Phenylmethylsulfonyl Fluoride → beta-Maltose

HRID 618967

Reaction equation

HRID 618967 reaction equation
C1C=CC(CS(F)(=O)=O)=CC=1.[CH2:12]([OH:45])[C@H:13]1[O:18][C@H:17]([O:19][C@H:20]2[C@H:25]([OH:26])[C@@H:24]([OH:27])[C@@H:23]([O:28][C@H:29]3[C@H:34]([OH:35])[C@@H:33]([OH:36])[C@@H:32]([OH:37])[O:31][C@@H:30]3[CH2:38][OH:39])[O:22][C@@H:21]2[CH2:40][OH:41])[C@H:16]([OH:42])[C@@H:15]([OH:43])[C@@H:14]1[OH:44]>>[CH2:12]([OH:45])[C@H:13]1[O:18][C@H:17]([O:19][C@H:20]2[C@H:25]([OH:26])[C@@H:24]([OH:27])[C@@H:23]([O:28][C@H:29]3[C@H:34]([OH:35])[C@@H:33]([OH:36])[C@@H:32]([OH:37])[O:31][C@@H:30]3[CH2:38][OH:39])[O:22][C@@H:21]2[CH2:40][OH:41])[C@H:16]([OH:42])[C@@H:15]([OH:43])[C@@H:14]1[OH:44].[O:19]=[CH:17][C@@H:16]([C@H:15]([C@@H:14]([C@@H:13]([CH2:12][OH:45])[OH:18])[OH:44])[OH:43])[OH:42].[CH2:12]([OH:45])[C@H:13]1[O:18][C@H:17]([O:19][C@H:20]2[C@H:25]([OH:26])[C@@H:24]([OH:27])[C@H:23]([OH:28])[O:22][C@@H:21]2[CH2:40][OH:41])[C@H:16]([OH:42])[C@@H:15]([OH:43])[C@@H:14]1[OH:44]

Participants

5

Reactant 2

HCID 4784Phenylmethylsulfonyl FluorideHCID 192826alpha-Maltotriose2×

Product 3

HCID 6255beta-MaltoseHCID 107526(2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanalHCID 192826alpha-Maltotriose

Procedure

Culture supernatants were sampled at approximately 50 and 100 hours, treated with 2 mM PMSF final concentration, and frozen. To estimate NOVAMYL™ expression, supernatants were mixed with an equal volume of 2X Laemmli loading buffer, immediately boiled, and loaded on 14% or 8-16% acrylamide TRIS-glycine gels purchased from a commercial source (NOVEX, San Diego, Calif.). Known amounts of a NOVAMYL™ standard were also loaded on the same gel to estimate the amount of NOVAMYL™ produced. In some cases, NOVAMYL™ titer was determined using maltotriose as substrate. Specifically, a sample of the enzyme is incubated with maltotriose at pH 5.0 and 37° C. for 30 minutes. The reaction is then stopped by adjusting the pH to approximately 11. The amount of glucose produced from the breakdown of maltotriose to glucose and maltose is then measured with glucose dehydrogenase and NADH at 340 nm under standard conditions. Known amounts of a NOVAMYL™ standard (Novo Nordisk A/S, Bagsvaerd, Denmark) are also run.

Workup

  1. aliquotCulture supernatants were sampled at approximately 50 and 100 hours
  2. concentrationconcentration
  3. temperaturefrozen
  4. additionwere mixed with an equal volume of 2X Laemmli loading buffer
  5. customproduced

Sources & evidence

ORD record
ord-c529e2b75e2941c58a098d04e164450e
Source dataset
uspto-grants-1999_09
Patent number
US05958728

Linked notes

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