CN113957115A - Fermentation method for high yield of gibberellic acid - Google Patents
Fermentation method for high yield of gibberellic acid Download PDFInfo
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- CN113957115A CN113957115A CN202111157633.XA CN202111157633A CN113957115A CN 113957115 A CN113957115 A CN 113957115A CN 202111157633 A CN202111157633 A CN 202111157633A CN 113957115 A CN113957115 A CN 113957115A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 103
- 230000004151 fermentation Effects 0.000 title claims abstract description 102
- 238000000034 method Methods 0.000 title claims abstract description 35
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 title claims abstract description 30
- IXORZMNAPKEEDV-OBDJNFEBSA-N gibberellin A3 Chemical compound C([C@@]1(O)C(=C)C[C@@]2(C1)[C@H]1C(O)=O)C[C@H]2[C@]2(C=C[C@@H]3O)[C@H]1[C@]3(C)C(=O)O2 IXORZMNAPKEEDV-OBDJNFEBSA-N 0.000 title claims abstract description 29
- 239000005980 Gibberellic acid Substances 0.000 title claims abstract description 28
- 239000007788 liquid Substances 0.000 claims abstract description 9
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 5
- 229910052799 carbon Inorganic materials 0.000 claims description 5
- 239000011573 trace mineral Substances 0.000 claims description 5
- 235000013619 trace mineral Nutrition 0.000 claims description 5
- 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 4
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 claims description 4
- 229910000365 copper sulfate Inorganic materials 0.000 claims description 4
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 4
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 4
- 239000011790 ferrous sulphate Substances 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 230000012010 growth Effects 0.000 claims description 4
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 4
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 4
- 229940099596 manganese sulfate Drugs 0.000 claims description 4
- 235000007079 manganese sulphate Nutrition 0.000 claims description 4
- 239000011702 manganese sulphate Substances 0.000 claims description 4
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 4
- 235000015393 sodium molybdate Nutrition 0.000 claims description 4
- 239000011684 sodium molybdate Substances 0.000 claims description 4
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 claims description 4
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 4
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 4
- 229960001763 zinc sulfate Drugs 0.000 claims description 4
- 240000008042 Zea mays Species 0.000 claims description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 3
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 3
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 3
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 3
- 235000005822 corn Nutrition 0.000 claims description 3
- 229960001781 ferrous sulfate Drugs 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 235000015112 vegetable and seed oil Nutrition 0.000 claims description 3
- 239000008158 vegetable oil Substances 0.000 claims description 3
- 229930006000 Sucrose Natural products 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 125000000185 sucrose group Chemical group 0.000 claims description 2
- 238000012546 transfer Methods 0.000 claims description 2
- 230000003111 delayed effect Effects 0.000 claims 1
- 238000003556 assay Methods 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 235000013399 edible fruits Nutrition 0.000 description 4
- 229930191978 Gibberellin Natural products 0.000 description 3
- 240000007594 Oryza sativa Species 0.000 description 3
- 235000007164 Oryza sativa Nutrition 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 239000003448 gibberellin Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 235000009566 rice Nutrition 0.000 description 3
- 230000024053 secondary metabolic process Effects 0.000 description 3
- 238000009825 accumulation Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 235000019198 oils Nutrition 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000019525 primary metabolic process Effects 0.000 description 2
- 229930000044 secondary metabolite Natural products 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 240000008415 Lactuca sativa Species 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 230000010152 pollination Effects 0.000 description 1
- 235000012045 salad Nutrition 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem 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
- C12P27/00—Preparation of compounds containing a gibbane ring system, e.g. gibberellin
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- 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 application discloses a fermentation method for high yield of gibberellic acid, which comprises transferring seed liquid of gibberellic acid into a fermentation tank for fermentation, wherein different air flow rates are adopted according to different fermentation stages in the fermentation, and the average air flow rate in the early stage of fermentation is<The average air flow in the middle stage of fermentation is more than the average air flow in the later stage of fermentation. The fermentation method can improve GA in the fermentation liquor3The content of (a).
Description
Technical Field
The application relates to the field of gibberellic acid, in particular to a fermentation method for high-yield gibberellic acid.
Background
Gibberellic acid is a secondary metabolite obtained by fermentation culture and metabolism of a kind of gibberellin, 116 kinds of gibberellin are separated, identified and named, wherein GA3 is the most widely applied in agriculture, and plays a great role in agricultural production in China.
GA3Has very high plant regulating activity and obvious regulating effect on the growth and development of various crops. In the north, GA3The gibberellic acid solution with a certain concentration is sprayed in the flowering phase, so that the fruit setting rate of fruit trees can be effectively improved, the fruit growth and development are promoted, and the fruit yield is improved by about 20-30%. In the south, gibberellic acid is mainly applied to hybrid rice seed production, and male and female parents of rice are regulated to bloom at the same time, so that the pollination rate of the female parents is greatly improved, the yield of hybrid rice seeds is further improved, the production cost of the seeds is reduced, and the burden of farmers is reduced.
CN201810048112.2 discloses a fermentation method of gibberellic acid, wherein the fermentation method discloses a fermentation method, and a fermentation medium of the fermentation method consists of the following components: 10-40 g/L of corn protein powder; 1-10 g/L potassium dihydrogen phosphate; 5-30 g/L of a small-molecular organic carbon source; 0.5-5 g/L of vegetable oil; 0.5-2 g/L magnesium sulfate; 0.5-2 g/L of ammonium sulfate; 0.1-1 g/L of trace elements; the air flow rate of the fermentation is 2000-3000 Nm3The fermentation temperature is 28-30 ℃; the fermentation time is 8-10 days, and the highest GA of the fermentation method3Up to 5372ppm, but still further GA improvement is required3The content of (a).
Disclosure of Invention
In view of the above-mentioned drawbacks, the present application provides a fermentation method for high yield of gibberellic acid, which can increase GA in fermentation broth3The content of (a).
The technical scheme is as follows: a fermentation method for high-yield gibberellic acid by high-yield gibberellic acid fermentation includes transferring the seed liquid of gibberellic acid into a fermentation tank for fermentation, and adopting different air flow rates according to different fermentation stages in the fermentation, wherein the average air flow rate in the early stage of fermentation is smaller than the air flow rate in the middle stage of fermentation and larger than the average air flow rate in the later stage of fermentation.
In one or more specific embodiments herein, the temperature of fermentation in the fermentation is from 28 ℃ to 30 ℃.
In one or more specific embodiments of the present application, the fermentation time in the fermentation is 8 to 10 days.
In one or more specific embodiments of the present application, the fermentation pressure in the fermentation is 0.01 to 0.05 MPa.
In one or more specific embodiments of the present application, the early fermentation stage is a lag phase and a logarithmic growth phase, the middle fermentation stage is a stationary phase, and the late fermentation stage is a decline phase.
In one or more specific embodiments of the present application, the pre-fermentation average air flow rate is 3500-3Flow rate 3000-3H; average air flow rate of 4200-4400Nm in middle fermentation period3/h, air flow rate 3500-4600Nm3H; average air flow 3900-3/h。
In one or more specific embodiments of the present application, the fermentation method for high yield of gibberellic acid produces ≧ 6.7 g/L.
In one or more specific embodiments of the present application, the fermentation medium in the fermentation of the method comprises:
1-9 parts of corn protein powder;
1-10 parts by weight of monopotassium phosphate;
1-30 parts by weight of a small-molecular organic carbon source;
0.5-5 parts by weight of vegetable oil;
0.5-2 parts by weight of magnesium sulfate;
0.5-2 parts by weight of ammonium sulfate;
0.1 to 1 part by weight of trace elements.
In one or more specific embodiments of the present application, the small molecule organic carbon source is sucrose and/or glucose.
In one or more specific embodiments of the present application, the trace elements are ferrous sulfate, zinc sulfate, manganese sulfate, sodium molybdate, copper sulfate and cobalt chloride, wherein the weight ratio of the ferrous sulfate, the zinc sulfate, the manganese sulfate, the sodium molybdate, the copper sulfate and the cobalt chloride is 0.5-1.5: 0.5-1.5: 0.5-1.5: 0.1-0.5: 0.1-0.5: 0.1 to 0.5.
The invention principle and the beneficial effects are as follows:
gibberellic acid is a secondary metabolite in the fermentation process, needs to be converted from primary metabolism to secondary metabolism, the accumulation of bacterial load is mainly carried out in the primary metabolism process, the required air quantity is less than that of the secondary metabolism, a large amount of oxygen is needed to participate in the metabolism in the secondary metabolism process, and a large amount of objective product gibberellic acid is accumulated. Therefore, the air flow is low in the early fermentation stage (delay period and logarithmic growth period), the air flow is high in the middle fermentation stage (stationary stage), and the air flow is low in the later fermentation stage (recession stage).
The present application surprisingly achieves the goal of increasing acid production by increasing the rapid accumulation of gibberellins by changing the constant air flow to a variable speed air flow.
Detailed Description
The present application will be further explained below.
Example 1 preparation of seed liquid before fermentation.
Preparing seed solution before fermentation according to CN201810048112.2 example 1.1-1.3, wherein GA3The strains, the process and the culture medium are completely the same.
The test data of the prepared seed liquid are shown in the following table 1.
TABLE 1 Pre-fermentation seed liquid assay data
The seed solutions obtained in example 1 were subjected to the fermentation liquid preparation in the following examples 2 to 6, respectively.
In examples 2 to 6, the concentrations and the amounts of the seed solutions were equal.
Example 2
The seed liquid prepared in example 1 was fermented according to the fermentation method 1.4 in example 1 of CN201810048112.2 to obtain a fermentation liquid, and during the fermentation process, samples were taken respectively for detection, and the results are shown in table 2 below.
The fermentation process comprises the following steps:
fermentation volume: the volume is fixed to 82t, the volume after the extinction is 90t, and the volume after the rotation is 100 t.
Fermentation temperature: 29. + -. 0.2 ℃.
Operating the tank pressure: 0.035 MPa.
Air flow rate: 2200Nm3/h。
Controlling the pH: after the operation is started, the PH is gradually increased, the PH is increased by 0.1/half hour each time, and the PH is controlled to be 5.0-5.2 by adding ammonia water in a flowing mode.
And (3) supplementary material control: the dissolved oxygen rebounds by 20 percent, and the automatic feeding of glucose, salad oil and glucose (45 percent) solution is started: oil (3.2L:1L), dissolved oxygen control point 20% -30%.
And (3) fermentation period: 9 days (total fermentation time was 198 hours, since the time of transfer to the fermenter was 6 hours in the afternoon, day 1 in the fermenter).
TABLE 2 fermentation broth assay data
In this example, the fermentation medium is shown in Table 3 below.
TABLE 3 fermentation Medium composition content and sources
Example 3
This example was the same as example 2 except that the air flow rate was different. Air flow Rate of 4600Nm in this example3/h。
In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 4 below.
TABLE 4 fermentation broth assay data
Example 4
This example was the same as example 2 except that the air flow rate was different. In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 5 below.
TABLE 5 fermentation broth assay data
In this example, the air flow process parameters are shown in Table 6 below.
TABLE 6 air flow Process parameters
Example 5
This example was the same as example 2 except that the air flow rate was different.
In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 7 below.
TABLE 7 fermentation broth assay data
In this example, the air flow process parameters are shown in Table 8 below.
TABLE 8 air flow Process parameters
Example 6
This example was the same as example 2 except that the air flow rate was different.
In the fermentation process of this example, samples were taken for detection, and the results are shown in Table 9 below.
TABLE 9 fermentation broth assay data
In this example, the air flow process parameters are shown in Table 10 below.
TABLE 10 air flow Process parameters
In this application, unless otherwise specified, all are prior art.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (10)
1. A fermentation method for high-yield gibberellic acid transfers the seed liquid of gibberellic acid into a fermentation tank for fermentation, the fermentation air flow is variable speed air flow, wherein the average air flow in the early stage of fermentation is smaller than the average air flow in the middle stage of fermentation and larger than the average air flow in the later stage of fermentation.
2. The fermentation method of high yield gibberellic acid according to claim 1, wherein the fermentation temperature is 28-30 ℃.
3. The fermentation method of high yield gibberellic acid according to any one of claims 1-2, wherein the fermentation time during the fermentation is 8-10 days.
4. The fermentation method of high yield gibberellic acid according to any one of claims 1 to 3, wherein the fermentation pressure is 0.01 to 0.05 MPa.
5. The fermentation method of high yield gibberellic acid according to any one of claims 1 to 4, wherein the early fermentation stage is a delayed fermentation stage and a logarithmic growth stage, the middle fermentation stage is a stationary stage, and the late fermentation stage is a declining stage.
6. The fermentation method for high yield of gibberellic acid according to claim 5, wherein the average air flow rate in the early stage of fermentation is 3500-3700Nm3Flow rate 3000-3H; average air flow rate of 4200-4400Nm in middle fermentation period3/h, air flow rate 3500-4600Nm3H; average air flow 3900-3/h。
7. The fermentation method of high yield gibberellic acid according to 1-6, wherein the fermentation method of high yield gibberellic acid produces an acid amount of 6.7g/L or more.
8. The fermentation method of high yield gibberellic acid according to any one of claims 1 to 7, wherein the fermentation medium used in the fermentation process comprises:
1-9 parts of corn protein powder;
1-10 parts by weight of monopotassium phosphate;
1-30 parts by weight of a small-molecular organic carbon source;
0.5-5 parts by weight of vegetable oil;
0.5-2 parts by weight of magnesium sulfate;
0.5-2 parts by weight of ammonium sulfate;
0.1 to 1 part by weight of trace elements.
9. The method of claim 8, wherein the small organic carbon source is sucrose and/or glucose.
10. The fermentation method of high yield gibberellic acid as claimed in claim 8-9, wherein the trace elements are ferrous sulfate, zinc sulfate, manganese sulfate, sodium molybdate, copper sulfate and cobalt chloride, wherein the weight ratio of ferrous sulfate, zinc sulfate, manganese sulfate, sodium molybdate, copper sulfate and cobalt chloride is 0.5-1.5: 0.5-1.5: 0.5-1.5: 0.1-0.5: 0.1-0.5: 0.1 to 0.5.
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| AU2013204308A1 (en) * | 2010-06-28 | 2013-05-09 | Commonwealth Scientific And Industrial Research Organisation | Methods of producing lipids |
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