Increasing interest in using food and agricultural wastes for low cost enzyme production continues in fermentation industry. Bacteria, especially Bacillus subtilis, are known to produce pectinases in synthetic media and media amended with agricultural wastes as a carbon source, whereas the potential of co-culturing Bacillus species to produce pectinases has not been fully studied yet. In this study, co-culture of B. subtilis and B. pumilus was optimized for solid load and pH to maximize pectinase production in submerged fermentation using apple pomace as the carbon source. Optimal pectinase activity was achieved as 11.25IU/mL with 15% apple pomace (solid load), pH 9.0, and 1/4 of culture ratio (B. subtilis/B. pumilus) at 30 degrees C after 24h of fermentation period according to the Box-Behnken response surface methodology. Thus, it was concluded that a twofold-increase in pectinase production was achieved by co-culturing of Bacillus spp. compared to our previous monoculture Bacillus study.