CN115141769A - Culture medium for improving thallus abundance of achromobacter or enterobacter and fermentation method thereof - Google Patents
Culture medium for improving thallus abundance of achromobacter or enterobacter and fermentation method thereof Download PDFInfo
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- 241000590020 Achromobacter Species 0.000 title claims abstract description 57
- 239000001963 growth medium Substances 0.000 title claims abstract description 54
- 241000588914 Enterobacter Species 0.000 title claims abstract description 31
- 238000000855 fermentation Methods 0.000 title claims abstract description 20
- 230000004151 fermentation Effects 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 title claims abstract description 13
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 32
- 239000000843 powder Substances 0.000 claims abstract description 32
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims abstract description 17
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims abstract description 17
- 229910052700 potassium Inorganic materials 0.000 claims abstract description 17
- 239000011591 potassium Substances 0.000 claims abstract description 17
- 235000019764 Soybean Meal Nutrition 0.000 claims abstract description 15
- 239000008367 deionised water Substances 0.000 claims abstract description 15
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 15
- 239000004455 soybean meal Substances 0.000 claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000001888 Peptone Substances 0.000 claims description 27
- 108010080698 Peptones Proteins 0.000 claims description 27
- 235000019319 peptone Nutrition 0.000 claims description 27
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 16
- 239000002609 medium Substances 0.000 claims description 12
- 241000982938 Enterobacter cancerogenus Species 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 5
- 241000588921 Enterobacteriaceae Species 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 241001052560 Thallis Species 0.000 claims description 2
- 238000009472 formulation Methods 0.000 claims 2
- 244000005700 microbiome Species 0.000 abstract description 5
- 230000000052 comparative effect Effects 0.000 description 34
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 28
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 22
- 239000007788 liquid Substances 0.000 description 20
- 230000001580 bacterial effect Effects 0.000 description 18
- 239000011780 sodium chloride Substances 0.000 description 14
- 244000046052 Phaseolus vulgaris Species 0.000 description 12
- 235000010627 Phaseolus vulgaris Nutrition 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
- 238000011081 inoculation Methods 0.000 description 11
- 229910052757 nitrogen Inorganic materials 0.000 description 11
- 229910017053 inorganic salt Inorganic materials 0.000 description 10
- 238000012360 testing method Methods 0.000 description 9
- 241000305071 Enterobacterales Species 0.000 description 8
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 7
- 229920002261 Corn starch Polymers 0.000 description 7
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 7
- 229920002472 Starch Polymers 0.000 description 7
- 229930006000 Sucrose Natural products 0.000 description 7
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 7
- 239000008120 corn starch Substances 0.000 description 7
- 239000008107 starch Substances 0.000 description 7
- 235000019698 starch Nutrition 0.000 description 7
- 239000005720 sucrose Substances 0.000 description 7
- 230000003287 optical effect Effects 0.000 description 6
- 238000013461 design Methods 0.000 description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 description 5
- 238000005457 optimization Methods 0.000 description 5
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 5
- 239000012880 LB liquid culture medium Substances 0.000 description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 241000193830 Bacillus <bacterium> 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
- 230000000694 effects Effects 0.000 description 2
- 238000013401 experimental design Methods 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 241000932047 Achromobacter sp. Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000147019 Enterobacter sp. Species 0.000 description 1
- 241000881812 Kluyvera intermedia Species 0.000 description 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229910052793 cadmium Inorganic materials 0.000 description 1
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 description 1
- 235000010216 calcium carbonate Nutrition 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- SDCJMBBHNJPYGW-UHFFFAOYSA-L disodium;hydrogen carbonate;chloride Chemical compound [Na+].[Na+].Cl.[O-]C([O-])=O SDCJMBBHNJPYGW-UHFFFAOYSA-L 0.000 description 1
- 229910052564 epsomite Inorganic materials 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
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- 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
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/025—Achromobacter
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Abstract
The invention relates to the technical field of microorganisms, and discloses a fermentation method for improving the abundance of achromobacter: based on a culture medium containing 0.4-0.5% of yeast powder, 0.5-1.5% of soybean meal, 0.5-1.5% of potassium dihydrogen and the balance of deionized water, the pH is adjusted to 8-8.5, the bottling amount is 10-20%, the temperature is 26-28 ℃, and the rotating speed is 160-220rpm for shake cultivation. Also disclosed is a fermentation process for providing enterobacter abundance: based on a culture medium containing 0.4-0.5% of yeast powder, 0.5-1.5% of soybean meal, 0.5-1.5% of potassium dihydrogen and the balance of deionized water, the pH is adjusted to 8-8.5, the bottling amount is 10-20%, the temperature is 26-28 ℃, and the rotating speed is 160-220rpm for shake cultivation. Can realize the rapid and large-scale expanded culture of the achromobacter or the enterobacter and provide a foundation for the industrial application of the achromobacter or the enterobacter.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a culture medium for improving the abundance of achromobacter or enterobacter and a fermentation method thereof.
Background
The Chinese invention patent (CN 109266575A) discloses a cadmium-tolerant Enterobacter (Enterobacter sp.) FM-1 which is subjected to amplification culture by adopting an LB liquid culture medium, wherein the LB liquid culture medium is prepared by containing 5g of beef extract, 10g of peptone and 5g of NaCl per liter of culture medium, and is sterilized by high-pressure steam at 121 ℃ for 20min and has natural pH.
The Chinese invention patent (CN 105062926A) discloses Achromobacter (Achromobacter sp.) H380 for heavy metal cadmium pollution control and application thereof, wherein an H380 strain is inoculated into a fresh 0.5-time LB culture medium for activation overnight, then the H380 strain is inoculated into the fresh 0.5-time LB culture medium in an inoculation amount of 1% (v/v), and the H380 strain is cultured in a constant-temperature shaking table at 28 ℃ until OD600 =0.5.
In the practical application of functional microorganisms such as enterobacteria, achromobacter and the like, the demand for bacteria is huge, and the common LB liquid culture medium is difficult to meet the demand for rapidly expanding and culturing the enterobacteria and the achromobacter.
Disclosure of Invention
In order to solve the technical problem that the rapid expansion culture is difficult in the practical application of functional microorganisms such as enterobacteria or achromobacter and the like, the invention provides the following technical scheme:
a culture medium for improving the abundance of Achromobacter thalli, which comprises the following components: 0.4-0.5% of yeast powder, 0.5-1.5% of soybean meal, 0.5-1.5% of potassium dihydrogen and the balance of deionized water.
Preferably, the medium comprises the following components: 0.4 percent of yeast powder, 1.0 percent of soybean meal, 1.5 percent of potassium dihydrogen and the balance of deionized water.
Preferably, the Achromobacter is Achromobacter (Achromobacter inuavis) SL8 with the deposit number: CCTCC M2022286.
The invention also provides a fermentation method for improving the abundance of achromobacter, which comprises the following steps: inoculating Achromobacter bacterium solution into the culture medium of any one of claims 1-3, adjusting the pH of the culture medium to 8-10, bottling at 20-40%, culturing at 26-28 deg.C and 160-220 rpm.
Preferably, in the fermentation method of the achromobacter abundance, the pH of the culture medium is 9, the bottling amount is 30%, the temperature is 27 ℃, and the rotating speed is 180rpm.
A culture medium for improving the abundance of enterobacter strains, which comprises the following components: 0.4-0.6% of yeast powder, 1.0-1.5% of peptone, 1.0-1.5% of dipotassium hydrogen phosphate and the balance of deionized water.
Preferably, the medium comprises the following components: 0.6 percent of yeast powder, 1.5 percent of peptone, 1.5 percent of dipotassium hydrogen phosphate and the balance of deionized water
Preferably, the enterobacteria is enterobacteria (Enterobacter cancerogenus) SL12 with a deposit number: CCTCC M2022288.
A fermentation method for improving the abundance of enterobacteriaceae, which comprises the following steps: inoculating the bacterial liquid of enterobacter to a culture medium containing 0.4-0.6% of yeast powder, 1.0-1.5% of peptone, 1.0-1.5% of dipotassium hydrogen phosphate and the balance of deionized water, adjusting the pH of the culture medium to 8-8.5, bottling the mixture in a shake bed at the temperature of 26-28 ℃ and the rotating speed of 160-220rpm, and culturing.
Preferably, in the fermentation method for increasing the abundance of enterobacteriaceae, the pH of the culture medium is 8.2, the bottling amount is 15%, the temperature is 26 ℃, and the rotating speed is 180rpm.
Compared with the prior art, the invention has the beneficial effects that:
compared with the common LB liquid culture medium and the conventional fermentation method thereof, the culture medium for improving the thallus abundance of the achromobacter or the enterobacter can quickly realize the expanded culture of the achromobacter or the enterobacter and provide a basis for the industrial application of the achromobacter or the enterobacter.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are a part of the embodiments of the present invention, but not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The Achromobacter in the invention is Achromobacter (Achromobacter inuavis) SL8, and the nucleotide sequence of 16SrDNA is shown as SE Q ID NO: 1; the other strain is Enterobacter (Enterobacter cancerogenus) SL12, and the nucleotide sequence of 16SrDNA is shown as SEQ ID NO. 2. Accession number of Enterobacter (Enterobacter cancerogenus) SL 12: CCTCC M2022288; achromobacter (Achromobacter angualis) SL8 deposit number: CCTCC M2022286. The 2 microorganisms are preserved in China Center for Type Culture Collection (CCTCC) No. 18 at No. 3 month 2022, and the preservation addresses are as follows: wuhan university, wuhan, china.
Test one: medium composition optimization
The basic conditions for strain culture are as follows: the seed solution was inoculated into LB liquid medium at an inoculum size of 0.5%, liquid content 100% (V/V), and cultured on a shaker at 30 ℃ and 160rpm for 24 hours, with 4 replicates for each treatment. Fixing other elements, and replacing the carbon source, the nitrogen source and the inorganic salt in the basic culture medium with different carbon sources, nitrogen sources and inorganic salts respectively. The replacement carbon source and the addition amount are as follows: 0.5% of yeast powder, corn starch, sucrose, soluble starch, maltose and NaHCO 3 (ii) a The replacement nitrogen source and the addition amount are as follows: 1.0% peptone, soybean meal, KNO 3 (ii) a The replacing inorganic salt and the addition amount are as follows: 1.0% of NaCl, dihydropotassium and K 2 HPO 4 、 MgSO 4 ·7H 2 O、CaCO 3 . In the present invention, (%) represents mass percent.
The 36 groups of media were designed as shown in table 1.
TABLE 1 experimental group design
| Numbering | Nitrogen source | Carbon source | Inorganic salt |
| 1 | KNO 3 | Sucrose | NaCl |
| 2 | Bean pulp | Yeast powder | MgSO 4 ·7H 2 O |
| 3 | Peptone | Soluble starch | Potassium dihydrogen phosphate |
| 4 | KNO 3 | NaHCO 3 | CaCO 3 |
| 5 | Bean pulp | Corn starch | CaCO 3 |
| 6 | KNO 3 | Corn starch | Potassium dihydrogen |
| 7 | Peptone | Yeast powder | CaCO 3 |
| 8 | Peptone | Maltose | NaCl |
| 9 | Bean pulp | Sucrose | MgSO 4 ·7H 2 O |
| 10 | Bean pulp | NaHCO 3 | Potassium dihydrogen |
| 11 | Peptone | Sucrose | Dipotassium hydrogen phosphate |
| 12 | KNO 3 | Yeast powder | NaCl |
| 13 | Bean pulp | Maltose | Dipotassium hydrogen phosphate |
| 14 | Peptone | Corn starch | NaCl |
| 15 | Bean pulp | Soluble starch | Dipotassium hydrogen phosphate |
| 16 | KNO 3 | Soluble starch | NaCl |
| 17 | KNO 3 | Maltose | MgSO 4 ·7H 2 O |
| 18 | Peptone | NaHCO 3 | NaCl |
| 19 | Peptone | Maltose | Potassium dihydrogen phosphate |
| 20 | Bean pulp | NaHCO3 | NaCl |
| 21 | KNO 3 | Soluble starch | MgSO4·7H2O |
| 22 | Bean pulp | Maltose | NaCl |
| 23 | KNO 3 | Corn starch | Dipotassium hydrogen phosphate |
| 24 | Peptone | Yeast powder | Dipotassium hydrogen phosphate |
| 25 | Peptone | Soluble starch | NaCl |
| 26 | KNO 3 | NaHCO 3 | Dipotassium hydrogen phosphate |
| 27 | Bean pulp | Sucrose | NaCl |
| 28 | KNO3 | Yeast powder | Potassium dihydrogen phosphate |
| 29 | Peptone | Sucrose | CaCO 3 |
| 30 | Peptone | Corn starch | MgSO 4 ·7H 2 O |
| 31 | Bean pulp | Soluble starch | CaCO3 |
| 32 | Peptone | NaHCO 3 | MgSO 4 ·7H 2 O |
| 33 | Bean pulp | Corn starch | NaCl |
| 34 | KNO 3 | Maltose | CaCO 3 |
| 35 | KNO 3 | Sucrose | Potassium dihydrogen phosphate |
| 36 | Bean pulp | Yeast powder | Potassium dihydrogen |
Respectively selecting the experimental design in the table 1 as a culture medium, inoculating the bacterial liquid into a 100mL triangular flask with adjusted pH according to the inoculation amount of 0.5%, performing shake culture at 30 ℃ and 160rpm for 24 hours, and then determining OD (optical density) of the bacterial liquid 600 The value is obtained. HAc, naOH/ammonia was used for pH adjustment. The results are shown in table 2:
TABLE 2 Achromobacter and Enterobacter strains abundance after experimental design in TABLE 1
| Numbering | Achromobacter abundance (hundred million CFU/mL) | Enterobacter abundance (hundred million CFU/mL) |
| 1 | 0.0 | 0.1 |
| 2 | 10.7 | 13.0 |
| 3 | 6.4 | 6.0 |
| 4 | 0.0 | 0.1 |
| 5 | 12.6 | 10.4 |
| 6 | 4.1 | 3.8 |
| 7 | 15.2 | 12.4 |
| 8 | 6.5 | 20.8 |
| 9 | 3.8 | 5.3 |
| 10 | 4.6 | 4.2 |
| 11 | 8.8 | 11.8 |
| 12 | 7.9 | 10.4 |
| 13 | 10.6 | 13.9 |
| 14 | 8.4 | 14.3 |
| 15 | 6.1 | 15.8 |
| 16 | 0.4 | 0.5 |
| 17 | 0.3 | 0.3 |
| 18 | 5.4 | 9.8 |
| 19 | 6.9 | 15.8 |
| 20 | 3.7 | 15.6 |
| 21 | 6.6 | 0.3 |
| 22 | 7.6 | 10.1 |
| 23 | 3.3 | 3.1 |
| 24 | 9.2 | 20.7 |
| 25 | 1.4 | 10.6 |
| 26 | 0.8 | 0.0 |
| 27 | 4.2 | 6.5 |
| 28 | 19.6 | 8.9 |
| 29 | 5.4 | 7.3 |
| 30 | 10.1 | 5.8 |
| 31 | 10.0 | 7.9 |
| 32 | 2.7 | 0.1 |
| 33 | 7.1 | 7.7 |
| 34 | 0.1 | 0.5 |
| 35 | 2.1 | 0.2 |
| 36 | 24.7 | 26.4 |
As shown in Table 2, the abundance of Achromobacter after culture with the treatment 7 medium was 15.2 hundred million CFU/mL; the abundance of Achromobacter after the culture with the treated 28 medium was 19.6 hundred million CFU/mL, and the abundance of Achromobacter after the culture with the treated 36 medium was 24.7 hundred million CFU/mL, and thus it was confirmed that the carbon source of Achromobacter was yeast powder, and the nitrogen source was soybean meal, peptone, and KNO 3 (ii) a The inorganic salt is NaCl, potassium dihydrogen or CaCO 3 。
With the culture medium of treatment 8The abundance of the cultured enterobacteria is 20.8 hundred million CFU/mL; the abundance of enterobacter by the culture medium of treatment 24 was 20.7 hundred million CFU/mL, and the abundance of achromobacter by the culture medium of treatment 36 was 26.4 hundred million CFU/mL, so it was confirmed that the carbon source of the achromobacter was yeast powder, and the nitrogen source was soybean meal and peptone; the inorganic salt is NaCl, potassium dihydrogen or K 2 HPO 4 。
(3) Orthogonal test optimization
Test factors and levels are shown in table 3, and an orthogonal test was designed to further determine the optimum ratio. The orthogonal test level design is shown in table 4.
TABLE 3 orthogonal test factors and levels
TABLE 4 orthogonal experimental treatment design
| Treatment of | Carbon source | Nitrogen source | Inorganic salt |
| 1 | 0.4％ | 0.5％ | 0.5％ |
| 2 | 0.4％ | 1.0％ | 1.5％ |
| 3 | 0.4％ | 1.5％ | 1.0％ |
| 4 | 0.5％ | 0.5％ | 1.5％ |
| 5 | 0.5％ | 1.0％ | 1.0％ |
| 6 | 0.5％ | 1.5％ | 0.5％ |
| 7 | 0.6％ | 0.5％ | 1.0％ |
| 8 | 0.6％ | 1.0％ | 0.5％ |
| 9 | 0.6％ | 1.5％ | 1.5％ |
(4) The colorless bacillus adopts the orthogonal test design in table 4, the bacterial liquid is inoculated into a 100mL triangular flask with natural pH according to the inoculation amount of 0.5%, shaking culture is carried out at 30 ℃ and 160rpm for 24h, the bacterial abundance of the colorless bacillus is detected, and the result is shown in table 5:
TABLE 5 abundance data of Achromobacter in different ratios of carbon source, nitrogen source and inorganic salt
In Table 5, the cell abundance of Achromobacter in the 6-ratio treated medium was 22.1 hundred million CFU/mL; the thallus abundance of the achromobacter culture medium with the treatment ratio of 19 is 22.9 hundred million CFU/mL; the thallus abundance of the culture medium Achromobacter with the processing ratio of 20 is 26.6 hundred million CFU/mL; the thallus abundance of the achromobacter in the culture medium with the processing ratio of 24 is 22.4 hundred million CFU/mL; therefore, the ratio of the culture medium suitable for the achromobacter is as follows: 0.4-0.5% of yeast powder, 0.5-1.5% of soybean meal, 0.5-1.5% of potassium dihydrogen and the balance of deionized water. The optimal culture medium ratio is as follows: 0.4% of yeast powder, 1.0% of soybean meal, 1.5% of potassium dihydrogen and the balance of deionized water.
(5) The enterobacter adopts the orthogonal test design in Table 4, the bacterial liquid is inoculated into a 100mL triangular flask with adjusted pH according to the inoculum concentration of 0.5 percent, shaking culture is carried out for 24 hours at 30 ℃ and 160rpm, the bacterial abundance of the achromobacter is detected, and the result is shown in Table 6:
TABLE 6 abundance data of Enterobacter under different ratios of carbon source, nitrogen source and inorganic salt
In Table 6, the cell abundance of Enterobacter bacteria treated with the medium at 12 ratios was 18.7 hundred million CFU/mL; the thallus abundance of the achromobacter in the culture medium with the treatment ratio of 18 is 23.0 hundred million CFU/mL; the thallus abundance of the enterobacter in the culture medium with the treated ratio of 20 is 26.6 hundred million CFU/mL; the thallus abundance of the enterobacter by adopting a culture medium with a treated 26-proportion ratio is 17.8 hundred million CFU/mL; therefore, the ratio of the culture medium suitable for the enterobacteria is as follows: 0.4-0.6% of yeast powder, 1.0-1.5% of peptone, 1.0-1.5% of dipotassium hydrogen phosphate and the balance of deionized water. The optimal culture medium ratio is as follows: 0.6 percent of yeast powder, 1.5 percent of peptone, 1.5 percent of dipotassium hydrogen phosphate and the balance of deionized water.
And (2) test II: achromobacter and enterobacter fermentation condition optimization
(1) Fermentation condition optimization of achromobacter
Example 1
Selecting 0.4% yeast powder, 1.0% soybean meal and 1.5% potassium dihydrogen as initial values of a culture medium, inoculating the bacterial liquid into a 100mL triangular flask with the adjusted pH value of 8 according to the inoculation amount of 0.5%, bottling the bacterial liquid into the flask in an amount of 20%, performing shake culture at the temperature of 26 ℃ and the rotating speed of 160rpm for 24 hours, and then measuring OD (optical density) after shaking culture 600 The value is obtained.
Example 2
Selecting 0.4% yeast powder, 1.0% soybean meal and 1.5% potassium dihydrogen as initial values of a culture medium, inoculating the bacterial liquid into a 100mL triangular flask with the adjusted pH of 9 according to the inoculation amount of 0.5%, bottling the bacterial liquid in the amount of 30%, performing shake culture at the temperature of 27 ℃ and the rotation speed of 180rpm for 24 hours, and determining OD (optical density) after shaking culture 600 The value is obtained.
Example 3
Selecting 0.4% yeast powder, 1.0% soybean meal and 1.5% potassium dihydrogen as initial values of a culture medium, inoculating the bacterial liquid into a 100mL triangular flask with the adjusted pH value of 10 according to the inoculation amount of 0.5%, bottling the bacterial liquid at the bottling amount of 40%, performing shake culture at the temperature of 28 ℃ and the rotation speed of 220rpm for 24 hours, and then measuring OD (optical density) after shake culture 600 The value is obtained.
The difference between comparative example 1 and example 1 is: the bottling amount is 10%.
The difference between comparative example 2 and example 1 is: the bottling amount is 50%.
The difference between comparative example 3 and example 1 is: the pH was 6.
The comparative example 4 differs from example 1 in that: the pH was 9.
The comparative example 5 differs from example 1 in that: the temperature was 23 ℃.
The difference between comparative example 6 and example 1 is: the temperature was 30 ℃.
The difference between comparative example 7 and example 1 is: the rotation speed was 140rpm.
The difference between comparative example 8 and example 1 is: the rotation speed was 250rpm.
The culture medium of comparative example 9 was a liquid LB medium.
TABLE 7 abundance of Achromobacter in examples 1 to 3 and comparative examples 1 to 9
| Treatment of | Abundance of Achromobacter (hundred million CFU/mL) |
| Example 1 | 72.1 |
| Example 2 | 75.6 |
| Example 3 | 73.8 |
| Comparative example 1 | 15.6 |
| Comparative example 2 | 23.4 |
| Comparative example 3 | 26.1 |
| Comparative example 4 | 29.8 |
| Comparative example 5 | 31.2 |
| Comparative example 6 | 32.8 |
| Comparative example 7 | 28.6 |
| Comparative example 8 | 29.7 |
| Comparative example 9 | 2.9 |
Table 7 shows that in the expanded culture of achromobacter, yeast powder is selected as a carbon source, soybean meal is selected as a nitrogen source, and potassium dihydrogen is used as an inorganic salt, so that the bottling amount, the pH, the temperature and the rotating speed of a culture medium in the fermentation culture are optimized, and compared with a comparative ratio of 9, the thallus abundance of the achromobacter is remarkably improved, and an unexpected technical effect is achieved; in addition, the data of comparative examples 1-8 show that the technical scheme of the invention is that the bottling amount, the pH value, the temperature and the rotating speed of the culture medium are continuously optimized, the thallus abundance is synergistically improved, and the simple conventional regulation is not performed.
The abundance of the achromobacter in the technical schemes of examples 1-3 is significantly higher than that of the data of comparative examples 1-9,
(2) Fermentation condition optimization of enterobacteria
Example 1
Selecting 0.6% yeast powder, 1.5% peptone and 1.5% dipotassium hydrogen phosphate as initial culture medium values, inoculating the bacterial liquid into a 100mL triangular flask with the adjusted pH value of 8 according to the inoculation amount of 0.5%, and packagingThe OD is measured after shake culture at 26 ℃ and 160rpm for 24h with a bottle amount of 10% 600 The value is obtained.
Example 2
Selecting 0.6% yeast powder, 1.5% peptone and 1.5% dipotassium hydrogen phosphate as initial culture medium values, inoculating the bacterial liquid into a 100mL triangular flask with the adjusted pH of 8.2 according to the inoculation amount of 0.5%, bottling the bacterial liquid into a bottle with the inoculation amount of 15%, performing shake culture at 26 ℃ and 180rpm for 24h, and determining OD (optical density) after shaking culture 600 The value is obtained.
Example 3
Selecting 0.6% yeast powder, 1.5% peptone and 1.5% dipotassium hydrogen phosphate as initial culture medium values, inoculating the bacterial liquid into a 100mL triangular flask with the adjusted pH value of 8.5 according to the inoculation amount of 0.5%, bottling the bacterial liquid with the inoculation amount of 30%, performing shake culture at 26 ℃ and 220rpm for 24h, and determining OD (optical density) after shaking culture 600 The value is obtained.
The difference between comparative example 1 and example 1 is: the bottling amount is 5%.
The difference between comparative example 2 and example 1 is: the bottling amount is 30 percent.
The difference between comparative example 3 and example 1 is: the pH was 7.
The difference between comparative example 4 and example 1 is: the pH was 9.
The difference between comparative example 5 and example 1 is: the temperature was 23 ℃.
The difference between comparative example 6 and example 1 is: the temperature was 30 ℃.
The comparative example 7 differs from example 1 in that: the rotation speed was 140rpm.
The comparative example 8 differs from example 1 in that: the rotation speed was 250rpm.
The culture medium of comparative example 9 was a liquid LB medium.
TABLE 8 cell abundance of Enterobacter intermedius in examples 1 to 3 and comparative examples 1 to 9
Table 8 shows that in the expanded culture of enterobacter, yeast powder is selected as a carbon source, peptone is selected as a nitrogen source, and potassium dihydrogen phosphate is used as an inorganic salt, so that the bottling amount, the pH value, the temperature and the rotating speed of a culture medium in fermentation culture are optimized, compared with comparative example 9, the thallus abundance of the enterobacter is obviously improved, and an unexpected technical effect is achieved; in addition, the data of comparative examples 1-8 show that the technical scheme of the invention is that the bottling amount, the pH value, the temperature and the rotating speed of the culture medium are continuously optimized, the thallus abundance is synergistically improved, and the method is not simple conventional selection.
Sequence listing
<110> vegetable research institute of Hunan province
<120> culture medium for increasing abundance of Achromobacter or Enterobacter, and fermentation method thereof
<141> 2022-06-21
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1398
<212> DNA
<213> Achromobacter (Achromobacter inuavis)
<400> 1
ggtatcgccc cccttgcggt taggctaact acttctggta aaacccactc ccatggtgtg 60
acgggcggtg tgtacaagac ccgggaacgt attcaccgcg acatgctgat ccgcgattac 120
tagcgattcc gacttcacgc agtcgagttg cagactgcga tccggactac gatcgggttt 180
ctgggattgg ctccccctcg cgggttggcg accctctgtc ccgaccattg tatgacgtgt 240
gaagccctac ccataagggc catgaggact tgacgtcatc cccaccttcc tccggtttgt 300
caccggcagt ctcattagag tgccctttcg tagcaactaa tgacaagggt tgcgctcgtt 360
gcgggactta acccaacatc tcacgacacg agctgacgac agccatgcag cacctgtgtt 420
ccggttctct tgcgagcact gccaaatctc ttcagcattc cagacatgtc aagggtaggt 480
aaggtttttc gcgttgcatc gaattaatcc acatcatcca ccgcttgtgc gggtccccgt 540
caattccttt gagttttaat cttgcgaccg tactccccag gcggtcaact tcacgcgtta 600
gctgcgctac caaggcccga aggccccaac agctagttga catcgtttag ggcgtggact 660
accagggtat ctaatcctgt ttgctcccca cgctttcgtg catgagcgtc agtgttatcc 720
caggaggctg ccttcgccat cggtgttcct ccgcatatct acgcatttca ctgctacacg 780
cggaattcca cctccctctg acacactcta gcccggtagt taaaaatgca gttccaaagt 840
taagctctgg gatttcacat ctttctttcc gaaccgcctg cgcacgcttt acgcccagta 900
attccgatta acgcttgcac cctacgtatt accgcggctg ctggcacgta gttagccggt 960
gcttattctg caggtaccgt cagtttcgcg gggtattaac ccacgacgtt tctttcctgc 1020
caaaagtgct ttacaacccg aaggccttca tcgcacacgc gggatggctg gatcagggtt 1080
tcccccattg tccaaaattc cccactgctg cctcccgtag gagtctgggc cgtgtctcag 1140
tcccagtgtg gctggtcgtc ctctcaaacc agctacggat cgtcgccttg gtgagccgtt 1200
accccaccaa ctagctaatc cgatatcggc cgctccaata gtgcaaggtc ttgcgatccc 1260
ctgctttccc ccgtagggcg tatgcggtat tagctacgct ttcgcgtagt tatcccccgc 1320
tactgggcac gttccgatac attactcacc cgttcgccac tcgccaccag accgaagtcc 1380
gtgctgccgt tcgactgc 1398
<210> 2
<211> 1393
<212> DNA
<213> Enterobacter (Enterobacter cancerogenus)
<400> 2
ccctcccgaa ggttaagcta cctacttctt ttgcaaccca ctcccatggt gtgacgggcg 60
gtgtgtacaa ggcccgggaa cgtattcacc gtagcattct gatctacgat tactagcgat 120
tccgacttca tggagtcgag ttgcagactc caatccggac tacgacgcac tttatgaggt 180
ccgcttgctc tcgcgaggtc gcttctcttt gtatgcgcca ttgtagcacg tgtgtagccc 240
tactcgtaag ggccatgatg acttgacgtc atccccacct tcctccagtt tatcactggc 300
agtctccttt gagttcccgg ccggaccgct ggcaacaaag gataagggtt gcgctcgttg 360
cgggacttaa cccaacattt cacaacacga gctgacgaca gccatgcagc acctgtctca 420
gagttcccga aggcaccaat ccatctctgg aaagttctct ggatgtcaag agtaggtaag 480
gttcttcgcg ttgcatcgaa ttaaaccaca tgctccaccg cttgtgcggg cccccgtcaa 540
ttcatttgag ttttaacctt gcggccgtac tccccaggcg gtcgacttaa cgcgttagct 600
ccggaagcca cgcctcaagg gcacaacctc caagtcgaca tcgtttacgg cgtggactac 660
cagggtatct aatcctgttt gctccccacg ctttcgcacc tgagcgtcag tctttgtcca 720
gggggccgcc ttcgccaccg gtattcctcc agatctctac gcatttcacc gctacacctg 780
gaattctacc cccctctaca agactctagc ctgccagttt cgaatgcagt tcccaggttg 840
agcccgggga tttcacatcc gacttgacag accgcctgcg tgcgctttac gcccagtaat 900
tccgattaac gcttgcaccc tccgtattac cgcggctgct ggcacggagt tagccggtgc 960
ttcttctgcg ggtaacgtca atcgatgagg ttattaacct caccgccttc ctccccgctg 1020
aaagtacttt acaacccgaa ggccttcttc atacacgcgg catggctgca tcaggcttgc 1080
gcccattgtg caatattccc cactgctgcc tcccgtagga gtctggaccg tgtctcagtt 1140
ccagtgtggc tggtcatcct ctcagaccag ctagggatcg tcgcctaggt gagccgttac 1200
cccacctact agctaatccc atctgggcac atctgatggc aagaggcccg aaggtccccc 1260
tctttggtct tgcgacgtta tgcggtatta gctaccgttt ccagtagtta tccccctcca 1320
tcaggcagtt tcccagacat tactcacccg tccgccgctc gtcacccgag agcaagctct 1380
ctgtgctacc gct 1393
Claims (10)
1. A culture medium for improving the abundance of Achromobacter thalli, which is characterized by comprising the following components: 0.4-0.5% of yeast powder, 0.5-1.5% of soybean meal, 0.5-1.5% of potassium dihydrogen and the balance of deionized water.
2. The media formulation of claim 1, wherein the media comprises the following components: 0.4% of yeast powder, 1.0% of soybean meal, 1.5% of potassium dihydrogen and the balance of deionized water.
3. The culture medium formulation according to claim 1 or 2, wherein the Achromobacter is Achromobacter (Achromobacter inuavis) SL8 with a deposit number of: CCTCC M2022286.
4. A fermentation method for improving the abundance of achromobacter is characterized by comprising the following steps: inoculating Achromobacter bacterium solution into the culture medium of any one of claims 1-3, adjusting the pH of the culture medium to 8-10, bottling at 20-40%, culturing at 26-28 deg.C and 160-220 rpm.
5. The fermentation process according to claim 4, wherein the culture medium has a pH of 9 and a bottling capacity of 30%, a temperature of 27 ℃ and a rotation speed of 180rpm.
6. A culture medium for improving the abundance of enterobacter strains is characterized by comprising the following components: 0.4-0.6% of yeast powder, 1.0-1.5% of peptone, 1.0-1.5% of dipotassium hydrogen phosphate and the balance of deionized water.
7. The culture medium according to claim 6, comprising the following components: 0.6 percent of yeast powder, 1.5 percent of peptone, 1.5 percent of dipotassium hydrogen phosphate and the balance of deionized water.
8. The medium according to claim 6 or 7, wherein the Enterobacter is Enterobacter (Enterobacter cancerogenus) SL12 with a deposit number: CCTCC M2022288.
9. A fermentation method for improving the abundance of enterobacteriaceae is characterized by comprising the following steps: inoculating Enterobacter bacteria solution into the culture medium of any one of claims 6-8, adjusting pH of the culture medium to 8-8.5, bottling 10-20%, culturing at 26-28 deg.C and 160-220 rpm.
10. Fermentation process according to claim 9, characterised in that the medium has a pH of 8.2 and a bottling quantity of 15%, a temperature of 26 ℃ and a rotation speed of 180rpm.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116179494A (en) * | 2022-10-13 | 2023-05-30 | 武汉生物制品研究所有限责任公司 | MDCK cell strain with low neoplasia, construction method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009005677A (en) * | 2007-05-11 | 2009-01-15 | Sumitomo Chemical Co Ltd | Method for producing malic acid |
| CN104531565A (en) * | 2014-12-10 | 2015-04-22 | 浙江工业大学 | Achromobacter denitrificans zjut1104 and application thereof |
| CN107988096A (en) * | 2017-03-10 | 2018-05-04 | 厦门天邦润生物科技有限公司 | A kind of multifunctional microbial soil-repairing agent and its preparation and application |
| CN110194708A (en) * | 2019-07-05 | 2019-09-03 | 湖南泰谷生态工程有限公司 | A kind of ferment slag soil remediation microbial inoculum and the preparation method and application thereof |
| CN110627582A (en) * | 2019-10-09 | 2019-12-31 | 山东省科学院能源研究所 | Preparation method of solid carbon-based fertilizer and liquid pesticide-fertilizer integrated preparation |
| CN110699291A (en) * | 2019-11-04 | 2020-01-17 | 浙江树人学院（浙江树人大学） | Achromobacter xylosoxidans with sulfide degradation performance and application thereof |
-
2022
- 2022-06-21 CN CN202210702104.1A patent/CN115141769A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009005677A (en) * | 2007-05-11 | 2009-01-15 | Sumitomo Chemical Co Ltd | Method for producing malic acid |
| CN104531565A (en) * | 2014-12-10 | 2015-04-22 | 浙江工业大学 | Achromobacter denitrificans zjut1104 and application thereof |
| CN107988096A (en) * | 2017-03-10 | 2018-05-04 | 厦门天邦润生物科技有限公司 | A kind of multifunctional microbial soil-repairing agent and its preparation and application |
| CN110194708A (en) * | 2019-07-05 | 2019-09-03 | 湖南泰谷生态工程有限公司 | A kind of ferment slag soil remediation microbial inoculum and the preparation method and application thereof |
| CN110627582A (en) * | 2019-10-09 | 2019-12-31 | 山东省科学院能源研究所 | Preparation method of solid carbon-based fertilizer and liquid pesticide-fertilizer integrated preparation |
| CN110699291A (en) * | 2019-11-04 | 2020-01-17 | 浙江树人学院（浙江树人大学） | Achromobacter xylosoxidans with sulfide degradation performance and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 陈雄 等: "响应面法优化地衣芽孢杆菌A2发酵培养基", 《中国饲料》, no. 7, pages 18 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116179494A (en) * | 2022-10-13 | 2023-05-30 | 武汉生物制品研究所有限责任公司 | MDCK cell strain with low neoplasia, construction method and application thereof |
| CN116179494B (en) * | 2022-10-13 | 2024-01-26 | 武汉生物制品研究所有限责任公司 | MDCK cell strain with low neoplasia, construction method and application thereof |
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Application publication date: 20221004 |