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Comparative transcriptomics and proteomics of p-hydroxybenzoate producing Pseudomonas putida S12: novel responses and implications for strain improvement.

Verhoef S, Ballerstedt H, Volkers RJ, de Winde JH, Ruijssenaars HJ - Appl. Microbiol. Biotechnol. (2010)

Bottom Line: Also many transporters were differentially expressed, one of which--a previously uncharacterized multidrug efflux transporter with locus tags PP1271-PP1273--was found to be associated with p-hydroxybenzoate export.Eliminating the most prominent of these resulted in a 22% p-hydroxybenzoate yield improvement.Remarkably, the upregulation of genes contributing to p-hydroxybenzoate formation was much higher in glucose than in glycerol-cultured cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628, BC, Delft, the Netherlands.

ABSTRACT
A transcriptomics and proteomics approach was employed to study the expression changes associated with p-hydroxybenzoate production by the engineered Pseudomonas putida strain S12palB1. To establish p-hydroxybenzoate production, phenylalanine-tyrosine ammonia lyase (pal/tal) was introduced to connect the tyrosine biosynthetic and p-coumarate degradation pathways. In agreement with the efficient p-hydroxybenzoate production, the tyrosine biosynthetic and p-coumarate catabolic pathways were upregulated. Also many transporters were differentially expressed, one of which--a previously uncharacterized multidrug efflux transporter with locus tags PP1271-PP1273--was found to be associated with p-hydroxybenzoate export. In addition to tyrosine biosynthesis, also tyrosine degradative pathways were upregulated. Eliminating the most prominent of these resulted in a 22% p-hydroxybenzoate yield improvement. Remarkably, the upregulation of genes contributing to p-hydroxybenzoate formation was much higher in glucose than in glycerol-cultured cells.

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Schematic overview of differentially expressed genes involved in relevant pathways of p-hydroxybenzoate biosynthesis, as derived from transcriptomics analysis of P. putida S12palB1. Gene names are indicated in italic or when absent, indicated by locus tags from P. putida KT2440. Red lines indicate upregulation and green lines indicate downregulation in P. putida S12palB1. The text boxes describe the proposed role in the enhanced p-hydroxybenzoate production of the differentially expressed genes
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Fig3: Schematic overview of differentially expressed genes involved in relevant pathways of p-hydroxybenzoate biosynthesis, as derived from transcriptomics analysis of P. putida S12palB1. Gene names are indicated in italic or when absent, indicated by locus tags from P. putida KT2440. Red lines indicate upregulation and green lines indicate downregulation in P. putida S12palB1. The text boxes describe the proposed role in the enhanced p-hydroxybenzoate production of the differentially expressed genes

Mentions: For the production of p-hydroxybenzoate, tyrosine biosynthesis in P. putida S12_427 was connected to the endogenous p-coumarate catabolic pathway by introducing the pal/tal gene from Rhodosporidium toruloides (Fig. 1; Verhoef et al. 2007). This gene encodes phenylalanine/tyrosine ammonia lyase (Pal/Tal) that converts tyrosine into p-coumarate. The resulting synthetic pathway could be employed to accumulate the p-coumarate catabolic intermediate p-hydroxybenzoate, after eliminating the pobA gene encoding p-hydroxybenzoate hydroxylase. Figure 3 presents an overview of p-hydroxybenzoate biosynthesis in the resulting p-hydroxybenzoate producing strain, P. putida S12palB1 (Verhoef et al. 2007).


Comparative transcriptomics and proteomics of p-hydroxybenzoate producing Pseudomonas putida S12: novel responses and implications for strain improvement.

Verhoef S, Ballerstedt H, Volkers RJ, de Winde JH, Ruijssenaars HJ - Appl. Microbiol. Biotechnol. (2010)

Schematic overview of differentially expressed genes involved in relevant pathways of p-hydroxybenzoate biosynthesis, as derived from transcriptomics analysis of P. putida S12palB1. Gene names are indicated in italic or when absent, indicated by locus tags from P. putida KT2440. Red lines indicate upregulation and green lines indicate downregulation in P. putida S12palB1. The text boxes describe the proposed role in the enhanced p-hydroxybenzoate production of the differentially expressed genes
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2874742&req=5

Fig3: Schematic overview of differentially expressed genes involved in relevant pathways of p-hydroxybenzoate biosynthesis, as derived from transcriptomics analysis of P. putida S12palB1. Gene names are indicated in italic or when absent, indicated by locus tags from P. putida KT2440. Red lines indicate upregulation and green lines indicate downregulation in P. putida S12palB1. The text boxes describe the proposed role in the enhanced p-hydroxybenzoate production of the differentially expressed genes
Mentions: For the production of p-hydroxybenzoate, tyrosine biosynthesis in P. putida S12_427 was connected to the endogenous p-coumarate catabolic pathway by introducing the pal/tal gene from Rhodosporidium toruloides (Fig. 1; Verhoef et al. 2007). This gene encodes phenylalanine/tyrosine ammonia lyase (Pal/Tal) that converts tyrosine into p-coumarate. The resulting synthetic pathway could be employed to accumulate the p-coumarate catabolic intermediate p-hydroxybenzoate, after eliminating the pobA gene encoding p-hydroxybenzoate hydroxylase. Figure 3 presents an overview of p-hydroxybenzoate biosynthesis in the resulting p-hydroxybenzoate producing strain, P. putida S12palB1 (Verhoef et al. 2007).

Bottom Line: Also many transporters were differentially expressed, one of which--a previously uncharacterized multidrug efflux transporter with locus tags PP1271-PP1273--was found to be associated with p-hydroxybenzoate export.Eliminating the most prominent of these resulted in a 22% p-hydroxybenzoate yield improvement.Remarkably, the upregulation of genes contributing to p-hydroxybenzoate formation was much higher in glucose than in glycerol-cultured cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628, BC, Delft, the Netherlands.

ABSTRACT
A transcriptomics and proteomics approach was employed to study the expression changes associated with p-hydroxybenzoate production by the engineered Pseudomonas putida strain S12palB1. To establish p-hydroxybenzoate production, phenylalanine-tyrosine ammonia lyase (pal/tal) was introduced to connect the tyrosine biosynthetic and p-coumarate degradation pathways. In agreement with the efficient p-hydroxybenzoate production, the tyrosine biosynthetic and p-coumarate catabolic pathways were upregulated. Also many transporters were differentially expressed, one of which--a previously uncharacterized multidrug efflux transporter with locus tags PP1271-PP1273--was found to be associated with p-hydroxybenzoate export. In addition to tyrosine biosynthesis, also tyrosine degradative pathways were upregulated. Eliminating the most prominent of these resulted in a 22% p-hydroxybenzoate yield improvement. Remarkably, the upregulation of genes contributing to p-hydroxybenzoate formation was much higher in glucose than in glycerol-cultured cells.

Show MeSH
Related in: MedlinePlus