Ammonia → L-Tryptophan

HRID 1955506Notes

Reaction equation

HRID 1955506 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.[Cl-].[K+].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 4873Potassium 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 YK3005 (FERM BP-1736) prepared in Preparation Example 4 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 3.8 g of KCl, and then water was added to the suspension to make its total amount 1,000 ml. This suspension was fed into a 3-liter fermentor (mfd. by Able Inc.) and reacted with stirring at 30° C. During the reaction, pH of the reaction solution was adjusted to maintain at 9.5 with 25% ammonia. Indole was added continuously or intermittently so that the indole concentration in the reaction solution wouldn't exceed 2 mmol/l while monitoring the indole concentration by a high-performance liquid chromatograph (mfd. by Shimazu Corp.). Further, glycine and formaldehyde were added continuously or intermittently so that the L-serine concentration wouldn't exceed the designated levels for the respective test sections shown in Table 2 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 yields of L-tryptophan based on glycine are shown in Table 2.

Workup

  1. customprepared in Preparation Example 4
  2. customreacted
  3. customDuring the reaction, pH of the reaction solution
  4. concentrationthe indole concentration by a high-performance liquid chromatograph (mfd. by Shimazu Corp.)
  5. additionFurther, glycine and formaldehyde were added continuously or intermittently so that the L-serine concentration
  6. concentrationthe L-serine concentration in the reaction solution by a high-performance liquid chromatograph(mfd
  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-956d9ef600514b80ba83238f857fcc7b
Source dataset
uspto-grants-1998_07
Patent number
US05776740

Reaction notes

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