with unique properties and wide applicability. However, a standard medium used for the cultivation of bacterial cellulose, the Hestrin–Schramm medium, is expensive and prevents wide scale extension of bacterial cellulose applications.In this research, a relatively low-cost culture media was successfully developed from wood hot water extracts for the Acetobacter xylinus 23769 strain. Hot water extract is a residual material originating from pulp mills and lignocellulosic biorefineries and consists of mainly monomeric sugars, organic acids and organics. The effects of different pH (5, 6, 7 and 8) and temperatures (26, 28 and 30◦C) were alsoexamined in this research. There were no significant differences in the crystallinity and the recorded I˛fraction of cellulose produced between Hestrin–Schramm and the Hot water extract medium. The maximum production of 0.15 g/l of bacterial cellulose was obtained at a pH of 8 and temperature of 28◦C. Glucose and xylose in the Hot water extract were the main nutrient sources utilized in all BC cultivations based on high-pressure liquid chromatography results. Hot water extract was shown to be a suitable carbon source for bacterial cellulose production, and a processwas established for bacterial production from lignocellulosic feedstocks without using any modification of the Hot water extract . Hot water extract is an abundant and relatively inexpensive forest by-product. Using Hot water extract for bacterial cellulose production could reduce burdens on the environment and also, achieve the goal of large scale bacterial cellulose production at lowcost without using added culture nutrients.