The invention discloses a process for realizing natural gas preparation with coke oven gas by using anaerobic microbes. The process comprises the steps that: a, coke oven gas in a coking furnace and CO2 gas in a CO2 storage tank are mixed, such that mixed gas is obtained; b, the mixed gas is delivered into an anaerobic reactor with high-activity anaerobic microbes; in the anaerobic reactor, CO2, H2 and CO in the original coke oven gas are utilized, and CH4 is produced; H2S in the original coke oven gas is produced into elemental S through sulfur bacteria in the high-activity anaerobic microbes; and c, N2 and other impurities are separated with a pressure-swing adsorption separation device or a cryogenic separation device, such that natural gas with high methane content is obtained. According to the invention, with a microbial process, CO2 is supplemented, and natural gas is prepared. The process is highly economical. With the process, catalyst is not needed, and high-temperature and high-pressure are not needed. Most components except methane in the coke oven gas can be converted into methane, such that CO2 emission is reduced, and effects of energy saving and environment protection are achieved.