The protective culture(HOLDBACTM-YM-C) was applied in the making of a new yoghurt brand that features inhibitory function for yeast and mould growth control.Using the orthogonal test on the inoculations of protective culture and yoghurt starter culture,fermentation temperature and sugar adding amounts,we have determined the best fermentation condition.The best fermentation parameter combinations,based on the evaluation of fermentation parameters,change of pH value,number of viable lactic acid bacteria,and texture characters during the storage,were: protective culture 10DCU/100L,yoghurt starter culture 7.5DCU/100L,fermentation temperature 37℃,sugar adding 8%,and yoghurt had very good antimicrobial property.