Discovery of Novel Tyrosine Ammonia Lyases for the Enzymatic Synthesis of p‐Coumaric Acid

p‐Coumaric acid (p‐CA) is a key precursor for the biosynthesis of flavonoids. Tyrosine ammonia lyases (TALs) specifically catalyze the synthesis of p‐CA from l‐tyrosine, which is a convenient enzymatic pathway. To explore novel and highly active TALs, a phylogenetic tree‐building approach was conducted including 875 putative TALs and 46 putative phenylalanine/tyrosine ammonia lyases (PTALs). Among them, 5 TALs and 3 PTALs were successfully characterized and found to exhibit the proposed enzymatic activity. The TAL from Chryseobacterium luteum sp. nov (TAL( clu )) has the highest affinity (K ( m )=0.019 mm) and conversion efficiency (k (cat)/K ( m= )1631 s(−1) ⋅ mm (−1)) towards l‐tyrosine. The reaction conditions for two purified enzymes and their E. coli recombinant cells were optimized and p‐CA yields of 2.03 g/L after 8 hours by TAL( clu ) and 2.35 g/L after 24 h by TAL from Rivularia sp. PCC 7116 (TAL( rpc )) in whole cells were achieved. These TALs are thus candidates for the construction of whole‐cell systems to produce the flavonoid precursor p‐CA.