From the new perspective of the dynamic isotopic fractionation of microbes and the mixture of secondary biogenic gas and thermogenic gas in coalbeds, this study evaluates the geochemical characteristics of mixed coalbed gas in Enhong. Additionally, by using isotopes, the study traces the microbial activities and the results of those activities, such as the methane production of bacteria, and reveals the formation mechanism and genetic types of coalbed gas. The measured delta C-13(1) values of the coalbed gas samples are from -54.5 parts per thousand to -32.0 parts per thousand. Based on the values of Ro and delta C-13(2), by calculation, the delta C-13(1) values of thermogenic methane range from -30.6 parts per thousand to -28.1 parts per thousand and from -30.7 parts per thousand to -28.3 parts per thousand, respectively. The results of the two calculations are highly consistent, and both are approximately 20 parts per thousand higher than the measured values. The measured delta DCH4 values are from -217 parts per thousand to -196 parts per thousand, being between thermogenic methane and microbiogenic methane; the delta(13)CCO2 values are from -30.5 parts per thousand to -23.9 parts per thousand, growing heavier with respect to the original thermogenic gas; the measured delta C-13(2) values are from -25.7 parts per thousand to -22.6 parts per thousand, and the estimated delta C-13(2) values are from -21.8 parts per thousand to -21.2 parts per thousand, also indicating growing heavier. The delta C-13(1) and delta C-13(2) values are negatively correlated; both the Delta delta(CC2)-C-13-C-1 and Delta delta(13)CCO2-C-1 values are increasing. All of the above characteristics indicate that CO2 is reduced into microbial genetic methane by methane-producing bacteria and it mixes with thermogenic gas, which is new evidence showing the existence of secondary biogenic gas. Via calculations using a variety of data, such as the values of delta C-13(1), delta C-13(2) and Ro, it is found that thermogenic methane accounts for approximately 38% to 58% of the total amount, and microbial genetic methane accounts for approximately 42% to 62%. The proportion of micro-biogenic methane reduced from the top down, which occupied more than 50% of that in the coalbed buried within 1000 m deep. It increased the content of coalbed gas by more than 1 times. Within 1000 m deep, coalbed temperatures are generally lower than 40 degrees C, which is the most appropriate section for methanogenic bacteria activity and secondary biogenic gas generation. Coalbed uplift to the shallow parts in the late stage is the basic geological condition for the formation of secondary biogenic gas, which has significant resource value.