Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing oxygen-containing organic compound
Patent
1997-06-12
1999-02-02
Saucier, Sandra E.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing oxygen-containing organic compound
435148, 435911, C12P 742, C12P 726
Patent
active
058663800
DESCRIPTION:
BRIEF SUMMARY
This application is a 371 of PCT/FR95/01173 filed Sep. 13, 1995.
The present invention relates to the obtaining oz vanillic acid and vanillin by bioconversion.
At the present time, vanillin is the flavouring most widely used in the agri-foodstuffs industries. However, the production of natural vanillin from vanilla pods covers only 20% of the market requirements, and its production cost is of the order of 25,000 FF/kg.
Vanillin may also be obtained by chemical synthesis; however, this method of obtaining the product, while suitable for the manufacture of perfumes and cosmetics, may give rise to legislative problems in the agri-foodstuffs industries. In addition, synthetic flavourings tend, moreover, to be less well liked by consumers than flavourings of natural origin.
Accordingly, an effort is being made to obtain aromatic compounds produced by means of biological processes, which employ microorganisms (bacteria, yeasts, fungi), animal or plant cells or their enzyme systems.
Vanillin is produced by certain plants and microorganisms, especially fungi, where it constitutes one of the degradation products of precursors containing an aromatic ring (ferulic acid and vanillic acid).
European Patent Application 453 368 in the name of the company PERNOD-RICARD describes the production of natural vanillin by bioconversion of ferulic acid or vanillic acid in the presence of a filamentous fungus of the Basidiomycetes group, Pycnoporus cinnabarinus.
In this fungus, four metabolic pathways of conversion of ferulic acid have been identified: coniferyl alcohol; thereafter different dimers are formed from this compound. This is a minor pathway. with the loss of two carbons and formation of vanillic acid. vanillin by the reductase I. The vanillin produced may then be reduced by the reductase II to vanillyl alcohol. decarboxylated to methoxyhydroquinone by the action of an intracellular vanillate hydroxylase.
Under the conditions described in Application EP 453 368, the production of vanillin by P. cinnabarinus MIC11 from 300 mg/l of ferulic acid is at most of the order of 45 mg/l (molar conversion yield of 20.5%), and from 300 mg/l of vanillic acid this production is at most of the order of 81.4 mg/l (molar conversion yield of 31%).
The object of the present invention is to improve the yield of the production of natural vanillin by bioconversion from precursors containing an aromatic ring (ferulic acid and vanillic acid). To this end, the inventors have sought to improve the yield of conversion of ferulic acid to vanillic acid, and the yield of conversion of vanillic acid to vanillin.
The subject of the present invention is a process for obtaining vanillic acid by bioconversion from ferulic acid, which process is characterized in that the said bioconversion is performed using a culture comprising at least one strain of a filamentous fungus chosen from the group consisting of Ascomycetes and Basidiomycetes, or at least one actinomycete strain.
The subject of the present invention is also a process for obtaining vanillin by bioconversion from ferulic acid, which process is characterized in that it comprises: defined above; filamentous fungus of the class Basidiomycetes.
According to a first variant of the process for obtaining vanillin according to the invention, the two steps are performed sequentially. In this case, the process according to the invention comprises: one strain of a filamentous fungus chosen from the group consisting of Ascomycetes and Basidiomycetes, or at least one actinomycete strain, and the vanillic acid produced is collected; added to a culture comprising at least one strain of a filamentous fungus of the class Basidiomycetes, and the vanillin produced is collected.
To carry out this variant, the fungi used in each of the steps are cultured separately until a biomass is obtained possessing the capacities needed for the bioconversion (that is to say a biomass of at least 0.5 g of dry matter per liter of culture), and the cultures obtained are then used successively.
According to a second variant of the
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patent: 5279950 (1994-01-01), Labuda et al.
Middelhoven, Wouter J., "Catabolism of benzene compounds by ascomycetous and basidiomycetous yeasts and yeastlike fungi: a literature review and an experimental approach", Chemical Abstracts,vol. 120, No. 19, May 9, 1994, Abstract No. 239831, pp. 498-499.
Falconnier, B. et al., "Vanillin as a product of ferulic acid biotransformation by the white-rot fungus Pycnoporus cinnabarinus I-937: Identification of metabolic pathways", Chemical Abstracts, vol. 121, No. 23, Dec. 5, 1994, Abstract No. 276417, p. 590.
Asther Marcel
Delattre Michel
Haon Mireille
Lesage-Meessen Laurence
Institut National De La Recherche Agronomique-I.N.R.A.
Saucier Sandra E.
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