WO2004029265A2 - Production de 2-kga - Google Patents

Production de 2-kga Download PDF

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WO2004029265A2
WO2004029265A2 PCT/EP2003/010488 EP0310488W WO2004029265A2 WO 2004029265 A2 WO2004029265 A2 WO 2004029265A2 EP 0310488 W EP0310488 W EP 0310488W WO 2004029265 A2 WO2004029265 A2 WO 2004029265A2
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fermentation
sorbose
kga
continuous
yeast
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PCT/EP2003/010488
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WO2004029265A3 (fr
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Tatsuo Hoshino
Teruhide Sugisawa
Yoshinori Takagi
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Dsm Ip Assets B.V.
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Publication of WO2004029265A3 publication Critical patent/WO2004029265A3/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
    • C12P7/58—Aldonic, ketoaldonic or saccharic acids
    • C12P7/60—2-Ketogulonic 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
    • C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously

Definitions

  • the present invention relates to a process for the continuous production of 2-keto- L-gulonic acid or a salt thereof from L-sorbose comprising cultivating a microorganism belonging to the genus Gluconohacter in the presence of second component.
  • 2-Keto-L-gulonic acid (hereinafter referred to as 2-KGA) is an important intermediate for the production of L-ascorbic acid.
  • This compound can be converted to L-ascorbic acid according to the well-known Reichstein method.
  • Examples for attempts to microbiologically produce 2-KGA directly from either L-sorbose or D-sorbitol as starting materials include trials with a mixed cultivation of microorganisms, specifically Pseudomonas striata and Gluconohacter oxydans.
  • the product concentration of this process is 30 g/L from a starting concentration of 70 g/L of L-sorbose, and 37 g/L from a starting concentration of 100 g/L of L-sorbose.
  • EP 0 221 707 Bl discloses the production of 2-KGA from L-sorbose by Pseudogluconobacter saccharoketogenes with and without concomitant bacteria. However, the product concentration of this process is at most 55.3 to 87.6 g/L (conversion ratio: 34.2 to 54.1%).
  • the present invention relates to a process for the production of 2-KGA or a salt thereof from L-sorbose by a continuous or semi-continuous fermentation using a microorganism selected from the group consisting of Gluconohacter oxydans DSM 4025 (FERM BP-3812), a microorganism belonging to the genus Gluconohacter having the identifying characteristics of G. oxydans DSM 4025 (FERM BP-3812) and mutants thereof; using a medium containing yeast or a yeast product, or in the presence of a second microorganism component.
  • a microorganism selected from the group consisting of Gluconohacter oxydans DSM 4025 (FERM BP-3812), a microorganism belonging to the genus Gluconohacter having the identifying characteristics of G. oxydans DSM 4025 (FERM BP-3812) and mutants thereof; using a medium containing yeast or a yeast product, or
  • a microorganism selected from the group consisting of Gluconohacter oxydans DSM 4025 (FERM BP-3812), a microorganism belonging to the genus Gluconohacter having the identifying characteristics of G. oxydans DSM 4025 (FERM BP-3812) and mutants thereof; in the presence of a second microorgan
  • a process for the production of 2-KGA or a salt thereof from L-sorbose by a continuous or semi-continuous fermentation is provided using a microorganism selected from the group consisting of Gluconohacter oxydans DSM 4025 (FERM BP-3812), a microorganism belonging to the genus Gluconohacter having the identifying characteristics of G. oxydans DSM 4025 (FERM BP- 3812) and mutants thereof; whereby instead of a second microorganism component the fermentation medium is supplemented with a yeast or yeast product.
  • a microorganism selected from the group consisting of Gluconohacter oxydans DSM 4025 (FERM BP-3812), a microorganism belonging to the genus Gluconohacter having the identifying characteristics of G. oxydans DSM 4025 (FERM BP- 3812) and mutants thereof; whereby instead of a second microorganism component the fermentation medium is supplemente
  • the present invention provides a process for the production of 2-
  • KGA from L-sorbose by a continuous or semi-continuous fermentation comprising the steps of:
  • steps (a) to (d) mentioned above refers to a continuous fermentation process, which is characterized in that nutrient medium is continuously fed to the fermentation vessel (step (b) as above) with a continuous withdrawal of fermentation broth (step (c) as above), so that the working volume in the fermentor may be kept about constant.
  • the process for the continuous production of 2-KGA as described above is carried out in one fermentation vessel.
  • the second microorganism component as used in the process of the present invention may be, for example, bacteria of the genera Bacillus and Xanthomonas.
  • strains like B. megate ⁇ um DSM 4026 and X maltophilia IFO 12692 are used. Any of these strains maybe incubated in an appropriate medium at 20°C to 40°C for 1 to 4 days and the resulting culture is used as an inoculum for cultivation in the presence of said concomitant bacteria.
  • the yeast or yeast product belongs to the genus Saccharomyces, such as for example S. cerevisiae.
  • yeast products also includes fresh yeast, dried yeast, and yeast extracts.
  • the incubation of step (a) above is carried out by the multistage continuous fermentation.
  • the present invention is directed to a process for the production of 2-KGA as described above using a multi-stage continuous fermentation.
  • a two-stage continuous fermentation is one embodiment of such a multi- stage fermentation mode.
  • the multi-stage continuous fermentation system can be applied to the 2-KGA fermentation process to create higher productivity and concentrations of 2-KGA without L-sorbose left, a situation preferable for the isolation step in preparing sodium 2-KGA.
  • Xanthomonas colonies to Gluconohacter colonies at the beginning of the fermentation process are not critical. This ratio may be in the range between about 1:10 and about 1:300 (Bacillus : Gluconohacter, and Xanthomonas : Gluconohacter). This ratio adjusts itself automatically, in the course of the fermentation process, to an optimal value.
  • microorganisms are cultured in a medium containing L-sorbose as well as appropriate nutrients
  • the microorganisms are conveniently cultured in an aqueous medium under aerobic conditions.
  • any conventional fermentation conditions can be utilized in carrying out the process of this invention.
  • the instant fermentation process maybe carried out at a pH between about 4.0 and about 9.0, preferably at a pH between about 6.0 and about 8.0.
  • a preferred temperature range for carrying out the instant fermentation process is between about 13°C and about 36°C. More preferably, the instant fermentation process maybe carried out at a temperature in the range of between about 18°C and about 33°C.
  • the fermentation time may be between about 100 h and about 1300 h.
  • the concentration of the L-sorbose used as starting material in the instant process may vary between about 20 and about 250 g/L, preferably between about 50 and about 200 g/L.
  • the culture medium used in the instant fermentation process usually contains such nutrients for the microorganisms as assimilable carbon sources, digestible nitrogen sources and inorganic substances, vitamins, trace elements and other growth promoting factors.
  • other substances which are carbon sources may also be added, such as glycerol, glucose, mannitol, fructose, D-arabitol and the like.
  • organic or inorganic substances may also be used as nitrogen sources, such as meat-extract, peptone, casein, corn steep liquor, urea, amino acids, nitrates, ammonium salts and the like.
  • inorganic substances magnesium sulfate, potassium phosphate, ferrous and ferric chlorides, calcium carbonate and the like maybe used.
  • the continuous fermentation system may consist of a medium reservoir, one or more fermentor(s) and harvesting vessel.
  • the nutrient medium containing L-sorbose may be fed to the fermentor during the fermentation process and the supply of nutrient medium to the fermentor may be adjusted to the rate of discharge of fermentation broth from the fermentor, so that the working volume in the fermentor may be kept about constant.
  • the fermentation in the fermentor may be carried out at a dilution rate (D) of liquid flow of about 0.01 to about 0.05 h "1 , more preferably of about 0.02 to about 0.045 h "1 .
  • 2-KGA from L-sorbose at a substantially improved yield, namely a product concentration of more than 112.2 g/L (molar conversion yield: 91.0%) when starting from an L-sorbose concentration of 120 g/L, and at higher yields when starting from higher concentrations.
  • the productivity of 2-KGA is calculated to be 3.9 to 4.8 g/L/h at the steady state of the continuous mode. Conversion yield is calculated based on the amounts of 2-KGA produced and L-sorbose consumed.
  • G. oxydans DSM 4025 in the presence of the second microorganism component, such as a microorganism of the genera Bacillus and Xanthomonas or yeast, can be applied by semi-continuous (repeated fed-batch) fermentation.
  • This semi-continuous mode in which a part of the culture broth is used as the inoculum for the next fermentation, is one of the methods to achieve high 2-KGA productivity.
  • oxydans DSM 4025 and the second microorganism component is achieved by, for example, retaining 10% (v/v) of the whole culture broth, which is obtained in 48 h of cultivation, in the fermentor, using it as the seed for the 2 n step, and filling the fermentor with fresh nutrient medium. According to the same manner as described with respect to the above 1 st step, a part of the 2 n step is used as seed for preparation of the 3 r step.
  • the biocatalysts such as biomass are discarded after each batch resulting in higher production cost and time required for reactor clean-up and start-up (inoculation) resulting in the loss of reactor productivity.
  • the semi-continuous mode can utilize the cultivation broth as the seed for the next cultivation. Therefore, the operation can be applied for the fermentation of 2-KGA without the several seed steps from the small scale seed and the time for the preparation of the culture such as the clean-up and start-up time. Furthermore, the higher reactor productivity can be kept for a long fermentation period.
  • the present invention relates, in part, to a method for the 2-KGA production from
  • L-sorbose by a multi-stage continuous fermentation process using a culture of G. oxydans DSM 4025 in the presence of the second microorganism component.
  • the multi-stage continuous fermentation system consists of two or more fermentation vessels. In order to optimize the yield of the process, the fermentation can be carried out by using more than one fermentation vessel in the entire process. For example, the fermentation of L- sorbose to 2-KGA can be carried out in two or more fermentation vessels that are positioned in consecutive order.
  • the present invention relates, in part, to a process for 2-KGA production from D- sorbitol by a multi-step fermentation system connecting the "D-sorbitol to L-sorbose fermentation process" and the "L-sorbose to 2-KGA fermentation process” in tandem.
  • This process uses the broth containing L-sorbose converted from D-sorbitol by the L- sorbose-producing bacterium, which is not specifically limited.
  • Various Gluconohacter strains such as G. xylinum and G. suhoxydans are known to produce L-sorbose from D- sorbitol efficiently.
  • the harvested broth containing the cells of the strain Gluconohacter maybe aseptically mixed with the sterilized nutrient medium, and L-sorbose concentration may be adjusted to 13%.
  • the L-sorbose broth may be directly used as a substrate without sterilization by heating. Therefore, when the fermentative production system of 2-KGA from D-sorbitol is constructed, L-sorbose fermentation system from D-sorbitol can be directly connected with the continuous fermentation system of 2-KGA from L-sorbose without sterilization of the L-sorbose fermented broth.
  • the 2-KGA fermentation in the two-stage continuous fermentation mode can be carried out in jar fermentors using the mixed cultivation G. oxydans DSM 4025 and X. maltophilia IFO 12692.
  • G. oxydans DSM 4025 and X. maltophilia IFO 12692 In the steady state of the continuous mode, it is possible to produce more than 128.9 g/L of 2-KGA and obtain more than 83.4% of the molar conversion yield .
  • L-sorbose in the mixed culture broth can be consumed completely.
  • Gluconohacter oxydans "Gluconohacter xylinum”, “Gluconohacter suhoxydans”, “Bacillus megaterium”, and "Xanthomonas maltophilia” also include synonyms or basonyms of such species having the same physico-chemical properties, as defined by the International Code of Nomenclature of Prokaryotes.
  • the fermentation is carried out at a dissolved oxygen concentration of from about 0.1 to about 200% air saturation, preferably of from about 5 to about 150% air saturation.
  • the fermentation is preferably carried out at oxygen concentrations in the gas flow of from about 0.1 to about 100% and at gas rates of from about 0.01 to about 2.0 v/v/min. (volume of gas per volume of reactor per minute).
  • the present invention is related to a process for continuous production of 2- KGA or a salt thereof from L-sorbose as described above, wherein the continuous production is carried out at a dissolved oxygen concentration of about 5 to 100% saturation in the fermentation vessel or at oxygen concentrations in the gas flow of about 0.1 to 100%.
  • fermentation according to the present invention may be carried out at normal pressure, i.e., at about 1 bar, it is generally preferred to work under the pressure from about 1 to about 5 bar, more preferably of about 1 to about 3 bar.
  • 2-KGA from L-sorbose without immobilization methods for microorganisms such as chemical bonding, or physical methods for cell retention, e.g., matrix entrapment, and without biomass hold back or retention such as membrane system.
  • the 2-KGA obtained according to the present process can be isolated from the reaction mixture, e.g. by the formation of a salt or by using differences in properties between the product and the surrounding impurities, such as solubility, absorbability and distribution coefficient between the solvents. Adsorption, e.g., on ion exchange resins constitutes a convenient means for isolating the product.
  • the thus obtained product ma be further purified in a conventional manner, e.g., by recrystallization or chromatography.
  • reaction mixture can be used directly for conversion to L-ascorbic acid by esterification, followed by enolization and lactonization.
  • the present invention is illustrated by the following Examples.
  • Example 1 Single-stage 2-KGA continuous fermentation from 12% L-sorbose by G. oxydans DSM 4025 in yeast-supplemented medium
  • the fermentor has a total volume of 3 L with a top drive system and temperature, pH, DO (dissolved oxygen) and exhaust gas monitor. Its working volume was 2 L and the temperature was controlled at 30°C. The agitation speed and aeration rate were set at 800 rpm and 1.0 L/min, respectively. The pH was controlled at 7.0 with sodium hydroxide solution. Furthermore, 10 L of the feeding medium were prepared to supply the fresh substrate continuously. Fermentation was started with an inoculum size of 10% (v/v). The continuous feeding rate was controlled with a peristaltic pump, and the broth containing 2-KGA was continuously withdrawn with another peristaltic pump from the fermentor to the harvesting reservoir while the working volume was kept at 2 L.
  • the production medium (2 L) and the continuous feeding medium (10 L) consisted of 12.0% L-sorbose (sterilized separately), 0.05% glycerol, 0.25% MgSO 4 -7H 2 O, 3.0% corn steep liquor, 7.5% baker's yeast, 0.15% antifoam CA-115.
  • the medium was sterilized at 121°C for 20 minutes.
  • the pH was controlled at 7.0 with NaOH during the fermentation.
  • the inoculum size of seed culture was 10% (v/v).
  • the culture mode was switched over to the continuous mode by fine tuning the continuous feeding rate to 81.0 ml/h, and the dilution rate (D) was set at 0.0405 h "1 .
  • the continuous fermentation was carried out for 120 h and the average 2-KGA product concentration was 112.2 g/L.
  • the molar conversion yield of 2-KGA was 91.3% in average.
  • the productivity of 2-KGA was calculated to be 3.9 to 4.8 g/L/h at the steady state of the continuous mode.
  • Example 2 Single-stage 2-KGA continuous fermentation from 14% L-sorbose by G. oxydans DSM 4025 in yeast-supplemented medium
  • Example 2 200 ml of the seed culture was prepared and inoculated into 2 L of the production medium as in Example 1. After the batch fermentation was done for 30 h, the culture mode was switched over to the continuous mode.
  • the continuous feeding medium (10 L) consisted of 14.0% L-sorbose (sterilized separately), 0.05% glycerol, 0.25% MgSO 4 -7H 2 O, 3.0% corn steep liquor, 7.5% baker's yeast and 0.15% antifoam CA-115.
  • the pH was controlled at 7.0 with NaOH during the fermentation.
  • the continuous fermentation was carried out for 100 h, and in the steady state of continuous mode, 134.3 g/L of 2-KGA was produced.
  • the D was calculated to be 0.0356 h "1 in average.
  • a 93.2% average molar conversion yield of 2-KGA was obtained.
  • the productivity of 2-KGA was calculated to be 4.78 g/L/hour at the steady state of the continuous mode.
  • Example 3 2-KGA fermentation from L-sorbose for repeated fed-batch (semi- continuous) mode by using the mixed culture of G. oxydans DSM 4025 and B. megate ⁇ um DSM 4026
  • B. megaterium DSM 4026 was grown on an agar medium containing 0.5% D- glucose, 0.5% beef extract, 0.5% polypepton (Nippon Seiyaku), 0.30% NaCl (pH 7.0 before sterilization), and 2.0% agar at 27°C for 1 day.
  • megate ⁇ um DSM 4026 and two to three loopfuls of the agar culture of G. oxydans DSM 4025 were simultaneously inoculated into 100 ml of the medium containing 2% L-sorbose, 0.3% beef extract, 0.3% yeast extract, 0.3% corn steep liquor, 1% polypepton (Nippon Seiyaku), 0.1% urea, 0.1% KH 2 PO , 0.02% MgSO 4 -7H 2 O (pH 6.7 before addition of CaCO 3 and sterilization), 0.10% CaCO 3) and 0.03% antifoam CA-115 in a 500 ml Erlenmeyer flask, and incubated at 30°C for 1 day. 10 ml of this culture were transferred into 100 ml of the same medium in a 500 ml
  • 200 ml of the seed culture as mentioned above was inoculated into 2 L of the production medium containing 8% L-sorbose (sterilized separately), 2.5% corn steep liquor, 0.0086% MgSO 4 -7H 2 0, 0.086% KH 2 PO 4 and 0.067% antifoam CA-115.
  • 1.8 L of the culture broth were harvested.
  • 0.2 L of the culture broth were left in the fermentor and used as the seed for the next fermentation.
  • the fermentor was filled with fresh medium containing 10% L-sorbose (sterilized separately), 2.5% corn steep liquor, 0.0086% MgSO 4 -7H 2 O, 0.086% KH 2 PO 4 , and 0.067% antifoam CA-115.
  • Example 4 Single-stage 2-KGA continuous fermentation from L-sorbose by using the mixed culture of G. oxydans DSM 4025 and B. megate ⁇ um DSM 4026
  • the production medium (2 L) and the continuous feeding medium (10 L) consisted of 10% L-sorbose (sterilized separately ), 2.5% corn steep liquor, 0.0086% MgSO 4 -7H 2 O, 0.086% KH 2 PO 4 , and 0.067% antifoam CA-115. These media were sterilized at 121°C for 20 minutes. The pH was controlled at 7.0 with NaOH during the fermentation. Into these production media, 200 ml of the seed culture prepared by the same manner as described in Example 3 was inoculated into 2 L of the production medium.
  • the culture mode was switched to the continuous mode by fine tuning the continuous feeding rate to 61.0 ml/h, and D was varied in the range of 0.0305 to 0.0407 h "1 .
  • the average 2-KGA concentration was 41.2 g/L and a 92.7% average molar conversion yield of 2-KGA was obtained.
  • the productivity of 2- KGA was calculated to be 1.51 g/L/h at the steady state of the continuous mode.
  • Example 5 Two-stage 2-KGA continuous fermentation from L-sorbose by using the mixed culture of G. oxydans DSM 4025 and maltophilia IFO 12692 in the 3 L fermentors
  • Example 2 In the two-stage continuous fermentation system a glass jar fermentor was equipped with the same apparatus as described in Example 2. This fermentation system consisted of two fermentors. Fermentation was usually started with an inoculum size of 10% (v/v). Both fermentors had a working volume of 2 L, the continuous feeding rate was controlled by the same rules as that in the single-stage mode with a peristaltic pump. The broth withdrawn from the 1 st fermentor was further transferred to the 2 n stage fermentor with a second pump. Finally, the broth was withdrawn to the harvest reservoir with a third peristaltic pump. The temperature was controlled at 30°C and the agitation speed and aeration rate were set at 800 rpm and 1.0 L/min, respectively. The pH was controlled at 7.0 with sodium hydroxide solution.
  • X. maltophilia IFO 12692 was grown on an agar medium containing 1% polypepton, 0.2% yeast extract, 0.1% MgSO 4 -7H 2 O (pH 7.0 before sterilization), and 2% agar at 27°C for 2 days.
  • One loopful of the agar culture of X maltophilia IFO 12692 and two to three loopfuls of the agar culture of G. oxydans DSM 4025 were simultaneously inoculated into and B. megaterium DSM 4026 in a 500 ml Erlenmeyer flask, and incubated at 30°C for 1 day. 10 ml of this culture were transferred into 100 ml of the same medium in a 500 ml Erlenmeyer flask and incubated at 30°C for 1 day. This culture was used for inoculation into the medium for jar fermentation.
  • the production medium (2 L) consisted of 12% L-sorbose (sterilized separately),
  • the two-stage continuous fermentation system consisted of the 1 st and 2 n stage fermentors.
  • the fermentation broth obtained in the 1 st stage fermentor was transferred to the 2 n stage fermentor to convert L-sorbose to 2-KGA completely.
  • D was varied in the range of 0.0277 to 0.070 h "1 and 0.0314 to 0.0549 h "1 for the 1 st and 2 nd fermentors, respectively, at appropriate times to investigate the D-value necessary to convert L- sorbose to 2-KGA completely without L-sorbose left in the final harvest tank.
  • the fermentation was stopped after 1,331.5 h.
  • the fermentation activity of the mixture of G was stopped after 1,331.5 h.
  • oxydans DSM 4025 and X maltophilia IFO 12692 by the two-stage continuous mode is summarized in Table 1.
  • the broth containing 113.1 g/L of 2-KGA was actually obtained at an apparent D of 0.0380 h "1 in the two-stage continuous mode, and the overall dilution rate is calculated to be 0.019 h "1 as shown in the second column.
  • the overall 2-KGA molar conversion yield was 90.1%.
  • the productivity of 2-KGA in the 1 st fermentor and the overall productivity of 2-KGA were calculated to be 3.34 and 2.15 g/L/h at the steady state of the continuous mode, respectively.
  • Example 6 Two-stage 2-KGA continuous fermentation from L-sorbose by using the mixed culture of G. oxydans DSM 4025 and maltophilia IFO 12692 in the 30 L fermentors
  • the same continuous culture mode as that applied in the 3 L fermentors was carried out using 30 L fermentors.
  • the continuous fermentation was carried out for more than 180 h and the stable 2-KGA production in the 2 n stage was observed without L-sorbose left, when D was set at 0.041 h "1 .
  • the 2-KGA productivity obtained in 3 L jar fermentors was reproduced in 30 L scale fermentors.
  • Table 2 summarizes the fermentation activities.
  • the productivity of 2-KGA in the 1 st fermentor was calculated to be 4.72 g/L/h at the steady state of the continuous mode.
  • the overall productivity was calculated to be 2.55 g/L/hour and the overall 2-KGA molar conversion yield was 89.6%.
  • Example 7 Two-stage 2-KGA continuous fermentation from L-sorbose broth converted from D-sorbitol by using the mixed culture of G. oxydans DSM 4025 andX maltophilia IFO 12692 in the 3 L jar fermentors
  • the continuous fermentation of 2-KGA from D-sorbitol was carried out, where a multi-step fermentation system connecting the "D-sorbitol to L-sorbose fermentation process" and the "L-sorbose to 2-KGA fermentation process” in tandem was constructed and the fermentation activity of 2-KGA from D-sorbitol was investigated.
  • the broth containing L-sorbose converted from D-sorbitol by the L-sorbose-producing bacterium, for example the strain Gluconohacter was applied as a source of L-sorbose.
  • the harvested broth containing Gluconohacter cells was aseptically mixed with the sterilized medium containing 3% corn steep liquor, 0.4% yeast extract, 0.25% MgSO -7H O, 0.05% glycerol and 0.067% antifoam CA-115, and L-sorbose concentration was adjusted to 13%.
  • the 2- KGA fermentation in the two-stage continuous culture mode was carried out in 3 L jar fermentors using the mixed cultivation G. oxydans DSM 4025 and X maltophilia IFO The fermentation was carried out for 290 h. The relatively stable fermentation activity was kept for the period and the average D was 0.0410 and 0.0404 h "1 in the 1 st and

Abstract

Procédé de production de 2-KGA à partir de L-sorbose selon un mode de fermentation continue ou semi-continue, selon lequel une culture mélangée de Gluconobacter oxydans DSM 4025 et d'un second microorganisme est utilisée ou selon lequel le second microorganisme est remplacé par une levure ou un produit à base de levure ajouté au milieu de fermentation.
PCT/EP2003/010488 2002-09-27 2003-09-22 Production de 2-kga WO2004029265A2 (fr)

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CN102757928B (zh) * 2012-08-09 2014-04-16 山东天力药业有限公司 一种2-酮基-l-古龙酸高耐受型氧化葡糖酸杆菌及其在维生素c发酵生产中的应用
CN103789387B (zh) * 2012-10-30 2016-03-30 山东天力药业有限公司 一种提高维生素c中间体2-酮基-l-古龙酸生产效率的方法
CN103290071B (zh) * 2013-06-09 2015-02-18 山东天力药业有限公司 一种2-酮基-l-古龙酸的制备方法

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EP0278447A2 (fr) * 1987-02-07 1988-08-17 Institute of Microbiology Academia Sinica Procédé de fermentation pour la production de l'acide-2-céto-L-gulonique
EP0366922A1 (fr) * 1988-09-30 1990-05-09 F. Hoffmann-La Roche Ag Procédé de fermentation pour la production de l'acide-2-céto-L-gulonique
EP0518136A2 (fr) * 1991-06-13 1992-12-16 F. Hoffmann-La Roche Ag Procédé de fermentation pour la production de l'acide-2-céto-L-gulonique
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Publication number Priority date Publication date Assignee Title
EP0221707A2 (fr) * 1985-10-22 1987-05-13 Takeda Chemical Industries, Ltd. Procédé de production de l'acide 2-céto-L-gulonique
EP0278447A2 (fr) * 1987-02-07 1988-08-17 Institute of Microbiology Academia Sinica Procédé de fermentation pour la production de l'acide-2-céto-L-gulonique
EP0366922A1 (fr) * 1988-09-30 1990-05-09 F. Hoffmann-La Roche Ag Procédé de fermentation pour la production de l'acide-2-céto-L-gulonique
EP0518136A2 (fr) * 1991-06-13 1992-12-16 F. Hoffmann-La Roche Ag Procédé de fermentation pour la production de l'acide-2-céto-L-gulonique
EP0972843A1 (fr) * 1998-07-17 2000-01-19 F. Hoffmann-La Roche Ag Procédé de fermentation en continu

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WO2004029265A3 (fr) 2004-05-27
AU2003283253A1 (en) 2004-04-19
CN100560728C (zh) 2009-11-18
CN1723287A (zh) 2006-01-18
AU2003283253A8 (en) 2004-04-19

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