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Using discarded tobacco leaves as raw materials, through aroma-bacterial screening and aroma components extracting, tobacco endogenous derived flavor was prepared and comparison was made on chemical components and sensory with tobacco leaf extract. Result indicated that: 1) 11 pure aroma-bacteria and 2 pure aroma-yeast were screened from 65 aroma-bacteria and 17 aroma-yeast. An Aureobasidium pullulans OF-01 was gotten by the style character evaluation eventually, and used for tobacco leaf flavor preparation; 2) A tobacco derived flavor was prepared by OF-01 strains through fermenting extracting, and concentrating. By comparison of the tobacco water extract, acids (586.18%), phenols (235.04%), and the alcoholes aldehydes (308.12%) in tobacco derived flavor promote greatly, and the quantities of scholar ketone, 3-hydroxy-2-butanone, and benzaldehyde incense increased over 1000%; 3) The tobacco derived flavor could increase armor, smooth smoke, reduce irritation, hide offensive and improve the effects of cigarette smoking. It had better compatibility with Yunnan cigarette, and could be used as flavor in cigarette production. [ABSTRACT FROM AUTHOR]