Tourism development can benefit from integrating biology, ecology, and cultural elements. Bio-Eco culture in tourism development emphasizes preserving and showcasing a destination’s unique natural and cultural assets. This approach recognizes that ecosystems and cultural assets are often intertwined and should be considered in developing sustainable tourism. However, tourism development activities often conflict with these efforts. A recent study shows that tourism development has been pursued independently, leading to an imbalance in tourism development. The situation in the study area is identical. Therefore, this study aims to enhance the relationship between cultural ecology and human behaviour as a tool for developing sustainable tourism. Jerejak Island, Penang was selected as a study area. A quantitative approach was applied, involving 150 tourists as respondents. The research identified the potential of tourism resources in the study area and investigated tourist typology through their behaviour. The study revealed a gap between conserving the island’s ecological and biological quality, neglecting the protection of heritage resources, and excessively meeting the needs of tourists. To achieve sustainable tourism development, it is necessary to address these three areas in parallel. It makes the Bio-Eco Cultural in tourism development approach suitable for the study area. Overall, the Bio-Eco cultural approach is also an effort to conserve local tourism resources.