Hydrochloric Acid → (6R,7R)-3-(carbamoyloxymethyl)-7-[[2-(2-furanyl)-2-methoxyimino-1-oxoethyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid

HRID 261148ノート

反応式

HRID 261148 の反応式
[OH:1][CH2:2][C:3]1[CH2:4][S:5][C@@H:6]2[C@H:13]([NH:14][C:15](=[O:25])[C:16](=[N:22][O:23][CH3:24])[C:17]3[O:18][CH:19]=[CH:20][CH:21]=3)[C:12](=[O:26])[N:7]2[C:8]=1[C:9]([OH:11])=[O:10].P([O-])([O-])([O:29][C:30](=O)[NH2:31])=O.P(OC[C@H]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H](O)[C@@H]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.[Cl-].[Mg+2].[Cl-].[OH-].[K+].Cl>[Cl-].[Mn+2].[Cl-].C(O)(=O)C>[C:30]([O:1][CH2:2][C:3]1[CH2:4][S:5][C@@H:6]2[C@H:13]([NH:14][C:15](=[O:25])[C:16](=[N:22][O:23][CH3:24])[C:17]3[O:18][CH:19]=[CH:20][CH:21]=3)[C:12](=[O:26])[N:7]2[C:8]=1[C:9]([OH:11])=[O:10])(=[O:29])[NH2:31]

参加物

9

反応物 6

HCID 313Hydrochloric AcidHCID 59575'-AtpHCID 14797Potassium HydroxideHCID 3423467Carbamic acid, monoanhydride with phosphoric acid, ion(2-)HCID 5360315Magnesium ChlorideHCID 54170549(6R,7R)-3-Hydroxymethyl-7-[2-(fur-2-yl)-2-methoxyiminoacetamido]ceph-3-em-4-carboxylic acid

生成物 1

HCID 41375(6R,7R)-3-(carbamoyloxymethyl)-7-[[2-(2-furanyl)-2-methoxyimino-1-oxoethyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid

試薬・触媒・溶媒 2

HCID 10313134Manganese(2+) dichloride触媒HCID 176Acetic Acid溶媒

実験手順

Concentrated enzyme (0.5 ml) was incubated at 28° in a final volume of 1.0 ml water containing (6R,7R)-3-hydroxymethyl-7-[2-methoxyimino-2-(fur-2-yl) acetamido]ceph-3-em-4-carboxylic acid (syn-isomer) (0,1 μmole), carbamoyl phosphate (3 μmoles), ATP (1 μmole), magnesium chloride (1 μmole), manganese chloride (1 μmole) & pipes buffer (50 μmoles) adjusted to pH 6.0 with potassium hydroxide. After 2 hr the reaction was stopped by the addition of acetic acid (20 μl). The pH was then adjusted to 2.0 with M hydrochloric acid and the precipitated protein was centrifuged off. The supernatant was extracted with ethylacetate (2.5 ml). The organic phase was collected and evaporated to dryness. The cephalosporins it contained were then separated by hplc [100 mm×5 diam. column Partisil 5 eluted at 1500 psi with chloroform:methanol:90% formic acid (25:2:1 v/v) saturated with water] and were detected by their absorption at 270 nm. Their retention time was compared with standard samples. The size of the U.V. absorbing peaks showed that 20% of the cephalosporin starting material had been converted to title compound.

後処理

  1. concentrationConcentrated enzyme (0.5 ml)
  2. customwas incubated at 28°
  3. extractionThe supernatant was extracted with ethylacetate (2.5 ml)
  4. customThe organic phase was collected
  5. customevaporated to dryness
  6. customThe cephalosporins it contained were then separated by hplc [100 mm×5 diam. column Partisil 5 eluted at 1500 psi with chloroform:methanol:90% formic acid (25:2:1 v/v) saturated with water]
  7. customwere detected by their absorption at 270 nm

出典と根拠

ORDレコード
ord-deb97bd49c524110a64ea3ae885506b7
出典データセット
uspto-grants-1979_08
特許番号
US04164447

反応ノート

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