CN108796026A - Production, extraction and the purifying process of lysine - Google Patents
Production, extraction and the purifying process of lysine Download PDFInfo
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- CN108796026A CN108796026A CN201810695127.8A CN201810695127A CN108796026A CN 108796026 A CN108796026 A CN 108796026A CN 201810695127 A CN201810695127 A CN 201810695127A CN 108796026 A CN108796026 A CN 108796026A
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- seed liquor
- lysine
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- 239000004472 Lysine Substances 0.000 title claims abstract description 38
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 32
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 23
- 238000000605 extraction Methods 0.000 title claims abstract description 16
- 230000008569 process Effects 0.000 title claims abstract description 12
- 238000000855 fermentation Methods 0.000 claims abstract description 55
- 230000004151 fermentation Effects 0.000 claims abstract description 55
- 239000007788 liquid Substances 0.000 claims description 30
- 239000002609 medium Substances 0.000 claims description 24
- 241000194108 Bacillus licheniformis Species 0.000 claims description 20
- 239000001963 growth medium Substances 0.000 claims description 20
- 239000000654 additive Substances 0.000 claims description 19
- 230000000996 additive effect Effects 0.000 claims description 19
- 241000894006 Bacteria Species 0.000 claims description 18
- 241000223261 Trichoderma viride Species 0.000 claims description 17
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 16
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 15
- 229930195725 Mannitol Natural products 0.000 claims description 15
- 239000000594 mannitol Substances 0.000 claims description 15
- 235000010355 mannitol Nutrition 0.000 claims description 15
- 239000002253 acid Substances 0.000 claims description 14
- VSGNNIFQASZAOI-UHFFFAOYSA-L calcium acetate Chemical compound [Ca+2].CC([O-])=O.CC([O-])=O VSGNNIFQASZAOI-UHFFFAOYSA-L 0.000 claims description 14
- 235000011092 calcium acetate Nutrition 0.000 claims description 14
- 239000001639 calcium acetate Substances 0.000 claims description 14
- 229960005147 calcium acetate Drugs 0.000 claims description 14
- 238000002156 mixing Methods 0.000 claims description 14
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 12
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 150000001875 compounds Chemical class 0.000 claims description 11
- 239000012530 fluid Substances 0.000 claims description 11
- 238000002360 preparation method Methods 0.000 claims description 11
- 240000008042 Zea mays Species 0.000 claims description 10
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 10
- 102000016943 Muramidase Human genes 0.000 claims description 9
- 108010014251 Muramidase Proteins 0.000 claims description 9
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 claims description 9
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 9
- 235000005822 corn Nutrition 0.000 claims description 9
- 239000004325 lysozyme Substances 0.000 claims description 9
- 229960000274 lysozyme Drugs 0.000 claims description 9
- 235000010335 lysozyme Nutrition 0.000 claims description 9
- 108091005508 Acid proteases Proteins 0.000 claims description 8
- 230000001580 bacterial effect Effects 0.000 claims description 8
- 238000011081 inoculation Methods 0.000 claims description 8
- 239000002054 inoculum Substances 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 8
- 238000011282 treatment Methods 0.000 claims description 8
- 102000004190 Enzymes Human genes 0.000 claims description 7
- 108090000790 Enzymes Proteins 0.000 claims description 7
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 7
- 229940088598 enzyme Drugs 0.000 claims description 7
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 claims description 7
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 7
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 7
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 7
- 238000011218 seed culture Methods 0.000 claims description 7
- 238000000926 separation method Methods 0.000 claims description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 241000209140 Triticum Species 0.000 claims description 6
- 235000021307 Triticum Nutrition 0.000 claims description 6
- 229940085298 biotin 10 mg Drugs 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 239000011691 vitamin B1 Substances 0.000 claims description 6
- 235000015099 wheat brans Nutrition 0.000 claims description 6
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims description 5
- 239000005864 Sulphur Substances 0.000 claims description 5
- 229940041514 candida albicans extract Drugs 0.000 claims description 5
- 239000012138 yeast extract Substances 0.000 claims description 5
- 241000223259 Trichoderma Species 0.000 claims description 4
- 238000010564 aerobic fermentation Methods 0.000 claims description 4
- 235000013339 cereals Nutrition 0.000 claims description 4
- 230000009849 deactivation Effects 0.000 claims description 4
- 238000004042 decolorization Methods 0.000 claims description 4
- 238000001704 evaporation Methods 0.000 claims description 4
- 230000008020 evaporation Effects 0.000 claims description 4
- 238000011534 incubation Methods 0.000 claims description 4
- 239000000047 product Substances 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 238000010792 warming Methods 0.000 claims description 4
- 239000003610 charcoal Substances 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 claims description 2
- 238000012549 training Methods 0.000 claims description 2
- 238000009630 liquid culture Methods 0.000 claims 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 30
- 235000018977 lysine Nutrition 0.000 description 28
- 210000004027 cell Anatomy 0.000 description 9
- 241000186226 Corynebacterium glutamicum Species 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 6
- 235000001014 amino acid Nutrition 0.000 description 6
- 150000001413 amino acids Chemical class 0.000 description 6
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 229910021529 ammonia Inorganic materials 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 239000011347 resin Substances 0.000 description 3
- 229920005989 resin Polymers 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- ZSLZBFCDCINBPY-ZSJPKINUSA-N acetyl-CoA Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCSC(=O)C)O[C@H]1N1C2=NC=NC(N)=C2N=C1 ZSLZBFCDCINBPY-ZSJPKINUSA-N 0.000 description 2
- 235000019270 ammonium chloride Nutrition 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000007599 discharging Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 210000001082 somatic cell Anatomy 0.000 description 2
- WPLOVIFNBMNBPD-ATHMIXSHSA-N subtilin Chemical compound CC1SCC(NC2=O)C(=O)NC(CC(N)=O)C(=O)NC(C(=O)NC(CCCCN)C(=O)NC(C(C)CC)C(=O)NC(=C)C(=O)NC(CCCCN)C(O)=O)CSC(C)C2NC(=O)C(CC(C)C)NC(=O)C1NC(=O)C(CCC(N)=O)NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C1NC(=O)C(=C/C)/NC(=O)C(CCC(N)=O)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)CNC(=O)C(NC(=O)C(NC(=O)C2NC(=O)CNC(=O)C3CCCN3C(=O)C(NC(=O)C3NC(=O)C(CC(C)C)NC(=O)C(=C)NC(=O)C(CCC(O)=O)NC(=O)C(NC(=O)C(CCCCN)NC(=O)C(N)CC=4C5=CC=CC=C5NC=4)CSC3)C(C)SC2)C(C)C)C(C)SC1)CC1=CC=CC=C1 WPLOVIFNBMNBPD-ATHMIXSHSA-N 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- 239000002351 wastewater Substances 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 108010059892 Cellulase Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- 102000018997 Growth Hormone Human genes 0.000 description 1
- 108010051696 Growth Hormone Proteins 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 235000019766 L-Lysine Nutrition 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- NPYPAHLBTDXSSS-UHFFFAOYSA-N Potassium ion Chemical compound [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 208000013738 Sleep Initiation and Maintenance disease Diseases 0.000 description 1
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 1
- 241000319304 [Brevibacterium] flavum Species 0.000 description 1
- HFNUUHLSQPLBQI-UHFFFAOYSA-N acetic acid;calcium Chemical compound [Ca].CC(O)=O HFNUUHLSQPLBQI-UHFFFAOYSA-N 0.000 description 1
- 229940100228 acetyl coenzyme a Drugs 0.000 description 1
- 239000002154 agricultural waste Substances 0.000 description 1
- 239000000908 ammonium hydroxide Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 235000019789 appetite Nutrition 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 235000021245 dietary protein Nutrition 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 229910001448 ferrous ion Inorganic materials 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 206010022437 insomnia Diseases 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000002075 main ingredient Substances 0.000 description 1
- 235000009973 maize Nutrition 0.000 description 1
- 229960001855 mannitol Drugs 0.000 description 1
- 239000012092 media component Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 210000001428 peripheral nervous system Anatomy 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229910001414 potassium ion Inorganic materials 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- -1 saccharide compound Chemical class 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 150000003626 triacylglycerols Chemical class 0.000 description 1
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to fermentation technical fields, disclose the production, extraction and purifying process of lysine comprising following steps：Step 1）Prepare seed liquor；Step 2）Prepare fermentation medium；Step 3）Fermenting and producing lysine；Step 4）Extraction and purifying.Present invention process is of low cost, and lysine production and purity are high.
Description
Technical field
The invention belongs to amino acid fermentation technical fields, and in particular to production, extraction and the purifying process of lysine.
Background technology
Lysine is a kind of amino acid that human body needs, a kind of indispensable nutriment.Protein is to constitute human body
The main ingredient of cell, the protein in food enter after human body by the first breaks down into amino acids of digestion, and then human body utilizes again
These amino acid synthesize new human body protein again, such as immune antiboidy, digestive ferment, plasma protein, growth hormone are all synthesis
Human body protein afterwards.In the various amino acid of synthetic protein, L-lysine is most important one kind, lacks lysine,
Other amino acid are just restricted or are not used, scientist it be referred to as the first essential amino acid of human body.Scientist also found,
It is also most required composition that L-lysine, which is most important in the long element of important substance suppression for control growth in humans, to the maincenter god of people
Through and peripheral nervous system all play an important role.Human body itself cannot synthesize L-lysine, it is necessary to be drawn from food and rely ammonia
Acid is to aid in other nutriments by the key substance that human body fully absorbs and utilizes, and human body only relies ammonia supplemented with enough L-
Acid could improve the absorption and utilization of food protein, reach balanced nutritious, enhancing development.Its effect mainly has：It improves
Intelligence promotes growth, builds up health；It improves a poor appetite, improve malnutritive situation；Improve insomnia, improves memory；Help generates
Antibody, hormone and enzyme improve immunity, increase hemochrome；The absorption of calcium, treatment is helped to prevent osteoporosis；It reduces in blood
The level of triglycerides prevents the generation of cardiovascular and cerebrovascular disease.
The method of production lysine mainly has fermentation method and enzyme process at present.Wherein, fermentation method is widest production method,
The major microorganisms of fermentation method production lysine have corynebacterium glutamicum, brevibacterium flavum etc..Most high acid in industrial production
Rate has been increased to every liter of zymotic fluid 100g or more, extraction rate reached to 80~90% or so.But the cost of culture medium restricts enterprise
Development, cost of material is high, and the profit of enterprise can accordingly reduce.
A kind of achievement in research " CN2018103846072, fermenting lysine culture medium " of applicant early period comprising corn
Skin processed material, corn steep liquor, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, ferrous sulfate heptahydrate, epsom salt, calcium acetate, mannitol,
Biotin, vitamin, water.This method is handled by primary raw material of maize peel, and acid production rate is high, reduce fermentation medium at
This；However, there remains addition other components, including corn steep liquor etc..
In the technique of existing extraction purification lysine, when adjusting the pH of lysine clear liquid, need to consume a large amount of sulphur
Acid consumes a large amount of liquefied ammonia again when being eluted with ammonium hydroxide, will produce a large amount of waste water again when carrying out water washing to resin, increases
The burden of environmental protection is added, and has caused the waste of resource, and can cause resin is broken to be lost in separation process, separation has been set
Standby performance requirement is high.This process route is long, and consumption of raw and auxiliary materials is big, of high cost, constrains the development of lysine industry.
Invention content
On the basis of research " a kind of technique of preparing lysine through fermentation " before applicant, the present invention continue to lysine into
It has gone and has extracted and purify.
To achieve the goals above, the present invention is achieved by the following technical solution：
Production, extraction and the purifying process of lysine comprising following steps：
Step 1）Prepare seed liquor；
Step 2）Prepare fermentation medium；
Step 3）Fermenting and producing lysine；
Step 4）Extraction and purifying.
Further, the step 1）Seed liquor is prepared, is included the following steps：Production lysine bacterial strain is accessed into seed culture
In base, culture to density is 3-5 × 108The seed liquor of cfu/mL；The group of the seed culture medium is divided into：Glucose 80g/L, ferment
Female cream 30g/L, corn steep liquor 10g/L, potassium dihydrogen phosphate 2g/L, dipotassium hydrogen phosphate 2g/L, ferrous sulfate heptahydrate 0.2g/L, seven water sulphur
Sour magnesium 0.2g/L, biotin 10mg/L, vitamin B1 2mg/L。
Further, the step 2）Fermentation medium is prepared, is included the following steps：
（1）Wheat stalk is crushed, then with wheat bran according to 2：3 mass ratio mixing, then it is 50 mesh to be crushed to grain size, is then added
It is added in the water of 4-5 times of weight, with the mixing speed warming while stirring of 200rpm to 60 DEG C, under heat-retaining condition, is ultrasonically treated
20-30min, ultrasonic power 500W, then 121 DEG C of steam treatment 10min, cooled to room temperature is to get compound；
（2）By Trichoderma viride seed liquor and bacillus licheniformis seed liquor according to 1-2:The volume ratio of 2-3 mixes, and obtains mixing and connects
Kind liquid, by combined inoculation liquid according to 6-8%（Account for step 1）The volume ratio of gained compound）Inoculum concentration be linked into and contain step 1）
It is cultivated in the fermentation tank of gained compound, temperature is 30 DEG C, and incubation time 72-96h obtains culture solution；
（3）It is ultrasonically treated culture solution, ultrasonic time 30-60min, ultrasonic power 500W, then it is 55 DEG C to adjust temperature, adds sulphur
Acid adjustment pH is 6, is separately added into lysozyme and acid protease, the additive amount of lysozyme is 20,000 U：1L solution, acid protease
Additive amount be 10,000 U：1L solution under heat-retaining condition, is digested 12-24 hours, is then filtered by filter screen, filter screen
Aperture is 5-10 microns, removes floccule, collects filtered solution, then 100 DEG C of enzyme deactivation 5min, cooled to room temperature, then add
The water of same volume, appropriate calcium acetate and mannitol, stir evenly, 121 DEG C of steam treatment 5min, cooled to room temperature, i.e.,
Obtain fermentation medium.
Further, the step 3）Fermenting and producing lysine, includes the following steps：By seed liquor connecing according to 5-10%
Kind amount is seeded in the fermentation tank containing fermentation medium, carries out aerobic fermentation, and fermentation temperature is 32-35 DEG C, and tank pressure is
0.05MPa, fermentation obtain zymotic fluid in 48-60 hours.
Further, the step 4）Extraction and purifying, include the following steps：
Add hydrochloric acid tune pH to 5.5 into zymotic fluid, be separated by solid-liquid separation using disk plate centrifuge, collects liquid, be pumped into bleacher
Decolorization is carried out, the activated carbon for accounting for liquid quality 0.5-0.8% is added in bleacher, decolourize 30min, is then centrifuged for removal and lives
Property charcoal, collect supernatant, then add hydrochloric acid tune pH to 4.9, it is 40 DEG C of temperature of control, true into being concentrated by evaporation in condensing crystallizing device
Reciprocal of duty cycle is that -0.09Mpa is centrifuged when the one third to be evaporated for being concentrated into original volume is to a quarter, collects wet crystalline substance
Body, drying.
Preferably, the preparation method of the Trichoderma viride seed liquor is：By Trichoderma viride streak inoculation in PDA culture medium
Culture, obtains single bacterium colony；Picking single bacterium colony is inoculated into PDA liquid medium and is cultivated, and obtains Trichoderma viride seed liquor.
Preferably, the preparation method of the bacillus licheniformis seed liquor is：Bacillus licheniformis is inoculated into LB solids
It is cultivated on culture medium, obtains single bacterium colony；Picking single bacterium colony is inoculated into LB liquid medium and is cultivated, and obtains lichens gemma bar
Bacterium seed liquor.
Preferably, the additive amount of the calcium acetate is 100-200mg/L
Preferably, the additive amount of the mannitol is 50-100mg/L.
Specifically used bacterial strain is corynebacterium glutamicum ATCC14997, Trichoderma viride in embodiment of the present invention
ATCC9275, bacillus licheniformis ATCC12759；Other functionally similar bacterial strains in same kind can also be used.The present invention
Bacterial strain seed liquor can also be prepared according to other training methods that the prior art is recorded, and only need to reach suitable inoculum density i.e.
It can.
Compared with prior art, the advantages of culture medium that prepared by the present invention includes mainly the following aspects：
Extraction and purification process of the present invention is simple, does not use resin, and cost is relatively low, and waste water yield is low；
The present invention to wheat stalk and wheat bran by carrying out heating immersion and supersound process, the cavitation of the ultrasonic wave utilized,
It generates partial high pressure high temperature to be impacted, contributes to the separation of fiber, albumen and carbohydrate, it is easier to by bacterial strain profit
With；
Trichoderma viride can generate cellulase, and bacillus licheniformis can generate protease and amylase, and the present invention is using green
Color trichoderma and bacillus licheniformis are combined carries out fermentation enzymolysis processing to wheat stalk and wheat bran processed material, then digests thalline,
The culture medium containing saccharide compound, mycoprotein, polypeptide and inorganic mineral is obtained, uses, improves for Escherichia coli
Fermentation efficiency, and agricultural wastes are utilized, reduce cost.The cavitation for the ultrasonic wave that the present invention utilizes, generation office
Portion's high pressure-temperature, impacts somatic cells, and cytomorphosis and rupture, auxiliary lysozyme is caused to carry out broken wall dissolving, and
It is assisted using acid protease, improves the enzymolysis efficiency of mycoprotein.
Portion takes part in the synthesis of acetyl coenzyme A to calcium acetate in the cell, can participate in tricarboxylic acid cycle, to increase
The metabolism degree of cell, suitable calcium acetate can promote bar bacterium somatic cells to grow, to promoting the fermentation of lysine；It is sweet
Dew alcohol is able to maintain that osmotic pressure, while preventing the outflow of cell moisture and the invasion of salinity, improves cell to water shortage, high temperature is high
The tolerance of salt and hypertonic environment, stablizes the function of enzymatic activity and large biological molecule, and thalline can absorb the sweet dew in culture medium
Alcohol resists the pressure of Thief zone, and in lysine fermentation process, the addition of appropriate mannitol can play quickening growth
The effect of rate, sugar consumption rate and rate of producing acid.
Description of the drawings
Fig. 1：Influence of the calcium acetate additive amount to fermentation production lysine amount；
Fig. 2：Influence of the mannitol additive amount to fermentation production lysine amount.
Specific implementation mode
Those skilled in the art can use for reference present disclosure, be suitably modified technological parameter realization.In particular, it should be pointed out that
All similar substitutions and modifications are apparent to those skilled in the art, they are considered as being included in this hair
It is bright.The product and method of the present invention is described by preferred embodiment, and related personnel can obviously not depart from this hair
Product as described herein and method are modified or are suitably changed and combined in bright content, spirit and scope, to realize and answer
Use the technology of the present invention.For a further understanding of the present invention, the following describes the present invention in detail with reference to examples.
Embodiment 1
Production, extraction and the purifying process of lysine comprising following steps：
Corynebacterium glutamicum ATCC14997 is chosen as fermentation strain；Corynebacterium glutamicum is accessed in seed culture medium,
Culture to density is 3 × 108The seed liquor of cfu/mL；The group of the seed culture medium is divided into：Glucose 80g/L, yeast extract
30g/L, corn steep liquor 10g/L, potassium dihydrogen phosphate 2g/L, dipotassium hydrogen phosphate 2g/L, ferrous sulfate heptahydrate 0.2g/L, epsom salt
0.2g/L, biotin 10mg/L, vitamin B12mg/L；
Seed liquor is seeded to according to 10% inoculum concentration in the fermentation tank containing fermentation medium, aerobic fermentation, fermentation are carried out
Temperature is 32 DEG C, and tank pressure is 0.05MPa, and fermentation obtains zymotic fluid for 48 hours, persistently adds ammonium chloride as fermentation in fermentation process
Liquid neutralizer makes zymotic fluid discharging pH controls 7.2；
Add hydrochloric acid tune pH to 5.5 into zymotic fluid, be separated by solid-liquid separation using disk plate centrifuge, collects liquid, be pumped into bleacher
Decolorization is carried out, the activated carbon for accounting for liquid quality 0.5% is added in bleacher, decolourize 30min, is then centrifuged for removal activated carbon,
Supernatant is collected, then adds hydrochloric acid tune pH to 4.9, into being concentrated by evaporation in condensing crystallizing device, controls 40 DEG C of temperature, vacuum degree
For -0.09Mpa, when the one third to be evaporated for being concentrated into original volume, it is centrifuged, collects wet crystal, dried at 100 DEG C
It does to obtain the final product；After testing：Purity is 98.7%, yield 73.5%.
The preparation method of the fermentation tank culture medium includes the following steps：
Step 1）Wheat stalk is crushed, then with wheat bran according to 2：3 mass ratio mixing, then it is 50 mesh to be crushed to grain size, so
It is added to afterwards in the water of 4 times of weight, with the mixing speed warming while stirring of 200rpm to 60 DEG C, under heat-retaining condition, at ultrasound
30min, ultrasonic power 500W are managed, then 121 DEG C of steam treatment 10min, cooled to room temperature is to get compound；
Step 2）By Trichoderma viride seed liquor and bacillus licheniformis seed liquor according to 2:3 volume ratio mixing, obtains mixing and connects
Kind liquid, by combined inoculation liquid according to 8%（Account for step 1）The volume ratio of gained compound）Inoculum concentration be linked into and contain step 1）Institute
It obtains and is cultivated in the fermentation tank of compound, temperature is 30 DEG C, and incubation time 72h obtains culture solution；
Step 3）It is ultrasonically treated culture solution, ultrasonic time 60min, ultrasonic power 500W, then it is 55 DEG C to adjust temperature, adds sulphur
Acid adjustment pH is 6, is separately added into lysozyme and acid protease, the additive amount of lysozyme is 20,000 U：1L solution, acid protease
Additive amount be 10,000 U：1L solution under heat-retaining condition, is digested 12 hours, is then filtered by filter screen, filter screen aperture
It is 5 microns, removes floccule, collects filtered solution, then 100 DEG C of enzyme deactivation 5min, cooled to room temperature, then add same volume
Water, the calcium acetate of final concentration of 200mg/L and the mannitol of 100mg/L, stir evenly, 121 DEG C of steam treatment 5min, it is natural
It is cooled to room temperature to get fermentation medium.
The preparation method of the Trichoderma viride seed liquor is：Trichoderma viride streak inoculation is cultivated in PDA culture medium, is obtained
To single bacterium colony；Picking single bacterium colony is inoculated into PDA liquid medium and is cultivated, and it is 1 × 10 to obtain cell concentration8Cfu/ml's is green
Color trichoderma seed liquor；
The preparation method of the bacillus licheniformis seed liquor is：Bacillus licheniformis is inoculated on LB solid mediums and is trained
It supports, obtains single bacterium colony；Picking single bacterium colony is inoculated into LB liquid medium and is cultivated, and it is 1 × 10 to obtain cell concentration8cfu/
The bacillus licheniformis seed liquor of ml.
Embodiment 2
Production, extraction and the purifying process of lysine comprising following steps：
Corynebacterium glutamicum ATCC14997 is chosen as fermentation strain；Corynebacterium glutamicum is accessed in seed culture medium,
Culture to density is 5 × 108The seed liquor of cfu/mL；The group of the seed culture medium is divided into：Glucose 80g/L, yeast extract
30g/L, corn steep liquor 10g/L, potassium dihydrogen phosphate 2g/L, dipotassium hydrogen phosphate 2g/L, ferrous sulfate heptahydrate 0.2g/L, epsom salt
0.2g/L, biotin 10mg/L, vitamin B12mg/L；
Seed liquor is seeded to according to 10% inoculum concentration in the fermentation tank containing fermentation medium, aerobic fermentation, fermentation are carried out
Temperature is 35 DEG C, and tank pressure is 0.05MPa, and fermentation obtains zymotic fluid for 60 hours, persistently adds ammonium chloride as fermentation in fermentation process
Liquid neutralizer makes zymotic fluid discharging pH controls 7.2；
Add hydrochloric acid tune pH to 5.5 into zymotic fluid, be separated by solid-liquid separation using disk plate centrifuge, collects liquid, be pumped into bleacher
Decolorization is carried out, the activated carbon for accounting for liquid quality 0.8% is added in bleacher, decolourize 30min, is then centrifuged for removal activated carbon,
Supernatant is collected, then adds hydrochloric acid tune pH to 4.9, into being concentrated by evaporation in condensing crystallizing device, controls 40 DEG C of temperature, vacuum degree
For -0.09Mpa, when a quarter to be evaporated for being concentrated into original volume, it is centrifuged, collects wet crystal, dried at 100 DEG C
It does to obtain the final product；After testing：Purity is 98.1%, yield 70.8%.
The preparation method of the fermentation tank culture medium includes the following steps：
Step 1）Wheat stalk is crushed, then with wheat bran according to 2：3 mass ratio mixing, then it is 50 mesh to be crushed to grain size, so
It is added to afterwards in the water of 5 times of weight, with the mixing speed warming while stirring of 200rpm to 60 DEG C, under heat-retaining condition, at ultrasound
20min, ultrasonic power 500W are managed, then 121 DEG C of steam treatment 10min, cooled to room temperature is to get compound；
Step 2）By Trichoderma viride seed liquor and bacillus licheniformis seed liquor according to 1:2 volume ratio mixing, obtains mixing and connects
Kind liquid, by combined inoculation liquid according to 6%（Account for step 1）The volume ratio of gained compound）Inoculum concentration be linked into and contain step 1）Institute
It obtains and is cultivated in the fermentation tank of compound, temperature is 30 DEG C, and incubation time 96h obtains culture solution；
Step 3）It is ultrasonically treated culture solution, ultrasonic time 30-60min, ultrasonic power 500W, then it is 55 DEG C to adjust temperature,
It is 6 to add sulfuric acid adjustment pH, is separately added into lysozyme and acid protease, the additive amount of lysozyme is 20,000 U：1L solution, acid egg
The additive amount of white enzyme is 10,000 U：1L solution under heat-retaining condition, is digested 24 hours, is then filtered by filter screen, filter screen
Aperture is 10 microns, removes floccule, collects filtered solution, then 100 DEG C of enzyme deactivation 5min, cooled to room temperature, then add phase
The mannitol of the water of same volume, the calcium acetate of final concentration of 100mg/L and 50mg/L, stirs evenly, 121 DEG C of steam treatments
5min, cooled to room temperature is to get fermentation medium.
The preparation method of the Trichoderma viride seed liquor is：Trichoderma viride streak inoculation is cultivated in PDA culture medium, is obtained
To single bacterium colony；Picking single bacterium colony is inoculated into PDA liquid medium and is cultivated, and it is 1 × 10 to obtain cell concentration8Cfu/ml's is green
Color trichoderma seed liquor；
The preparation method of the bacillus licheniformis seed liquor is：Bacillus licheniformis is inoculated on LB solid mediums and is trained
It supports, obtains single bacterium colony；Picking single bacterium colony is inoculated into LB liquid medium and is cultivated, and it is 1 × 10 to obtain cell concentration8cfu/
The bacillus licheniformis seed liquor of ml.
Embodiment 3
Each Contents of Main Components in fermentation medium of the present invention：
Control group is set to examine influence of the bacterial strain to culture medium Contents of Main Components, control group 1：Only with Trichoderma viride；It is right
According to group 2：Only with bacillus licheniformis.Concrete outcome is shown in Table 1：
Table 1
| Component target | Saccharic composition g/L | High molecular weight protein（50000 Da or more） | Small molecular protein（50000 Da or less） | Calcium ion mg/L | Potassium ion mg/L | Ferrous ion mg/L |
| Embodiment 1 | 72.6 | 19.1 | 25.4 | 126.7 | 309.5 | 60.8 |
| Control group 1 | 61.5 | 14.5 | 14.1 | 85.8 | 221.6 | 41.8 |
| Control group 2 | 37.9 | 20.5 | 11.7 | 66.4 | 148.8 | 32.5 |
As shown in table 1, compared with the control group 1-2 using single bacterial strain, combined using Trichoderma viride and bacillus licheniformis
It is more rich comprehensively to handle the nutrient media components obtained, meets the use standard of Corynebacterium glutamicum fermentation and acid.
Embodiment 4
Fermentation medium product acid activity is tested：
Control group 1：Glucose 80g/L, yeast extract 30g/L, corn steep liquor 10g/L, potassium dihydrogen phosphate 2g/L, dipotassium hydrogen phosphate 2g/
L, ferrous sulfate heptahydrate 0.2g/L, epsom salt 0.2g/L, biotin 10mg/L, vitamin B12mg/L；
Control group 2：Glucose 80g/L, yeast extract 30g/L, corn steep liquor 10g/L, potassium dihydrogen phosphate 2g/L, dipotassium hydrogen phosphate 2g/
L, ferrous sulfate heptahydrate 0.2g/L, epsom salt 0.2g/L, calcium acetate 200mg/L, mannitol 50mg/L, biotin 10mg/
L, vitamin B12mg/L；
For specific zymotechnique with reference to embodiment 1, specific fermentation and acid amount is shown in Table 2：
Table 2
| Group | Embodiment 1 | Comparative example 1 | Comparative example 2 |
| Lysine production mg/100ml | 14.1 | 11.9 | 12.7 |
| Biomass g/L（Dry weight） | 14.7 | 13.1 | 13.5 |
As shown in table 2, control group 1 is common fermentation medium, and control group 2 is added to calcium acetate on the basis of common culture medium
And mannitol, compared with control group 1-2, the biomass and production acid of embodiment 1 measure equal highest, illustrate 1 fermentation medium of embodiment
Culture effect it is more preferable.
Embodiment 5
The influence of calcium acetate and mannitol additive amount to fermentation production lysine amount：
It is experimental example with embodiment 1, under the premise of maintaining other components constant, the concentration gradient that sets calcium acetate is respectively 0,
50,100,200,400,800mg/L, as shown in Figure 1, with the increase of acetic acid calcium concentration, lysine production is continuously improved, when adding
After dosage increases to 100mg/L, yield amplification is not obvious, and after 200mg/L, yield decreases, therefore, present invention selection
The additive amount of 100-200mg/L.
It is experimental example with embodiment 1, under the premise of maintaining other components constant, the concentration gradient for setting mannitol is respectively
0,25,50,100,200,400mg/L, as shown in Fig. 2, with the increase of mannitol concentration, lysine production also with raising,
When additive amount increases to 100mg/L, lysine production reaches maximum value, with the increase of mannitol additive amount, lysine production
Amount decreases, and therefore, the present invention selects the additive amount of selection 50-100mg/L.
The explanation of above example is only intended to facilitate the understanding of the method and its core concept of the invention.It should be pointed out that for this
For the those of ordinary skill of technical field, without departing from the principle of the present invention, the present invention can also be carried out several
Improvement and modification, these improvement and modification are also fallen within the protection scope of the claims of the present invention.
Claims (10)
1. the production of lysine, extraction and purifying process comprising following steps：
Step 1）Prepare seed liquor；
Step 2）Prepare fermentation medium；
Step 3）Fermenting and producing lysine；
Step 4）Extraction and purifying.
2. technique according to claim 1, which is characterized in that the step 1）Seed liquor is prepared, is included the following steps：It will
It produces in lysine bacterial strain access seed culture medium, culture to density is（3-5）×108The seed liquor of cfu/mL；The seed training
The group for supporting base is divided into：Glucose 80g/L, yeast extract 30g/L, corn steep liquor 10g/L, potassium dihydrogen phosphate 2g/L, dipotassium hydrogen phosphate 2g/
L, ferrous sulfate heptahydrate 0.2g/L, epsom salt 0.2g/L, biotin 10mg/L, vitamin B1 2mg/L。
3. technique according to claim 2, which is characterized in that the step 2）Fermentation medium is prepared, including is walked as follows
Suddenly：
（1）Wheat stalk is crushed, then with wheat bran according to 2：3 mass ratio mixing, then it is 50 mesh to be crushed to grain size, is then added
It is added in the water of 4-5 times of weight, with the mixing speed warming while stirring of 200rpm to 60 DEG C, under heat-retaining condition, is ultrasonically treated
20-30min, ultrasonic power 500W, then 121 DEG C of steam treatment 10min, cooled to room temperature is to get compound；
（2）By Trichoderma viride seed liquor and bacillus licheniformis seed liquor according to 1-2:The volume ratio of 2-3 mixes, and obtains mixing and connects
Kind liquid, step is linked by combined inoculation liquid according to the inoculum concentration of 6-8%（1）It is cultivated in gained compound, temperature 30
DEG C, incubation time 72-96h obtains culture solution；
（3）It is ultrasonically treated culture solution, ultrasonic time 30-60min, ultrasonic power 500W, then it is 55 DEG C to adjust temperature, adds sulphur
Acid adjustment pH is 6, is separately added into lysozyme and acid protease, the additive amount of lysozyme is 20,000 U：1L solution, acid protease
Additive amount be 10,000 U：1L solution under heat-retaining condition, is digested 12-24 hour, is then filtered, collection filtered solution, 100 DEG C of enzyme deactivations
5min, cooled to room temperature, then the water, appropriate calcium acetate and mannitol of same volume are added, it stirs evenly, 121 DEG C of steam
5min is handled, cooled to room temperature is to get fermentation medium.
4. technique according to claim 2 or 3, which is characterized in that the step 3）Fermenting and producing lysine, including it is as follows
Step：Seed liquor is seeded to according to the inoculum concentration of 5-10% in the fermentation tank containing fermentation medium, aerobic fermentation, hair are carried out
Ferment temperature is 32-35 DEG C, and tank pressure is 0.05MPa, and fermentation obtains zymotic fluid in 48-60 hours.
5. technique according to claim 4, which is characterized in that the step 4）Extraction and purifying, include the following steps：
Add hydrochloric acid tune pH to 5.5 into zymotic fluid, be separated by solid-liquid separation using disk plate centrifuge, collects liquid, be pumped into bleacher
Decolorization is carried out, the activated carbon for accounting for liquid quality 0.5-0.8% is added in bleacher, decolourize 30min, is then centrifuged for removal and lives
Property charcoal, collect supernatant, then add hydrochloric acid tune pH to 4.9, it is 40 DEG C of temperature of control, true into being concentrated by evaporation in condensing crystallizing device
Reciprocal of duty cycle is that -0.09Mpa is centrifuged when the one third to be evaporated for being concentrated into original volume is to a quarter, collects wet crystalline substance
Body, drying.
6. technique according to claim 3, which is characterized in that the preparation method of the Trichoderma viride seed liquor is：It will be green
Color trichoderma streak inoculation is cultivated in PDA culture medium, obtains single bacterium colony；Picking single bacterium colony is inoculated into PDA liquid medium progress
Culture, obtains Trichoderma viride seed liquor.
7. technique according to claim 3, which is characterized in that the preparation method of the bacillus licheniformis seed liquor is：
Bacillus licheniformis is inoculated on LB solid mediums and is cultivated, single bacterium colony is obtained；Picking single bacterium colony is inoculated into LB Liquid Cultures
It is cultivated on base, obtains bacillus licheniformis seed liquor.
8. technique according to claim 3, which is characterized in that the additive amount of the calcium acetate is 100-200mg/L.
9. technique according to claim 3, which is characterized in that the additive amount of the mannitol is 50-100mg/L.
10. being allowed to the lysine product of the preparation of the technique described in one according to claim 1-9.
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|---|---|---|---|---|
| CN110055296A (en) * | 2019-05-13 | 2019-07-26 | 南京牧奇亚生物科技有限公司 | The preparation method and applications of lysine |
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| CN110499261A (en) * | 2018-05-17 | 2019-11-26 | 卢松 | The preparation process of protein enzymatic hydrolyzate and its application in fermented and cultured |
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| CN112662713A (en) * | 2020-12-28 | 2021-04-16 | 天津科技大学 | Culture medium for producing L-lysine by high-density fermentation and method thereof |
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Citations (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ATA238075A (en) * | 1974-04-01 | 1977-05-15 | Mueller Hans | PROCESS FOR THE PRODUCTION OF PROTEIN FROM PROTEIN-CONTAINING STARCH AND CELLULOSE PRODUCTS |
| JPS55108291A (en) * | 1979-02-13 | 1980-08-20 | Mitsubishi Petrochem Co Ltd | Production of l-lysine through fermentation process |
| CN1139520A (en) * | 1996-03-05 | 1997-01-08 | 高银相 | Production method for biological multi-bacterial fermented fodder made from stalks |
| WO2007087815A2 (en) * | 2004-12-17 | 2007-08-09 | Metanomics Gmbh | Process for the control of production of fine chemicals |
| US20090226571A1 (en) * | 2005-09-07 | 2009-09-10 | Basf Se | Fermentative Production of Non-Volatile Microbial Metabolism Products in Solid Form |
| WO2011057083A1 (en) * | 2009-11-06 | 2011-05-12 | Novozymes, Inc. | Polypeptides having xylanase activity and polynucleotides encoding same |
| CN103080306A (en) * | 2010-08-20 | 2013-05-01 | 科德克希思公司 | Use of glycoside hydrolase 61 family proteins in processing of cellulose |
| WO2013067305A1 (en) * | 2011-11-02 | 2013-05-10 | Texas Tech University System | Media compositions for promoting bacterial and fungal growth |
| CN104212870A (en) * | 2014-09-28 | 2014-12-17 | 呼伦贝尔东北阜丰生物科技有限公司 | Process for fermentation production of lysine hydrochloride |
| CN105886556A (en) * | 2016-06-14 | 2016-08-24 | 徐州生物工程职业技术学院 | Method for producing citric acid through mixed-strain fermentation crop straw |
| CN106544373A (en) * | 2016-11-21 | 2017-03-29 | 长沙道勤生物科技有限公司 | A kind of method for producing lysine and feedstuff |
| CN106755156A (en) * | 2016-12-28 | 2017-05-31 | 安徽丰原发酵技术工程研究有限公司 | A kind of fermentation process of L lysines |
| CN106800435A (en) * | 2016-12-27 | 2017-06-06 | 深圳市芭田生态工程股份有限公司 | A kind of synergistic urea and preparation method thereof |
| CN107164450A (en) * | 2017-06-30 | 2017-09-15 | 齐齐哈尔龙江阜丰生物科技有限公司 | A kind of recoverying and utilizing method of fermentation waste liquid of lysine |
| CN107267427A (en) * | 2017-08-17 | 2017-10-20 | 齐齐哈尔龙江阜丰生物科技有限公司 | A kind of threonine mother liquor recycling method |
| JP2018085962A (en) * | 2016-11-29 | 2018-06-07 | 国立大学法人 岡山大学 | METHOD OF PRODUCING L-LYSINE α-OXIDASE |
| CN108796004A (en) * | 2018-06-20 | 2018-11-13 | 齐齐哈尔龙江阜丰生物科技有限公司 | A kind of technique of preparing lysine through fermentation |
| CN112048526A (en) * | 2020-09-21 | 2020-12-08 | 山东柠檬生化有限公司 | High-conversion-rate production method of fermented citric acid |
-
2018
- 2018-06-29 CN CN201810695127.8A patent/CN108796026A/en active Pending
Patent Citations (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ATA238075A (en) * | 1974-04-01 | 1977-05-15 | Mueller Hans | PROCESS FOR THE PRODUCTION OF PROTEIN FROM PROTEIN-CONTAINING STARCH AND CELLULOSE PRODUCTS |
| JPS55108291A (en) * | 1979-02-13 | 1980-08-20 | Mitsubishi Petrochem Co Ltd | Production of l-lysine through fermentation process |
| CN1139520A (en) * | 1996-03-05 | 1997-01-08 | 高银相 | Production method for biological multi-bacterial fermented fodder made from stalks |
| WO2007087815A2 (en) * | 2004-12-17 | 2007-08-09 | Metanomics Gmbh | Process for the control of production of fine chemicals |
| US20090226571A1 (en) * | 2005-09-07 | 2009-09-10 | Basf Se | Fermentative Production of Non-Volatile Microbial Metabolism Products in Solid Form |
| WO2011057083A1 (en) * | 2009-11-06 | 2011-05-12 | Novozymes, Inc. | Polypeptides having xylanase activity and polynucleotides encoding same |
| CN103080306A (en) * | 2010-08-20 | 2013-05-01 | 科德克希思公司 | Use of glycoside hydrolase 61 family proteins in processing of cellulose |
| WO2013067305A1 (en) * | 2011-11-02 | 2013-05-10 | Texas Tech University System | Media compositions for promoting bacterial and fungal growth |
| CN104212870A (en) * | 2014-09-28 | 2014-12-17 | 呼伦贝尔东北阜丰生物科技有限公司 | Process for fermentation production of lysine hydrochloride |
| CN105886556A (en) * | 2016-06-14 | 2016-08-24 | 徐州生物工程职业技术学院 | Method for producing citric acid through mixed-strain fermentation crop straw |
| CN106544373A (en) * | 2016-11-21 | 2017-03-29 | 长沙道勤生物科技有限公司 | A kind of method for producing lysine and feedstuff |
| JP2018085962A (en) * | 2016-11-29 | 2018-06-07 | 国立大学法人 岡山大学 | METHOD OF PRODUCING L-LYSINE α-OXIDASE |
| CN106800435A (en) * | 2016-12-27 | 2017-06-06 | 深圳市芭田生态工程股份有限公司 | A kind of synergistic urea and preparation method thereof |
| CN106755156A (en) * | 2016-12-28 | 2017-05-31 | 安徽丰原发酵技术工程研究有限公司 | A kind of fermentation process of L lysines |
| CN107164450A (en) * | 2017-06-30 | 2017-09-15 | 齐齐哈尔龙江阜丰生物科技有限公司 | A kind of recoverying and utilizing method of fermentation waste liquid of lysine |
| CN107267427A (en) * | 2017-08-17 | 2017-10-20 | 齐齐哈尔龙江阜丰生物科技有限公司 | A kind of threonine mother liquor recycling method |
| CN108796004A (en) * | 2018-06-20 | 2018-11-13 | 齐齐哈尔龙江阜丰生物科技有限公司 | A kind of technique of preparing lysine through fermentation |
| CN112048526A (en) * | 2020-09-21 | 2020-12-08 | 山东柠檬生化有限公司 | High-conversion-rate production method of fermented citric acid |
Non-Patent Citations (3)
| Title |
|---|
| CHUNDAKKADU KRISHNA: "Solid-State Fermentation Systems—An Overview" * |
| 周小秋: "合成赖氨酸在鱼类饲料中的应用研究进展" * |
| 施庆珊: "发酵法生产均聚氨基酸研究进展" * |
Cited By (8)
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|---|---|---|---|---|
| CN110499261A (en) * | 2018-05-17 | 2019-11-26 | 卢松 | The preparation process of protein enzymatic hydrolyzate and its application in fermented and cultured |
| CN110055296A (en) * | 2019-05-13 | 2019-07-26 | 南京牧奇亚生物科技有限公司 | The preparation method and applications of lysine |
| CN110358797A (en) * | 2019-08-13 | 2019-10-22 | 南京牧奇亚生物科技有限公司 | A kind of fermentation method production method of vitamin B12 |
| CN110791535A (en) * | 2019-12-02 | 2020-02-14 | 齐齐哈尔龙江阜丰生物科技有限公司 | Process for producing and extracting lysine |
| CN112662713A (en) * | 2020-12-28 | 2021-04-16 | 天津科技大学 | Culture medium for producing L-lysine by high-density fermentation and method thereof |
| CN112662713B (en) * | 2020-12-28 | 2022-08-19 | 天津科技大学 | Culture medium for producing L-lysine by high-density fermentation and method thereof |
| CN116496950A (en) * | 2022-09-27 | 2023-07-28 | 欧铭庄生物科技(天津)有限公司滨海新区分公司 | Lysine production strain and application thereof, and lysine production method |
| CN116496950B (en) * | 2022-09-27 | 2023-10-24 | 欧铭庄生物科技(天津)有限公司滨海新区分公司 | Lysine production strain and application thereof, and lysine production method |
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