Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
  • C07C45/78—Separation; Purification; Stabilisation; Use of additives
  • C07C45/80—Separation; Purification; Stabilisation; Use of additives by liquid-liquid treatment
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
  • C07C45/78—Separation; Purification; Stabilisation; Use of additives
  • C07C45/79—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
  • C07C45/78—Separation; Purification; Stabilisation; Use of additives
  • C07C45/81—Separation; Purification; Stabilisation; Use of additives by change in the physical state, e.g. crystallisation
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
  • C07C45/78—Separation; Purification; Stabilisation; Use of additives
  • C07C45/81—Separation; Purification; Stabilisation; Use of additives by change in the physical state, e.g. crystallisation
  • C07C45/82—Separation; Purification; Stabilisation; Use of additives by change in the physical state, e.g. crystallisation by distillation
  • C07C45/83—Separation; Purification; Stabilisation; Use of additives by change in the physical state, e.g. crystallisation by distillation by extractive distillation

Definitions

• the present invention relates to a process for purifying vanillin or a vanillin derivative obtained by a biotechnological process.
• Vanillin can be obtained by various methods known to those skilled in the art, and in particular by the following two routes:
• a so-called natural route based on a biotechnological process comprising in particular the culture of a microorganism capable of allowing the biotransformation of a fermentation substrate into vanillin. It is in particular known from application EP0885968 such a process in which the fermentation substrate is ferulic acid. US patent 5017388 describes a process in which the fermentation substrate is eugenol and/or isoeugenol. These processes lead to the preparation of a vanillin called natural vanillin.
• vanillin can also be prepared according to a method described as "bio-based” in which the vanillin is derived from lignin, mention may be made in particular of documents US 2745796, DE1132113 and the article entitled “Preparation of lignin from wood dust as vanillin source and comparison of different extraction method” by Azadbakht et al in International Journal of Biology and Biotechnology, 2004, vol 1, No 4, pp 535-537.
• natural vanillin can be purified according to the process described in application EP 2791098 which includes a liquid/liquid extraction step of impurities having a higher pKa than that of vanillin.
• the yield of this process is good, generally greater than 80%, however in order to obtain improved qualities in terms of organoleptic properties such as the smell and/or the color of the vanillin, several additional purification steps are necessary, thus reducing the overall yield of this process.
• the energy efficiency of this process is also globally degraded due to the use of large quantities of solvents.
• vanillin or its derivatives obtained by a biotechnological process may contain certain impurities having boiling points very close to vanillin or its derivatives. It is therefore necessary to size the equipment appropriately in order to allow effective separation of vanillin or its derivatives from these products. This generally implies a lengthening of the residence time in the distillation equipment, which can generate new impurities due to the instability at high temperature of the vanillin and/or impurities.
• the present invention relates to a process for the purification of a fermentation must (M), obtained by a biotechnological process, comprising biomass and vanillin or its derivatives, the said purification process comprising:
• FIGS. 1 and 2 schematize various processes for the purification of vanillin or its derivatives obtained by a biotechnological process according to the present invention.
• the expression “natural vanillin” designates a vanillin obtained by a biotechnological process.
• a process for the preparation of natural vanillin denotes here a biotechnological process comprising the culture of a microorganism capable of allowing the transformation of a fermentation substrate into vanillin.
• the microorganism may be of wild origin or be a genetically modified microorganism (GMM) obtained by molecular biology
• GMM genetically modified microorganism
• vanillin can be produced by a fermentation process of glucose or protocatechuic acid as described in patent application WO 2013/022881.
• vanillin derivative refers to any compound which may be derived from vanillin and in particular to vanillin in salified form or to glucovanillin.
• the term “derivative of vanillic alcohol” refers to any compound which may be derived from vanillic alcohol and in particular to vanillic alcohol in salified form or to the glucoside of vanillic alcohol.
• crystallization refers to a process in which a substance changes to the solid state by a physical process, such as in particular lowering the temperature.
• the present invention relates to a process for the purification of a fermentation must (M), obtained by a biotechnological process, comprising biomass and vanillin or its derivatives, the said purification process comprising:
• the vanillin or its derivatives are in aqueous solution in a fermentation broth (M).
• the process of the present invention consists in purifying a fermentation broth (M) comprising vanillin or its derivatives.
• concentration of vanillin or its derivatives in the fermentation broth (M) is between 0.2 and 5% by weight, preferably between 0.8 and 2.5% by weight, very preferably between 1 .0 and 1.8% by weight.
• the aqueous solution further comprises biomass, in general, the amount of biomass is between 0.5 and 5% by weight of dry matter, preferably between 1.0 and 2.0% by weight of dry matter.
• the fermentation must (M) can also comprise other compounds such as ferulic acid, and derivatives of ferulic acid, vanillic acid, vanillic alcohol or its derivatives, coumaric acid, para-hydroxybenzaldehyde, guaiacol. These compounds can be separated from vanillin or its derivatives according to the method of the present invention.
• at least one compound chosen from ferulic acid, ferulic acid derivatives, vanillic acid, vanillic alcohol or its derivatives, coumaric acid, para-hydroxybenzaldehyde or guaiacol can be used in particular as as a synthetic intermediate.
• the purification process of the present invention comprises a step (a) of separating the biomass of a fermentation broth (M) from an aqueous stream (Al) comprising vanillin or its derivatives.
• This step is a separation step of a solid phase: the biomass, from a liquid phase.
• the liquid phase obtained at the end of the biomass separation step is an aqueous phase (Al) comprising vanillin or its derivatives.
• the biomass separation step can be carried out by filtration such as frontal filtration or tangential filtration, in particular membrane filtration, such as microfiltration, ultrafiltration, nanofiltration, or reverse osmosis.
• Membrane filtration can be carried out in concentration or in diafiltration.
• the biomass separation step is a frontal filtration, it is possible to add adjuvants in order to improve the efficiency of the filtration.
• the biomass separation step can be carried out by one or more microfiltration membranes, in particular having a cutting limit of approximately 0.2 ⁇ m, followed by one or more ultrafiltrations having a limit of smaller cut than microfiltration.
• the vanillin or its derivatives are dissolved in the aqueous phase and the biomass is retained by the membranes.
• ultrafiltration also allows the separation of molecules dissolved in the fermentation must.
• a solvent preferably water
• the amount of solvent added is between 0.5 and 5 equivalents by volume of fermentation broth.
• microfiltration, ultrafiltration or diafiltration can be coupled with a nanofiltration or reverse osmosis step.
• nanofiltration makes it possible to increase the vanillin concentration in the retentate from the nanofiltration step while the water passes through the membrane (permeate from the nanofiltration step).
• the permeate from this nanofiltration step can advantageously be recycled.
• reverse osmosis has a cut limit of less than or equal to 100 Da.
• nanofiltration has a cut limit of less than or equal to 400 Da.
• nanofiltration has a cutting limit greater than or equal to 100 Da, for example cutting limit between 100 and 250 Da.
• the method of the present invention may optionally comprise a step (aO) of stabilizing the fermentation broth (M).
• Step (a) of the process according to the present invention can be carried out on a stabilized fermentation broth (M).
• stabilization refers to any method making it possible to avoid the degradation, in particular by reduction, of the vanillin or its derivatives between the end of the fermentation and the purification process.
• the stabilization can be carried out by adding at least one compound.
• the compound is preferably chosen from sodium benzoate, ascorbic acid and its salts, potassium, calcium or sodium sorbate, zinc sulphate, propanoic acid, acetic acid or its salts, sodium diacetate.
• the amount of compound added is between 0.2 g/L and 6 g/L.
• the stabilization can be carried out by changing the temperature of the fermentation broth.
• the temperature is controlled so as to reach a temperature between 15°C and 23°C, preferably between 18°C and 21°C.
• the stabilization can be carried out by changing the pH of the fermentation broth.
• the pH is controlled so as to reach a pH less than or equal to 7.5, preferably less than or equal to 7, very preferably less than or equal to 6.8.
• the pH is controlled so as to achieve a pH greater than or equal to 5.0, preferably greater than or equal to 6.
• the fermentation broth can also be pasteurized.
• the fermentation broth is then heated to a temperature between 50°C and 90°C, preferably between 60°C and 80°C.
• the heating is generally maintained for a period comprised between 10 min and 120 min, preferably comprised between 15 min and 45 min, for example for 20 min.
• the fermentation must can be stabilized by ultrasound.
• ultrasound is emitted in the fermentation must for a period of between 10 min and 120 min.
• these aspects can be carried out separately or jointly, thus, by way of non-limiting example, it is possible to modify the temperature, the pH and to add a compound according to the conditions described above.
• the purification process generally comprises at least one stage of liquid/liquid extraction of the aqueous stream (Al) making it possible to separate an aqueous stream (A2) and an organic stream (O1) comprising vanillin or its derivatives.
• the liquid/liquid extraction step advantageously makes it possible to transfer the vanillin or its derivatives in aqueous solution in the stream (Al) into an organic phase (01).
• an organic phase (01) comprising vanillin or its derivatives is obtained.
• the liquid/liquid extraction step advantageously allows the separation of vanillin or its derivatives from water-soluble compounds such as vanillic acid, ferulic acid in salified and/or protonated form.
• the pH of the aqueous solution is between 5 and 8, preferably between 5.5 and 7.5, very preferably between 6 and 7.
• the extraction solvent is chosen for these solubilization properties of vanillin or its derivatives, advantageously the solvent can be of biosourced origin.
• the chosen solvent is compatible with the standards of the food industry, in particular FEMA GRAS, immiscible with water.
• the solvent is chosen from methyl ethyl ketone, methyl isobutyl ketone, ethyl acetate, isopropyl acetate, cyclohexane or mixtures thereof.
• the liquid/liquid extraction step can be a discontinuous liquid-liquid extraction.
• the volume ratio of solvent relative to the aqueous solution (Al) of vanillin or its derivatives is between 1:5 and 5:1, from preferably comprised between 1:1 and 5:1, preferably comprised between 1.5:1 and 3:1.
• the liquid/liquid extraction step can be carried out continuously.
• the volume ratio of solvent relative to the aqueous solution (Al) of vanillin or its derivatives is between 5:1 and 1:2, preferably between 3:1 and 1:1.
• the yield of vanillin or its derivatives from the liquid/liquid extraction step is generally greater than or equal to 95%, preferably greater than or equal to 97%, very preferably greater than or equal to 98%.
• the stages of separation of the biomass and of liquid/liquid extraction can be carried out simultaneously.
• This particular aspect makes it possible to extract vanillin or its derivatives in the organic solvent (01) from the fermentation must (M).
• the biomass is separated from the two-phase system. This process is advantageous in that the loss of vanillin or its derivatives in the biomass is reduced.
• This separation step can in particular be carried out continuously by centrifugation, in particular by using a two- or three-phase centrifuge or a counter-current extractor.
• the organic stream (01) comprising the vanillin or its derivatives obtained at the end of the liquid/liquid extraction step generally has a concentration of vanillin or its derivatives of between 0.1% by weight and 10% by weight.
• the organic stream (01) further includes the solvent used during the liquid-liquid extraction.
• the flux (01) can also include other compounds such as ferulic acid, and derivatives of ferulic acid, vanillic acid, vanillic alcohol or its derivatives, coumaric acid, para-hydroxybenzaldehyde, guaiacol.
• a stripping step can be carried out on the aqueous stream (A2) obtained at the end of the liquid-liquid extraction step.
• the stripping step allows the recovery of organic compounds contained in the aqueous stream (A2), in particular vanillin or its derivatives.
• stripping makes it possible to improve the yield of vanillin or its derivatives.
• the stripping step also makes it possible to facilitate the treatment of the aqueous effluents.
• the method of the present invention comprises at least one step (c) of concentrating an organic stream (01) to obtain an organic stream (02) comprising vanillin or its derivatives, in which the compounds having a lower boiling point higher than that of vanillin or its derivatives are separated from said organic stream (01).
• step (c) of concentration makes it possible to evaporate the solvent used during the liquid-liquid extraction.
• the vanillin or its derivatives are in liquid or solid form depending on the temperature. In general, at least 95% of the solvent has been separated from the vanillin or its derivatives.
• the concentration step can consist of one or more concentration steps.
• At least 95% of the quantity of solvent present in the organic flow (01) of vanillin can be evaporated.
• the solvent contained in the organic flux (01) of vanillin is partially evaporated to obtain an organic flux (02).
• the vanillin concentration in the organic stream (02) is greater than the vanillin concentration in the organic stream (01).
• the organic stream (01) of vanillin or its derivatives is concentrated so as to obtain an organic stream (02) of vanillin having a concentration of between 10% by weight and 95% by weight.
• the organic stream (02) can be subjected to one or more concentration steps so as to obtain vanillin or its derivatives in liquid or solid form depending on the temperature.
• the concentration step can be carried out in a continuous process or in a batch process.
• the concentration step can be carried out in a stirred reactor or in a distillation column.
• the concentration step is carried out under vacuum, preferably at a pressure of between 5 mbar and 300 mbar, preferably between 50 mbar and 250 mbar.
• the concentration step is generally carried out at a temperature between 25°C and 100°C.
• the concentration step can be carried out using tray or packed distillation columns, falling film evaporators, wiped film evaporators or partition columns.
• the concentration step is carried out at a pressure at the distillation head of between 0.5 bar and 5 bar, preferably between 1 bar and 3 bar.
• the temperature at the distillation head is generally between 75°C and 150°C, preferably between 85°C and 120°C and very preferably between 90°C and 110°C.
• the concentration step is carried out in such a way as to reduce the transit time of the vanillin or of its derivatives in the reactors or columns.
• the concentration step makes it possible to recover the organic stream (02) comprising vanillin or its derivatives at the bottom of the column while the solvent, and more generally the compounds with less boiling lower than that of vanillin or its derivatives are collected at the top of the column.
• the organic stream (02) comprising vanillin or its derivatives generally has a purity greater than or equal to 90%.
• organic flux (02) may be present in the organic flux (02) in particular vanillic alcohol or its derivatives, vanillic acid, acetovanillone, 4-((4-hydroxy-3-methoxy-benzyl)oxy)-3-methoxybenzaldehyde and 4 - hydroxy-3-(4-hydroxy-3-methoxybenzyl)-5-methoxybenzaldehyde.
• the process of the present invention may comprise at least one step in which the compounds having a boiling point higher than that of vanillin or its derivatives are separated from an organic stream (02) comprising vanillin or its derivatives to obtain a flux (03) comprising vanillin or its derivatives.
• This step can be a tar removal step.
• Step (d) can be carried out in a distillation column, a partition column, a falling film evaporator, or a wiped film evaporator.
• step (d) can be carried out in the presence of a technical adjuvant, for example a plasticizer.
• a plasticizer preferably a plasticizer authorized by the regulations on food products, for example polyethylene glycol, can be added.
• the method of the present invention comprises at least one step of crystallizing the vanillin or its derivatives contained in the stream (03).
• the crystallization step makes it possible to purify a flux (03) comprising vanillin or its derivatives.
• the stream (03) has a purity of between 85% and 99%, preferably greater than or equal to 90%, very preferably greater than or equal to 95%.
• the crystallization step makes it possible to produce a vanillin or its purified derivatives having a purity greater than or equal to 95%, preferably greater than or equal to 97%, very preferably greater than or equal to 99%.
• Vanillin or its derivatives having a purity of between 85% and 99% may also comprise at least one other compound chosen from vanillic alcohol or its derivatives, vanillic acid, guaiacol, acetovanillone, 4-((4-hydroxy-3-methoxy -benzyl)oxy)-3-methoxybenzaldehyde and 4-hydroxy-3-(4-hydroxy-3-methoxybenzyl)-5-methoxybenzaldehyde.
• Crystallization is usually carried out in an alcoholic solution.
• the solvent used for the crystallization can be a water-soluble alcohol, preferably ethanol.
• the solvent used for the crystallization can be a water/alcohol mixture.
• the amount of alcohol is between 2% and 40% by weight, preferably between 5% and 35% by weight, and very preferably between 15% and 25% by weight.
• the vanillin concentration, at the start of crystallization is between 5% and 60% by weight, preferably between 10% and 50% by weight, advantageously between 15% and 35% by weight, and even more preferably between 15% and 25%.
• the crystallization allows the separation of the vanillin or its derivatives from the impurities contained in the stream (03), this separation is advantageously carried out without degradation of the vanillin or its derivatives.
• the crystallization is carried out at a temperature between 0°C and 50°C.
• the crystallization yield is generally greater than or equal to 80%.
• the vanillin or its derivatives obtained at the end of the crystallization step generally has a color in ethanolic solution at 10% by weight less than or equal to 150 Hazen, preferably less than or equal to 100 Hazen, and very preferably less than or equal at 50 Hazen. Moreover, the vanillin or its derivatives obtained at the end of the purification process according to the present invention has consistent organoleptic properties.
• the vanillin is extracted with 6 kg of ethyl acetate.
• the solvent of the upper phase is completely distilled in a rotavapor at 50° C. under vacuum at 100 mbar, to obtain a medium concentrated in vanillin at 78%.
• this latter medium is crystallized by dissolving it in a stirred reactor containing a water/ethanol mixture (9/2) at 40° C. to obtain a vanillin concentration of 20%. The whole is cooled with stirring to 0°C. The crystallized vanillin is filtered through a Büchner filter, washed with cold water and then dried in an oven at 50° C. under 200 mbar for 20 hours. The purity of the vanillin obtained is 99.7% with a color in 10% by weight ethanolic solution of 19 Hazen. Crystallized vanillin also has good organoleptic properties.

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• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Crystallography & Structural Chemistry (AREA)
• Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
• Preparation Of Compounds By Using Micro-Organisms (AREA)
• Extraction Or Liquid Replacement (AREA)

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• 2021
  • 2021-03-15 FR FR2102535A patent/FR3120627B1/fr active Active
• 2022
  • 2022-03-14 BR BR112023018033A patent/BR112023018033A2/pt unknown
  • 2022-03-14 EP EP22714427.6A patent/EP4308535A1/de active Pending
  • 2022-03-14 WO PCT/EP2022/056528 patent/WO2022194772A1/fr active Application Filing
  • 2022-03-14 US US18/550,941 patent/US20240140896A1/en active Pending
  • 2022-03-14 CN CN202280021205.0A patent/CN117062797A/zh active Pending

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