BioChem - DOS

Highly active and specific tyrosine ammonia-lyases from diverse origins enable enhanced production of aromatic compounds in bacteria and Saccharomyces cerevisiae

Applied and environmental microbiology

Jendresen, Christian Bille; Stahlhut, Steen Gustav; Li, Mingji; Gaspar, Paula; Siedler, Solvej; Fö rster, Jochen; Maury, Jé rô me; Borodina, Irina; Nielsen, Alex Toftgaard; Pettinari, M. J.

<P><B>ABSTRACT</B><P> Phenylalanine and tyrosine ammonia-lyases form cinnamic acid and <I>p</I> -coumaric acid, which are precursors of a wide range of aromatic compounds of biotechnological interest. Lack of highly active and specific tyrosine ammonia-lyases has previously been a limitation in metabolic engineering approaches. We therefore identified 22 sequences <I>in silico</I> using synteny information and aiming for sequence divergence. We performed a comparative <I>in vivo</I> study, expressing the genes intracellularly in bacteria and yeast. When produced heterologously, some enzymes resulted in significantly higher production of <I>p</I> -coumaric acid in several different industrially important production organisms. Three novel enzymes were found to have activity exclusively for phenylalanine, including an enzyme from the low-GC Gram-positive bacterium Brevibacillus laterosporus , a bacterial-type enzyme from the amoeba Dictyostelium discoideum , and a phenylalanine ammonia-lyase from the moss Physcomitrella patens (producing 230 &mu;M cinnamic acid per unit of optical density at 600 nm [OD 600 ]) in the medium using Escherichia coli as the heterologous host). Novel tyrosine ammonia-lyases having higher reported substrate specificity than previously characterized enzymes were also identified. Enzymes from Herpetosiphon aurantiacus and Flavobacterium johnsoniae resulted in high production of <I>p</I> -coumaric acid in Escherichia coli (producing 440 &mu;M <I>p</I> -coumaric acid OD 600 unit <SUP>&#x2212;1</SUP> in the medium) and in Lactococcus lactis . The enzymes were also efficient in Saccharomyces cerevisiae , where <I>p</I> -coumaric acid accumulation was improved 5-fold over that in strains expressing previously characterized tyrosine ammonia-lyases. </P></P>

2015

American Society for Microbiology

0099-2240

1098-5336

81

13

pp.4458-4476

10.1128/aem.00405-15

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART94456023

https://aem.asm.org/content/aem/81/13/4458.full.pdf