A central composite design (CCD) was used to evaluate the effects of fermentation temperature (20-30ºC) and kefir grains amount (2-8%w/v) on total phenolic content and antioxidant activities of apple juice and whey based novel beverage fermented by kefir grains. The response surface methodology (RSM) showed that the significant secondorder polynomial regression equation with high R 2 (>0.86) was successfully fitted for all response as function of independent variable. The overall optimum region was found to be at the combined level of 7.56%w/v kefir grains and temperature of 24.82ºC with the highest value for total phenolic content (TPC) and antioxidant activities. At this optimum point TPC, 1, 1-Diphenyl-2-picrylhydrazyl radical scavenging, metal chelating effect, reducing power, inhibition of linoleic acid autoxidation and inhibition of ascorbate autoxidation were 165.02 mgGA/l, 0.38 ml/1 ml, 0.757 (absorbance at 700 nm), 46.12 %, 65.33 % and 21 %, respectively. No significant difference (p<0.05) was found between actual values and predicated values.