Abstract: The enzymes and the characteristics of the community of the petroleum-degrading bacteria play a crucial role in the crude oil biodegradation. The prediction of kinetics of the key groups of hydrocarbons in crude oil was important to evaluate the bioremediation speed and constant. Most of the n-alkanes (C9–C29) were degraded in 25 days, and the average degradation rates of C18∼C27 higher than 100 μg g-1d-1. The hopanes, such as H30, had a biodegradation rate more than 10 μg g-1d-1. The related enzymes activities changed along with the crude oil biodegradation, especially dehydrogenase. The 16S rRNA gene amplicon sequencing revealed that Proteobacteria, Firmcutes, Bacteroidetes, Actinobacteria, Acidobacteria were the main petroleum hydrocarbon degraders during the crude oil biodegradation, and the top two highest relative abundance of the genera were Alcaligenes and Acinetobacter. Acinetobacter presented positive correlation to biodegradation of n-alkanes and PAHs. Based on COG analysis, the largest group involved in the general function was amino acid transport and metabolism. The functional categories of bacterial communities were mainly focused on the carbohydrate and amino acid metabolism, xenobiotics biodegradation and metabolism, membrane transport, and so on. Overall, these findings highlight the potential guideline for more adequate monitoring of microbial degradation of crude oil.