CN113957115B - Fermentation method for high yield of gibberellic acid - Google Patents
Fermentation method for high yield of gibberellic acid Download PDFInfo
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- 238000000855 fermentation Methods 0.000 title claims abstract description 98
- 230000004151 fermentation Effects 0.000 title claims abstract description 95
- 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 29
- 239000005980 Gibberellic acid Substances 0.000 title claims abstract description 28
- 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 28
- 238000000034 method Methods 0.000 title claims abstract description 27
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- 238000004519 manufacturing process Methods 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
- 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
- 229960001781 ferrous sulfate Drugs 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
- 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
- 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
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 3
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 3
- 239000011573 trace mineral Substances 0.000 claims description 3
- 235000013619 trace mineral Nutrition 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
- 108010068370 Glutens Proteins 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 235000021312 gluten Nutrition 0.000 claims description 2
- 235000012054 meals Nutrition 0.000 claims description 2
- 150000003384 small molecules Chemical class 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 125000000185 sucrose group Chemical group 0.000 claims description 2
- 230000003698 anagen phase Effects 0.000 claims 1
- 239000000203 mixture Substances 0.000 abstract description 4
- 235000010633 broth Nutrition 0.000 description 8
- 238000001514 detection method Methods 0.000 description 7
- 238000012360 testing method Methods 0.000 description 5
- 235000013399 edible fruits Nutrition 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 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
- 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
- 229930000044 secondary metabolite Natural products 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
- 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
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 235000014493 Crataegus Nutrition 0.000 description 1
- 241001092040 Crataegus Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 229930191978 Gibberellin Natural products 0.000 description 1
- 240000008415 Lactuca sativa Species 0.000 description 1
- 235000006029 Prunus persica var nucipersica Nutrition 0.000 description 1
- 244000017714 Prunus persica var. nucipersica Species 0.000 description 1
- 241000219094 Vitaceae Species 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 239000003448 gibberellin Substances 0.000 description 1
- 235000021021 grapes Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000010152 pollination Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 235000012045 salad Nutrition 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000012546 transfer 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 gibberellic acid, which comprises transferring gibberellic acid seed solution into a fermentation tank for fermentation, wherein different air flow rates are adopted in the fermentation according to different fermentation stages, and the average air flow rate in the fermentation stage<The average air flow in the middle fermentation period is greater than the average air flow in the later fermentation period. The fermentation method can improve GA in fermentation liquor 3 Is contained in the composition.
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 fermenting, culturing and metabolizing gibberellic acid, and 116 kinds of secondary metabolites are separated, identified and named, wherein GA3 which is the most widely used in agriculture plays a great role in agricultural production in China.
GA 3 Has very high plant regulating activity and obvious regulating effect on the growth and development of various crops. In the north, GA 3 Is mainly applied to the commercial crops such as grapes, red dates, hawthorns, nectarines and the like, spraying gibberellic acid solution with a certain concentration in the flowering phase can effectively improve the fruit setting rate of the fruit trees, promote the growth and development of the fruit, and improve the yield of the fruit trees by 20 percentAbout 30%. In the south, gibberellic acid is mainly applied to hybrid rice seed production, and the parent and parent of rice are regulated to bloom at the same time, so that the pollination rate of the parent is greatly improved, the yield of hybrid rice seeds is further improved, the production cost of the seeds is reduced, and the burden is lightened for farmers.
CN201810048112.2 discloses a fermentation method of gibberellic acid, in which a fermentation method is disclosed, and the fermentation medium of the fermentation method is composed of the following components: 10-40 g/L of corn protein powder; 1-10 g/L of monopotassium phosphate; 5-30 g/L of small molecular organic carbon source; vegetable oil 0.5-5 g/L; 0.5-2 g/L of magnesium sulfate; 0.5-2 g/L of ammonium sulfate; trace elements 0.1-1 g/L; the air flow rate of fermentation is 2000-3000 Nm 3 And/h, fermenting at 28-30 ℃; the fermentation time is 8-10 days, and the highest GA of the fermentation method 3 Reaching 5372ppm, still further GA elevation is required 3 Is contained in the composition.
Disclosure of Invention
In view of the above-mentioned drawbacks, in one aspect, the present application provides a fermentation method for high yield gibberellic acid, which can increase GA in fermentation broth 3 Is contained in the composition.
The technical proposal is as follows: a fermentation method for producing gibberellic acid with high yield comprises transferring gibberellic acid seed solution into a fermentation tank for fermentation, wherein different air flow rates are adopted according to different fermentation stages, and the average air flow rate in the earlier stage of fermentation is less than the average air flow rate in the middle stage of fermentation.
In one or more specific embodiments of the present application, the temperature of the fermentation in the fermentation is 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 from 0.01 to 0.05MPa.
In one or more specific embodiments of the present application, the pre-fermentation period is a delay period and a logarithmic growth period, the mid-fermentation period is a stationary period, and the post-fermentation period is a decay period.
In one or more specific embodiments of the present application, the average air flow rate in the pre-fermentation period is 3500-3700Nm 3 Per h, air flow 3000-4000Nm 3 /h; average air flow rate in mid-fermentation period is 4200-4400Nm 3 /h, air flow 3500-4600Nm 3 /h; the average air flow rate in the late fermentation period is 3900-4100 and the air flow rate is 3800-4200Nm 3 /h。
In one or more specific embodiments of the present application, the fermentation process for high gibberellic acid production yields ∈6.7g/L.
In one or more specific embodiments of the present application, the fermentation medium in the fermentation of the method comprises:
1-9 parts by weight of corn gluten meal;
1-10 parts by weight of monopotassium phosphate;
1-30 parts by weight of small molecular organic carbon source;
vegetable oil 0.5-5 weight portions;
0.5 to 2 parts by weight of magnesium sulfate;
0.5 to 2 parts by weight of ammonium sulfate;
0.1 to 1 weight portion of microelements.
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 ferrous sulfate, zinc sulfate, manganese sulfate, sodium molybdate, copper sulfate to cobalt chloride is 0.5-1.5: 0.5 to 1.5:0.5 to 1.5:0.1 to 0.5:0.1 to 0.5:0.1 to 0.5.
The invention has the following principle and beneficial effects:
gibberellic acid is a secondary metabolite in a fermentation process, the gibberellic acid is required to be converted from primary metabolism to secondary metabolism, bacterial amount accumulation is carried out in the primary metabolism for the main purpose, the required air amount is relatively small compared with the secondary metabolism, and a large amount of oxygen is required to participate in metabolism in the secondary metabolism, so that a large amount of target gibberellic acid is accumulated. Therefore, the air flow rate in the early fermentation stage (delay stage and logarithmic growth stage) is low, the air flow rate in the middle fermentation stage (stationary stage) is high, and the air flow rate in the late fermentation stage (decay stage) is low.
The present application surprisingly achieves the objective of improving acid production by changing the constant air flow to a variable speed air flow, increasing the rapid accumulation of gibberellin.
Detailed Description
The present application will be further described below.
Example 1 preparation of seed liquid before fermentation.
Preparation of seed liquid before fermentation according to the procedure of 1.1-1.3 in example 1 of CN201810048112.2, wherein GA 3 The strains, the process, the culture medium and the like are identical.
The seed liquid test data obtained are shown in Table 1 below.
TABLE 1 seed liquid detection data before fermentation
The seed solutions prepared in example 1 were prepared by the fermentation broths of examples 2 to 6 below, respectively.
In examples 2 to 6, the seed solution was used in the same concentration and amount.
Example 2
The seed solution prepared in example 1 was fermented according to the fermentation method of CN201810048112.2 example 1, 1.4 to obtain a fermentation broth, and during the fermentation, samples were taken separately for detection, and the results are shown in table 2 below.
The fermentation process comprises the following steps:
fermentation volume: the volume is fixed by 82t, the volume after elimination is 90t, and the volume after rotation is 100 t.
Fermentation temperature: 29.+ -. 0.2 ℃.
Operating the tank pressure: 0.035MPa.
Air flow rate: 2200Nm 3 /h。
PH control: after the operation is started, the PH is gradually increased, and the PH is controlled to be 5.0-5.2 by adding ammonia water after each time of 0.1/half hour.
And (3) material supplementing control: dissolved oxygen rebound 20%, automatic feeding glucose, salad oil and glucose (45%) solution: oil (3.2L: 1L), dissolved oxygen control point 20% -30%.
Fermentation period: 9 days (since the transfer time to the fermenter was in the afternoon, the fermentation time in the fermenter was only 6 hours on day 1, and the total fermentation time was 198 hours).
TABLE 2 fermentation broth detection data
In this example, the fermentation medium is shown in Table 3 below.
TABLE 3 composition and sources of fermentation Medium
Example 3
This example is the same as example 2 except that the air flow rate is different. The air flow rate in this example was 4600Nm 3 /h。
During the fermentation process of this example, samples were taken separately for detection, and the results are shown in Table 4 below.
TABLE 4 fermentation broth test data
Example 4
This example is the same as example 2 except that the air flow rate is different. During the fermentation process of this example, samples were taken separately for detection, and the results are shown in Table 5 below.
TABLE 5 fermentation broth test 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 is the same as example 2 except that the air flow rate is different.
During the fermentation process of this example, samples were taken separately for detection, and the results are shown in Table 7 below.
TABLE 7 fermentation broth test 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 is the same as example 2 except that the air flow rate is different.
During the fermentation process of this example, samples were taken separately for detection, and the results are shown in Table 9 below.
TABLE 9 fermentation broth test 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 indicated, they are all prior art.
The foregoing description is only of the preferred embodiments of the present application and is not intended to limit the same, but rather, various modifications and variations may be made by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.
Claims (8)
1. A fermentation method for high yield gibberellic acid, which comprises transferring gibberellic acid seed solution into a fermentation tank for fermentation, wherein the fermentation air flow is variable speed air flow, and the average air flow in the middle fermentation period is greater than the average air flow in the later fermentation period and greater than the average air flow in the earlier fermentation period;
the fermentation early stage is a fermentation delay period and a logarithmic growth phase, the fermentation middle stage is a stable period, and the fermentation later stage is a decay period;
the average air flow rate of the fermentation earlier stage is 3500-3700Nm, the air flow rate is 3000-4000Nm h; average air flow rate in the middle fermentation period is 4200-4400Nm h, and air flow rate is 3500-4600Nm h; the average air flow rate in the later fermentation period is 3900-4100 and the air flow rate is 3800-4200 Nm.
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 in the fermentation is 8-10 days.
4. The fermentation method of high yield gibberellic acid according to any one of claims 1-2, wherein the fermentation pressure is 0.01-0.05 MPa.
5. The fermentation process for high-yield gibberellic acid according to claims 1-2, wherein the fermentation process for high-yield gibberellic acid produces acid +.6.7 g/L.
6. The fermentation process for high gibberellic acid production according to any one of claims 1 to 2, wherein the fermentation medium in the fermentation of the process comprises:
1-9 parts of corn gluten meal;
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;
and 0.1-1 parts by weight of microelements.
7. The method for fermenting gibberellic acid with high yield according to claim 6, wherein the small molecule organic carbon source is sucrose and/or glucose.
8. The fermentation method of high yield gibberellic acid according to claim 6, 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 to 1.5:0.5 to 1.5:0.1 to 0.5:0.1 to 0.5:0.1 to 0.5.
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