Adipose tissue is one of the main regulative sites for energy metabolism. Excess lipid storage and expansion of white adipose tissue (WAT) is the primary contributor to obesity, a strong predisposing factor for development of insulin resistance. Sentrin-specific protease (SENP) 2 has been shown to play a role in metabolism in murine fat and skeletal muscle cells, and we have previously demonstrated its role in energy metabolism of human skeletal muscle cells. In the present work, we have investigated the impact of SENP2 on fatty acid and glucose metabolism in primary human fat cells by using cultured primary human adipocytes to knock down the SENP2 gene. Glucose uptake and oxidation, as well as accumulation and distribution of oleic acid into complex lipids were decreased while oleic acid oxidation was increased in SENP2-knockdown cells compared to control cells. Furthermore, de novo lipogenesis was reduced by SENP2-knockdown in the adipocytes and expression of several metabolically important regulators and markers of browning of WAT were upregulated both on gene and protein level. In conclusion, SENP2 is an important regulator of energy metabolism in primary human adipocytes, and the results indicates a potential role in browning of the cells as well.