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Applied and Environmental Microbiology, June 2005, p. 2962-2969, Vol. 71, No. 6
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.6.2962-2969.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Metabolic Engineering of the Phenylpropanoid Pathway in Saccharomyces cerevisiae

Hanxiao Jiang,1 Karl V. Wood,2 and John A. Morgan1*

School of Chemical Engineering,1 Department of Chemistry, Purdue University, West Lafayette, Indiana 479072

Received 7 July 2004/ Accepted 18 December 2004

Flavonoids are valuable natural products derived from the phenylpropanoid pathway. The objective of this study was to create a host for the biosynthesis of naringenin, the central precursor of many flavonoids. This was accomplished by introducing the phenylpropanoid pathway with the genes for phenylalanine ammonia lyase (PAL) from Rhodosporidium toruloides, 4-coumarate:coenzyme A (CoA) ligase (4CL) from Arabidopsis thaliana, and chalcone synthase (CHS) from Hypericum androsaemum into two Saccharomyces cerevisiae strains, namely, AH22 and a pad1 knockout mutant. Each gene was cloned and inserted into an expression vector under the control of a separate individual GAL10 promoter. Besides its PAL activity, the recombinant PAL enzyme showed tyrosine ammonia lyase activity, which enabled the biosynthesis of naringenin without introducing cinnamate 4-hydroxylase (C4H). 4CL catalyzed the conversion of both trans-cinnamic acid and p-coumaric acid to their corresponding CoA products, which were further converted to pinocembrin chalcone and naringenin chalcone by CHS. These chalcones were cyclized to pinocembrin and naringenin. The yeast AH22 strain coexpressing PAL, 4CL, and CHS produced approximately 7 mg liter–1 of naringenin and 0.8 mg liter–1 of pinocembrin. Several by-products, such as 2',4',6'-trihydroxydihydrochalcone and phloretin, were also identified. Precursor feeding studies indicated that metabolic flux to the engineered flavonoid pathway was limited by the flux to the precursor L-tyrosine.

* Corresponding author. Mailing address: School of Chemical Engineering, Purdue University, FRNY Hall, West Lafayette, IN 47907-2050. Phone: (765) 494-4088. Fax: (765) 494-0805. E-mail: jamorgan{at}purdue.edu.

Applied and Environmental Microbiology, June 2005, p. 2962-2969, Vol. 71, No. 6
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.6.2962-2969.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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