To improve the capacity of L-Phenylalanine( L-Phe) production with Escherichia coli BR-42 δpykA( pAP-B03),a pyruvate kinase mutant strain,the effects of different induction temperature( 36℃,38℃ and 40℃) on cell growth and L-Phe production were investigated. Based on the analysis of fermentation parameters,a threestage induction temperature controlled strategy was proposed,in which,the induction temperature was controlled at 40℃ during 13. 5 ~ 20 h,38℃ during 21 ~ 37 h,and 36℃ after 38 h,respectively. With the strategy,the cell growth,plasmid maintenance,and the key enzyme activities in late induction phase were improved,resulting in an enhanced L-Phe production. At 48 h,L-Phe titer reached 52. 70 g / L,which was 12. 01%,13. 77%,and 22. 39% higher than that at 36℃,38℃,and 40℃,respectively,which indicated that the three-stage induction temperature controlled strategy based on the analysis of fermentation parameters was favorable to L-Phe production in fed-batch culture.