CS238800B1 - Method of fermenting of l-lysine - Google Patents
Method of fermenting of l-lysine Download PDFInfo
- Publication number
- CS238800B1 CS238800B1 CS841138A CS113884A CS238800B1 CS 238800 B1 CS238800 B1 CS 238800B1 CS 841138 A CS841138 A CS 841138A CS 113884 A CS113884 A CS 113884A CS 238800 B1 CS238800 B1 CS 238800B1
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- CS
- Czechoslovakia
- Prior art keywords
- lysine
- hydrolyzate
- fermentation
- flour
- nitrogen source
- Prior art date
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- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 title claims description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 24
- 238000000855 fermentation Methods 0.000 claims abstract description 16
- 230000004151 fermentation Effects 0.000 claims abstract description 16
- 235000013312 flour Nutrition 0.000 claims abstract description 12
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 12
- 235000019766 L-Lysine Nutrition 0.000 claims abstract description 10
- 239000004472 Lysine Substances 0.000 claims abstract description 10
- 238000004519 manufacturing process Methods 0.000 claims abstract description 7
- 244000046095 Psophocarpus tetragonolobus Species 0.000 claims abstract description 5
- 235000010580 Psophocarpus tetragonolobus Nutrition 0.000 claims abstract description 5
- 238000002360 preparation method Methods 0.000 claims abstract description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 4
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 4
- 230000002378 acidificating effect Effects 0.000 claims abstract description 3
- 244000005700 microbiome Species 0.000 claims abstract description 3
- 241000319304 [Brevibacterium] flavum Species 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 5
- 229910052500 inorganic mineral Inorganic materials 0.000 claims description 4
- 239000011707 mineral Substances 0.000 claims description 4
- 241000186226 Corynebacterium glutamicum Species 0.000 claims description 2
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 2
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 235000015097 nutrients Nutrition 0.000 claims description 2
- 150000007513 acids Chemical class 0.000 claims 1
- 238000012258 culturing Methods 0.000 claims 1
- 238000000227 grinding Methods 0.000 claims 1
- 239000003102 growth factor Substances 0.000 claims 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 abstract description 4
- 229930006000 Sucrose Natural products 0.000 abstract description 4
- 239000005720 sucrose Substances 0.000 abstract description 4
- 235000017060 Arachis glabrata Nutrition 0.000 abstract description 3
- 244000105624 Arachis hypogaea Species 0.000 abstract description 3
- 235000010777 Arachis hypogaea Nutrition 0.000 abstract description 3
- 235000018262 Arachis monticola Nutrition 0.000 abstract description 3
- 235000001014 amino acid Nutrition 0.000 abstract description 3
- 150000001413 amino acids Chemical class 0.000 abstract description 3
- 235000020776 essential amino acid Nutrition 0.000 abstract description 3
- 239000003797 essential amino acid Substances 0.000 abstract description 3
- 239000000413 hydrolysate Substances 0.000 abstract description 3
- 235000020232 peanut Nutrition 0.000 abstract description 3
- 239000001963 growth medium Substances 0.000 abstract description 2
- 235000003276 Apios tuberosa Nutrition 0.000 abstract 1
- 241000186146 Brevibacterium Species 0.000 abstract 1
- 244000170226 Voandzeia subterranea Species 0.000 abstract 1
- 235000013030 Voandzeia subterranea Nutrition 0.000 abstract 1
- 235000007924 ground bean Nutrition 0.000 abstract 1
- 230000007062 hydrolysis Effects 0.000 abstract 1
- 238000006460 hydrolysis reaction Methods 0.000 abstract 1
- 238000009630 liquid culture Methods 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 6
- 239000000203 mixture Substances 0.000 description 4
- UKAUYVFTDYCKQA-UHFFFAOYSA-N -2-Amino-4-hydroxybutanoic acid Natural products OC(=O)C(N)CCO UKAUYVFTDYCKQA-UHFFFAOYSA-N 0.000 description 3
- UKAUYVFTDYCKQA-VKHMYHEASA-N L-homoserine Chemical compound OC(=O)[C@@H](N)CCO UKAUYVFTDYCKQA-VKHMYHEASA-N 0.000 description 3
- 240000008042 Zea mays Species 0.000 description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 3
- 238000009835 boiling Methods 0.000 description 3
- 235000005822 corn Nutrition 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 235000010755 mineral Nutrition 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonium chloride Substances [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 241000186216 Corynebacterium Species 0.000 description 1
- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 108010073771 Soybean Proteins Proteins 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 235000019797 dipotassium phosphate Nutrition 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000012499 inoculation medium Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 239000003531 protein hydrolysate Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 229940001941 soy protein Drugs 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Vynález se týká způsobu fermentační přípravy L-lysinu v tekutých kultivačních médiích, které obsahují jako zdroj uhlíku sacharosu a jako hlavní zdroj dusíku kyselý hydrolysát profokarpové mouky (jemně mleté boby Psophocarpus tetragonolobus). Tímto hydtolysátem je možno nahradit standardně používaný hydrolysát arašídové mouky jako zdroj dusíku ve fsrmentačních médiích. Jako produkční mikroorganismy jsou používány mutantní kmeny Brevibacterium flavum a Corynebacteriura glutamicum, která se vyznačují rezistencí k analogům aminokyselin a dále vyžadují ke svému růstu některé esenciální aminokyseliny.The invention relates to a fermentation process the preparation of L-lysine in liquid culture media that contain as a carbon source sucrose and as the main nitrogen source is acidic profocarp flour hydrolysate (finely ground bean Psophocarpus tetragonolobus). This by hydrolysis can be replaced by standard used peanut flour hydrolysate as nitrogen source in the fermentation media. They are used as production microorganisms mutant Brevibacterium strains flavum and Corynebacteriura glutamicum, which are characterized by resistance to amino acid analogs and further require growth some essential amino acids.
Description
Vynález se týká způsobu fermentačni přípravy .L-lysinu v tekutých živných médiích, která obsahují jako zdroj uhlíku sacharosu a jako komplexní zdroj dusíku hydrolysét psofokarpové mouky, dále kukuřičný extrakt, minerální soli, vitaminy a další růstové látky. Uvedený postup umožňuje použití nového typu komplexního zdroje dusíku za dosud používaný hydrolysét arašídové mouky. K přípravě uvedeného hydrolysátu se použije jemně mletá mouka z bobů Psophocarpus tetragonolobus a tato se dále hydrolysuje standardním postupem minerální kyselinou. Aminokyselinová složeni hydrolysátu psofokarpové mouky je analogické hydrolysátu sojové mouky; dále vyěěím obsahem některých esenciálních aminokyselin předčí běžně používaný hydrolysét arašídová mouky.The present invention relates to a process for the fermentation of L-lysine in liquid nutrient media containing sucrose as a carbon source and a psofocarp flour hydrolyzate as a carbon source, corn extract, mineral salts, vitamins and other growth agents. This process allows the use of a new type of complex nitrogen source for the peanut flour hydrolyzate hitherto used. To prepare said hydrolysate, finely ground Psophocarpus tetragonolobus bean flour is used and is further hydrolyzed by a mineral acid standard procedure. The amino acid composition of psofocarp flour hydrolyzate is analogous to soy flour hydrolyzate; furthermore, the content of some essential amino acids exceeds the commonly used peanut flour hydrolysate.
Jako produkčních mikroorganismů se používá mutantních kmenů Brevibacterium flavum a Corynebacterium glutamicum, které se vyznačují resistencí na analogy aminokyselin a dále vyžadují k růstu některá esenciální aminokyseliny. V patentových spisech jsou jako zdroje dusíku udávány různá typy hydrolysátů dusíkatých substrátů; např. frano. patent 2 033 119 popisuje jako zdroj dusíku hydrolysét sojového proteinu, stejný zdroj dusíku uvádějí déle patentové spiey NSR 2 100 159 a 2 321 461; jako další zdroje dusíku se uvádějí hydrolysét kaseinu v USA patent, spisu 3 595 751, dále japonský patent, spis 515 75 popisuje jako zdroj dusíku NZ-amin; jako vhodný zdroj dusíku se používají také hydrolysáty kvasničné biomasy a další dusíkaté látky.Mutant strains of Brevibacterium flavum and Corynebacterium glutamicum are used as production microorganisms, which are characterized by resistance to amino acid analogues and further require some essential amino acids to grow. Various types of nitrogen substrate hydrolysates are mentioned as nitrogen sources in the patents; eg frano. U.S. Pat. No. 2,033,119 discloses a soy protein hydrolyzate as the nitrogen source; the same nitrogen source has been reported for a longer time by the German Patent Specifications 2,100,159 and 2,321,461; other nitrogen sources include casein hydrolyzate in U.S. Pat. No. 3,595,751; Japanese Patent 515,775 discloses NZ-amine as the nitrogen source; Yeast biomass hydrolysates and other nitrogenous substances are also used as a suitable nitrogen source.
Fermentačni způsob přípravy L-lysinu podle vynálezu uváděj! následující příklady.The fermentation process for the preparation of L-lysine according to the invention discloses: the following examples.
Přiklad 1Example 1
Kmenem Brevibacterium flavum (homoserin”,AECr) se zaočkuje 500ml varná baňka, která obsahuje 50 ml inokulačniho média o tomto složení: sacharosa 30 g, octan sodný kryst.A 500 ml boiling flask containing 50 ml of an inoculum medium of the following composition is inoculated with Brevibacterium flavum (homoserine ”, AEC r ): sucrose 30 g, sodium acetate crystals.
g, kukuřičný extrakt 30 g, voda dest. ad 1 litr; pH media 7,0. Po zaočkování se baňky kultivují na rotační třepačce (6,7Hz) po dobu 18 až 24 hodin při teplotě 29 °C.g, corn extract 30 g, water dest. ad 1 liter; pH media 7.0. After seeding, the flasks were cultured on a rotary shaker (6.7 Hz) for 18-24 hours at 29 ° C.
Vyrostlou kulturou se v množství 10 % obj. zaočkují 500ml varné baňky, které obsahuji 20 ml fermentačního média o tomto složení: sacharosa 180 g, kyselý hydrolysét psofokarpové mouky (7,0 až 8,1 mg celk.N/ml) 200 ml, kukuřičný extrakt 10 g, hydrogenfosforečnan draselný 1 g, síran hořečnatý kryst. 0,1 g,uhličitan vápenatý mikromletý 30 g, voda dest. ad 1 litr; pH media 7,0; kultivace déle probíhá na rotační třepačce při 29 °C po dobu 96 hodin.A 500 ml boiling flask containing 20 ml of a fermentation medium of the following composition is inoculated at 10% by volume of the grown culture: sucrose 180 g, psofocarp acid acid hydrolyzate (7.0 to 8.1 mg total N / ml) 200 ml, corn extract 10 g, potassium hydrogen phosphate 1 g, magnesium sulfate crystals. 0.1 g, micronized calcium carbonate 30 g, water dest. ad 1 liter; pH media 7.0; cultivation takes place on a rotary shaker at 29 ° C for 96 hours.
V průběhu fermentace se upravuje pH kultury pomoci 10% roztoku amoniaku na hodnotu 7,0 až 7,2. Produkce L-lysinu dosahuje 42,4 g/litr fermentačního média za 96 hodin kultivace.During fermentation, the pH of the culture is adjusted to 7.0-7.2 with 10% ammonia solution. L-lysine production reaches 42.4 g / liter of fermentation medium in 96 hours of culture.
Příklad 2Example 2
Kmenem Corynebacterium glufcamicum(homoserin~,AECr) se zaočkuje inokulační baňka jak se uvádí v příkladu 1 a další postup kultivace a fermentačni přípravy L-lysinu je stejný jak.je popsáno v příkladu 1. Za 96 hodin kultivace je dosahováno produkce 39,8 g L-lysinu/litr fermentačního média.The Corynebacterium glufcamicum (homoserine®, AEC r ) strain is seeded with an inoculation flask as described in Example 1 and the further culture and fermentation preparation of L-lysine is the same as described in Example 1. Production of 39.8 hours is achieved in 96 hours of culture g of L-lysine / liter of fermentation medium.
Příklad 3Example 3
Kmenem Brevibacterium flavum(homoserin~,AECr) se zaočkuje 500ml varná baňka,, která obsahuje 50 ml inokulačniho média o složení jak se uvádí v příkladu 1. Vyrostlou kulturou se zaočkuje dvoulitrový laboratorní fermentačni tank, který obsahuje 800 ml fermentačního média, jehož složení je popsáno v příkladu 1. Jako komplexního zdroje dusíku se použije kyselý hydrolysét psofokarpové mouky z bobů Psophocarpus tetragonolobus, var.Binh-Minh. Další kultivací za odpovídajícího míchání a vzdušněni, úpravy pH a teploty 29 °C je po 96 hodinách fermentace dosahováno produkce 70,9 g L-lysinu/litr kultivačního média.A 500 ml boiling flask containing 50 ml of inoculation medium of the composition as described in Example 1 is inoculated with Brevibacterium flavum (homoserine®, AEC r ). A 2-liter laboratory fermentation tank containing 800 ml of fermentation medium containing is described in Example 1. An acid hydrolyzate of psopocarp flour from Psophocarpus tetragonolobus, var.Binh-Minh is used as a complex nitrogen source. By further cultivation with adequate stirring and air, pH adjustment and temperature of 29 ° C, after 96 hours of fermentation, 70.9 g of L-lysine / liter of culture medium are produced.
Příklad 4Example 4
Postup fermentační přípravy L-lysinu kmenem Brevibacterium flavum je stejný jak je uvedeno v příkladu 3 s tím rozdílem, že jako zdroje dusíku je použito kyselého hydrolysátu psofokarpové mouky z bobů Psophocarpus tetragonolobus, var. Chiabu. Při výše uvedených parametrech fermentace je dosahováno produkce 71,2 g L-lysinu/litr fermentačního média za 96 hodin kultivace.The procedure for the fermentative preparation of L-lysine by Brevibacterium flavum is the same as described in Example 3 except that the acidic hydrolyzate of psofocarp flour from Psophocarpus tetragonolobus, var. Chiabu. With the above fermentation parameters, production of 71.2 g of L-lysine / liter of fermentation medium is achieved in 96 hours of culture.
Claims (2)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS841138A CS238800B1 (en) | 1984-02-17 | 1984-02-17 | Method of fermenting of l-lysine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS841138A CS238800B1 (en) | 1984-02-17 | 1984-02-17 | Method of fermenting of l-lysine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CS113884A1 CS113884A1 (en) | 1985-05-15 |
| CS238800B1 true CS238800B1 (en) | 1985-12-16 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS841138A CS238800B1 (en) | 1984-02-17 | 1984-02-17 | Method of fermenting of l-lysine |
Country Status (1)
| Country | Link |
|---|---|
| CS (1) | CS238800B1 (en) |
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1984
- 1984-02-17 CS CS841138A patent/CS238800B1/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| CS113884A1 (en) | 1985-05-15 |
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