WO2013120924A2 - Procédés de production biotechnologique d'acides organiques par l'intermédiaire d'au moins une souche de levure - Google Patents

Procédés de production biotechnologique d'acides organiques par l'intermédiaire d'au moins une souche de levure Download PDF

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Publication number
WO2013120924A2
WO2013120924A2 PCT/EP2013/052922 EP2013052922W WO2013120924A2 WO 2013120924 A2 WO2013120924 A2 WO 2013120924A2 EP 2013052922 W EP2013052922 W EP 2013052922W WO 2013120924 A2 WO2013120924 A2 WO 2013120924A2
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WO
WIPO (PCT)
Prior art keywords
organic acids
yeast strain
production
candida
fumarate
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PCT/EP2013/052922
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German (de)
English (en)
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WO2013120924A3 (fr
Inventor
Andreas Aurich
Roland Arno MÜLLER
Ina Ludwig
Isabel WAENGLER
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Thyssenkrupp Uhde Gmbh
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Publication of WO2013120924A2 publication Critical patent/WO2013120924A2/fr
Publication of WO2013120924A3 publication Critical patent/WO2013120924A3/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/44—Polycarboxylic acids
    • C12P7/46—Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/44—Polycarboxylic acids
    • C12P7/48—Tricarboxylic acids, e.g. citric acid

Definitions

  • Dicarboxylic acids such as fumarate (literally: salt of fumaric acid, in the context of the present invention, the terms “fumarate” and “fumaric acid” are used interchangeably), which occur in the tricarboxylic acid cycle of living organisms and play an important role in the metabolism of many organisms, have a variety of industrial applications.
  • Fumaric acid is of interest for numerous applications in the food industry and is used as an acidulant E 297, as a substitute for tartaric acid in beverages and baking powder. Fumaric acid can be used as a building block as a starting material for polymer production, as a mordant in dyeing, in the production of unsaturated polyester resins, paints, coatings and for the production of polyhydric alcohols and paper sizes. Likewise, the chemical conversion of fumaric acid into succinic acid or maleic acid is possible. The monoethyl and dimethyl esters of fumaric acid are used in medicines for psoriasis (psoriasis).
  • Fumaric acid is currently produced industrially by the isomerization of maleic acid, which in turn is obtained from maleic anhydride.
  • the starting material for maleic anhydride is primarily petrochemical-derived benzene, but also mixtures of n-butanes.
  • fumarate is characterized mainly by chemical transformation Since the 1940s, great efforts have been made to replace these processes by microbial processes, using renewable raw materials and vegetable or animal by-products from petrochemical reactants.
  • filamentous fungi of the genus Rhizopus were experimented with, which can produce up to 97.7 g of fumarate per liter of medium when using glucose as a substrate (Murray 2011).
  • yeasts have properties that are particularly advantageous for biotechnological fermentation processes. The following aspects are worth mentioning:
  • yeast Candida hydrocarbofumarica (later taxonomically classified as Candida blankii) was identified as being suitable for fumarate production (Furukawa et al., 1978).
  • Studies with the strain C. hydrocarbofumarica Et-15-2 resulted in product concentrations of 30-50 g / l fumarate at productivities of 0.2-0.3 g / l * h when using paraffins as substrate.
  • the production of fumarate as intermediate for the production of L-aspartic acid in a two-stage process is described, wherein the realized in the first stage Fumarat raw with C.
  • hydrocarbofumarica ATCC20473 the highest product concentrations and productivities described for yeasts, with up to 57 g / 1 fumarate and 0.8 g / l * h (Table 2).
  • the substrate used is n paraffins having up to 20 carbon atoms, preferably having from 13 to 18 carbon atoms. Later work with various C. blankii strains led to rather moderate results (33-36 g / 1 fumarate).
  • An object of the invention is to provide alternative processes for producing organic acids, preferably fumarate. Another object of the invention is to provide processes for the production of organic acids, preferably fumarate, in which renewable raw materials can be used. Another object of the invention is to provide the advantages of at least one yeast strain for to make available the production of organic acids, preferably fumarate.
  • a process for the biotechnological production of organic acids by at least one yeast strain wherein at least one renewable raw material is used as the substrate.
  • Yeasts are unicellular fungi that multiply by budding or division (cleavage). Most belong to the section of ascomycota.
  • the term "yeast” for unicellular fungi is often used in contrast to the filamentous fungi, which include, for example, molds such as Rhizopus.
  • yeast strains is intended to include strains of the genera Saccharomyces; Schizosaccharomyces; Wickerhamia; Debayomyces; Hansenula; Hanseniaspora; Pichia; Kloeckera; Candida; Zygo saccharomyces; Ogataea; Kuraishia; Komagataeüa; Yarrowia; Metschnikowia; Wiüiopsis; Nakazawaea; Kluyveromyces; Cryptococcus; Torulaspora; Torulopsis; Bullera; Rhodotorula; Sorobolomyces; Pseudozyma; Saccharomycopsis; Saccharomycodes; Trichosporon and / or Trichoderma are understood.
  • renewable raw material refers to organic raw materials which are at least indirectly derived from agricultural and forestry production or which are biogenic waste products, in contrast to which in the known yeast-based processes for the production of dicarboxylic acids only finite Resources in the form of petrochemical products (eg n-paraffins) are used with the term “organic acids” are preferred here organic acids of the Citrateyklus meaning citrate, isocitrate, ⁇ -ketoglutarate, succinate, fumarate, malate and oxaloacetate.
  • the substrate is selected from the group comprising glucose, fructose, sucrose, galactose, lactose, maltose, sorbose, xylose, arabinose, trehalose, ethanol, glycerol, erythritol, oleic acid, rapeseed, soybean and / or sunflower oil ,
  • the substrate is very particularly preferably glucose, which can be produced, for example, by enzymatic degradation of starch or cellulose.
  • glycerol which is obtained, for example, by transesterification in the production of biodiesel (about 100 kg per ton of biodiesel).
  • the production of the organic acids takes place under submerged culture under aerobic conditions.
  • Cultivation is strain-specific at pH values of 3-8 and a temperature of 20 ° C to 40 ° C, preferably at pH 5-7 and temperatures of 26 ° C to 33 ° C.
  • the production of the organic acids takes place as submerged culture in batch mode, in fed-batch mode, in repeated-fed batch mode or in continuous mode.
  • the organic acids to be produced are at least one C4-dicarboxylic acid. Very particular preference is given to fumarate.
  • At least one yeast strain belongs to the genus Candida. It is particularly preferred that at least one yeast strain belongs to the species Candida rugosa and / or Candida blankii. Preferred embodiments relate to the strains Candida rugosa IFO 0731, Candida spec. MUCL 29842, Candida blankii MUCL 30066, Candida blankii MUCL 30304, Candida blankii MUCL 30304, Candida blankii IF01973, Candida blankii IHEM 4004, Candida blankii MYA 3435, Candida blankii CBS 6788, and Candida blankii CBS 6833.
  • culturing is carried out in a strain-specific manner at pH values of 3-8 and a temperature of 20.degree. C. to 40.degree. C., preferably at pH 5-7, and at temperatures of 26.degree. C. to 33.degree. Therefore, a neutralizing agent is required to control the pH decrease caused by the organic acids produced.
  • At least one yeast strain is genetically modified.
  • Such a genetic modification may preferably be chosen such that it does not a) have at least one homologous protein, less strongly or in dysfunctional form
  • At least one neutralizing agent selected from the group containing CaC0 3 , Ca (OH) 2 , KOH, NaOH, Na 2 C0 3 , NaHC0 3 , NH 4 OH, (NH 4 ) 2 C0 3 , NH 4 HC0 3 and mixtures of these substances is used.
  • aqueous form neutralizing agents selected from KOH, NaOH, Na 2 C0 3 , NaHC0 3 , NH 4 OH, (NH 4 ) 2 C0 3 and NH 4 HC0 3 provided.
  • Such neutralizing agents have particular advantages. Thus, they are easier to handle, have a simple metering because they are liquid metered and no inaccuracies evoke by not or poorly soluble components, and are easier to keep sterile.
  • the neutralizing agent is Na 2 C0 3 .
  • the invention provides for the use of at least one yeast strain for the biotechnological production of organic acids, wherein at least one renewable raw material is used as the substrate.
  • Example 1 The wild-type strain Candida blankii IFO 1973 was cultivated on YPD agar plates of the following composition for 24 to 48 hours at 30 ° C. in each case:
  • the preculture was cultured in 500 ml Erlenmeyer flasks with baffles, at 30 ° C., at pH 5.5-7, for 48 h on a shaking apparatus at a shaking frequency of 160 rpm.
  • the precultivation was carried out by inoculating from 5 ml of inoculum to 50 ml of sterilized nutrient medium prepared with distilled water (composition see above). By regular Substratnachdostechnik the glucose concentration was maintained during the cultivation in a range of 40 to 60 g / 1.
  • the cultivation was carried out under sterile conditions at a temperature of 30 ° C and a shaking frequency of 160 rpm.
  • the pH was in a range below pH 5.5 with 15% NaOH, and above pH 7.0 with HCl (2N ) corrected.
  • the strain Candida rugosa IFO 0731 was cultivated on a shake flask scale as described in Example 1.
  • Example 1 the YNB medium with vitamins and without amino acids (No. 291940, BD Difco) was used in the pre- and main culture. Unlike Example 1, refined rapeseed oil was used as the substrate.
  • the cultivation was carried out under sterile conditions at a temperature of 30 ° C. and a shaking frequency of 140 rpm. The pH was corrected in a range below pH 5.5 with 15% NaOH. After 214 hours, the amount of fumaric acid formed was 22.7 g / l. Other acids were 2.6 g / l ketoglutarate; 5.2 g / 1 malate and 5.8 g / 1 succinate formed. This corresponded to 62% selectivity for fumaric acid. The productivity was 0, 1 g / l * h. In the cultivation period 29 g / 1 biomass were formed.

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  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

La présente invention concerne des procédés de production biotechnologique d'acides organiques par l'intermédiaire d'au moins une souche de levure, au moins une matière première renouvelable étant utilisée comme substrat.
PCT/EP2013/052922 2012-02-14 2013-02-14 Procédés de production biotechnologique d'acides organiques par l'intermédiaire d'au moins une souche de levure WO2013120924A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102012101153A DE102012101153A1 (de) 2012-02-14 2012-02-14 Verfahren zur biotechnologischen herstellung von organischen säuren durch mindestens einen hefestamm
DE102012101153.2 2012-02-14

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WO2013120924A2 true WO2013120924A2 (fr) 2013-08-22
WO2013120924A3 WO2013120924A3 (fr) 2014-02-20

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102015216815A1 (de) 2015-09-02 2017-03-02 Thyssenkrupp Ag Verfahren und Anlage zur Gewinnung einer in einem Fermentationsprozess hergestellten Carbonsäure
CN111218484A (zh) * 2020-01-16 2020-06-02 江南大学 一种通过外源添加无机碳源对己酸发酵强化的方法

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4013508A (en) 1974-11-21 1977-03-22 Liquichimica S.P.A. Process for the production of 1-aspartic acid by fermentation of hydrocarbons

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE123305T1 (de) * 1987-12-22 1995-06-15 Willem Hemmo Kampen Verfahren zur produktion von ethanol, glycerin und bernsteinsäure.
US6066480A (en) * 1998-09-21 2000-05-23 General Electric Company Method for high specific bioproductivity of α,ω-alkanedicarboxylic acids
DE102008002715A1 (de) * 2008-06-27 2009-12-31 Evonik Röhm Gmbh 2-Hydroxyisobuttersäure produzierende rekombinante Zelle
DE102010029973A1 (de) * 2010-06-11 2011-12-15 Evonik Degussa Gmbh Mikrobiologische Herstellung von C4-Körpern aus Saccharose und Kohlendioxid
WO2012103263A2 (fr) * 2011-01-25 2012-08-02 Finley Kenneth R Compositions et méthodes pour la production de malate et de fumarate
DE102011056290A1 (de) * 2011-12-12 2013-06-13 Thyssenkrupp Uhde Gmbh Pilzstämme mit genetischer modifikation betreffend einen carbonsäure-transporter

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4013508A (en) 1974-11-21 1977-03-22 Liquichimica S.P.A. Process for the production of 1-aspartic acid by fermentation of hydrocarbons

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
FURUKAWA T; DE MIRANDA LR; MATSUYOSHI T: "Fermentative Production ofFumaric Acid from n-Paraffins by Candida blankii", J FERMENT TECHNOL, vol. 56, 1978, pages 546 - 549
KACLIKOVA E; LACHOWICZ TM; GBELSKA Y; SUBIK J: "Fumaric acid overproduction in yeast deficient in fumarase", FEMS MICROBIOLOGY LETTERS, vol. 91, 1992, pages 101 - 106, XP023921886, DOI: doi:10.1111/j.1574-6968.1992.tb05192.x
MURRAY, M.-Y.: "Industrial Biotechnology and Commodity Products", vol. 3, 2011, ELSEVIER, article "Comprehensive Biotechnology"

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102015216815A1 (de) 2015-09-02 2017-03-02 Thyssenkrupp Ag Verfahren und Anlage zur Gewinnung einer in einem Fermentationsprozess hergestellten Carbonsäure
WO2017036934A1 (fr) 2015-09-02 2017-03-09 Thyssenkrupp Industrial Solutions Ag Procédé et installation de récupération de l'acide carboxylique généré au cours d'un processus de fermentation
CN111218484A (zh) * 2020-01-16 2020-06-02 江南大学 一种通过外源添加无机碳源对己酸发酵强化的方法
CN111218484B (zh) * 2020-01-16 2021-10-22 江南大学 一种通过外源添加无机碳源对己酸发酵强化的方法

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