In this research, the protein was extracted from leather waste by using alkaline protease produced by Bacillus amyloliquefaciens. The purpose of this research is to investigate the method of protein extraction from leather waste by chemical method associated with alkaline protease enzyme. Two extraction methods were compared here: 1) protein extraction with acid-base and enzyme; 2) protein extraction with base and enzyme. The results show that the protein extraction methods with base-enzyme can produce protein extracts of 92.23±9.61 mg/L at the first extraction. And the second extraction, the protein extracts were 84.57±6.11 mg/L. For protein extraction methods with acid-base and enzyme, it gave the protein extracts of 18.23±4.16 mg/L and then second extraction gave the protein extracts of 10.90±7.21 mg/L. It can be seen that the protein extraction method with base-enzyme is the most effective method for extracting protein with significant statistical difference.