In this research,protoplast was prepared with the initial wine yeast of a kind of automatic separated pineapple yeast H10,which was used to make UV mutagenesis to breed fine mutant strain.Process conditions and Ultraviolet irradiation time of yeast protoplast of customize pineapple wine were studied and mutant strains were primarily and secondly screened.The results had shown that a cell age of 10h,a enzymolysis temperature of 32℃,a enzymatic hydrolysate pH of 6.2 and a enzyme digestion time of 90min were the best process conditions for protoplast preparing.The best Ultraviolet irradiation time was 150s.After primary and second screens of mutant strains,a fine yeast mutant strain NO H10-15 which suits tropical pineapple wine production with good fermentability,aroma producing ability,alcohol and SO2 resistance and genetic stability will be produced.