Method of enzymatically synthesizing 3′-phosphoadenosine-5′-phosphosulfate

摘要

The invention provides a method for producing 3′-phosphoadenosine 5′-phosphosulfate (PAPS), the method including subjecting ATP to sulfation and phosphorylation by use of adenosine 5′-triphosphate sulfurylase (ATPS) and adenosine 5′-phosphosulfate kinase (APSK), wherein an adenosine 5′-triphosphate (ATP) supply/regeneration system including adenosine 5′-monophosphate (AMP), polyphosphate, polyphosphate-driven nucleoside 5′-diphosphate kinase (PNDK), and polyphosphate:AMP phosphotransferase (PAP), or an adenosine 5′-triphosphate (ATP) supply/regeneration system including adenosine 5′-monophosphate (AMP), polyphosphate, polyphosphate-driven nucleoside 5′-diphosphate kinase (PNDK), and adenylate kinase (ADK) is employed instead of ATP.

主权项

1. A method for producing a substance by an enzymatic reaction that uses ATP as an energy source and/or a substrate, the method comprising:
providing components of an enzymatic reaction and an ATP supply/regeneration system, wherein the ATP supply/regeneration system produces ATP, and performing the enzymatic reaction in the presence of the ATP supply/regeneration system to produce the substance, wherein the ATP supply/regeneration system comprises adenosine 5′-monophosphate (AMP), polyphosphate:AMP phosphotransferase (PAP), polyphosphate, and a polyphosphate-driven nucleoside 5′-diphosphate kinase (PNDK) that consists of the amino acid sequence of amino acids 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, or 87 to 357 of SEQ ID NO:10 and PNDK activity.