Limits...
Precursor-Directed Combinatorial Biosynthesis of Cinnamoyl, Dihydrocinnamoyl, and Benzoyl Anthranilates in Saccharomyces cerevisiae.

Eudes A, Teixeira Benites V, Wang G, Baidoo EE, Lee TS, Keasling JD, Loqué D - PLoS ONE (2015)

Bottom Line: We recently demonstrated the potential of using yeast (Saccharomyces cerevisiae) for the biological production of a few cinnamoyl anthranilates by heterologous co-expression of 4-coumaroyl:CoA ligase from Arabidopsis thaliana (4CL5) and HCBT.Our results demonstrate the use of enzyme promiscuity in biological synthesis to achieve high chemical diversity within a defined class of molecules.This work also points to the potential for the combinatorial biosynthesis of diverse and valuable cinnamoylated, dihydrocinnamoylated, and benzoylated products by using the versatile biological enzyme 4CL5 along with characterized cinnamoyl-CoA- and benzoyl-CoA-utilizing transferases.

View Article: PubMed Central - PubMed

Affiliation: Joint BioEnergy Institute, Emery Station East, 5885 Hollis St, 4th Floor, Emeryville, California, 94608, United States of America; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, 94720, United States of America.

ABSTRACT
Biological synthesis of pharmaceuticals and biochemicals offers an environmentally friendly alternative to conventional chemical synthesis. These alternative methods require the design of metabolic pathways and the identification of enzymes exhibiting adequate activities. Cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates are natural metabolites which possess beneficial activities for human health, and the search is expanding for novel derivatives that might have enhanced biological activity. For example, biosynthesis in Dianthus caryophyllus is catalyzed by hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/ benzoyltransferase (HCBT), which couples hydroxycinnamoyl-CoAs and benzoyl-CoAs to anthranilate. We recently demonstrated the potential of using yeast (Saccharomyces cerevisiae) for the biological production of a few cinnamoyl anthranilates by heterologous co-expression of 4-coumaroyl:CoA ligase from Arabidopsis thaliana (4CL5) and HCBT. Here we report that, by exploiting the substrate flexibility of both 4CL5 and HCBT, we achieved rapid biosynthesis of more than 160 cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates in yeast upon feeding with both natural and non-natural cinnamates, dihydrocinnamates, benzoates, and anthranilates. Our results demonstrate the use of enzyme promiscuity in biological synthesis to achieve high chemical diversity within a defined class of molecules. This work also points to the potential for the combinatorial biosynthesis of diverse and valuable cinnamoylated, dihydrocinnamoylated, and benzoylated products by using the versatile biological enzyme 4CL5 along with characterized cinnamoyl-CoA- and benzoyl-CoA-utilizing transferases.

No MeSH data available.


Structure of cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates.(A) Cinnamoyl anthranilates. Tranilast: R1 = R2 = R3 = R6 = H, R4 = R5 = OMe. (B) Dihydrocinnamoyl anthranilates. DHavnD: R1 = R2 = R3 = R4 = R6 = H, R5 = OH. (C) Benzoyl anthranilates. Dianthramide B from D. caryophyllus: R1-6 = H.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4591981&req=5

pone.0138972.g001: Structure of cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates.(A) Cinnamoyl anthranilates. Tranilast: R1 = R2 = R3 = R6 = H, R4 = R5 = OMe. (B) Dihydrocinnamoyl anthranilates. DHavnD: R1 = R2 = R3 = R4 = R6 = H, R5 = OH. (C) Benzoyl anthranilates. Dianthramide B from D. caryophyllus: R1-6 = H.

Mentions: Cinnamoyl and benzoyl anthranilates are bipartite molecules consisting of cinnamate or benzoate moieties amide-linked to anthranilic acids (Fig 1). The beneficial pharmacological effects of these molecules on human health have been well-documented over the past few years. For example, avenanthramides are natural cinnamoyl anthranilates found in oats and possess antioxidant, anti-inflammatory, and antiproliferative bioactivities [1,2]. Tranilast ([N-(3’,4’-dimethoxycinnamoyl)-anthranilic acid], Fig 1A) is a synthetic cinnamoyl anthranilate marketed in Japan for the treatment of allergic diseases, scleroderma, and hypertrophic scars associated with excessive fibrotic response [3]. In particular, tranilast is an antifibrotic agent that inhibits several profibrotic growth factors [4–6]. Recent efforts have been made for the development of tranilast analogs to optimize the antifibrotic effects and reduce toxicity at higher doses [7]. For instance, modification of functional groups on the cinnamoyl ring and the introduction of halogens resulted in cinnamoyl anthranilates with higher bioavailability and enhanced inhibitory effects on fibrosis [8–12]. Other structure optimizations have included double bond saturation resulting in dihydrocinnamoyl anthranilates such as dihydroavenanthramide D (DHavnD, Fig 1B), which is an anti-inflammatory used for the treatment of skin disorders and is currently evaluated for its antidiabetic and anticancer effects [13–15]. Benzoyl anthranilates (Fig 1C) are found in some plant species such as D. caryophyllus [16]; and several analogs were shown to inhibit human aldo-keto reductases involved in different pathophysiological conditions such as prostate cancer [17], as well as to possess cytotoxic activity toward cancer cell lines [18]. Moreover, certain halogenated benzoyl anthranilates are candidates for the treatment of infectious diseases because of their inhibitory effects on the malaria agent Plasmodium falciparum [19], the human African trypanosomiasis agent Trypanosoma brucei [20,21], and the opportunistic pathogenic bacterium Pseudomonas aeruginosa [22,23].


Precursor-Directed Combinatorial Biosynthesis of Cinnamoyl, Dihydrocinnamoyl, and Benzoyl Anthranilates in Saccharomyces cerevisiae.

Eudes A, Teixeira Benites V, Wang G, Baidoo EE, Lee TS, Keasling JD, Loqué D - PLoS ONE (2015)

Structure of cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates.(A) Cinnamoyl anthranilates. Tranilast: R1 = R2 = R3 = R6 = H, R4 = R5 = OMe. (B) Dihydrocinnamoyl anthranilates. DHavnD: R1 = R2 = R3 = R4 = R6 = H, R5 = OH. (C) Benzoyl anthranilates. Dianthramide B from D. caryophyllus: R1-6 = H.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591981&req=5

pone.0138972.g001: Structure of cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates.(A) Cinnamoyl anthranilates. Tranilast: R1 = R2 = R3 = R6 = H, R4 = R5 = OMe. (B) Dihydrocinnamoyl anthranilates. DHavnD: R1 = R2 = R3 = R4 = R6 = H, R5 = OH. (C) Benzoyl anthranilates. Dianthramide B from D. caryophyllus: R1-6 = H.
Mentions: Cinnamoyl and benzoyl anthranilates are bipartite molecules consisting of cinnamate or benzoate moieties amide-linked to anthranilic acids (Fig 1). The beneficial pharmacological effects of these molecules on human health have been well-documented over the past few years. For example, avenanthramides are natural cinnamoyl anthranilates found in oats and possess antioxidant, anti-inflammatory, and antiproliferative bioactivities [1,2]. Tranilast ([N-(3’,4’-dimethoxycinnamoyl)-anthranilic acid], Fig 1A) is a synthetic cinnamoyl anthranilate marketed in Japan for the treatment of allergic diseases, scleroderma, and hypertrophic scars associated with excessive fibrotic response [3]. In particular, tranilast is an antifibrotic agent that inhibits several profibrotic growth factors [4–6]. Recent efforts have been made for the development of tranilast analogs to optimize the antifibrotic effects and reduce toxicity at higher doses [7]. For instance, modification of functional groups on the cinnamoyl ring and the introduction of halogens resulted in cinnamoyl anthranilates with higher bioavailability and enhanced inhibitory effects on fibrosis [8–12]. Other structure optimizations have included double bond saturation resulting in dihydrocinnamoyl anthranilates such as dihydroavenanthramide D (DHavnD, Fig 1B), which is an anti-inflammatory used for the treatment of skin disorders and is currently evaluated for its antidiabetic and anticancer effects [13–15]. Benzoyl anthranilates (Fig 1C) are found in some plant species such as D. caryophyllus [16]; and several analogs were shown to inhibit human aldo-keto reductases involved in different pathophysiological conditions such as prostate cancer [17], as well as to possess cytotoxic activity toward cancer cell lines [18]. Moreover, certain halogenated benzoyl anthranilates are candidates for the treatment of infectious diseases because of their inhibitory effects on the malaria agent Plasmodium falciparum [19], the human African trypanosomiasis agent Trypanosoma brucei [20,21], and the opportunistic pathogenic bacterium Pseudomonas aeruginosa [22,23].

Bottom Line: We recently demonstrated the potential of using yeast (Saccharomyces cerevisiae) for the biological production of a few cinnamoyl anthranilates by heterologous co-expression of 4-coumaroyl:CoA ligase from Arabidopsis thaliana (4CL5) and HCBT.Our results demonstrate the use of enzyme promiscuity in biological synthesis to achieve high chemical diversity within a defined class of molecules.This work also points to the potential for the combinatorial biosynthesis of diverse and valuable cinnamoylated, dihydrocinnamoylated, and benzoylated products by using the versatile biological enzyme 4CL5 along with characterized cinnamoyl-CoA- and benzoyl-CoA-utilizing transferases.

View Article: PubMed Central - PubMed

Affiliation: Joint BioEnergy Institute, Emery Station East, 5885 Hollis St, 4th Floor, Emeryville, California, 94608, United States of America; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, 94720, United States of America.

ABSTRACT
Biological synthesis of pharmaceuticals and biochemicals offers an environmentally friendly alternative to conventional chemical synthesis. These alternative methods require the design of metabolic pathways and the identification of enzymes exhibiting adequate activities. Cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates are natural metabolites which possess beneficial activities for human health, and the search is expanding for novel derivatives that might have enhanced biological activity. For example, biosynthesis in Dianthus caryophyllus is catalyzed by hydroxycinnamoyl/benzoyl-CoA:anthranilate N-hydroxycinnamoyl/ benzoyltransferase (HCBT), which couples hydroxycinnamoyl-CoAs and benzoyl-CoAs to anthranilate. We recently demonstrated the potential of using yeast (Saccharomyces cerevisiae) for the biological production of a few cinnamoyl anthranilates by heterologous co-expression of 4-coumaroyl:CoA ligase from Arabidopsis thaliana (4CL5) and HCBT. Here we report that, by exploiting the substrate flexibility of both 4CL5 and HCBT, we achieved rapid biosynthesis of more than 160 cinnamoyl, dihydrocinnamoyl, and benzoyl anthranilates in yeast upon feeding with both natural and non-natural cinnamates, dihydrocinnamates, benzoates, and anthranilates. Our results demonstrate the use of enzyme promiscuity in biological synthesis to achieve high chemical diversity within a defined class of molecules. This work also points to the potential for the combinatorial biosynthesis of diverse and valuable cinnamoylated, dihydrocinnamoylated, and benzoylated products by using the versatile biological enzyme 4CL5 along with characterized cinnamoyl-CoA- and benzoyl-CoA-utilizing transferases.

No MeSH data available.