Cotton is produced in over 50 countries worldwide, averaging 20-24 million tons per year. India is one of the largest consumers of cotton, accounting for about 60% of the total consumption of cotton. Demands in handloom fabrics are increasing throughout the world due to their comfort in wearing particularly in the tropical countries. Hence, to make this sector internationally and globally competitive, it requires maintaining standard quality parameters. Fortunately, in the present time, commercially viable alternative methods for preparing and finishing cotton fibre substrates based on the use of enzymes have emerged. Such methods will ensure the supremacy of cotton over other fibers for decades to come. The present research was planned by keeping in view the emerging trend of bio-processing, use of swelling agents and dyeing with natural dyes. The present study was conducted to optimize the pretreatment process with enzymes and swelling agents on handloom cotton fabric. Experiments were conducted to determine optimum values of four variables for acid and neutral cellulase enzyme treatment, namely, pH, concentration, treatment time and temperature. Concentration, treatment time and temperature were the three variables optimized for the swelling agents. Dyeing variables i.e. concentration of dye material, extraction time, dyeing time, mordant concentration and method of mordanting with natural mordant were optimized.