Pulmonary delivery of anti-tubercular drugs can be an effective treatment for tuberculosis. The main objective of the present work was to prepare rifabutin loaded xyloglucan microspheres as dry powder inhaler for pulmonary delivery. Xyloglucan is natural polysaccharide with mucoadhesive property and temperature responsive gelling ability allows its application in microspheres development. Xyloglucan microspheres were prepared by using single step spray drying process using lactose. All microsphere formulations were evaluated for various physical properties such as density and flow. On the basis of results of evaluation parameters such as entrapment efficiency, mucoadhesion, swelling and in vitro drug release, microspheres with 2% xyloglucan and 1% lactose monohydrate were found to be most favorable. They possessed morphology and particle size distribution suitable for pulmonary administration. XRD studies reveal amorphous nature of microspheres. In vitro DPI performance demonstrates suitability of xyloglucan based microspheres for pulmonary delivery. In conclusion, it is suggested that this natural polymer based microspheres containing rifabutin DPI formulation could be used as a significant enhanced treatment for TB.