CN109355325A - The symbiosis production. art of particle threonine and granule protein - Google Patents
The symbiosis production. art of particle threonine and granule protein Download PDFInfo
- Publication number
- CN109355325A CN109355325A CN201811213884.3A CN201811213884A CN109355325A CN 109355325 A CN109355325 A CN 109355325A CN 201811213884 A CN201811213884 A CN 201811213884A CN 109355325 A CN109355325 A CN 109355325A
- Authority
- CN
- China
- Prior art keywords
- threonine
- fermentation
- particle
- granule protein
- centrifugation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000004473 Threonine Substances 0.000 title claims abstract description 55
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 title claims abstract description 53
- 239000008187 granular material Substances 0.000 title claims abstract description 22
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 22
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 22
- 239000002245 particle Substances 0.000 title claims abstract description 21
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 17
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 title claims abstract description 9
- 238000000855 fermentation Methods 0.000 claims abstract description 52
- 230000004151 fermentation Effects 0.000 claims abstract description 52
- 238000000034 method Methods 0.000 claims abstract description 21
- 238000005119 centrifugation Methods 0.000 claims abstract description 16
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 45
- 239000007788 liquid Substances 0.000 claims description 21
- 239000006228 supernatant Substances 0.000 claims description 16
- 241000195597 Chlamydomonas reinhardtii Species 0.000 claims description 13
- 238000001035 drying Methods 0.000 claims description 12
- 206010013786 Dry skin Diseases 0.000 claims description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 8
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 8
- 239000002054 inoculum Substances 0.000 claims description 8
- 230000001376 precipitating effect Effects 0.000 claims description 8
- 238000000108 ultra-filtration Methods 0.000 claims description 6
- 240000008042 Zea mays Species 0.000 claims description 5
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 5
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 5
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 5
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 5
- 235000005822 corn Nutrition 0.000 claims description 5
- 230000009471 action Effects 0.000 claims description 4
- 210000002421 cell wall Anatomy 0.000 claims description 4
- 239000000919 ceramic Substances 0.000 claims description 4
- 235000013339 cereals Nutrition 0.000 claims description 4
- 239000013078 crystal Substances 0.000 claims description 4
- 239000012467 final product Substances 0.000 claims description 4
- 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 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 4
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 4
- 238000005453 pelletization Methods 0.000 claims description 4
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 4
- 238000005507 spraying Methods 0.000 claims description 4
- 230000003519 ventilatory effect Effects 0.000 claims description 4
- 235000020985 whole grains Nutrition 0.000 claims description 4
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 claims description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims 2
- 235000009754 Vitis X bourquina Nutrition 0.000 claims 1
- 235000012333 Vitis X labruscana Nutrition 0.000 claims 1
- 240000006365 Vitis vinifera Species 0.000 claims 1
- 235000014787 Vitis vinifera Nutrition 0.000 claims 1
- 239000002253 acid Substances 0.000 claims 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims 1
- 229910021529 ammonia Inorganic materials 0.000 claims 1
- KCIDZIIHRGYJAE-YGFYJFDDSA-L dipotassium;[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate Chemical compound [K+].[K+].OC[C@H]1O[C@H](OP([O-])([O-])=O)[C@H](O)[C@@H](O)[C@H]1O KCIDZIIHRGYJAE-YGFYJFDDSA-L 0.000 claims 1
- 229910052739 hydrogen Inorganic materials 0.000 claims 1
- 239000001257 hydrogen Substances 0.000 claims 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims 1
- 238000007873 sieving Methods 0.000 claims 1
- 230000008569 process Effects 0.000 abstract description 9
- 150000001413 amino acids Chemical class 0.000 abstract description 8
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 81
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 53
- 229960002898 threonine Drugs 0.000 description 48
- 235000018102 proteins Nutrition 0.000 description 15
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 12
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 11
- 229910052799 carbon Inorganic materials 0.000 description 11
- 239000008103 glucose Substances 0.000 description 11
- 241000588724 Escherichia coli Species 0.000 description 10
- 229940024606 amino acid Drugs 0.000 description 8
- 235000001014 amino acid Nutrition 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 8
- 238000009825 accumulation Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 239000000047 product Substances 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000005469 granulation Methods 0.000 description 3
- 230000003179 granulation Effects 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- RZVAJINKPMORJF-UHFFFAOYSA-N Acetaminophen Chemical compound CC(=O)NC1=CC=C(O)C=C1 RZVAJINKPMORJF-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 101100465553 Dictyostelium discoideum psmB6 gene Proteins 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 101100169519 Pyrococcus abyssi (strain GE5 / Orsay) dapAL gene Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 238000002425 crystallisation Methods 0.000 description 2
- 230000008025 crystallization Effects 0.000 description 2
- 101150011371 dapA gene Proteins 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000037353 metabolic pathway Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000029553 photosynthesis Effects 0.000 description 2
- 238000010672 photosynthesis Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 241000195585 Chlamydomonas Species 0.000 description 1
- 241000186226 Corynebacterium glutamicum Species 0.000 description 1
- YTBSYETUWUMLBZ-QWWZWVQMSA-N D-threose Chemical compound OC[C@@H](O)[C@H](O)C=O YTBSYETUWUMLBZ-QWWZWVQMSA-N 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical class C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 241000319304 [Brevibacterium] flavum Species 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- -1 aliphatic amino acid Chemical class 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 230000006860 carbon metabolism Effects 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 230000034659 glycolysis Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 238000009655 industrial fermentation Methods 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- VSEWZMQQQHLHQT-UHFFFAOYSA-N manganese;sulfuric acid;hydrate Chemical compound O.[Mn].OS(O)(=O)=O VSEWZMQQQHLHQT-UHFFFAOYSA-N 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000012092 media component Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 230000021332 multicellular organism growth Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 238000005303 weighing Methods 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
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/08—Lysine; Diaminopimelic acid; Threonine; Valine
-
- 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
- C12P21/00—Preparation of peptides or proteins
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention belongs to technical field of amino acid production, disclose the symbiosis production. art of particle threonine and granule protein comprising following steps: step 1) fermentation, step 2 centrifugation, and step 3) prepares particle threonine, and step 4) prepares granule protein.Present invention process can be improved the content of particle threonine and granule protein, and simple process is feasible, have a extensive future.
Description
Technical field
The invention belongs to amino acids production fields, and in particular to the symbiosis production. art of particle threonine and granule protein.
Background technique
Threonine (Threonine is abbreviated as Thr), scientific name 2 amino 3 hydroxybutyric acid, nineteen thirty-five is by W.C.Rose
It separates and identifies in fibrin hydrolysate and, Meger studies its space structure within 1936, because of it
Structure is similar to threose, therefore is named as threonine.Threonine belongs to aliphatic amino acid, slightly sweet, is to constitute people and dynamic plant
A kind of essential amino acid of object protein, is mainly used for medicine, chemical reagent, nutrition fortifier, can strengthen dairy products, have
Restore human-body fatigue, the effect of enhancing development.In recent years, with the development of economy, market continues surely threonine requirement
It is fixed to increase, it is most fast one of the amino acid kind of demand growth, especially in chemistry and biochemistry, food additives, feed addition
The dosage rapid development of agent etc., big substituted tryptophan and the development that becomes in addition to lysine, methionine is most rapid
The third-largest amino acid.L-threonine is added in mixed feed, has the characteristics that as follows: 1. the amino acid of adjustable feed is flat
Weighing apparatus promotes poultry growth；2. meat can be improved；3. the nutritive value of the low feed of amino acid digestibility can be improved；4. can reduce
Feedstuff cost；Therefore in China, European Union member countries and American States, feedstuff industry has been widely used in it.
Currently, the production method of threonine mainly has fermentation method, protein Hydrolyze method and 3 kinds of chemical synthesis, microorganism
Fermentation method produces threonine, because the advantages that its simple process and low cost has become current main stream approach.L-threonine it is main
Production bacterial strain has Corynebacterium glutamicum, brevibacterium flavum and Escherichia coli.Threonine Fermentation technology is primarily present fermentation effect at present
The defect that rate is low and purity is not up to standard.Bacterial strain is transformed to improve the yield of amino acid, early all the time by various methods in people
The phase most widely used mutation breeding under the conditions of various, and with the exposition of amino acid bio metabolic pathway, it is purposive
Metabolic pathway is transformed on a molecular scale and also appears its advantage gradually, in addition, the fermentation condition optimization process in middle reaches with
And the reclaiming clean process in downstream is also an emphasis.
Colibacillus engineering strain is the main bacterial strain of microbe industrial fermentation production threonine, generates L- Soviet Union in fermentation
While propylhomoserin, the metabolic by-products such as acetic acid, alanine, valine and arginine can be also generated, influence bacterium to a certain extent
Body growth and the synthesis and accumulation of L-threonine.Wherein, the inhibitory effect of acetic acid is particularly evident, when Acetic Acid Accumulation is to certain dense
When spending, the specific growth rate of thallus declines rapidly, and Product formation substantially reduces, and forms vicious circle, while foreign gene
Expression is also heavily affected.Acetic Acid Accumulation is adversely affected caused by cell metabolism, is using Escherichia coli as host strain
Express a very important problem of foreign protein.How the content of acetic acid is reduced, to improve biomass and threonine
Yield is the emphasis that we study.Document " control of acetic acid in L-threonine fermentation process, science and technology of fermenting communication 2012 " exists
During Escherichia coli fermentation prepares threonine, the generation of its by-product acetic acid is controlled by selecting suitable fermentation condition,
It can reduce the generation of acetic acid, but reduce amplitude and be reduced to unobvious, the yield of threonine cannot be greatly improved, there is no industry
Change the value promoted.The culture medium that patented technology " a kind of ultralow moisture content threonine production method " before applicant is recorded: Portugal
Grape sugar 80g/L, corn pulp 20g/L, ammonium sulfate 2g/L, calcium carbonate 0.75g/L, KH2PO40.2g/L, K2HPO4 0.2g/L, NaCl
0.2g/L, pH value 6.5；The content of threonine can reach 10g/100ml in fermentation liquid, but the phase is dead after fermentation for bacterial strain
Rate is higher, and glucose utilization is larger, also there is to be hoisted threonine yield in fermentation liquid.
Summary of the invention
In order to solve the defects of prior art, the invention proposes the symbiosis production. arts of particle threonine and granule protein.
Present invention process can be improved the content of particle threonine and granule protein, and simple process is feasible, have a extensive future.
The present invention is achieved by the following technical solution:
The symbiosis production. art of particle threonine and granule protein comprising following steps: step 1) fermentation, step 2 centrifugation, step
Rapid 3) to prepare particle threonine, step 4) prepares granule protein.
Further, the step 1) fermentation, includes the following steps: the colibacillus engineering that will produce threonine according to 6-
10% inoculum concentration, which is linked into the fermentor containing fermentation medium, ferments, and 30-32 DEG C of temperature, tank pressure is 0.04-
0.05MPa, ventilatory capacity 0.3-0.5vvm, revolving speed 100rpm, fermentation time culture is 36-48h, then according to 6-10%'s
Inoculum concentration accesses Chlamydomonas reinhardtii, continues fermented and cultured 36-48h, stops fermentation, collects fermentation liquid.
Further, step 2 centrifugation, include the following steps: fermentation liquid first pass around disk plate centrifuge with
4000rpm is centrifuged 5min, collects supernatant liquid and precipitating.
Further, the step 3) prepares particle threonine, includes the following steps: the warp of supernatant liquid obtained by step 2
Ceramic membrane filter is crossed, filtered solution is collected, filtered solution is separated through decanter centrifuge, centrifugal speed 5000rpm, centrifugation time is
3min collects supernatant；Then pass through ultrafiltration membrance filter, collect filtered solution for the intermittent single-action condensing crystallizing pot knot of filtered solution
Crystal is collected by centrifugation in crystalline substance, then 120 DEG C of dryings, compresses slabbing, then put into granulation tower, in boiling under the action of thermal current
Rise state；65 DEG C of fluidized bed dryings, then through broken whole grain, pass sequentially through 20 mesh and 50 meshes, weeded out thick, meticulous particle,
The granule for collecting target grain size, is packed to obtain the final product.
Further, the step 4) prepares granule protein, includes the following steps: precipitating obtained by step 2 being added to 3
In the hydrochloric acid solution of the 5M of times weight, 80 DEG C are heated to, 12h is handled under heat-retaining condition, then with disc separator with 3000rpm
Revolving speed centrifugation 5min remove cell wall, collect supernatant, be condensed into paste, finally by slurry-spraying pelletizing fluidized bed drying,
Obtain granule protein.
Further, the component of the fermentation medium are as follows: glucose 20-30g/L, glycerol 20-30g/L, corn pulp 20-
30g/L, ammonium sulfate 2-3g/L, potassium dihydrogen phosphate 0.2-0.3g/L, dipotassium hydrogen phosphate 0.2-0.3g/L, epsom salt 0.1-
0.2g/L, ferrous sulfate heptahydrate 0.01-0.02g/L, manganese sulfate monohydrate 0.01-0.02g/L, pH value 6.5-6.8.
The beneficial effect of starting point and acquirement that the present invention studies mainly includes but is not limited to the following aspects:
When the aerobic culture of Escherichia coli, oxygen molecule is electron transmission final receptor, and as cell is grown, oxygen consumption constantly increases
Add, occur for hypoxgia, so that TCA circulation is obstructed, threonine yield is reduced, and carbon metabolism approach more flows to acetic acid way
Diameter, Acetic Acid Accumulation increase sharply.
Cell concentration has a direct impact the generation of acetic acid, and earlier fermentation, cell density is not high, and oxygen demand is relatively fewer,
It ferments the middle and later periods, when cell concentration increases, oxygen demand increases, and is easy to cause the too fast generation of acetic acid, when cell concentration is too low, again
The synthesis of purpose product can be reduced.
Fermenting carbon source selection glucose and glycerol of the present invention, earlier fermentation, cell density is low, and oxygen-supplying amount is sufficient, large intestine bar
Bacterium preferentially uses glucose as carbon source, can promote the generation of growing microorganism and threonine；It ferments the middle and later periods, glucose is consumed
To the greatest extent, Escherichia coli use glycerol as carbon source at this time, since the rate that cell absorbs glycerol is lower, under the carbon flow of glycolysis
Drop, to reduce the accumulation of acetic acid, while improving the yield of threonine；
The present invention can carry out non-light and work as carbon source using the acetic acid in fermentation liquid by being inoculated with Chlamydomonas reinhardtii in fermentation
With, and it is more difficult use glycerol as carbon source, thus relieve to Escherichia coli produce threonine inhibiting effect, additionally it is possible to carry out micro-
The photosynthesis of amount discharges oxygen, for Escherichia coli fermentation produce threonine come using.By adding Chlamydomonas reinhardtii, it is not only able to mention
The yield of high threonine, and mycoprotein yield also correspondinglys increase.
Powdered threonine is prepared into graininess threonine by the present invention, can improve mobility, convenient for preservation and transport,
Solubility is controlled, the quality and added value of threonine are improved；Thallus is prepared into particle mycoprotein, is convenient for preservation and fortune
It is defeated, yeast powder can be substituted as nutrient media components, feed addictive is can be also used for, improve added value of product.
Figure of description
Fig. 1: the acetic acid content of each group in different time points.
Specific embodiment
Those skilled in the art can use for reference present disclosure, be suitably modified realization of process parameters.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.Product and method of the invention is described by preferred embodiment, and related personnel can obviously not depart from this hair
Product as described herein and method are modified in bright content, spirit and scope or appropriate changes and combinations, 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
The symbiosis production. art of particle threonine and granule protein comprising following steps:
By colibacillus engineering K12 △ dapA seed liquor, (concentration of seed liquor is 1 × 10 to step 1)8Cfu/mL) according to 10%
Inoculum concentration be linked into the fermentor containing fermentation medium and ferment, 30 DEG C of temperature, tank pressure is 0.04MPa, ventilatory capacity
For 0.5vvm, revolving speed 100rpm, fermentation time culture is 36h, then accesses Chlamydomonas reinhardtii (Rhein according to 10% inoculum concentration
The concentration of chlamydomonas is 1 × 105Cfu/mL), continue fermented and cultured 48h, stop fermentation, collect fermentation liquid；
The component of the fermentation medium are as follows: glucose 20g/L, glycerol 20g/L, corn pulp 20g/L, ammonium sulfate 2g/L, phosphoric acid
Potassium dihydrogen 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, epsom salt 0.1g/L, ferrous sulfate heptahydrate 0.01g/L, sulfuric acid monohydrate
Manganese 0.01g/L, pH value 6.5；
Step 2 fermentation liquid first passes around disk plate centrifuge and is centrifuged 5min with 4000rpm, collects supernatant liquid and precipitating；
Step 3) supernatant liquid is filtered by ceramic membrane (10,000 Da molecular cut off), collects filtered solution, by filtered solution through sleeping spiral shell from
Scheming separation, centrifugal speed 5000rpm, centrifugation time 3min are collected supernatant (protein content is less than 0.5%)；Then it passes through
Ultrafiltration membrance filter is crossed, filtered solution is collected, ultrafiltration retaining molecular weight is 300Da；By the intermittent single-action condensing crystallizing of filtered solution
Pot crystallization, is collected by centrifugation crystal, and then 120 DEG C of dryings to moisture content are 0.8%, compresses slabbing, then put into granulation tower,
It is in fluidized state under the action of thermal current；65 DEG C of fluidized bed dryings, then through broken whole grain, 20 mesh and 50 meshes are passed sequentially through,
It weeded out thick, meticulous particle, and collected the granule of target grain size, pack to obtain the final product；
Step 4) is heated to 80 DEG C, under heat-retaining condition in the hydrochloric acid solution for the 5M that precipitating is added to 3 times of weight obtained by step 2
12h is handled, cell wall is then removed with the revolving speed centrifugation 5min of 3000r/min with disc separator, supernatant is collected, is condensed into
Paste obtains granule protein finally by slurry-spraying pelletizing fluidized bed drying.
Embodiment 2
The symbiosis production. art of particle threonine and granule protein comprising following steps:
By colibacillus engineering K12 △ dapA seed liquor, (concentration of seed liquor is 1 × 10 to step 1)8Cfu/mL) according to 6%
Inoculum concentration, which is linked into the fermentor containing fermentation medium, ferments, and 32 DEG C of temperature, tank pressure is 0.04MPa, and ventilatory capacity is
0.4vvm, revolving speed 100rpm, fermentation time culture are 48h, then access Chlamydomonas reinhardtii (Chlamydomonas reinhardtii according to 8% inoculum concentration
Concentration be 1 × 105Cfu/mL), continue fermented and cultured 48h, stop fermentation, collect fermentation liquid；
The component of the fermentation medium are as follows: glucose 30g/L, glycerol 30g/L, corn pulp 30g/L, ammonium sulfate 3g/L, phosphoric acid
Potassium dihydrogen 0.3g/L, dipotassium hydrogen phosphate 0.3g/L, epsom salt 0.2g/L, ferrous sulfate heptahydrate 0.02g/L, manganese sulfate monohydrate
0.02g/L, pH value 6.8；
Step 2 fermentation liquid first passes around disk plate centrifuge and is centrifuged 5min with 4000rpm, collects supernatant liquid and precipitating；
Step 3) supernatant liquid is filtered by ceramic membrane (10,000 Da molecular cut off), collects filtered solution, by filtered solution through sleeping spiral shell from
Scheming separation, centrifugal speed 5000rpm, centrifugation time 3min are collected supernatant (protein content is less than 0.5%)；Then it passes through
Ultrafiltration membrance filter is crossed, filtered solution is collected, ultrafiltration retaining molecular weight is 300Da；By the intermittent single-action condensing crystallizing of filtered solution
Pot crystallization, is collected by centrifugation crystal, and then 120 DEG C of dryings to moisture content are 0.8%, compresses slabbing, then put into granulation tower,
It is in fluidized state under the action of thermal current；65 DEG C of fluidized bed dryings, then through broken whole grain, 20 mesh and 50 meshes are passed sequentially through,
It weeded out thick, meticulous particle, and collected the granule of target grain size, pack to obtain the final product；
Step 4) is heated to 80 DEG C, under heat-retaining condition in the hydrochloric acid solution for the 5M that precipitating is added to 3 times of weight obtained by step 2
12h is handled, cell wall is then removed with the revolving speed centrifugation 5min of 3000r/min with disc separator, supernatant is collected, is condensed into
Paste obtains granule protein finally by slurry-spraying pelletizing fluidized bed drying.
Embodiment 3
Influence of the different factors to production amount of threonine and yield of acetic acid in present invention process:
Group is set:
Experimental group: embodiment 1；
Control group 1: not adding Chlamydomonas reinhardtii, remaining is the same as embodiment 1；
Glycerol: being replaced with the glucose of equal quality by control group 2, remaining is the same as embodiment 1；
Control group 3: not adding Chlamydomonas reinhardtii, while glycerol being replaced with to the glucose of equal quality, remaining is the same as embodiment 1.
The content of threonine and acetic acid is shown in Table 1 in each final fermentation liquid of group:
Table 1
| Group | Threonine g/L | Acetic acid g/L |
| Experimental group | 137.1 | 0.6 |
| Control group 1 | 102.5 | 13.9 |
| Control group 2 | 118.2 | 4.7 |
| Control group 3 | 96.4 | 15.6 |
Conclusion: experimental group can utilize the acetic acid in threonine fermentation liquid by carrying out assisted fermentation processing to Chlamydomonas reinhardtii
Non- light and effect are carried out as carbon source, to relieve the inhibiting effect that Escherichia coli are produced with threonine, additionally it is possible to carry out micro
Photosynthesis discharge oxygen, for Escherichia coli fermentation produce threonine come using；Part glucose is substituted by glycerol simultaneously, with
The consumption of glucose, Escherichia coli use glycerol as carbon source, due to cell absorb glycerol rate it is lower, reduce acetic acid
Accumulation, while improving the yield of threonine, pass through each group comparative test and find, compared with control group 1-3, the present invention
The production amount of threonine of experimental group is respectively increased 33.76%, 15.99%, 42.22%；And acetic acid content is only 0.6g/L, is equivalent to pair
According to the 3.85% of group 1.
The present invention also has detected the yield of acetic acid of each group in different time points, by taking embodiment 1 as an example, chooses fermentation respectively
Afterwards, 36h, 48h, 60h, 72h, 84h, totally 5 time points are detected, and concrete outcome is shown in Fig. 1.In experimental group,
With the increase of fermentation time, acetic acid content is reduced rapidly, and is relieved the synthesis to threonine and is inhibited, to improve threonine
Secretory volume；And in control group 1 and 3, due to there is no addition Chlamydomonas reinhardtii, acetic acid content is caused to continue growing；In control group 2,
Due to being added to Chlamydomonas reinhardtii, so that acetic acid content gradually declines, but fall is lower than experimental group, it may be possible to because of experiment
Group is added to glycerol as carbon source, and the carbon source into acetate pathway reduces, and Chlamydomonas reinhardtii is more difficult uses glycerol as carbon source,
It is only capable of using acetic acid, and then causes the fall of experimental group acetic acid more obvious.
The above described is only a preferred embodiment of the present invention, be not intended to limit the present invention in any form, though
The right present invention has been described by way of example and in terms of the preferred embodiments, however, being not intended to limit the invention, any technology people for being familiar with this profession
Member can make a little change or modification using the technology contents disclosed certainly without departing from the scope of the present invention, at
For the equivalent embodiment of equivalent variations, but anything that does not depart from the technical scheme of the invention content, according to the technical essence of the invention
Any simple modification, equivalent change and modification to the above embodiments, belong in the range of technical solution of the present invention.
Claims (6)
1. the symbiosis production. art of particle threonine and granule protein comprising following steps: step 1) fermentation, step 2 centrifugation,
Step 3) prepares particle threonine, and step 4) prepares granule protein.
2. technique according to claim 1, which is characterized in that the step 1) fermentation includes the following steps: that Soviet Union's ammonia will be produced
The colibacillus engineering of acid is linked into the fermentor containing fermentation medium according to the inoculum concentration of 6-10% to ferment, temperature
30-32 DEG C of degree, tank pressure are 0.04-0.05MPa, ventilatory capacity 0.3-0.5vvm, revolving speed 100rpm, fermentation time 36-
Then 48h accesses Chlamydomonas reinhardtii according to the inoculum concentration of 6-10%, continue fermented and cultured 36-48h, stops fermentation, collects fermentation liquid.
3. technique according to claim 2, which is characterized in that the step 2 centrifugation includes the following steps: that fermentation liquid passes through
It crosses disk plate centrifuge and 5min is centrifuged with 4000rpm, collect supernatant liquid and precipitating.
4. technique according to claim 3, which is characterized in that the step 3) prepares particle threonine, including walks as follows
It is rapid: supernatant liquid obtained by step 2 being passed through into ceramic membrane filter, filtered solution is collected, filtered solution is centrifuged, collects supernatant；Then
By ultrafiltration membrance filter, then condensing crystallizing is carried out, crystal is collected by centrifugation, then 120 DEG C of dryings, compresses slabbing, then put into and make
It is in fluidized state under the action of thermal current in grain tower；65 DEG C of fluidized bed dryings, then through broken whole grain, sieving is packed to obtain the final product.
5. technique according to claim 4, which is characterized in that the step 4) prepares granule protein, includes the following steps:
By in the hydrochloric acid solution for the 5M that precipitating is added to 3 times of weight obtained by step 2,80 DEG C are heated to, 12h is handled under heat-retaining condition, so
Centrifugation removal cell wall afterwards, collects supernatant, is condensed into paste, finally by slurry-spraying pelletizing fluidized bed drying, obtains particle
Albumen.
6. according to claim 2-5 be allowed to one described in technique, which is characterized in that the component of the fermentation medium are as follows: grape
Sugared 20-30g/L, glycerol 20-30g/L, corn pulp 20-30g/L, ammonium sulfate 2-3g/L, potassium dihydrogen phosphate 0.2-0.3g/L, phosphoric acid
Hydrogen dipotassium 0.2-0.3g/L, epsom salt 0.1-0.2g/L, ferrous sulfate heptahydrate 0.01-0.02g/L, manganese sulfate monohydrate
0.01-0.02g/L, pH value 6.5-6.8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811213884.3A CN109355325A (en) | 2018-10-18 | 2018-10-18 | The symbiosis production. art of particle threonine and granule protein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811213884.3A CN109355325A (en) | 2018-10-18 | 2018-10-18 | The symbiosis production. art of particle threonine and granule protein |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109355325A true CN109355325A (en) | 2019-02-19 |
Family
ID=65345710
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811213884.3A Pending CN109355325A (en) | 2018-10-18 | 2018-10-18 | The symbiosis production. art of particle threonine and granule protein |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109355325A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110051634A (en) * | 2019-05-29 | 2019-07-26 | 绥化象屿金谷生化科技有限公司 | A kind of L-threonine processes for wet granulation |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005014840A1 (en) * | 2003-08-01 | 2005-02-17 | Degussa Ag | Process for the preparation of l-threonine |
| CN101456822A (en) * | 2008-11-28 | 2009-06-17 | 山东恩贝生物工程有限公司 | Novel process for extracting threonine |
| CN104757273A (en) * | 2015-04-12 | 2015-07-08 | 呼伦贝尔东北阜丰生物科技有限公司 | Process for preparing granular type threonine product |
| CN104844468A (en) * | 2015-04-12 | 2015-08-19 | 呼伦贝尔东北阜丰生物科技有限公司 | Environment-friendly technology for treating threonine fermentation mother liquid |
| CN107208118A (en) * | 2014-09-18 | 2017-09-26 | 基因组股份公司 | The non-natural microorganism of energy efficiency with raising |
| CN108658798A (en) * | 2018-05-03 | 2018-10-16 | 齐齐哈尔龙江阜丰生物科技有限公司 | The fermentation preparation process of particle threonine |
-
2018
- 2018-10-18 CN CN201811213884.3A patent/CN109355325A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005014840A1 (en) * | 2003-08-01 | 2005-02-17 | Degussa Ag | Process for the preparation of l-threonine |
| CN101456822A (en) * | 2008-11-28 | 2009-06-17 | 山东恩贝生物工程有限公司 | Novel process for extracting threonine |
| CN107208118A (en) * | 2014-09-18 | 2017-09-26 | 基因组股份公司 | The non-natural microorganism of energy efficiency with raising |
| CN104757273A (en) * | 2015-04-12 | 2015-07-08 | 呼伦贝尔东北阜丰生物科技有限公司 | Process for preparing granular type threonine product |
| CN104844468A (en) * | 2015-04-12 | 2015-08-19 | 呼伦贝尔东北阜丰生物科技有限公司 | Environment-friendly technology for treating threonine fermentation mother liquid |
| CN108658798A (en) * | 2018-05-03 | 2018-10-16 | 齐齐哈尔龙江阜丰生物科技有限公司 | The fermentation preparation process of particle threonine |
Non-Patent Citations (3)
| Title |
|---|
| MOON, MYOUNGHOON等: "Mixotrophic growth with acetate or volatile fatty acids maximizes growth and lipid production inChlamydomonas reinhardtii", 《ALGAL RESEARCH-BIOMASS BIOFUELS AND BIOPRODUCTS》 * |
| ZHANG JIAN-GUO等: ""Valorization of Spent Escherichia coli Media Using Green Microalgae Chlamydomonas reinhardtii and Feedstock Production", 《FRONTIERS IN MICROBIOLOGY》 * |
| 胡丹 等: "L-苏氨酸发酵过程中乙酸的控制", 《发酵科技通讯》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110051634A (en) * | 2019-05-29 | 2019-07-26 | 绥化象屿金谷生化科技有限公司 | A kind of L-threonine processes for wet granulation |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP6913033B2 (en) | Gas fermentation for protein or feed production | |
| CN109182171B (en) | Mutagenic strain for high yield of gamma-aminobutyric acid and biological preparation thereof | |
| CN108658798A (en) | The fermentation preparation process of particle threonine | |
| JP2019531058A (en) | Single cell proteins from thermophilic fungi | |
| CN104686784B (en) | The technique that a kind of utilization waste water of mgs prepares feed | |
| CN108034599B (en) | One plant of Lactobacillus brevis for efficiently synthesizing γ-aminobutyric acid from brewed spirit system | |
| CN102796779A (en) | Biological method for preparing gamma-aminobutyric acid | |
| CN102559523A (en) | Selenium-rich yeast, selenium-rich yeast hydrolysate and preparation method of the hydrolysate | |
| CN109486876A (en) | A method of threonine is extracted and is purified in fermentation | |
| CN109439702A (en) | The technique for handling threonine high gravity fermentation waste water | |
| CN116496950B (en) | Lysine production strain and application thereof, and lysine production method | |
| CN114107073A (en) | Method for producing hypha protein by utilizing molasses | |
| CN110004192A (en) | A kind of method of preparing granular type threonine | |
| CN113046253B (en) | Culture method for improving heat resistance of kluyveromyces marxianus | |
| CN109136299B (en) | Method for preparing, extracting and purifying threonine | |
| CA3001675C (en) | Bio-based n-acetyl-l-methionine and use thereof | |
| CN110396530A (en) | A method of improving production amount of threonine and yield | |
| CN109355325A (en) | The symbiosis production. art of particle threonine and granule protein | |
| CN109706195B (en) | Production process of granular threonine | |
| CN101926412A (en) | Production method of compound amino acid feed additive rich in gamma-aminobutyric acid (GABA) | |
| CN107326052B (en) | Method for improving glutamate decarboxylase activity by using D101 macroporous adsorption resin | |
| CN112481321B (en) | Process for producing granular threonine | |
| CN104212869B (en) | A kind of ultralow moisture content threonine production method | |
| CN1807592A (en) | Method for producing lactic acid bacteria formulation and lactic acid bacteria bacteriocin using ethanol draff liquid | |
| CN109943604A (en) | A method of improving glutamic acid fermentation conversion ratio and recovery rate |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20190219 |