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One-pot enzymatic sulfation process using 3'-phosphoadenosine-5'-phosphosulfate and recycled phosphorylated adenosine intermediates

  • US 5,919,673 A
  • Filed: 03/22/1995
  • Issued: 07/06/1999
  • Est. Priority Date: 03/22/1995
  • Status: Expired due to Term
First Claim
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1. A process for using 3'"'"'-phosphoadenine-5'"'"'-phosphosulfate (PAPS) in an enzyme-catalyzed sulfation of an acceptor with recycling of phosphorylated adenosine intermediates that comprises the steps of:

  • (a) admixing the following ingredients in an aqueous medium containing magnesium and potassium ions within a single vessel to form an aqueous reaction medium(i) 3'"'"'-nucleotidase or 3'"'"'(2'"'"'),5'"'"'-bisphosphate nucleotidase;

    (ii) ATP sulfurylase;

    (iii) APS kinase;

    (iv) pyrophosphorylase;

    (v) a sulfotransferase;

    (vi) at least one adenine-containing compound selected from the group consisting of ATP, ADP, AMP, APS, PAPS and PAP;

    (vii) sulfate ion;

    (viii) an ATP-regenerating system comprising a phosphate donor and a phosphorylating enzyme; and

    (ix) a sulfate acceptor for said sulfotransferasethe concentration of said sulfate ion being greater than the concentration of all of said adenine-containing compound, and the activity of said 3'"'"'-nucleotidase being less than that of the enzymes of (ii)-(v); and

    maintaining said aqueous reaction medium at a pH value of about 5 to about 10 at a temperature of about zero degrees C to about 40°

    C. for a time period sufficient for said acceptor to be sulfated.

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