid chromatograph (mfd. by Shimazu Corp.). Also, glycine and formaldehyde were added continuously or intermittently so that the L-serine concentration in the reaction solution wouldn't exceed the designated levels in the respective test sections shown in Table 1 while monitoring the L-serine concentration in the reaction solution by a high-performance liquid chromatograph (mfd. by Shimazu Corp.). Glycine was added in a total amount of 22.5 g. The L-tryptophan concentration in the reaction solution was monitored by a high-performance liquid chromatograph (mfd. by Shimazu Corp.). The yield (mol %) of L-tryptophan after the end of the reaction, based on the glycine added in the reaction solution, is shown in Table 1.

Workup

  1. customprepared in Preparation Example 3
  2. additionThis suspension was poured into a 3-liter jar fermentor (mfd. by Able Inc.)
  3. customreacted
  4. concentrationthe indole concentration in the reaction solution by a high-performance liquid chromatograph (mfd. by Shimazu Corp.)
  5. additionAlso, glycine and formaldehyde were added continuously or intermittently so that the L-serine concentration in the reaction solution
  6. concentrationthe L-serine concentration in the reaction solution by a high-performance liquid chromatograph (mfd. by Shimazu Corp.)
  7. additionGlycine was added in a total amount of 22.5 g
  8. concentrationThe L-tryptophan concentration in the reaction solution

Sources & evidence

ORD record
ord-391cf22dba7243ad862c46470c99899d
Source dataset
uspto-grants-1998_07
Patent number
US05776740

Reaction notes

Loading…