WO2005021772A1 - Procede de preparation de l-lysine - Google Patents

Procede de preparation de l-lysine Download PDF

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Publication number
WO2005021772A1
WO2005021772A1 PCT/EP2004/008882 EP2004008882W WO2005021772A1 WO 2005021772 A1 WO2005021772 A1 WO 2005021772A1 EP 2004008882 W EP2004008882 W EP 2004008882W WO 2005021772 A1 WO2005021772 A1 WO 2005021772A1
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Prior art keywords
lysine
concentration
product
fermentation broth
led
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PCT/EP2004/008882
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English (en)
Inventor
Marc Gerigk
Thomas Hermann
Brigitte Bathe
Ralf Kelle
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Degussa Ag
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Priority claimed from DE2003139847 external-priority patent/DE10339847A1/de
Application filed by Degussa Ag filed Critical Degussa Ag
Publication of WO2005021772A1 publication Critical patent/WO2005021772A1/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
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/04—Alpha- or beta- amino acids
    • C12P13/08—Lysine; Diaminopimelic acid; Threonine; Valine

Definitions

  • the invention relates to an improved process for the fermentative preparation of L-lysine using coryneform bacteria.
  • L-Lysine is used in human medicine, in the pharmaceuticals industry, in the foodstuffs industry and very particularly in animal nutrition.
  • L-lysine can be prepared by fermentation of strains of coryneform bacteria, in particular Corynebacterium glutamicum. Because of the great importance of this amino acid, . work is constantly being undertaken to improve the preparation processes . Improvements to the process can relate to fermentation measures, such as e.g. stirring and supply of oxygen, or the composition of the nutrient media, such as e.g. the sugar concentration during the fermentation, or the working up to the product' form, by e.g. ion exchange chromatography, or the intrinsic output properties, i.e. those of genetic origin, of the bacterium itself.
  • fermentation measures such as e.g. stirring and supply of oxygen, or the composition of the nutrient media, such as e.g. the sugar concentration during the fermentation, or the working up to the product' form, by e.g. ion exchange chromatography, or the intrinsic output properties, i.e. those of genetic origin, of the bacterium itself.
  • the inventors had the object of providing new measures for improved fermentative preparation of L-lysine.
  • the invention provides a fermentation process, which comprises a procedure in which a) a coryneform bacterium which produces L-lysine is inoculated and cultured in at least a first nutrient medium, b) at least a further nutrient medium or several further nutrient media is/are then fed continuously to the culture in one or several feed streams, the futher nutrient medium or the further nutrient media comprising at least one source of carbon, at least one source of nitrogen and at least one source of phosphorus, under conditions which allow the formation of L-lysine, and at the same time culture broth is removed from the culture with at least one or several removal streams which substantially corresponds/correspond to the feed stream or the total of the feed streams, wherein c) over the entire period of time of step (b) a concentration of the source (s) of carbon of not more than 10 g/1.
  • the plant output of a fermentation unit which produces L-lysine can be increased by culturing by the batch process (batch) or feed process (fed batch) in the first culturing step (a) described above, at least one additional nutrient medium being employed if the feed process is used.
  • at least one further nutrient medium or several further nutrient media are fed continuously to the culture in one or several feed streams and at the same time culture broth is removed from the culture with at least one 5 or several removal streams, which substantially corresponds/correspond to the feed stream or the total of the feed streams .
  • the bacterium is inoculated 10 in at least a first nutrient medium and cultured by the batch process (batch) or feed process (fed batch) . If the feed process is used, an additional nutrient medium is fed in after more than 0 to not more than 10 hours, preferably after 1 to 10 hours, preferably after 2 to 10 hours and 15 particularly preferably after 3 to 7 hours.
  • the first nutrient medium comprises as the source of carbon one or more of the compounds chosen from the group consisting of sucrose, molasses from sugar beet or cane
  • Organic nitrogen-containing compounds such as peptones, yeast extract, meat extract, malt extract, corn steep 30 liquor, soya bean flour and urea
  • inorganic compounds such as ammonia, ammonium sulfate, ammonium chloride, ammonium phosphate, ammonium carbonate and ammonium nitrate, potassium nitrate and potassium sodium nitrate, can be used as the source of nitrogen in the first nutrient medium.
  • the sources of nitrogen can be used individually, or as a mixture in concentrations of 1 to 50 g/kg, preferably 3 to 40 g/kg, particularly preferably 5 to 30 g/kg.
  • Phosphoric acid alkali metal or alkaline earth metal salts of phosphoric acid, in particular potassium dihydrogen phosphate or dipotassium hydrogen phosphate or the corresponding sodium-containing salts, polymers of phosphoric acid or the hexaphosphoric acid ester of inositol, also called phytic acid, can be used as the source of phosphorus in the first nutrient medium in concentrations of 0.1 to 5 g/kg, preferably 0.3 to 3 g/kg, particularly preferably 0.5 to 2,0 g/kg.
  • the culture medium must furthermore comprise salts of metals, such as e.g. magnesium sulfate or iron sulfate, which are necessary for growth.
  • essential growth substances such as amino acids (e.g. homoserine) and vitamins (e.g. thi,.amine) , ,. can be employed in addition to the , abovementioned substances.
  • Antifoams such as e.g. fatty acid polyglycol esters, can be employed to control the development of foam.
  • the additional nutrient medium which is used in a feed process (fed batch) in general comprises merely as the source of carbon one or more of the compounds chosen from the group consisting of sucrose, molasses from sugar beet or cane sugar, fructose, glucose, starch hydrolysate, lactose, galactose, maltose, xylose, acetic acid, ethanol and methanol, in concentrations of 300 to 700 g/kg, preferably 400 to 650 g/kg, and optionally an inorganic source of nitrogen, such as e.g.
  • the constituents of the further nutrient medium can be fed to the culture in the form of a single further nutrient medium and in a plurality of further nutrient media.
  • the 10 further nutrient medium or the further nutrient media are fed to the culture in at least one (1) feed stream or in a plurality of feed streams of at least 2 to 10, preferably 2 to 7 or 2 to 5 feed streams .
  • The. further nutrient medium or the further nutrient media comprises/comprise as the source of carbon one or more of the compounds chosen from the group consisting of sucrose, molasses from sugar beet or cane sugar, fructose, glucose, starch hydrolysate, maltose, xylose, acetic acid, ethanol
  • the further nutrient medium or the further nutrient media furthermore comprises or comprise a source of nitrogen
  • organic nitrogen-containing compounds such as peptones, yeast extract,- meat extract, malt extract, corn steep liquor, soya bean flour and urea
  • inorganic compounds such as ammonia, ammonium sulfate, ammonium chloride, ammonium phosphate, ammonium carbonate, ammonium
  • the sources of nitrogen can be used individually or as a mixture in concentrations of 5 to 500 g/kg, preferably 25 to 400 g/kg.
  • the further nutrient medium or the further nutrient media furthermore comprises or comprise a source of phosphorus consisting of phosphoric acid, alkali metal or alkaline earth metal salts of phosphoric acid, in particular potassium dihydrogen phosphate or dipotassium hydrogen phosphate or the corresponding sodium-containing salts, polymers of phosphoric acid or the hexaphosphoric acid ester of inositol, also called phytic acid.
  • the sources of phosphorus can be used individually or as a mixture in concentrations of 0.3 to 3 g/kg, preferably 0.5 to 2 g/kg.
  • the culture medium must furthermore comprise salts of metals, such as e.g. magnesium sulfate or iron sulfate, which are necessary for growth, in concentrations of 0.0003 to 15 g/kg, preferably in concentrations of 0.008 to
  • a single further nutrient medium is used, this is typically fed to the culture in one feed stream. If a plurality of further nutrient media are used, these. are fed in a corresponding plurality of feed streams. If a plurality of further nutrient media are used, it should be noted that these in each case can comprise only one of the sources of carbon, nitrogen or phosphorus described, or also a mixture of the sources of carbon, nitrogen or • phosphorus described.
  • the feed stream or the sum of the feed streams in the process according to the invention are added at a rate corresponding to an average residence time of less than 60 hours, less than 55 hours, less than 50 hours, less than 45 hours, less than 40 hours, preferably less than 35 hours, very particularly preferably less than 30 hours.
  • the average residence time here is the theoretical time the particles remain in a continuously operated culture.
  • the average residence time is described by the ratio of the volume of liquid in the reactor and the amount flowing through (Biotechnologie [Biotechnology] ; H. Weide, J. Paca and . A. Knorre; Gustav Fischer Verlag Jena; 1991) .
  • Intensive growth at the start of culturing is usually a logarithmic growth phase.
  • the logarithmic growth phase is in general followed by a phase of less intensive cell growth than in the logarithmic phase.
  • At least one further nutrient medium or several further nutrient media are fed continuously .to the culture in. one . or several feed streams and at the same time culture broth is removed from the culture with at least one or several removal streams, which substantially corresponds/correspond , to the feed stream or the total of the feed streams .
  • the rate of the removal stream or the removal streams corresponds to 80% - 120%, 90% - 110% of the feed stream or of the sum of the feed streams.
  • the removal can be realized industrially by pumping off and/or by draining off the culture broth.
  • the concentration of the source of carbon over the entire period of time of step (b) and/or in the case of a prior culturing in the feed process (fed batch) , also during the feeding of the additional .nutrient medium is adjusted to not more than 10 g/1, not more than 5 g/1 * preferably not more than 3 g/1, particularly preferably not more than 1 g/1.
  • the concentration of the source of carbon is determined here with the aid of methods which are prior art.
  • ⁇ -D-Glucose is determined e.g. in a YSI 02700 Select glucose analyzer from Yellow Springs Instruments (Yellow Springs, Ohio, USA) .
  • the culture broth removed can be provided with oxygen or an oxygen-containing gas until the concentration of the source of carbon falls below 2 g/1; below 1 g/1; or below 0.5 g/1.
  • the yield (Yp / s) is at least. 43 wt.%; at least 45 wt.%; at least 48 wt.%; at least 50 wt.%; at least 52 wt.%.
  • the yield Y P/S is defined here as the ratio of the total amount of L-lysine formed in a culturing to the total amount of the source of carbon employed or• consumed.
  • L-lysine is formed with a space/time yield (STY) of at least 2.5 g/1, preferably at least 3.5 g/1, in particular at least 2.5 to 3.0 g/1 per h, at least 3.0 to more than 4.0 g/1, at least 4.0 to
  • the space/time yield is defined here as the ratio of the total amount of L-lysine formed in a culturing to the actively producing volume of the culture over the total period of time of culturing.
  • the space/time yield is also called the volumetric productivity.
  • the L-lysine concentration (c) based on lysine HCl, in the fermentation broth led off is at least 100 g/1, at least 110 g/1, at least 120 g/1, preferably more than 130 g/1, particularly preferably more than 140 g/1.
  • STY space/time yield
  • Yp / s yield
  • c L-lysine concentration
  • the output index based on the process according to the invention, reaches at least 130 g 2 /(l 2 *h), at least 140 g 2 ,/(l 2 ,*h), at least 150 g 2 ./(l 2 *h), at least ... 160 g a /(l 2 *h), at least 170 g 2 /(l 2 *h), at least 190 g 2 /(l 2 *h), at least 210 g 2 /(l 2 *h), at least 230 g 2 /(l 2 *h), at least 250 g 2 /(l 2 *h).
  • the temperature is adjusted in a range from 28°C to 40°C, preferably 30 to 35°C.
  • the fermentation can be carried out under normal pressure or optionally under increased pressure, preferably under an increased pressure of 0 to 2.5 bar, particularly preferably under 0 to 1.5 bar.
  • the oxygen partial pressure is regulated at 5 to 50%, preferably approx. 20% atmospheric saturation.. Regulation of the pH to a pH of approx. 6 to 8, preferably 6.5 to 7.5, can be effected with ammonia gas or 25% aqueous ammonia.
  • the culturing conditions can remain constant during the culturing or can be changed. Likewise, the culturing conditions in step (a) and (b) can be identical or can differ.
  • the process as claimed in claim 1 comprises a procedure in which the oxygen uptake rate (OUR) established in step b) is not more than 350 mmol/ (l*h) , not more than 325 mmol/(l*hj, not more than 300 mmol/ (l*h) , not more than 275 mmol/ (l*h) , not more than 250 mmol/ (l*h) , not more than 225 mmol/ (l*h) , not more than 200 mmol/ (l*h) , not more than 175 mmol/ (l*h) , not more than 150 mmol/ (l*h) .
  • the oxygen uptake rate (OUR) established in step b) is not more than 350 mmol/ (l*h) , not more than 325 mmol/(l*hj, not more than 300 mmol/ (l*h) , not more than 275 mmol/ (l*h) , not more than 250 mmol/ (l
  • the oxygen uptake rate OUR here describes the specific oxygen absorption rate by the microorganisms in mmol 0 2 per litre of fermentation broth and hour (Biotechnologie [Biotechnol.ogy]; D. Schlee and H.-P. Kleber, Gustav. Fischer Verlag Jena; 1991) .
  • the process according to the invention is operated for at least 100 hours, preferably more than 150 hours, in particular more than 200 hours, preferably more than 250 hours, particularly preferably more than 300 hours.
  • the volume of the culture is exchanged at least 1 time, at least 2 times, at least 4 times, at least 6 times, at least 8 times, at least 10 times, at least 12 times, at least 20 times.
  • the streams described for the first nutrient media and further nutrient media or the sum of the streams of first nutrient media and further nutrient media comprise, complex constituents.
  • Sources of carbon or nitrogen which have a purity of less than 95% in the form employed are described as complex constituents.
  • Such a complex constituent is one or more compounds from the group consisting of peptones, yeast extracts, meat extracts, malt extracts, corn steep liquor and soya bean flour.
  • the content of complex constituents in the nutrient media employed is less than 10 wt.%, less than 5 wt.%, less than 2.5 wt.%, less than 1.0 wt.%, less than 0.5 wt.%.
  • the osmolarity of the L-lysine- containing fermentation broth led off is less than 2,100 mosm/1, preferably less than 1,800 mosm/1, in particular less than 1,500 mosm/1, preferably less than 1,200 mosm/1.
  • the osmolarity is defined as the concentration of osmotically active particles in 1 litre of liquid volume.
  • a 1 molar glucose solution corresponds to 1,000 mosm/1 (Biotechnologie [Biotechnology] ;. H. Wei.de, J. Paca and W. A. Knorre; Gustav
  • coryneform bacteria in particular of the genus Corynebacterium, are suitable for carrying out the process according to the invention.
  • the coryneform bacteria are, in particular, the genus Corynebacterium.
  • the species Corynebacterium glutamicum and furthermore the species Brevibacterium flavum and Corynebacterium thermoaminogenes are to be mentioned in particular. These are known among experts for their ability to produce L-lysine.
  • Suitable strains of the genus Corynebacterium, in particular of the species Corynebacterium glutamicum, are in particular the known wild-type strains
  • the coryneform bacteria contain at least one copy of an lysC gene or allele, which codes for an aspartate kinase, which is insensitive towards the inhibition of lysine or mixtures of lysine and threonine (lysC fbr ) .
  • Such bacteria are typically resistant to the lysine analogue S-(2- aminoethyl) -cysteine . (AEC) .
  • L-Lysine-producing coryneform bacteria which one or more of the features chosen from the group consisting of lysC allele (lysC fbr ) , horn allele (hom leaky ) , zwf allele, coding for an NADPH-insensitive glucose 6-phosphate dehydrogenase, and the pyc allele which codes for pyruvate carboxylase are furthermore suitable.
  • the pyc allele is described in EP 1 108 790.
  • L-Lysine-producing coryneform bacteria which have one or more resistances chosen from the group consisting of azauracil r (Aza r ) , rifamycin r (Rif r ) , streptomycin r (Strep r ) are also suitable.
  • L-Lysine-producing coryneform bacteria which include. at least the following properties are furthermore suitable: two (2) copies of an lysC allele, which codes for a lysine- resistant aspartate kinase (lysC fbr ) , a hom allele, which codes for an attenuated homoserine dehydrogenase (hom leay ) and two (2) copies of a zwf allele, which codes for an NADPH-insensitive glucose 6-phosphate dehydrogenase.
  • L-Lysine-producing coryneform bacteria which contain one or more of the properties chosen from the group consisting of three (3) , four (4) or five (5) copies of an lysC allele (lysC fbr ) , two (2).. copies of an lysE gene, two (2) copiea ⁇ of a zwal gene are furthermore suitable.
  • diaminopimelic acid analogues includes compounds such as 4-fluoro-diaminopimelic acid, 4-hydroxy-diaminopimelic acid, 4-oxo-diaminopimelic acid or 2,4, 6-triaminopimelic acid.
  • Mutagenesis methods are used to produce the coryneform bacteria according to the invention which are sensitive towards 4-hydroxy-diaminopimelic acid.
  • Conventional in vivo mutagenesis methods using mutagenic substances such as, for example, N-methyl-N' -nitro-N-nitrosoguanidine, or ultraviolet light (Miller, J. H. : A Short Course in Bacterial Genetics. A Laboratory Manual and Handbook for Escherichia coli and Related Bacteria, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 1992) can be used for the mutagenesis.
  • the coryneform bacteria which are sensitive towards 4-hydroxy-diaminopimelic acid can be identified by plating out on nutrient medium plates containing 4-hydroxy- diaminopimelic acid. Final concentrations of approx. 5 to 15 g/1, for example approx. 10 g/1 4-hydroxy-diaminopimelic acid in the nutrient medium are particularly suitable for this. At this concentration, mutants which are sensitive towards 4-hydroxy-diaminopimelic acid can be distinguished from the unchanged parent strains by a slowed-down growth. After selection has taken place, the mutants which are sensitive towards 4-hydroxy-diaminopimelic acid show an improved L-lysine production. A . , ⁇ «-
  • the L-lysine-producing coryneform bacteria have the ability to divert the carbon flow through the oxidative pentose phosphate pathway with a percentage content of more than 75%, more than 85%, more than 95%, more than 105%, more than 115%, more, than 125%, more than 135%, more than 145%.
  • the L-lysine-producing coryneform bacteria furthermore have the ability to divert the carbon flow through the tricarboxylic acid cycle, based on the acetyl radicals which are transferred from acetyl-CoA to oxaloacetate by the citrate synthase reaction, with a percentage content of at least 1% but not more than 20%, at least 2% but. not more than 18%, at least 3% but not more than 16%..
  • the tricarboxylic acid cycle also serves for synthesis, of compounds which are essential precursors of the amino acid synthesis pathway.
  • Oxaloacetate serves, for example, as a precursor of lysine. synthesis.
  • the withdrawal of these precursors from the tricarboxylic acid cycle is compensated by replenishing reactions, so-called anaplerotic reactions- Depending on the nature of the source of carbon, growth rate and product formation of the coryneform bacteria, these reactions can proceed forwards or backwards. Forwards in this connection means that the carbon flow takes place from glycolysis in the direction of the tricarboxylic acid cycle (e.g.
  • L-Lysine-producing coryneform bacteria which are particularly suitable according to the invention are those which have the ability to divert the carbon flow through the anaplerotic reactions, based on the sum of pyruvate and phosphoenol pyruvate (PEP) , which are converted into oxaloacetate by PEP carboxylase and pyruvate carboxylase respectively, coded by ppc and pyc respectively, with a percentage content of more than 19%, more than 23%, more than 26%, more than 28%, more than 30%, more than 33%, more than 35%, more than 37%. According to the definition, this corresponds to a net flow through the anaplerotic reactions of more than 19%, more than 23%, more than 26%, more than 28%, more than 30%, more than 33%, more than 35%, more than 37%.
  • PEP phosphoenol pyruvate
  • L-Lysine-producing coryneform bacteria which are particularly suitable according to the invention are those which have the ability to divert the carbon flow through aspartate kinase, coded by lysC, with a percentage content of at least 28% but not more than 60%, at least 30% .but not more than 57%, at least 32% but not more than 53%, at least 33% but not more than 50%. .> . . .. - • .
  • L-Lysine-producing coryneform bacteria which are furthermore suitable according to the invention are those which have the ability to divert the carbon flow through diaminopimelate dehydrogenase, coded by ddh, with a percentage content of at least 49% but not more than 98%, at least 53% but not more than 95%, at least 56% but not more than 91%, at least 58% but not more than 87%.
  • the L-lysine containing fermentation broth from the process according to the invention which is led off has a solids content of at least 10 wt.%, at least 12.5 wt.%, at least 15 wt.%, at least 17.5 wt.%.
  • the L-lysine produced can then be purified from the fermentation broth. Separation methods such as, for example, centrifugation, filtration, decanting, flocculation or a combination thereof are employed for the removal or separating off of the biomass.
  • the L-lysine- containing broth is then purified by known methods, such as, for example, by ion exchange chromatography, ion exclusion chromatography, extraction, crystallization, precipitation or a combination thereof.
  • the L-lysine-containing fermentation broth led off can also be dewatered.
  • the fermentation broth is thickened or concentrated by known methods, such as, for example, with the aid of a rotary evaporator, thin film evaporator, falling film evaporator, by reverse osmosis, by nanofiltration or a combination thereof.
  • the water is removed by this means to the extent of 10% to 90%.
  • the pH can be changed into the acidic (pH 2 to 5) or alkaline (pH 9 to 12) range by addition of acid or alkali.
  • Separation methods such as, for example, centrifugation, filtration, decanting, flocculation or a combination thereof are employed for the removal or separating off of the biomass.
  • the broth obtained is then thickened or concentrated by known methods, such as, for example, with the aid of a rotary evaporator, thin film evaporator, falling film evaporator, by reverse osmosis, by nanofiltration or a combination thereof.
  • the water is removed by this means to the extent of 10% to 90%.
  • Concentrated broth furthermore can then be worked up by methods of freeze drying, spray drying, spray granulation or by other processes, to give a preferably free-flowing, finely divided powder.
  • This free-flowing, finely divided powder can then in turn by converted by suitable compacting or granulating processes into a coarse-grained, readily free-flowing, storable and largely dust-free product.
  • the water is then removed to the extent in total of more than 90% by this means, so that the water content in the product is less than 10%, less than 5%.
  • the process according to the invention is distinguished with respect to the conventional fed batch process above all by an increased space/time yield.
  • An L-lysine-containing product of the following composition is preferably obtained from the fermentation broth by removal of water: Lysine 35 - 80 wt Protein max. 7 wt . Carboxylic aci .ds ma . 7 wt . Total sugars ma . 9 wt . Fats and oils max. 5 wt . Minerals 3 -30 wt.%
  • this L-lysine- containing product after dewatering this L-lysine- containing product has a water content of at least 0.5 wt . %, but. not more than 5.0 wt . % .
  • the fermentation broth led off should furthermore adhere to the following by-product concentrations.
  • the L-lysine-containing fermentation broth for the preparation of an L-lysine-containing product it is preferable for the L-lysine-containing fermentation broth to have a total by-product concentration of not more than 5.0%, preferably not more than 4.0%, 3.0%, 2.5%, 2.0%, particularly preferably not more than 1.5%, 1.0% or 0.5%. It is particularly desirable for the L-lysine-containing fermentation broth to have a total by-product concentration of less than 0.5%.
  • the L-lysine-containing product after dewatering and subsequent granulation the L-lysine-containing product has an average particle size of > 0.1 to 1.0 mm, preferably in an amount of more than 97%, in particular more than 98%.
  • the L-lysine- containing product has a bulk density of at least 600 kg/m 3 , preferably 650 kg/m 3 , in particular 700 kg/m 3 , but preferably greater than 750 kg/m 3 .
  • an additive can be added to the L-lysine-containing product, in order to improve the properties of the product.
  • the content of additive added, in particular oil, here should be 0.02 - 2.0 wt.%, based on the total amount of L-lysine-containing product, on the surface.
  • the L-lysine-containing product comprises as an. additive one or more of. the oils, chosen— from the group consisting of mineral oil, vegetable oils, soya oil, olive oil, soya/lecithin mixtures, edible oils, mixtures of vegetable oils, on the surface.
  • the L-lysine-containing product has a lactate content of ⁇ 3 wt.%, ⁇ 2 wt.%, ⁇ 1 wt.%, ⁇ 0.5 wt.%, ⁇ 0.1 wt . % .
  • L-lysine and other amino acids can be carried out by anion exchange chromatography with subsequent ninhydrin derivatization, as described by Spackman et al . (Analytical Chemistry, 30: 1190-1206 (1958) ) or it can be carried out by reversed phase. HPLC, as described by Lindroth et al . (Analytical Chemistry 51: 1167-1174 (1979) ) .

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Abstract

L'invention se rapporte à un procédé amélioré de préparation fermentative de L-lysine au moyen des bactéries de coryneform qui fabriquent L-lysine.
PCT/EP2004/008882 2003-08-29 2004-08-07 Procede de preparation de l-lysine WO2005021772A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
DE2003139847 DE10339847A1 (de) 2003-08-29 2003-08-29 Verfahren zur Herstellung von L-Lysin
DE10339847.3 2003-08-29
US49971003P 2003-09-04 2003-09-04
US60/499,710 2003-09-04

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WO2007141111A2 (fr) * 2006-06-02 2007-12-13 Evonik Degussa Gmbh Procédé de production d'un additif alimentaire pour animaux contenant de la l-lysine
DE102007019643A1 (de) 2007-04-26 2008-10-30 Evonik Degussa Gmbh Verfahren zur Herstellung von zuckerhaltigen Hydrolysaten aus Lignocellulose
DE102008001874A1 (de) 2008-05-20 2009-11-26 Evonik Degussa Gmbh Verfahren zur Herstellung von L-Aminosäuren
US7785840B2 (en) 2006-07-13 2010-08-31 Evonik Degussa Gmbh Method of production of L-amino acids
WO2010149574A1 (fr) 2009-06-25 2010-12-29 Evonik Degussa Gmbh Procédé pour préparer des acides aminés l par fermentation
WO2011124477A2 (fr) 2010-03-30 2011-10-13 Evonik Degussa Gmbh Procédé de production par fermentation de l-ornithine
DE102010025124A1 (de) 2010-06-25 2011-12-29 Forschungszentrum Jülich GmbH Verfahren zur Herstellung von D-Aminosäuren, Mikroorganismus, sowie Vektor
DE102011006716A1 (de) 2011-04-04 2012-10-04 Evonik Degussa Gmbh Mikroorganismus und Verfahren zur fermentativen Herstellung einer organisch-chemischen Verbindung
DE102011118019A1 (de) 2011-06-28 2013-01-03 Evonik Degussa Gmbh Varianten des Promotors des für die Glyzerinaldehyd-3-phosphat-Dehydrogenase kodierenden gap-Gens
WO2013160124A1 (fr) 2012-04-27 2013-10-31 Evonik Industries Ag Aloha-isopropylmalate synthases résistant à la rétroaction
EP2762571A1 (fr) 2013-01-30 2014-08-06 Evonik Industries AG Microorganisme et procédé de fabrication par fermentation d'acides aminés
US8802400B2 (en) 2005-10-08 2014-08-12 Evonik Degussa Gmbh L-lysine-containing feed additives
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