SAM Regeneration Pathway Using Formic Acid for Faster Methylation
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Solution Overview
Problem
Existing methods for recycling S-adenosylmethionine (SAM) do not sufficiently accelerate the methylation reaction and are inefficient in producing methyl compounds.
Innovation Solution
A method utilizing formic acid or its salts to regenerate SAM by converting formaldehyde to formic acid, followed by a series of enzymatic reactions involving formate-tetrahydrofolate ligase, methenyl-tetrahydrofolate cyclohydrolase, and other enzymes to produce 5-methyltetrahydrofolate, which is then used to regenerate SAM and produce methyl compounds.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If methanol is added to a culture medium of a microorganism coexpressing methylase and methanol dehydrogenase to recycle SAM, then SAM can be regenerated, but the methylation reaction is not sufficiently accelerated
Solution Approach 1:
The invention changes the carbon source parameter from methanol to formic acid, and introduces a multi-enzyme system (FtfL, Fch, MtdA, MetF) to convert formic acid through formate-tetrahydrofolate ligase, methenyl-tetrahydrofolate cyclohydrolase, methylene-tetrahydrofolate dehydrogenase, and 5,10-methylene-tetrahydrofolate reductase pathways. This parameter change and enzyme system introduction significantly accelerates the methylation reaction while maintaining SAM regeneration
Solution Approach 2:
The invention introduces formic acid as an intermediary substance that is converted to formate, then to 5-methyltetrahydrofolate, which serves as a methyl group donor to regenerate SAM. This intermediary pathway (formic acid → formate → 5-methyl-THF → SAM) provides an efficient route that accelerates the overall methylation reaction compared to direct methanol addition
2Quantity of substance
If formaldehyde dehydrogenase activity is reduced to suppress conversion of formaldehyde to formic acid, then SAM recycling is improved, but the overall efficiency of methyl compound production is insufficient
Solution Approach 1:
Instead of suppressing formaldehyde to formic acid conversion (the conventional approach), the invention inverts the pathway by directly supplying formic acid as the starting material and using a controlled multi-enzyme system to convert it to 5-methyltetrahydrofolate and then to SAM. This inverted approach avoids the need to suppress formaldehyde dehydrogenase while achieving efficient SAM recycling and methyl compound production
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method efficiently regenerates SAM and accelerates the methylation reaction, enhancing the production of methyl compounds.
Implementation Method 1
the production of 5-methyltetrahydrofolate from formic acid or a salt thereof and tetrahydrofolate is performed in the presence of formate-tetrahydrofolate ligase (FtfL)
Implementation Method 2
methenyl-tetrahydrofolate cyclohydrolase (Fch)
Implementation Method 3
methylene-tetrahydrofolate dehydrogenase (MtdA)
Implementation Method 4
5,10-methylene-tetrahydrofolate reductase (MetF)
Implementation Method 5
the conversion of S-adenosylhomocysteine to homocysteine is performed in the presence of S-adenosylhomocysteine nucleosidase (Mtn) and S-ribosylhomocysteine lyase (LuxS)
Implementation Method 6
the production of methionine by transferring a methyl group of the 5-methyltetrahydrofolate to the homocysteine is performed in the presence of methionine synthase (MetH or MetE)
Implementation Method 7
the conversion of the methionine to S-adenosylmethionine is performed in the presence of methionine adenosyltransferase (MetK)
Implementation Method 8
the conversion of methanol to formaldehyde is performed in the presence of methanol dehydrogenase (Medh) and/or methanol oxidase (Mox)
Implementation Method 9
the conversion of formaldehyde to formic acid is performed in the presence of formaldehyde dehydrogenase (FdhA)
Data Source
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AI summary
An object of the present invention is to provide a method for producing a methyl compound. The present invention provides a method for producing a methyl compound using formic acid.