Adipic acid (AA), a key aliphatic dicarboxylic acid, is conventionally manufactured through energy-intensive, multi-step chemical processes with significant environmental impacts. In contrast, biological production methods offer more sustainable alternatives but are often limited by low productivity. To overcome these challenges, this study reports the engineering of a single Escherichia coli for efficient biosynthesis of AA starting from cyclohexanol (CHOL), KA oil (mixture of CHOL and cyclohexanone (CHONE)), or cyclohexane (CH). To start with, a comprehensive screening of rate-limiting enzymes is conducted, particularly focusing on cytochrome P450 monooxygenase, followed by the optimization of protein expression using strategies such as protein fusion, promoter replacement, and genome editing. Consequently, an engineered E. coli capable of efficiently converting either KA oil or CH into AA is obtained, achieving remarkable product titers of 110 and 22.6 g L