Thiamine Hydrochloride → L-Glutamic Acid

HRID 2076753Notes

Reaction equation

HRID 2076753 reaction equation
O=C[C@@H]([C@H]([C@@H]([C@@H](CO)O)O)O)O.S([O-])([O-])(=O)=O.[NH4+].[NH4+].OP([O-])(O)=O.[K+].CC1[N+:31](CC2C=NC(C)=NC=2N)=CSC=1CCO.Cl.[Cl-].[OH:46][C:47]([CH2:49][CH2:50][CH2:51][CH2:52][C@H:53]1[C@@H:61]2[C@@H:56]([NH:57][C:58]([NH:60]2)=[O:59])[CH2:55][S:54]1)=[O:48].[OH-].[K+].[C:64](=[O:67])([O-:66])[O-].[Ca+2]>[O-]S([O-])(=O)=O.[Mn+2]>[NH2:31][C@H:49]([C:47]([OH:46])=[O:48])[CH2:50][CH2:51][C:64]([OH:66])=[O:67].[OH:48][C:47]([CH2:49][CH2:50][CH2:51][CH2:52][C@H:53]1[C@@H:61]2[C@@H:56]([NH:57][C:58]([NH:60]2)=[O:59])[CH2:55][S:54]1)=[O:46]

Participants

10

Reactant 7

HCID 6202Thiamine HydrochlorideHCID 10112Calcium CarbonateHCID 14797Potassium HydroxideHCID 107526(2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanalHCID 171548BiotinHCID 516951Monobasic potassium phosphateHCID 6097028Ammonium Sulfate

Product 2

HCID 33032L-Glutamic AcidHCID 171548Biotin

Reagents, catalysts & solvents 1

HCID 24580Manganese SulfateCatalyst

Conditions

Temperature
31.5 °C

Procedure

The single cross-over recombinant thus obtained was named 2A-1 strain. The wild-type 13869 strain and 2A-1 strain were inoculated in 20 ml of a flask medium (30 g/l glucose, 15 g/l ammonium sulfate, 1 g/l KH2PO4, 0.4 g/l MgSO4.7H2O, 0.01 g/l FeSO4.7H2O, 0.01 g/l MnSO4.4-5H2O, 200 μg/l VB1 (vitamin B1), 300 μg/l Biotin, and 0.48 g/l soybean hydrolysates (T-N: total nitrogen), adjusted to pH 8.0 with KOH: autoclaved at 115° C. for 10 minutes), followed by addition of 1 g of heat-sterilized calcium carbonate, and each of the strains was cultured with shaking at 31.5° C. After the sugar was completely consumed, the concentration of L-glutamic acid which had accumulated in the medium was determined. The results are shown in Table 1 (OD620 is turbidity at 620 nm of culture solution diluted to 101 times, and indicates the cell amount, and Glu (g/L) indicates the amount of accumulated L-glutamic acid). It was found that the 2A-1 strain produced L-glutamic acid in the presence of an excess amount of biotin, whereas the parent strain ATTCC 13869 did not produce L-glutamic acid at all.

Workup

  1. customThe single cross-over recombinant thus obtained
  2. customThe wild-type 13869 strain and 2A-1 strain were inoculated in 20 ml of a flask
  3. customAfter the sugar was completely consumed
  4. additiondiluted to 101 times

Sources & evidence

ORD record
ord-51d155510dd0495691bbb8a2e8f1be58
Source dataset
uspto-grants-2011_04
Patent number
US07927844B2

Reaction notes

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