The invention discloses a two-step pH regulation method for producing ethanol by fermentation of synthesis gas, which is characterized by comprising the following steps: strictly deoxidizing a culture medium in an anaerobic fermentation bottle, and inoculating acetic acid-producing mixed strains into the strictly deoxidized anaerobic fermentation bottle containing the culture medium according to a certain volume ratio; setting the pH value of a culture solution in an anaerobic fermentation bottle according to the requirement of the acetic acid-producing mixed bacteria on pH in rapid growth, and adjusting and keeping the constant pH value through a pH value real-time measurement and feedback device so as to promote the growth of the acetic acid-producing bacteria and enable the acetic acid-producing bacteria to enter a rapid biomass enrichment stage; continuously introducing synthesis gas containing CO, CO2 and H2 into an anaerobic fermentation bottle as a gas substrate for fermentation, regulating the pH value of a culture solution in the anaerobic fermentation bottle to a proper range and keeping the pH value constant according to the pH value requirement of the strain for rapid ethanol production after the strain grows to a stable phase and is enriched with enough biomass, so that acetic acid-producing mixed bacteria enter a rapid ethanol production stage, and the method can be widely applied to the fields of energy conservation and emission reduction, environmental improvement, clean energy and the like.