Materials and methods for the biosynthesis of seven carbon chemicals in the presence of methanol oxidation

A method for regulating biosynthesis of a product in a host cell, wherein the product comprises pimelic acid, 7-aminoheptanoate, 7-hydroxyheptanoate, heptamethylenediamine, 7-aminoheptanol, or 1,7-heptanediol, or salts thereof, using a pathway having a pimeloyl-acyl carrier protein (pimeloyl-ACP) intermediate, the method comprising converting methanol to formate via enzymatic reaction, wherein the formate is used in the conversion of tetrahydrofolate to N1°-formyl-tetrahydrofolate, wherein:

• the methanol is produced during pimeloyl-ACP methyl ester esterase (BioH) enzyme activity in which BioH removes a methyl group from pimeloyl-ACP methyl ester during conversion of pimeloyl-ACP methyl ester to pimeloyl ACP, wherein the BioH activity is increased compared to an unmodified host cell:
• and the method comprises downregulating (a) bifunctional protein (FolD) activity by attenuating folD and/or (b) formate acetyltransferase 1 (PflB) activity and formate acetyltransferase-like enzyme (TdcE) activity:
• and the method comprises inserting into the host cell (i) a gene encoding an S-formylglutathione hydrolase (frmB), (ii) a gene encoding a formate-tetrahydrofolate ligase (fhs), (iii) a gene encoding an alcohol dehydrogenase (adh), and (iv) a gene encoding an S-(hydroxymethyl) glutathione dehydrogenase (frmA), wherein the S-formylglutathione hydrolase has at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs:
• 27-30, the formate-tetrahydrofolate ligase has at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs:
• 18-22, the alcohol dehydrogenase has at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:
• 31, and the S-(hydroxymethyl) glutathione dehydrogenase has at least 90% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs:
• 23-26.