CN107034249A - A kind of method that the useless bacteria residue of L ornithines is back to L ornithine fermenting and producings - Google Patents
A kind of method that the useless bacteria residue of L ornithines is back to L ornithine fermenting and producings Download PDFInfo
- Publication number
- CN107034249A CN107034249A CN201710492580.4A CN201710492580A CN107034249A CN 107034249 A CN107034249 A CN 107034249A CN 201710492580 A CN201710492580 A CN 201710492580A CN 107034249 A CN107034249 A CN 107034249A
- Authority
- CN
- China
- Prior art keywords
- orn
- bacteria residue
- fermentation
- fermenting
- useless
- 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
- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 title claims abstract description 78
- 241000894006 Bacteria Species 0.000 title claims abstract description 59
- 238000000034 method Methods 0.000 title claims abstract description 23
- AHLPHDHHMVZTML-BYPYZUCNSA-N ornithyl group Chemical class N[C@@H](CCCN)C(=O)O AHLPHDHHMVZTML-BYPYZUCNSA-N 0.000 title abstract 7
- UTJLXEIPEHZYQJ-UHFFFAOYSA-N Ornithine Natural products OC(=O)C(C)CCCN UTJLXEIPEHZYQJ-UHFFFAOYSA-N 0.000 title abstract 2
- 229960003104 ornithine Drugs 0.000 title abstract 2
- 238000000855 fermentation Methods 0.000 claims abstract description 65
- 230000004151 fermentation Effects 0.000 claims abstract description 65
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000004475 Arginine Substances 0.000 claims description 18
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 18
- 239000002893 slag Substances 0.000 claims description 13
- 230000007062 hydrolysis Effects 0.000 claims description 12
- 238000006460 hydrolysis reaction Methods 0.000 claims description 12
- 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 10
- 239000008103 glucose Substances 0.000 claims description 10
- 239000002609 medium Substances 0.000 claims description 10
- 241000186226 Corynebacterium glutamicum Species 0.000 claims description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 239000007787 solid Substances 0.000 claims description 8
- 230000002950 deficient Effects 0.000 claims description 7
- 239000001963 growth medium Substances 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 239000002054 inoculum Substances 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 235000002639 sodium chloride Nutrition 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 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 claims description 4
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 229960002668 sodium chloride Drugs 0.000 claims description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 3
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 3
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 3
- 244000005700 microbiome Species 0.000 claims description 3
- 229930003451 Vitamin B1 Natural products 0.000 claims description 2
- 229960002685 biotin Drugs 0.000 claims description 2
- 235000020958 biotin Nutrition 0.000 claims description 2
- 239000011616 biotin Substances 0.000 claims description 2
- 229910052799 carbon Inorganic materials 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 2
- 229920000053 polysorbate 80 Polymers 0.000 claims description 2
- 238000004321 preservation Methods 0.000 claims description 2
- 229960003495 thiamine Drugs 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
- 239000011691 vitamin B1 Substances 0.000 claims description 2
- 235000010374 vitamin B1 Nutrition 0.000 claims description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims 1
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims 1
- 239000012138 yeast extract Substances 0.000 abstract description 10
- 229940041514 candida albicans extract Drugs 0.000 abstract description 7
- 125000001477 organic nitrogen group Chemical group 0.000 abstract description 7
- 238000005119 centrifugation Methods 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 abstract description 6
- 241000233866 Fungi Species 0.000 abstract description 5
- 240000008042 Zea mays Species 0.000 abstract description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 abstract description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 abstract description 3
- 235000005822 corn Nutrition 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 abstract description 3
- 241001515965 unidentified phage Species 0.000 abstract description 3
- 238000004140 cleaning Methods 0.000 abstract description 2
- 238000007599 discharging Methods 0.000 abstract description 2
- 238000003912 environmental pollution Methods 0.000 abstract description 2
- 238000005903 acid hydrolysis reaction Methods 0.000 abstract 1
- 235000011149 sulphuric acid Nutrition 0.000 abstract 1
- 239000001117 sulphuric acid Substances 0.000 abstract 1
- 235000009697 arginine Nutrition 0.000 description 16
- 239000000243 solution Substances 0.000 description 7
- 239000012267 brine Substances 0.000 description 4
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 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 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 150000003839 salts Chemical class 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 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 1
- 229930064664 L-arginine Natural products 0.000 description 1
- 235000014852 L-arginine Nutrition 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
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 238000010564 aerobic fermentation Methods 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000002929 anti-fatigue Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- VTYYLEPIZMXCLO-UHFFFAOYSA-L calcium carbonate Substances [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 229910052564 epsomite Inorganic materials 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 230000037257 muscle growth Effects 0.000 description 1
- 229920000768 polyamine Polymers 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000004143 urea cycle Effects 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
- C12P13/10—Citrulline; Arginine; Ornithine
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a kind of method that the useless bacteria residue of L ornithines is back into L ornithine fermenting and producings.Replace yeast extract, corn steep liquor as organic nitrogen source with the hydrolyzate formed after sulphuric acid hydrolysis the L ornithines fermentation fungi residues being collected by centrifugation, be back to the fermenting and producing of L ornithines.Compared with prior art, the present invention can reduce fermentation method and prepare the production cost of L ornithines, and avoid discharging miscellaneous bacteria caused by useless bacteria residue, bacteriophage propagation risk and environmental pollution, be a kind of production technology of cleaning.
Description
Technical field
The invention belongs to biological chemical field, and in particular to a kind of that the useless bacteria residue of L-Orn is back into L-Orn fermentation
The method of production.
Background technology
L-Orn belongs to basic amino acid, is primarily involved in the urea cycle in organism, for citrulling in organism,
Arginine, proline, the biosynthesis of polyamines, have effects that protecting liver and detoxication delays growth of tumour cell, in addition L-Orn
Also have and promote muscle growth and antifatigue effect, therefore, L-Orn is increasingly extensive in the application of medicine, healthcare field,
With good market prospects.
Microorganism direct fermentation is the important sources for producing L-Orn, and strain is mainly the smart ammonia of Corynebacterium glutamicum
Thalline amount reproduction is just can guarantee that containing appropriate arginine in sour deficient strain, culture medium, and synthesizes L-Orn.Patent
ZL201010286236.8 discloses a kind of method that fermentation prepares L-Orn, using glucose as carbon source, L-Orn yield
Up to 35~45g/L.Patent ZL201510024099.3 discloses one kind and prepares L- birds using fibre bed reactor aerobic fermentation
The method of propylhomoserin, improves fermentation process efficiency.But these methods are using yeast extract or the corn of high concentration costly
Starch the organic nitrogen source as fermentation medium, therefore nitrogen source cost is higher, and after L-Orn fermentation ends aging glutamic acid
Bar bacterium is handled as solid slag, once not only pollution environment can also induce miscellaneous bacteria accidentally or bacteriophage is largely numerous for processing
Grow, serious risk is brought to fermenting and producing.A kind of method of the useless bacteria residue of reasonable utilization L-Orn is developed as can be seen here for reality
The clean manufacturing of existing L-Orn is most important.
The content of the invention
The useless bacteria residue of L-Orn is back to L-Orn fermentation life the technical problem to be solved in the present invention is to provide one kind
The method of production, risk larger the problems such as too high with the cost for solving prior art presence.
In order to solve the above technical problems, the technical solution adopted by the present invention is as follows：
A kind of method that the useless bacteria residue of L-Orn is back into L-Orn fermenting and producing, it comprises the following steps：
(1) will be standby after the useless bacteria residue washing of L-Orn；
(2) the useless bacteria residue of the L-Orn after being washed in step (1) is added and be hydrolyzed in aqueous sulfuric acid, hydrolysis is completed
After centrifuge, take supernatant, produce bacteria residue hydrolyzate；
(3) gained bacteria residue hydrolyzate in step (2) is prepared into L-Orn fermentation medium, and use ammoniacal liquor as nitrogen source
Adjust pH to 6.8~7.0；Strain is accessed after the sterilized processing of culture medium after regulation pH, for L-Orn fermentation.
In step (1), described L-Orn gives up bacteria residue for arginine-deficient type Corynebacterium glutamicum fermenting and producing L- birds
Obtained wet bacteria slag is collected after propylhomoserin, solid content is 23~25%.
Wherein, described arginine-deficient type Corynebacterium glutamicum is general in China Committee for Culture Collection of Microorganisms
Logical microorganism center preservation, deposit number is CGMCC No.3663.The bacterial strain patent " a kind of Corynebacterium glutamicum and
Disclosed in the method that L-Orn and its salt are prepared using the bacterium " (application number 201010286236.8).
In step (1), described washing refers to the useless bacteria residue of L-Orn with sodium-chloride water solution with 1：3 volume ratio is mixed
After conjunction, 15~20min, abandoning supernatant are centrifuged with 8000rpm rotating speed；Wherein, the concentration of described sodium-chloride water solution is
0.9wt%, washing times are 2~3 times.
In step (2), the amount ratio of the useless bacteria residue of L-Orn after being washed in aqueous sulfuric acid and step (1) is 1mL：2
~3g；Wherein, the concentration of described aqueous sulfuric acid is 4.0~8.0mol/L.
In step (2), described hydrolysis refers at 80~90 DEG C, under 100~200rpm speed of agitator, hydrolysis 18
~24h.
Wherein, the bacterial strain that fermentation obtains being inoculated with the bacterial strain and step (3) of the useless bacteria residue of L-Orn in step (1) needs phase
Together, it is preferably all the arginine-deficient type Corynebacterium glutamicum that deposit number is CGMCC No.3663.
In step (3), the composition of described L-Orn fermentation medium is：Bacteria residue hydrolyzate, glucose 90g/L,
K2HPO4·3H2O 1.5g/L, MgSO47H2O 3.7g/L, FeSO47H2O 20mg/L, MnSO4H2O 20mg/L,
(NH4)2SO440g/L, Tween 80 0.6g/L, the μ g/L of biotin 50, the μ g/L of vitamin B1 100, pH6.8~7.0；Wherein, its
In, the addition of bacteria residue hydrolyzate make it that arginic content is 200~300mg/L in L-Orn fermentation medium.Culture
121 DEG C of base, 15min sterilizings.Wherein glucose individually carries out moise-heat sterilization.
In step (3), the condition of L-Orn fermentation is：Inoculum concentration is 3~5%, and fermentation temperature is 30 DEG C, fermentation process
It is middle with ammoniacal liquor control pH be 6.8~7.0, be passed through filtrated air fermentation, throughput be 0.5~1.5VVM, mixing speed 300~
500rpm, when fermentation to residual glucose is less than 1g/L, terminates fermentation.
Wherein, the composition of seed culture medium used in L-Orn fermentation is：Glucose 25g/L, yeast extract 5g/L,
(NH4)2SO45g/L, K2HPO4·3H2O 0.5g/L, KH2PO41g/L, MgSO4·7H2O 2.5g/L, NaH2PO4·2H2O
0.5g/L, CaCO310g/L, pH 7.6~7.8.121 DEG C, 15min sterilizings.Seed liquor condition of culture is：Shaking flask liquid amount
10%, inoculum concentration 2%, 30 DEG C, 200rpm concussion and cultivates 10h.
Beneficial effect：
Compared with prior art, the present invention has following advantage：
Compared with the fermentation of existing L-Orn uses the organic nitrogen sources such as yeast extract, corn steep liquor, the hydrolyzate of useless bacteria residue is used
As organic nitrogen source, it can not only reduce fermentation method and prepare the cost of material of L-Orn, and avoid discharging useless bacteria residue and cause
Miscellaneous bacteria, bacteriophage propagates risk and environmental pollution, is a kind of production technology of cleaning.
Embodiment
According to following embodiments, the present invention may be better understood.However, as it will be easily appreciated by one skilled in the art that real
Apply the content described by example and be merely to illustrate the present invention, without should be also without limitation on sheet described in detail in claims
Invention.
The detection of arginine content uses slope opening reagent method in embodiment, is specifically shown in document (L-arginine in zymotic fluid
Detection method [J] food and medicine, 2007,9 (1):18-20.).
Embodiment 1
With reference to a kind of existing patent " Corynebacterium glutamicum and the method for preparing using the bacterium L-Orn and its salt " (application
Number 201010286236.8), prepare L-Orn with arginine-deficient type Corynebacterium glutamicum CGMCC No.3663 fermentations and finish
Afterwards, zymotic fluid is centrifuged into 20min with 8000rpm rotating speed as in supercentrifuge, takes out supernatant and produced for L-Orn
The extraction of product, the bacteria residue of collection is as raw material is utilized, and the solid content of wet bacteria slag is 23~25%.
Embodiment 2
By the L-Orn fermentation fungi residues brine that is collected by centrifugation 2 times, the wet bacteria slag of collection uses 4 respectively
~8.0mol/L sulfuric acid solutions, the volume mass ratio of itself and wet bacteria slag solid content is 1：2.5, it is hydrolyzed respectively, speed of agitator
For 200rpm, hydrolysis temperature is 90.0 DEG C, and hydrolysis time is 20h.The arginine content in supernatant is detected after centrifugation, 1 is shown in Table.
It can be seen that when sulfuric acid concentration is 6mol/L, the arginine content in hydrolyzate is higher, therefore sulfuric acid concentration during selective hydrolysis is
6mol/L。
Influence of the sulfuric acid concentration of table 1 to bacteria residue hydrolysis effect
| Sulfuric acid concentration (mol/L) | 4 | 5 | 6 | 7 | 8 |
| Total nitrogen content (mg/mL) | 27.53 | 31.22 | 33.32 | 34.27 | 34.76 |
| Arginine content (mg/mL) | 3.37 | 4.01 | 4.58 | 4.23 | 3.78 |
Embodiment 3
By the L-Orn fermentation fungi residues brine that is collected by centrifugation 3 times, the wet bacteria slag of collection is used
6.0mol/L sulfuric acid solutions are hydrolyzed, and the volume mass ratio of itself and wet bacteria slag solid content is 1：2.5, speed of agitator is 150rpm, water
It is 90 DEG C to solve temperature, and hydrolysis time is 20h.Arginine content is 4.43mg/mL in the hydrolyzate of acquisition, according to fermentation medium
Middle arginine concentrations are 200mg/L standard, and 46mL hydrolyzates replacement yeast extract is added to ferment in 1L fermentation mediums and made
Standby L-Orn, fermentation pair is carried out with the culture medium containing 20g/L yeast extracts (other compositions are identical) under identical fermentation condition
According to fermentation condition is：Inoculum concentration is 3%, 30 DEG C of fermentation temperature, and it is 6.8, the throughput of filtrated air that pH is controlled in fermentation process
For 0.5VVM, mixing speed 500rpm, when fermentation to residual glucose is less than 1g/L, fermentation is terminated.Finally with bacteria residue hydrolyzate
During for organic nitrogen source, L-Orn yield is 34.3g/L, and the L-Orn yield 35.2g/L compareed.
Embodiment 4
By the L-Orn fermentation fungi residues brine that is collected by centrifugation 2 times, the wet bacteria slag of collection is used
6.0mol/L sulfuric acid solutions are hydrolyzed, and the volume mass ratio of itself and wet bacteria slag solid content is 1：2.0, speed of agitator is 150rpm, water
It is 85 DEG C to solve temperature, and hydrolysis time is 24h.Arginine content is 4.21mg/mL in the hydrolyzate of acquisition, according to fermentation medium
Middle arginine concentrations are 250mg/L standard, and 60mL hydrolyzates replacement yeast extract is added to ferment in 1L fermentation mediums and made
Standby L-Orn, fermentation pair is carried out with the culture medium containing 20g/L yeast extracts (other compositions are identical) under identical fermentation condition
According to fermentation condition is：Inoculum concentration is 4%, 30 DEG C of fermentation temperature, and it is 6.9, the throughput of filtrated air that pH is controlled in fermentation process
For 1.0VVM, mixing speed 400rpm, when fermentation to residual glucose is less than 1g/L, fermentation is terminated.Finally with bacteria residue hydrolyzate
During for organic nitrogen source, L-Orn yield is 33.2g/L, and the L-Orn yield 34.5g/L compareed.
Embodiment 5
By the L-Orn fermentation fungi residues brine that is collected by centrifugation 2 times, the wet bacteria slag of collection is used
6.0mol/L sulfuric acid solutions are hydrolyzed, and the volume mass ratio of itself and wet bacteria slag solid content is 1：3.0, speed of agitator is 150rpm, water
It is 80 DEG C to solve temperature, and hydrolysis time is 18h.Arginine content is 4.11mg/mL in the hydrolyzate of acquisition, according to fermentation medium
Middle arginine concentrations are 300mg/L standard, and 73mL hydrolyzates replacement yeast extract is added to ferment in 1L fermentation mediums and made
Standby L-Orn, fermentation pair is carried out with the culture medium containing 45g/L yeast extracts (other compositions are identical) under identical fermentation condition
According to fermentation condition is：Inoculum concentration is 5%, 30 DEG C of fermentation temperature, and it is 7.0, the throughput of filtrated air that pH is controlled in fermentation process
For 1.5VVM, mixing speed 300rpm, when fermentation to residual glucose is less than 1g/L, fermentation is terminated.Finally with bacteria residue hydrolyzate
During for organic nitrogen source, L-Orn yield is 31.7g/L, and the L-Orn yield 23.5g/L compareed.
Claims (8)
1. a kind of method that the useless bacteria residue of L-Orn is back to L-Orn fermenting and producing, it is characterised in that it includes as follows
Step：
(1) will be standby after the useless bacteria residue washing of L-Orn；
(2) the useless bacteria residue of the L-Orn after being washed in step (1) is added and be hydrolyzed in aqueous sulfuric acid, centrifuged, take supernatant
Liquid, produces bacteria residue hydrolyzate；
(3) gained bacteria residue hydrolyzate in step (2) is prepared into L-Orn fermentation medium, and adjusted with ammoniacal liquor as nitrogen source
PH to 6.8~7.0；Strain is accessed after the sterilized processing of culture medium after regulation pH, for L-Orn fermentation.
2. the method according to claim 1 that the useless bacteria residue of L-Orn is back to L-Orn fermenting and producing, its feature
It is, in step (1), described L-Orn gives up bacteria residue for arginine-deficient type Corynebacterium glutamicum fermenting and producing L-Orn
Obtained wet bacteria slag is collected afterwards, and solid content is 23~25%.
3. the method according to claim 2 that the useless bacteria residue of L-Orn is back to L-Orn fermenting and producing, its feature
It is, described arginine-deficient type Corynebacterium glutamicum is in China Committee for Culture Collection of Microorganisms's common micro-organisms
Center preservation, deposit number is CGMCC No.3663.
4. the method according to claim 1 that the useless bacteria residue of L-Orn is back to L-Orn fermenting and producing, its feature
It is, in step (1), described washing refers to the useless bacteria residue of L-Orn with sodium-chloride water solution with 1：3 volume ratio mixing
Afterwards, 15~20min, abandoning supernatant are centrifuged with 8000rpm rotating speed；Wherein, the concentration of described sodium-chloride water solution is
0.9wt%, washing times are 2~3 times.
5. the method according to claim 1 that the useless bacteria residue of L-Orn is back to L-Orn fermenting and producing, its feature
It is, in step (2), the amount ratio of the useless bacteria residue of L-Orn after being washed in aqueous sulfuric acid and step (1) is 1mL：2~
3g；Wherein, the concentration of described aqueous sulfuric acid is 4.0~8.0mol/L.
6. the method according to claim 1 that the useless bacteria residue of L-Orn is back to L-Orn fermenting and producing, its feature
It is, in step (2), described hydrolysis refers at 80~90 DEG C, under 100~200rpm speed of agitator, hydrolysis 18~
24h。
7. the method according to claim 1 that the useless bacteria residue of L-Orn is back to L-Orn fermenting and producing, its feature
It is, in step (3), the composition of described L-Orn fermentation medium is：Bacteria residue hydrolyzate, glucose 90g/L,
K2HPO4·3H2O 1.5g/L, MgSO47H2O 3.7g/L, FeSO47H2O 20mg/L, MnSO4·H2O 20mg/L,
(NH4)2SO440g/L, Tween 80 0.6g/L, the μ g/L of biotin 50, the μ g/L of vitamin B1 100, pH6.8~7.0；Wherein, bacterium
The addition of slag hydrolyzate make it that arginic content is 200~300mg/L in L-Orn fermentation medium.
8. the method according to claim 1 that the useless bacteria residue of L-Orn is back to L-Orn fermenting and producing, its feature
It is, in step (3), the condition of L-Orn fermentation is：Inoculum concentration is 3~5%, and fermentation temperature is 30 DEG C, in fermentation process
With ammoniacal liquor control pH be 6.8~7.0, be passed through filtrated air fermentation, throughput be 0.5~1.5VVM, mixing speed 300~
500rpm, when fermentation to residual glucose is less than 1g/L, terminates fermentation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710492580.4A CN107034249A (en) | 2017-06-26 | 2017-06-26 | A kind of method that the useless bacteria residue of L ornithines is back to L ornithine fermenting and producings |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710492580.4A CN107034249A (en) | 2017-06-26 | 2017-06-26 | A kind of method that the useless bacteria residue of L ornithines is back to L ornithine fermenting and producings |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107034249A true CN107034249A (en) | 2017-08-11 |
Family
ID=59541276
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710492580.4A Pending CN107034249A (en) | 2017-06-26 | 2017-06-26 | A kind of method that the useless bacteria residue of L ornithines is back to L ornithine fermenting and producings |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107034249A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108048519A (en) * | 2017-12-25 | 2018-05-18 | 伊犁川宁生物技术有限公司 | A kind of cephalosporin bacteria residue processing method, hydrolyzate and its application |
| CN112725385A (en) * | 2019-10-28 | 2021-04-30 | 中国石油化工股份有限公司 | Method for preparing long-chain dicarboxylic acid by fermentation |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101955901A (en) * | 2010-09-19 | 2011-01-26 | 南京工业大学 | Corynebacterium glutamicum and method for preparing L-ornithine and salts thereof by using same |
-
2017
- 2017-06-26 CN CN201710492580.4A patent/CN107034249A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101955901A (en) * | 2010-09-19 | 2011-01-26 | 南京工业大学 | Corynebacterium glutamicum and method for preparing L-ornithine and salts thereof by using same |
Non-Patent Citations (1)
| Title |
|---|
| 耿晓玲等: "L-鸟氨酸发酵废菌渣水解固形物回用工艺的研究", 《食品研究与开发》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108048519A (en) * | 2017-12-25 | 2018-05-18 | 伊犁川宁生物技术有限公司 | A kind of cephalosporin bacteria residue processing method, hydrolyzate and its application |
| CN108048519B (en) * | 2017-12-25 | 2021-10-01 | 伊犁川宁生物技术有限公司 | Cephalosporin C fungus residue treatment method, hydrolysate and application thereof |
| CN112725385A (en) * | 2019-10-28 | 2021-04-30 | 中国石油化工股份有限公司 | Method for preparing long-chain dicarboxylic acid by fermentation |
| CN112725385B (en) * | 2019-10-28 | 2022-09-09 | 中国石油化工股份有限公司 | Method for preparing long-chain dicarboxylic acid by fermentation |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101544993B (en) | Method for producing L-lactic acid by Bacillus coagulans CGMCC No.2602 | |
| CN101560537B (en) | Fermentation method for producing high viscocity xanthan gum by xanthomonas campestris | |
| CN101979627A (en) | Method for preparing glutamic acid fermentation organic nitrogen additive from glutamic acid fermentation waste thalli | |
| CN101613726A (en) | Utilize microbial fermentation to prepare the method for transparent xanthan gum | |
| CN103409476A (en) | Fermentation, separation and purification method of L-isoleucine | |
| Pattanamanee et al. | Repeated-batch production of hydrogen using Rhodobacter sphaeroides S10 | |
| CN103088089B (en) | Method for fermenting acarbose | |
| CN106350558B (en) | Method for degrading feather by combining enzyme and bacteria | |
| CN107034249A (en) | A kind of method that the useless bacteria residue of L ornithines is back to L ornithine fermenting and producings | |
| CN101748162B (en) | Method for realizing nitrogen source recycling utilization by utilizing waste cell microbial fermentation | |
| CN104726381A (en) | Bacterial strain for producing L-lysine and L-lysine producing method thereof | |
| CN105219810B (en) | A kind of method of D-ALPHA-Hydroxypropionic acid and L-lysine Joint Production | |
| CN110129225A (en) | γ~polyglutamic acid producing strains and breeding prepare γ~polyglutamic acid method | |
| CN101886095A (en) | Method for producing high-concentration D-lactic acid by adopting synchronous enzymolysis and fermentation on peanut meal and special culture medium thereof | |
| CN108285914A (en) | A kind of zymotechnique of L-Trp | |
| CN105175275B (en) | A kind of isolation and purification method of L ornithine | |
| CN103952447A (en) | Method for producing succinic acid by virtue of fermentation under anaerobic conditions | |
| CN103305437B (en) | L-ammonium lactate tolerant bacterium and application thereof | |
| CN102191283B (en) | Method for preparing succinic acid through fermentation by cheap nitrogen sources | |
| CN101701206B (en) | Method for producing microorganism culture media nitrogen source from waste organisms | |
| CN110540982B (en) | Fermentation method for improving activity of Thermobacteroid cellulase | |
| CN107760734A (en) | A kind of method and its application for improving production amount of threonine | |
| CN103725724A (en) | Method of preparing cadaverine by virtue of immobilized hafnia alvei | |
| CN103911333B (en) | One strain is produced the bacterial strain of high yield phenylalanine and is produced the method for phenylalanine | |
| CN107058449A (en) | A kind of kitchen garbage bacillus amyloliquefaciens and the method for Lactobacillus rhamnosus mixed fermentation lactic acid producing |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170811 |
|
| RJ01 | Rejection of invention patent application after publication |