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Method for preparing adenosine triphosphate
| Details |
Inventors: Whitesides, George M.; Garrett, Patricia E.; Siegel, Merrell G.;
Assignee: Massachusetts Institute of Technology (Cambridge, MA)
Primary Examiner: Brown;; Johnnie R.
Assistant Examiner:
Attorney, Agent or Firm: Smith, Jr.; Arthur A., Conlin; David G.
Acyl phosphate salts suitable for use as phosphate donors in the production of cofactors, such as adenosine triphosphate, are prepared by acylation of phosphoric acid with a ketene, followed by reaction with ammonia or an amine. The reaction product, e.g. diammonium acetyl phosphate, can normally be used directly in enzymic production of cofactors without the necessity of prior conversion to the free acid or another salt, such as the sodium salt. The acyl phosphate salts are produced in high yields, and the method allows production of cofactors for use in biosynthesis from readily available materials. |
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DETAILED DESCRIPTION We claim: 1.
A method of preparing adenosine triphosphate in an enzyme-catalyzed reaction, said method comprising the step of reacting adenosine diphosphate in the presence of a phosphotransferase with an acetyl phosphate salt in which the cation is derived from the group of ammonia and primary, secondary and tertiary amines.
2.
The method of claim 1, wherein the acetyl phosphate salt is diammonium acetyl phosphate.
3.
The method of claim 2, in which the acetyl phosphate salt is dianilinium acetyl phosphate.
4.
The method of claim 1, wherein the reaction is carried out by mixing adenosine diphosphate and the acetyl phosphate salt in the presence of acetate kinase.
5.
The method of claim 4, wherein said adenosine diphosphate and said acetyl phosphate salt are mixed with from about 700 to about 2000 units of acetate kinase per mole of acetyl phosphate salt per day.
6.
The method of claim 5, wherein said acetyl phosphate salt is diammonium acetyl phosphate.
7.
The method of claim 4, wherein said acetyl phosphate salt is diammonium acetyl phosphate and said acetate kinase is present in an amount from about 1200 to about 1600 units per mole of diammonium acetyl phosphate per day.
8.
The method of claim 1, wherein said reaction is carried out in the presence of adenylate kinase.
9.
The method of claim 5, wherein said adenosine diphosphate and said acetyl phosphate salt are mixed with from about 200 to about 1400 units of adenylate kinase per mole of acetyl phosphate per day.
10.
The method of claim 8, further comprising producing at least a portion of said adenosine diphosphate by reacting adenosine monophosphate in the presence of said adenylate kinase.
Description:
BACKGROUND OF THE INVENTION This invention deals with the preparation of salts of acyl phosphoric acids, more particularly with the preparation of ammonium salts of acyl phosphates, such as diammonium acetyl phosphate.
Such compounds are useful in the production of phosphorylating agents such as adenosine triphosphate (ATP), a required cofactor in many reactions in the biosynthesis of materials such as protein, carbohydrates, nucleotides, nucleic acids, and terpenes
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Molecular Biology 
