Decarboxylation of Ferulic Acid to 4-Vinyl Guaiacol by Streptomyces setonii

Max, Belén; Carballo, Javier; Cortés, Sandra; Domínguez, José Manuel

doi:10.1007/s12010-011-9424-7

Cited by 20 publications

(11 citation statements)

References 26 publications

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“…in some studies (Max et al . ; Zhang ). On the basis of analysis results, 4‐vinylguaiacol was supposed to be an degradation product of Cβ‐Cγ bond cleavage of guaiacylglycerol.…”

Section: Resultsmentioning

confidence: 99%

Bacillus amyloliquefaciensCotA degradation of the lignin model compound guaiacylglycerol-β-guaiacyl ether

Yang

Gao

et al. 2018

Lett Appl Microbiol

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The cotA gene from Bacillus amyloliquefaciens MN-13 was cloned and expressed in Escherichia coli Transetta. Nucleotide sequence analysis showed an open reading frame of 1542 bp encoding a polypeptide comprised of 513 amino acids. The degradation of lignin model compounds by recombinant CotA was investigated by HPLC-MS with guaiacylglycerol-β-guaiacyl ether as the substrate. The compounds including guaiacol, 3-(4-hydro-3-methoxyphenyl)-3-oxo-propanol and 4-hydro-3-methoxy acetophenone detected by HPLC-MS verified the rupture of β-O-4 bond and oxidation Cα bond of guaiacylglycerol-β-guaiacyl ether by CotA. 4-vinylguaiacol and 1-(4-hydroxy-3-methoxyl phenyl)-1-(2-methoxyl) phenoxyl ethylene were first time found in the degradation products of guaiacylglycerol-β-guaiacyl ether. The appearance of 4-vinylguaiacol and 4-hydro-3-methoxy acetophenone confirmed the cleavage of Cβ-Cγ bond. 1-(4-hydroxy-3-methoxyl phenyl)-2-(2-methoxyl) phenoxyl ethylene was coupled by the radical reaction of 4-vinylguaiacol with guaiacol. Otherwise, no corresponding degradation product was found to give a proof of cleavage of Cα-Cβ bond in guaiacylglycerol-β-guaiacyl ether by CotA.

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“…in some studies (Max et al . ; Zhang ). On the basis of analysis results, 4‐vinylguaiacol was supposed to be an degradation product of Cβ‐Cγ bond cleavage of guaiacylglycerol.…”

Section: Resultsmentioning

confidence: 99%

Bacillus amyloliquefaciensCotA degradation of the lignin model compound guaiacylglycerol-β-guaiacyl ether

Yang

Gao

et al. 2018

Lett Appl Microbiol

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“…ATCC 39116 is an important organism for the production of natural vanilla flavor (6,9,41). Several laboratories have studied this strain, but most of them focused on the optimization of fermentation parameters (7,8,10,11). So far, only one knockout and one genomic integration have been described for this strain, which highlights the difficulties in genetically manipulating it (5,6).…”

Section: Discussionmentioning

confidence: 99%

“…The resulting mutant reached the currently highest described vanillin concentration obtained from ferulic acid without extraction of the fermentation broth (5). The other publications focused on the optimization of the fermentation conditions (7)(8)(9)(10) or on the use of adsorbing materials (11). The transformation of this strain has been established in 2002, the genome sequence has been available since 2012, and usable expression vectors with different promoters have existed since 2014 (12)(13)(14).…”

mentioning

confidence: 99%

Development of an Improved System for the Generation of Knockout Mutants of Amycolatopsis sp. Strain ATCC 39116

Meyer

Pupkes

Steinbüchel

2017

Appl Environ Microbiol

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The Gram-positive actinomycete Amycolatopsis sp. strain ATCC 39116 is used for the industrial production of natural vanillin. Previously, the only gene deletion performed in this strain targeted the gene vdh, coding for a vanillin dehydrogenase. The generation of this mutant suffered from a high number of illegitimate recombinations and a low rate of homologous recombination. To alleviate this, we constructed an optimized deletion system based on a modified suicide vector. Thereby, we were able to increase the rate of homologous integration from less than 1% of the analyzed clones to 20% or 50%, depending on the targeted gene. We were furthermore able to reduce the screening effort needed to identify homogenotes through the use of the rpsL gene from Saccharopolyspora erythraea, which confers streptomycin sensitivity on clones still carrying the suicide vector. The new suicide vector is p6SUI5ERPSL, and its applicability was demonstrated by the deletion of three Amycolatopsis gene clusters. The deletion of the first of the gene clusters, coding for an aldehyde oxidase (yagRST), led to no altered phenotype compared to the parent strain; deletion of the second, coding for a vanillic acid decarboxylase (vdcBCD), led to a phenotype that was strongly impaired in its growth with vanillic acid as the sole carbon source and also unable to form guaiacol; and deletion of the third, coding for a vanillate demethylase (vanAB), led to only a negligible impact in comparison. Therefore, we showed that decarboxylation of vanillic acid is the main degradation pathway in Amycolatopsis sp. ATCC 39116 while the demethylation plays only a minor role and does not compensate the deletion of vdcBCD.IMPORTANCE Amycolatopsis sp. ATCC 39116 is an important microorganism used for the production of natural vanillin from ferulic acid. In contrast to this importance, it has previously been shown that this strain is hard to manipulate on a genetic level. We therefore generated an optimized system to facilitate the deletion of genes in this strain. This allowed us to greatly reduce the time and work requirements for generating deletions. This could allow the improvement of vanillin production in the future and also the elucidation of metabolic pathways. To test our deletion system, we deleted three gene clusters in Amycolatopsis sp. ATCC 39116. One showed no involvement in the metabolism of vanillin, while the second proved to be the main pathway of vanillic acid degradation and completely stopped the formation of the off-flavor guaiacol. The third appeared to have only a negligible impact on the degradation of vanillic acid.

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“…This bacterium has an outstanding tolerance to the desired product, and it is currently the most promising microbial production platform, by far, for natural vanillin. Thus, several previous studies focused on optimization of the production conditions, aiming at higher yields and higher final product concentrations (24)(25)(26).…”

Section: Discussionmentioning

confidence: 99%

Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin

Fleige

Meyer

Steinbüchel

2016

Appl Environ Microbiol

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The Gram-positive bacterium Amycolatopsis sp. ATCC 39116 is used for the fermentative production of natural vanillin from ferulic acid on an industrial scale. The strain is known for its outstanding tolerance to this toxic product. In order to improve the productivity of the fermentation process, the strain's metabolism was engineered for higher final concentrations and molar yields. Degradation of vanillin could be decreased by more than 90% through deletion of the vdh gene, which codes for the central vanillin catabolism enzyme, vanillin dehydrogenase. This mutation resulted in improvement of the final concentration of vanillin by more than 2.2 g/liter, with a molar yield of 80.9%. Further improvement was achieved with constitutive expression of the vanillin anabolism genes ech and fcs, coding for the enzymes feruloyl-coenzyme A (CoA) synthetase (fcs) and enoyl-CoA hydratase/aldolase (ech). The transcription of both genes was shown to be induced by ferulic acid, which explains the unwanted adaptation phase in the fermentation process before vanillin was efficiently produced by the wild-type cells. Through the constitutive and enhanced expression of the two genes, the adaptation phase was eliminated and a final vanillin concentration of 19.3 g/liter, with a molar yield of 94.9%, was obtained. Moreover, an even higher final vanillin concentration of 22.3 g/liter was achieved, at the expense of a lower molar yield, by using an improved feeding strategy. This is the highest reported vanillin concentration reached in microbial fermentation processes without extraction of the product. Furthermore, the vanillin was produced almost without by-products, with a molar yield that nearly approached the theoretical maximum. IMPORTANCEMuch effort has been put into optimization of the biotechnological production of natural vanillin. The demand for this compound is growing due to increased consumer concerns regarding chemically produced food additives. Since this compound is toxic to most organisms, it has proven quite difficult to reach high concentrations and molar yields. This study shows that improvements in the final vanillin concentrations and molar yields can be made through a combination of modification of the fermentation parameters and molecular strain engineering, without the need for methods such as continuous extraction from the fermentation broth. Using this approach, we were able to reach a final vanillin concentration of 22.3 g/liter, which is the highest vanillin concentration reported to date that was generated with Amycolatopsis sp. ATCC 39116 without additional extraction of the toxic product. With an annual worldwide demand of more than 16,000 tons, vanillin is one of the most important flavor compounds (1). It is widely used in various applications, such as the production of fragrances, pharmaceuticals, foods, and beverages (2). The vast majority (99%) of the industrially used vanillin is chemically synthesized from lignin and crude oil derivatives, since the extraction of vanillin from the pods o...

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Decarboxylation of Ferulic Acid to 4-Vinyl Guaiacol by Streptomyces setonii

Cited by 20 publications

References 26 publications

Bacillus amyloliquefaciensCotA degradation of the lignin model compound guaiacylglycerol-β-guaiacyl ether

Bacillus amyloliquefaciensCotA degradation of the lignin model compound guaiacylglycerol-β-guaiacyl ether

Development of an Improved System for the Generation of Knockout Mutants of Amycolatopsis sp. Strain ATCC 39116

Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin

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