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Abstract
Phenylalanine and tyrosine ammonia-lyases form cinnamic acid and p-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 in silico using synteny information and aiming for sequence divergence. We performed a comparative in vivo study, expressing the genes intracellularly in bacteria and yeast. When produced heterologously, some enzymes resulted in significantly higher production of p-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 μM cinnamic acid per unit of optical density at 600 nm [OD600]) 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 p-coumaric acid in Escherichia coli (producing 440 μM p-coumaric acid OD600 unit−1 in the medium) and in Lactococcus lactis. The enzymes were also efficient in Saccharomyces cerevisiae, where p-coumaric acid accumulation was improved 5-fold over that in strains expressing previously characterized tyrosine ammonia-lyases.
Original language | English |
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Journal | Applied and Environmental Microbiology |
Volume | 81 |
Issue number | 13 |
Pages (from-to) | 4458-4476 |
Number of pages | 19 |
ISSN | 0099-2240 |
DOIs | |
Publication status | Published - 2015 |
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Dive into the research topics of 'Highly Active and Specific Tyrosine Ammonia-Lyases from Diverse Origins Enable Enhanced Production of Aromatic Compounds in Bacteria and Saccharomyces cerevisiae'. Together they form a unique fingerprint.Projects
- 1 Finished
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BacHBERRY: From BACterial Hosts for production of Bioactive phenolics from bERRY fruits to products
Förster, J. (Project Coordinator), Dudnik, A. (Project Manager), Gaspar, P. (Project Participant), Stahlhut, S. G. (Project Participant), Ögmundarson, Ó. (Project Participant), Rasmussen, B. K. (Project Manager) & Lohmann, R. (Project Manager)
European Commission- European Research Area
01/11/2013 → 31/10/2016
Project: Research