Despite the extensive study of two-dimensional layered KNb3O8 (KN), there still remains some vital problems need to be clarified for future applications in environmental purification. Here we demonstrated the successful preparation of interlayer-controlled KN nanoflakes using alkaline hydrothermal conditions by adjusting the amount of thiourea in the reaction. This process resulted in KN nanoflakes with a larger specific surface area than previously reported. Moreover, the initial pH of dye solution and discrepant preferential orientation of interlayer peak have been proved to significantly influence the adsorption and photocatalysis performances of KN. In addition, relevant photocatalysis mechanisms have been expounded, by combined the first-principles calculation. The present work could be helpful in revealing the intrinsic adsorption-photocatalysis features of KN and other similar niobates.