Bacterial cellulose had been proven to be a very versatile natural polymer produced by bacteria. One of the major constraints in producing bacterial cellulose is the high cost of fermentation medium. This study examines the potential use of papaya juice as a low-cost fermentation medium for the production of bacterial cellulose. The fermentation of Acetobacter xylinum 0416 was carried out under static fermentation with an initial pH of 5.5, a temperature of 28±1°C and a fermentation period of 5 days using different juice extracted from ripe and unripe papaya. The highest production of bacterial cellulose was detected in ripe papaya pulp juice with a total weight of 35.37 g/l. Juice obtained from various parts of the fruit produced bacterial cellulose between 3.33 g/l and 16.10 g/l. By referring to the standard medium (Hestrin-Schramm (HS) medium), ripe papaya pulp juice shows the highest potential to be used as an alternative for the production of bacterial cellulose. This is due to its high glucose concentration that provide suitable conditions for A.xylinum 0416 to grow and produce bacterial cellulose.