The presence of hydrogen sulfide (H2S) in biogas is one of the biggest factors limiting the use of biogas since related it can cause corrosion in internal combustion engines. This study investigated and compared the properties and sulfur oxidizing activities of Halothiobacillus neapolitanus (HTN) and Paracoccus pantotrophus (PCP) in their suitable conditions for apply in biotrickling filter to remove hydrogen sulfide in biogas. These bacteria were screened and characterized from different wastewater treatment plants. The results indicated that HTN had higher specific growth rate than PCP. However, the sulfate production rates of HTN and PCP are not significantly different, but HTN can produce higher sulfate concentration, and can tolerant high sulfide and sodium chloride concentration and low pH, which are advantages to apply in biotrickling filter in term of preventing contaminations. This study demonstrated that HTN is better option than PCP for application in the hydrogen sulfide removal in the biogas. However, PCP has challenge to apply for hydrogen sulfide removal in the other conditions such as denitrifying condition.