CN115927004B - Bacterial strain capable of producing high-content gibberellin GA7, application thereof and production method thereof - Google Patents
Bacterial strain capable of producing high-content gibberellin GA7, application thereof and production method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of bioengineering, and discloses a strain for producing high-content gibberellin GA7, application thereof and a production method thereof. A strain mainly accumulating gibberellin GA7 is classified and named as gibberella caner (Fusar i um fuj i kuro i) NRF02, and is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of No. M20221211 in the year 08 and 01. The strain is obtained by carrying out ARTP mutation breeding on a strain of a mixture of gibberellin GA4+7 mainly accumulated in the original strain. The method for producing gibberellin GA7 by fermenting the gibberella ampelina comprises the following steps: the bacterial strain preserved in the liquid nitrogen tank is activated by a liquid culture medium, and then is subjected to seed culture and fermentation culture to obtain a mixture of gibberellin GA4 and GA7 which are target products, wherein GA7 accounts for more than 90% of the mixture. The fermentation conditions are controlled in stages, the fermentation is carried out for 120 hours, the fermentation level reaches 1258mg/L, the gibberellin GA7 content is high, the fermentation level is stable, and the method is suitable for industrial production of high-content gibberellin GA7.
Description
Technical Field
The invention relates to the technical field of logistics devices, in particular to a strain for producing high-content gibberellin GA7, application thereof and a production method thereof.
Background
The gibberellins GA4 and GA7 related by the invention have the following structural formulas:
gibberellins (GAs) are a class of plant hormones belonging to the diterpenoid class, which are widely found in higher plants, fungi and bacteria. There are a wide variety of gibberellins, a total of 136 of which have been found. Among these are mainly GA1, GA3, GA4 and GA7, which are biologically active. GA3 is developed most successfully, and the production, application, research and development form a larger scale, so that the GA3 is one of main plant growth regulators. Gibberellin compounds have various regulatory functions as plant growth regulators. Gibberellins produced are different under different illumination and temperature conditions, at different periods of plant growth and even at different parts of the same plant. GA7 is mainly produced in the shoot tip and immature seeds, and GA4 is found in the roots, stems, leaves and seeds of plants. Gibberellins appear to be GA3 > GA7 > GA4 on activities that promote plant stem growth. GA3 has wide application at present, but because GA3 has high activity, overgrowth of hypocotyl can be promoted when plant dormancy is broken, lodging resistance of plants is reduced, rapid growth of epidermal cells can be promoted, so that epidermis horny layer is thinner, and fruits are easy to be broken by long spots. With the intensive research of gibberellins, other homologs of the family, particularly GA4 and GA7, are increasingly paid attention to, gibberellin GA4 products are available on the market, and gibberellin GA4+7 mixture products with different proportions (mainly a product with the ratio of 2:1, and the gibberellin GA4+7 mixture has the functions of breaking dormancy and not inducing the growth of hypocotyls, protecting flowers and fruits, improving the toughness of the epidermis and the horny layer of fruits, preventing brown spots and the like).
Gibberellin GA7 is currently mainly obtained by liquid submerged fermentation. However, there are two major problems in the production process, one is that the yield of GA7 is low; secondly, the mixture of GA4 and GA7 is coexistent (GA 4 is converted into GA7 under the action of dehydrogenase) and the separation of GA7 from the mixture is very difficult, and the two are only different by one double bond, so that the molecular weight is very close, the separation and purification cost of GA7 is high, and the large-scale commercial application of GA7 is seriously hindered. The research reports on gibberellin GA7 at home and abroad are very few, the biological chemical stock of Zhejiang river is controlled by changing the composition of a culture medium and the fermentation process, the metabolic pathway is changed to accumulating gibberellin GA7 (CN 201710316511.8), the fermentation period is 8-10 days, the yield of gibberellin GA7 reaches 1039mg/L, the content of gibberellin GA7 is more than 90%, and the fermentation period is long. Yang Wenge, et al (CN 201210464476.1), discloses a method for separating and purifying GA4 and/or GA7, wherein GA7 is separated from a mixture of GA4 and GA7, and either GA4 or GA7 is obtained from a mixture of GA4 and GA7.
From the viewpoint of overall productivity, these strains or processes are not suitable for large-scale industrial production of gibberellin GA7.
Disclosure of Invention
The invention aims to provide gibberella ampeloides and a method for producing gibberellin GA7 by using the gibberella ampelopsis, wherein the content of GA7 in fermentation liquor is more than 90% of the mixture of GA4 and GA7, fermentation conditions are controlled through two stages, and carbon sources are supplemented in the fermentation process to improve the yield of gibberellin GA7.
The aim of the invention can be achieved by the following technical scheme:
in a first aspect, a gibberellin GA 7-producing strain is classified under the name: hedyotis fujikuroi (Fusarium fujikuroi) NRF02 was preserved in the CCTCC-China center for type culture collection (China center for type culture Collection) at the accession number CCTCC NO. M20221211, university of Wuhan, hubei province, china. The strain adopts staged control of fermentation conditions, carbon sources are supplemented in the fermentation process to maintain the total sugar at 0.7-1.0%, the fermentation is carried out for 120 hours, the fermentation level reaches 1258mg/L, the gibberellin GA7 content of the strain is high, the fermentation level is stable, and the strain is suitable for industrial production.
The strain is obtained by screening a mixture strain of original mainly accumulated gibberellin GA4+7 through ARTP mutagenesis.
The specific process is as follows:
preparing tender hypha: selecting mycelium blocks with the length and width of 1cm respectively, and scattering in a glass bottle containing 20ml of 0.90% physiological saline for 10min; sterile microporous filters of diameter 80mm and pore size 0.45 μm were spread on plates with PDA medium (glucose 2%, potato 20%, agar 1.8%). Sucking 1ml of the scattered bacterial suspension, uniformly coating on a microporous filter membrane, and placing the microporous filter membrane in a mould incubator for culturing at 28 ℃ for about 36 hours.
Protoplast preparation: preparing 30ml of mixed enzyme solution, filtering and sterilizing the mixed enzyme solution for later use by using a suction filtration bottle with a microporous membrane of 0.22 mu m, wherein the mixed enzyme solution comprises 2.5% of cellulase, 0.3% of snailase and 2.0% of pectinase and 4.0% of NaCl; placing the cultured filter membrane with tender hyphae in a blank culture dish, sucking about 20ml of mixed enzyme solution, scraping the tender hyphae off by using a tool, taking out the filter membrane by using tweezers, and carrying out enzymolysis for about 4 hours at 28 ℃; and centrifuging the enzymolysis liquid to remove enzyme liquid, washing the enzymolysis liquid once with 4.0% NaCl steady-permeation solution, centrifuging again to remove supernatant, and fixing the volume to a corresponding volume with 4.0% NaCl steady-permeation solution to obtain protoplast bacterial suspension.
ARTP mutagenesis treatment: and (3) uniformly coating 10 mu L of protoplast bacterial suspension on the surface of a sterilizing slide, transferring a sterilizing culture dish with the slide into an ARTP operating room, placing the slide into a corresponding groove by using sterile forceps, adjusting a knob of a manual rotary sample carrier to ensure that the distance between the slide and a jet outlet of a plasma generator is about 2-3mm, setting the air quantity to 8-12SLM, setting the power to 100-200W, and controlling the time to 0-60s. The treated slide was placed in a centrifuge tube containing 1mL of sterile solution for shaking elution, and the eluent was diluted and spread in a regeneration medium for culture, the regeneration medium components (sucrose 2.0%, peptone 0.2%, yeast extract 0.2%, KH2PO40.05%, sodium chloride 3.6%, agar 1.5%).
And (3) fermenting culture and stability test, picking single colony on a regeneration plate, and detecting the concentration of gibberellin GA4 and gibberellin GA7 in fermentation broth after seed culture and fermenting culture. The initially screened 20 strains have gibberellin GA7 concentration higher than 800mg/L and GA7 content higher than 80%, and further re-screening to obtain 5 strains of high-yield strains. Further testing the stability of the strain, selecting a strain with good genetic stability, wherein the strain is named as fusarium gracilis (fusarium fujikuroi) NRF02, the concentration of gibberellin GA7 products is high and stable above 850mg/L, and the content of gibberellin GA7 is up to 94.8%.
The seed culture medium comprises 2.0-4.0% of glucose, 1.5-3.0% of soybean cake powder, 0.8-2.0% of peanut cake powder, KH2PO40.80-2.0%, (NH 4) 2SO40.005-0.015%, 0.05-0.15% of MgSO4.7H2O, and the temperature is 28-30 ℃, the rotation speed of a shaking table is 200-300rpm, and the seed liquid is obtained after culturing for 24-48 hours;
the fermentation medium comprises the following components: 4.0 to 12.0 percent of carbon source, 2.5 to 4.0 percent of nitrogen source, 0.08 to 0.16 percent of soybean oil, 0.08 to 0.12 percent of MgSO4.7H2O, 2SO40.02 to 0.05 percent of (NH 4), KH2PO40.15 to 0.35 percent of ZnSO and 40.005 to 0.01 percent of ZnSO. In the early stage of fermentation, the culture temperature is 28-30 ℃, the stirring rotation speed is 200-300rpm, the aeration rate is 4-6m < 3 >/h, and the culture time is 24-48 hours; in the later fermentation period, the culture temperature is 31-33 ℃, the stirring rotation speed is 160-220rpm, the aeration rate is 5-8m < 3 >/h, and the culture time is 72-96 hours.
In a second aspect, a method for producing gibberellin GA7 by fermenting gibberella ampelina includes steps of preserving seeds in a liquid nitrogen tank, activating the seeds by a liquid culture medium, culturing the seeds, fermenting the seeds, and controlling fermentation conditions in stages:
(1) Liquid seed activation: an activation culture medium, 2.0-4.0% of glucose, 1.5-3.0% of soybean cake powder, 0.8-2.0% of peanut cake powder, KH2PO40.80-2.0%, (NH 4) 2SO40.005-0.015%, mgSO4.7H2O0.05-0.15%, and the temperature of 28-30 ℃ and the rotation speed of a shaking table of 200-300rpm, and culturing for 24-48 hours to obtain an activation bacterial liquid;
(2) Seed culture: inoculating 100-150mL of activated bacterial liquid into a 50L fermentation tank (30L/50L) containing a seed culture medium, wherein the seed culture medium comprises 2.0-4.0% of glucose, 1.5-2.5% of soybean cake powder, 1.5-3.0% of peanut cake powder, 2.0-4.0% of dextrin, KH2PO40.08-0.16%, 2SO40.01-0.03% of (NH 4), 0.05-0.15% of MgSO4.7H2O, the temperature is 28-30 ℃, the stirring rotation speed is 250-350rpm, and the ventilation amount is 1.5-3.0m < 3 >/h for culturing for 18-36 hours to obtain the seed liquid;
(3) Fermentation culture: inoculating 8-12% (V/V) of seed solution into a 100L fermentation tank (70L/100L) containing a fermentation medium, wherein the fermentation medium comprises the following components: 4.0 to 12.0 percent of carbon source, 2.5 to 4.0 percent of nitrogen source, 0.08 to 0.16 percent of soybean oil, 0.08 to 0.12 percent of MgSO4.7H2O, 2SO40.02 to 0.05 percent of (NH 4), KH2PO40.15 to 0.35 percent of ZnSO and 40.005 to 0.01 percent of ZnSO. In the early stage of fermentation, the culture temperature is 28-30 ℃, the stirring rotation speed is 200-300rpm, the aeration rate is 4-6m < 3 >/h, and the culture time is 24-48 hours; in the later fermentation period, the culture temperature is 31-33 ℃, the stirring rotation speed is 160-220rpm, the aeration rate is 5-8m < 3 >/h, and the culture time is 72-96 hours.
(4) Gibberellin GA4, GA7 assay method: instrument: shimadzu LC-20AT, a 150mm×4.6mm (id) stainless steel column was used, and an ODS-C18 (5 μm packing) column was packed. Mobile phase: methanol+water+formic acid=67+33+0.05 (v/v) before use. Flow rate: 0.7mL/min. Detection wavelength: 210nm. Sample injection amount: 20. Mu.L. Sample solution preparation: the pH of the fermentation broth is regulated to about 7.0 by using 2% NaOH solution, the fermentation broth is filtered by filter paper, 5.0mL of filtrate is accurately sucked and diluted to a scale by using a mobile phase in a 25mL volumetric flask, and then the supernatant is centrifugally taken and filtered by using a 0.45 mu m filter membrane so as to prepare for sample injection detection. Retention time: gibberellic acid GA7 for about 9.4min and gibberellic acid GA4 for about 11.0min.
The invention has the beneficial effects that: the invention provides a gibberella (Fusarium fujikuroi) strain which can produce gibberellin GA7 with higher yield in a shorter fermentation period, and the gibberellin GA7 content can obtain a gibberellin GA7 product with high content. Through step-by-step culture of the strain, the yield of gibberellin GA7 can reach 850mg/L, and the content of gibberellin GA7 can reach more than 90%; when the fermentation conditions are controlled by adopting double-stage culture, the yield of the fed sugar gibberellin GA7 in the fermentation process can reach 1258mg/L, the content of gibberellin GA7 is more than 90%, the method has good application prospect, and the method has positive promotion effect on the industrial production of gibberellin GA7.
Drawings
FIG. 1 is a map of gibberellin product of gibberella caner (Fusarium fujikuroi) NRF 02.
Biological material preservation information:
the invention discloses a fusarium fujikuroi (Fusarium fujikuroi) NRF02 which is already in CCTCC-China center for type culture collection (CCTCC for short);
the classification is named: gibberella fusarium (fusarium fujikuroi) NRF02;
report of the preservability survival of gibberella (fusarium fujikuroi) NRF02;
the preservation date is 2022, 08 and 01;
the preservation number is CCTCCNO20221211.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
(1) Seed activation: the components of the activation culture medium are 3.0% of glucose, 1.8% of soybean cake powder, 1.5% of peanut cake powder, KH2PO41.5%, (NH 4) 2SO40.01%, mgSO4.7H2O0.10%, the temperature is 29 ℃, the rotation speed of a shaking table is 260rpm, and the culture is carried out for 36 hours to obtain an activated bacterial liquid;
(2) Seed culture: 100mL of activated bacterial liquid is inoculated into a 50L fermentation tank (30L/50L) containing a seed culture medium, the seed culture medium comprises 3.0% of glucose, 2.0% of soybean cake powder, 2.5% of peanut cake powder, 3.0% of dextrin, KH2PO40.15%, (NH 4) 2SO40.02%, mgSO4.7H2O0.10%, the temperature is 29 ℃, the stirring rotation speed is 300rpm, and the ventilation rate is 2.5m3/h for culturing for 24 hours, so as to obtain seed liquid;
(3) Fermentation culture: the fermentation medium comprises the following components: 10% of starch liquefaction liquid, 1.5% of peanut cake powder, 1.5% of soybean cake powder, 0.12% of soybean oil, 0.10% of MgSO4.7H2O, (NH 4) 2SO40.03%, KH2PO40.20% of ZnSO40.0075% and 70L/100L of liquid loading. 10% (V/V) of the seed solution was inoculated into a 100L fermenter containing a fermentation medium at 30℃and a rotation speed of 300rpm at an aeration rate of 6m3/h, and fermented for 120 hours to obtain a fermentation broth, and the yield of gibberellin GA7 was examined to determine that gibberellin GA7 was 850mg/L, and the gibberellin GA7 content was 94.8%.
Example 2
(1) Seed activation: as in example 1.
(2) Seed culture: as in example 1;
(3) The fermentation medium comprises the following components: 10% of starch liquefaction liquid, 1.5% of peanut cake powder, 1.5% of soybean cake powder, 0.12% of soybean oil, 0.10% of MgSO4.7H2O, (NH 4) 2SO40.03%, KH2PO40.20% of ZnSO40.0075% and 70L/100L of liquid loading. 10% (V/V) of the seed solution was inoculated into a 100L fermenter containing a fermentation medium at 28℃and a rotation speed of 200rpm at an aeration rate of 5m3/h, and fermented for 120 hours to obtain a fermentation broth, and the yield of gibberellin GA7 was 725mg/L and the gibberellin GA7 content was 85.4%.
Example 3
(1) Seed activation: as in example 1.
(2) Seed culture: as in example 1;
(3) The fermentation medium comprises the following components: 10% of starch liquefaction liquid, 1.5% of peanut cake powder, 1.5% of soybean cake powder, 0.12% of soybean oil, 0.10% of MgSO4.7H2O, (NH 4) 2SO40.03%, KH2PO40.20% of ZnSO40.0075% and 70L/100L of liquid loading. 10% (V/V) of the seed solution was inoculated into a 100L fermenter containing a fermentation medium at 32℃and a rotational speed of 300rpm at an aeration rate of 8m3/h, and fermented for 120 hours to obtain a fermentation broth, which was examined for the gibberellin GA7 yield of 875mg/L and gibberellin GA7 content of 90.5%.
Example 4
(1) Seed activation: as in example 1.
(2) Seed culture: as in example 1;
(3) The fermentation medium comprises the following components: 10% of starch liquefaction liquid, 1.5% of peanut cake powder, 1.5% of soybean cake powder, 0.12% of soybean oil, 0.10% of MgSO4.7H2O, (NH 4) 2SO40.03%, KH2PO40.20% of ZnSO40.0075% and 70L/100L of liquid loading. 10% (V/V) of the seed solution was inoculated into a 100L fermenter containing a fermentation medium. The fermentation conditions are controlled in stages, the fermentation is carried out for 0-48 hours at 29 ℃, the rotation speed is 300rpm, the fermentation is finished for 48 hours at 32 ℃, the rotation speed is 200rpm, the fermentation is carried out for 120 hours, the fermentation liquid is obtained, and the yield of the gibberellin GA7 is 1048mg/L and the content of the gibberellin GA7 is 92.8 percent after detection.
Example 5
(1) Seed activation: as in example 1.
(2) Seed culture: as in example 1;
(3) The fermentation medium comprises the following components: 10% of starch liquefaction liquid, 1.5% of peanut cake powder, 1.5% of soybean cake powder, 0.12% of soybean oil, 0.10% of MgSO4.7H2O, (NH 4) 2SO40.03%, KH2PO40.20% of ZnSO40.0075% and 70L/100L of liquid loading. 10% (V/V) of the seed solution was inoculated into a 100L fermenter containing a fermentation medium. The fermentation conditions are controlled in stages, the fermentation is carried out for 0-48 hours at 32 ℃, the rotating speed is 200rpm, and the ventilation rate is 6m3/h; after 48 hours, the fermentation is finished at 28 ℃, the rotating speed is 300rpm, the ventilation rate is 4m3/h, the fermentation is carried out for 120 hours, the fermentation liquor is obtained, and the yield of gibberellin GA7 is 986mg/L and the content of gibberellin GA7 is 91.2 percent after detection.
Example 6
(1) Seed activation: as in example 1.
(2) Seed culture: as in example 1;
(3) The fermentation medium comprises the following components: 5% of starch liquefied liquid, 1.5% of peanut cake powder, 1.5% of soybean cake powder, 0.12% of soybean oil, 0.10% of MgSO4.7H2O, (NH 4) 2SO40.03%, KH2PO40.20% of ZnSO40.0075% and 70L/100L of liquid loading. 10% (V/V) of the seed solution was inoculated into a 100L fermenter containing a fermentation medium. The fermentation conditions are controlled in stages, the fermentation is carried out for 0-48 hours at 29 ℃, the rotating speed is 300rpm, the rotating speed is 200rpm, the fermentation is finished at 32 ℃ after 48 hours, the maltose is supplemented when the total sugar is reduced to 0.7%, the concentration of the total sugar is controlled to be 0.7-0.8%, the fermentation is carried out for 120 hours, the fermentation liquor is obtained, and the yield of gibberellin GA7 is 1258mg/L and the content of gibberellin GA7 is 93.6% after detection.
Example 7
(1) Seed activation: as in example 1.
(2) Seed culture: as in example 1;
(3) The fermentation medium comprises the following components: 5% of starch liquefied liquid, 1.5% of peanut cake powder, 1.5% of soybean cake powder, 0.12% of soybean oil, 0.10% of MgSO4.7H2O, (NH 4) 2SO40.03%, KH2PO40.20% of ZnSO40.0075% and 70L/100L of liquid loading. 10% (V/V) of the seed solution was inoculated into a 100L fermenter containing a fermentation medium. The fermentation conditions are controlled in stages, the fermentation is carried out for 0-48 hours at 29 ℃, the rotating speed is 300rpm, the rotating speed is 200rpm, the fermentation is finished at 32 ℃ after 48 hours, the maltose is supplemented when the total sugar is reduced to 0.7%, the concentration of the total sugar is controlled to be 0.7-0.8%, the fermentation is carried out for 120 hours, the fermentation liquor is obtained, and the yield of gibberellin GA7 is 1258mg/L and the content of gibberellin GA7 is 94.5% after detection.
Finally, it should be noted that: the foregoing description is only illustrative of the preferred embodiments of the present invention, and although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described, or equivalents may be substituted for elements thereof, and any modifications, equivalents, improvements or changes may be made without departing from the spirit and principles of the present invention.
Claims (7)
1. A strain producing gibberellin GA7, which is classified and named: gibberella cappa (Fusarium fujikuroi) NRF02 was preserved in the CCTCC-chinese collection at the accession number cctcccno. m20221211 at the date of 2022, month 08 and 01.
2. The use of the strain according to claim 1 for fermentative production of gibberellin GA7, wherein fermentation conditions are controlled in stages, and when total sugar is reduced to less than 0.7%, carbon sources are added to control total sugar concentration, and fermentation is performed for 120 hours at a fermentation level of 1258mg/L to obtain a high GA 7-containing product.
3. A method for producing gibberellin GA7 by fermentation using the strain of claim 1, which comprises the steps of:
(1) The strain of claim 1 is inoculated to a liquid activation medium for activation, wherein the components of the activation medium are as follows: glucose 2.0-4.0%, soybean cake powder 1.5-3.0%, peanut cake powder 0.8-2.0%, KH 2 PO 4 0.80 -2.0%， (NH 4 ) 2 SO 4 0.005-0.015%，MgSO 4 .7H 2 O0.05-0.15%, temperature 28-30deg.C, shaking table rotation speed 200-300rpm, culturing for 24-48 hr to obtain activated bacterial liquid;
(2) Inoculating 100-150mL of activated bacterial liquid into a 50L fermentation tank containing seed culture medium, wherein the seed culture medium comprises glucose 2.0-4.0%, soybean cake powder 1.5-2.5%, peanut cake powder 1.5-3.0%, and aleurone 2.0%-4.0%， KH 2 PO 4 0.08-0.16% ， (NH 4 ) 2 SO 4 0.01-0.03% ， MgSO 4 .7H 2 O0.05-0.15%, temperature 28-30deg.C, stirring speed 250-350rpm, aeration rate 1.5-3.0m 3 Culturing for 18-36 hours to obtain seed solution;
(3) Inoculating 8-12% (V/V) of seed solution into a 100L fermentation tank containing a fermentation medium, wherein the fermentation medium comprises the following components: 4.0 to 12.0 percent of carbon source, 2.5 to 4.0 percent of nitrogen source, 0.08 to 0.16 percent of soybean oil and MgSO 4 .7H 2 O 0.08-0.12% ， (NH 4 ) 2 SO 4 0.02-0.05%,KH 2 PO 4 0.15-0.35%，ZnSO 4 0.005-0.01%。
4. A method according to claim 3, characterized by comprising the step of inoculating gibberella tabaci (Fusariumf ujikuroi) NRF02 to a fermentation medium, and culturing gibberella tabaci (Fusariumf ujikuroi) NRF02 by double-stage fermentation; the double-stage fermentation culture gibberella caner (fusarium fujikuroi) NRF02 comprises a fermentation early stage and a fermentation late stage; in the early stage of fermentation, the culture temperature is 28-30deg.C, the stirring rotation speed is 200-300rpm, and the aeration rate is 4-6m 3 And/h, culturing for 24-48 hours; in the late fermentation stage, the culture temperature is 31-33 ℃, the stirring rotation speed is 160-220rpm, and the ventilation rate is 5-8m 3 And/h, the culture time is 72-96 hours.
5. A method according to claim 3, wherein the carbon source in the fermentation medium is one or a combination of starch liquefact, dextrin, maltose, glucose.
6. A method according to claim 3, wherein the total carbon during the fermentation culture is reduced to less than 0.7% and the sugar is supplemented by one or more of dextrin, maltose and maltose, and the total carbon is maintained at 0.7-1.0%.
7. The method according to claim 3, wherein the nitrogen source in the fermentation medium is soybean cake powder or peanut cakeFlour, corn gluten meal, rice gluten meal, (NH) 4 ) 2 SO 4 、NH 4 COOH、NaNO 3 、KNO 3 One or a combination of several of the above.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102942547A (en) * | 2012-11-16 | 2013-02-27 | 南京工业大学 | Separation and purification method of GA4 (gibberellin A4) and/or GA7 (gibberellin A7) |
| CN107177659A (en) * | 2017-05-08 | 2017-09-19 | 浙江钱江生物化学股份有限公司 | A kind of fermentation method production gibberellin A7Method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102942547A (en) * | 2012-11-16 | 2013-02-27 | 南京工业大学 | Separation and purification method of GA4 (gibberellin A4) and/or GA7 (gibberellin A7) |
| CN107177659A (en) * | 2017-05-08 | 2017-09-19 | 浙江钱江生物化学股份有限公司 | A kind of fermentation method production gibberellin A7Method |
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