Metabolic engineering of Pseudomonas fluorescens for the production of vanillin from ferulic acid

Gioia, Diana Di; Luziatelli, Francesca; Negroni, Andrea; Ficca, Anna Grazia; Fava, Fabio; Ruzzi, Maurizio

doi:10.1016/j.jbiotec.2011.08.014

Cited by 110 publications

(65 citation statements)

References 23 publications

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“…Conversely, several workers have used Pseudomonas for the production of vanillin by making its recombinant strain using gene viz. feruloyl-CoA synthetase (fcs) and hydratase/aldolase (ech) to convert ferulic acid into vanillin (Gioia et al, 2011). Recombinant strains of E.coli and other bacteria were also developed by inserting the EhyAB, CalA, and CalB gene for vanillin production, which catalysed the same reaction as that of the HCHL gene (Kaur and Chakraborty, 2013).…”

Section: Introductionmentioning

confidence: 99%

Vanillin production in metabolically engineered Beta vulgaris hairy roots through heterologous expression of Pseudomonas fluorescens HCHL gene

Singh

Khan

Pandey

et al. 2015

Industrial Crops and Products

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Section: Introductionmentioning

confidence: 99%

Vanillin production in metabolically engineered Beta vulgaris hairy roots through heterologous expression of Pseudomonas fluorescens HCHL gene

Singh

Khan

Pandey

et al. 2015

Industrial Crops and Products

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“…ATCC 39116, whose 16S rRNA gene sequence is identical to that of Amycolatopsis sp. HR167, the whole genome was sequenced to identify enzymes involved in vanillin catabolism, as previously done for other vanillin-producing strains (6,8,(16)(17)(18). In order to promote a more efficient accumulation of vanillin by preventing its oxidation to vanillic acid, enzymes with homology to characterized vanillin dehydrogenases (VDHs) of other bacteria were screened.…”

mentioning

confidence: 99%

Investigation of the Amycolatopsis sp. Strain ATCC 39116 Vanillin Dehydrogenase and Its Impact on the Biotechnical Production of Vanillin

Fleige

Hansen

Krøll

et al. 2013

Appl Environ Microbiol

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bThe actinomycete Amycolatopsis sp. strain ATCC 39116 is capable of synthesizing large amounts of vanillin from ferulic acid, which is a natural cell wall component of higher plants. The desired intermediate vanillin is subject to undesired catabolism caused by the metabolic activity of a hitherto unknown vanillin dehydrogenase (VDH ATCC 39116 ). In order to prevent the oxidation of vanillin to vanillic acid and thereby to obtain higher yields and concentrations of vanillin, the responsible vanillin dehydrogenase in Amycolatopsis sp. ATCC 39116 was investigated for the first time by using data from our genome sequence analysis and further bioinformatic approaches. The vdh gene was heterologously expressed in Escherichia coli, and the encoded vanillin dehydrogenase was characterized in detail. VDH ATCC 39116 was purified to apparent electrophoretic homogeneity and exhibited NAD ؉ -dependent activity toward vanillin, coniferylaldehyde, cinnamaldehyde, and benzaldehyde. The enzyme showed its highest level of activity toward vanillin at pH 8.0 and at a temperature of 44°C. In a next step, a precise vdh deletion mutant of Amycolatopsis sp. ATCC 39116 was generated. The mutant lost its ability to grow on vanillin and did not show vanillin dehydrogenase activity. A 2.3-times-higher vanillin concentration and a substantially reduced amount of vanillic acid occurred with the Amycolatopsis sp. ATCC 39116 ⌬vdh::Km r mutant when ferulic acid was provided for biotransformation in a cultivation experiment on a 2-liter-bioreactor scale. Based on these results and taking further metabolic engineering into account, the Amycolatopsis sp. ATCC 39116 ⌬vdh::Km r mutant represents an optimized and industrially applicable platform for the biotechnological production of natural vanillin.

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“…Unlike CreG, CreC can only employ NADP ϩ as a cofactor, because no catalytic activity was detected in the presence of NAD ϩ . This feature is distinct to other benzylaldehyde dehydrogenases, which have been reported to have the capacity to utilize both NAD ϩ and NADP ϩ as cofactors (53)(54)(55). CreD Is a Universal Phosphohydrolase-Our previous study demonstrated that CreD displayed phosphohydrolase activity against the unnatural substrate 4-nitrophenylphosphate (22).…”

Section: Volume 291 • Number 12 • March 18 2016mentioning

confidence: 88%

Characterization of a Unique Pathway for 4-Cresol Catabolism Initiated by Phosphorylation in Corynebacterium glutamicum

et al. 2016

Journal of Biological Chemistry

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4-Cresol is not only a significant synthetic intermediate for production of many aromatic chemicals, but also a priority environmental pollutant because of its toxicity to higher organisms. In our previous studies, a gene cluster implicated to be involved in 4-cresol catabolism, creCDEFGHIR, was identified in Corynebacterium glutamicum and partially characterized in vivo. In this work, we report on the discovery of a novel 4-cresol biodegradation pathway that employs phosphorylated intermediates. This unique pathway initiates with the phosphorylation of the hydroxyl group of 4-cresol, which is catalyzed by a novel 4-methylbenzyl phosphate synthase, CreHI. Next, a unique class I P450 system, CreJEF, specifically recognizes phosphorylated intermediates and successively oxidizes the aromatic methyl group into carboxylic acid functionality via alcohol and aldehyde intermediates. Moreover, CreD (phosphohydrolase), CreC (alcohol dehydrogenase), and CreG (aldehyde dehydrogenase) were also found to be required for efficient oxidative transformations in this pathway. Steady-state kinetic parameters (K m and k cat ) for each catabolic step were determined, and these results suggest that kinetic controls serve a key role in directing the metabolic flux to the most energy effective route.

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Metabolic engineering of Pseudomonas fluorescens for the production of vanillin from ferulic acid

Cited by 110 publications

References 23 publications

Vanillin production in metabolically engineered Beta vulgaris hairy roots through heterologous expression of Pseudomonas fluorescens HCHL gene

Vanillin production in metabolically engineered Beta vulgaris hairy roots through heterologous expression of Pseudomonas fluorescens HCHL gene

Investigation of the Amycolatopsis sp. Strain ATCC 39116 Vanillin Dehydrogenase and Its Impact on the Biotechnical Production of Vanillin

Characterization of a Unique Pathway for 4-Cresol Catabolism Initiated by Phosphorylation in Corynebacterium glutamicum

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