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A central composite design (CCD) was used to evaluate the effects of fermentation temperature (20-30 ºC) 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 second-order polynomial regression equation with high R(2) (&gt;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 24.82 ºC 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 &lt; 0.05) was found between actual values and predicated values.

Optimization of processing conditions to improve antioxidant activities of apple juice and whey based novel beverage fermented by kefir grains