CN102899273B - Method for improving spore germination rate of micromonospora echinospora - Google Patents
Method for improving spore germination rate of micromonospora echinospora Download PDFInfo
- Publication number
- CN102899273B CN102899273B CN201210424789.4A CN201210424789A CN102899273B CN 102899273 B CN102899273 B CN 102899273B CN 201210424789 A CN201210424789 A CN 201210424789A CN 102899273 B CN102899273 B CN 102899273B
- Authority
- CN
- China
- Prior art keywords
- spore germination
- spore
- micromonospora echinospora
- germination rate
- improved
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Pretreatment Of Seeds And Plants (AREA)
- Mushroom Cultivation (AREA)
Abstract
The invention belongs to the technical field of fermentation, in particular to a method for improving the spore germination rate of a micromonospora echinospora, which specifically comprises the following steps that: firstly, a spore germination culture medium is prepared, fresh micromonospora echinospora spores are suspended in the spore germination culture medium, are thermally activated in a water bath of 50-65 DEG C for 5-20 minutes, and then are rapidly cooled, and placed on a shaker at a temperature of 35-37 DEG C and a shaking speed of 200-240rpm for pre-germination for 1-4 hours, so that the spore germination rate is increased from 10-20% to 30-50%, and simultaneously, the synchronization of spore germination and mycelial growth is improved. According to the method provided by the invention, the cost is reduced, the process is simplified, so that the original four-stage fermentation is improved into the three-stage fermentation; and the utilization rate of equipment is improved, so that the production efficiency is greatly improved, and the predictability of the fermentation process is increased.
Description
Technical field
The invention belongs to fermentation technical field, be specifically related to a kind of method that improves micromonospora echinospora spore germination rate.
Background technology
Isepamicin (Isepamicin) is as semisynthetic aminoglycoside antibiotics of new generation, the similar gentamicin of its antimicrobial spectrum, has very strong anti-microbial effect to escherichia coli, citrobacter, klebsiella bacillus, enterobacteria, serratia marcecens, Bacillus proteus, Pseudomonas aeruginosa etc.It is except ear, renal toxicity are low, also stable to bacteriogenic most aminoglycoside modifying enzymes, there is good post antibiotic effect, therefore the bacterial strain of many resistance to gentamicins and the resistance to amikacin of part is still responsive to it, it is all effective that septicemia, wound, scald, operation wound, superinfection, chronic bronchitis, bronchiectasis infection, pneumonia, pyelonephritis, urocystitis and the peritonitis particularly gentamicin sulphate Resistant strain being caused etc. infect disease, and therefore it has good development prospect.
As the isepamicin of semisynthetic antibiotics, it is on 1 bit amino of gentamicinB, to connect that isoserine is derivative to be formed.And gentamicinB is as the important precursor substance of synthetic isepamicin, be by micromonospora echinospora (
micromonospora echinospora) fermentation generation.The mode of production of gentamicinB is that micromonospora echinospora is accessed to seeding tank after spore preparation, shaking flask mycelium culture, and seed accesses fermentor tank through one-level, secondary, three grades of expansion levels after cultivating.Spore preparation is an indispensable link during fermentation industry is produced, and the quality of spore quality will directly have influence on the success or not of fermentative production.General agar slant culture-medium or the general solid medium of adopting of spore preparation cultivated and obtained after inoculation.Because the spore of micromonospora belongs to the aleuriospore of rudimentary type, there is thick internal layer cell walls, these factors have caused the spore of micromonospora to have 80～90%, and composing type statospore only has 10～20% spore to sprout.The feature of the low germination rate of micromonospora, impels our necessary inoculum size and the increase fermentation progression of strengthening on zymotechnique, and complex process, plant factor reduction, cost are raise.
Summary of the invention
The object of the present invention is to provide a kind of method that improves micromonospora echinospora spore germination rate.
In order to achieve the above object, the present invention is by the following technical solutions:
(1) preparation of spore germination substratum: get soybean cake powder 2-4% and Semen Maydis powder 1.5-3%, the constant volume that adds water, with 6N HCl tune pH2.0-3.0, boil and maintain 0.5-1.5 hour, in process, add the moisture evaporating, cooling rear readjustment pH6.0-7.0, filters out that to soak juice for subsequent use;
(2) add Zulkovsky starch 1-2% above-mentioned in soaking juice, glucose 0.05-0.1%, saltpetre 0.05-0.1%, peptone 0.05-0.1%, Tryptones 0.05-0.1%, cobalt chloride 0.0001-0.0003%, potassium primary phosphate 0.005-0.01%, dipotassium hydrogen phosphate 0.01-0.03%, adjusts pH7.2～7.5, makes spore germination substratum;
(1) micromonospora echinospora, after slant activation, adds spore germination substratum to inclined-plane scraping spore, obtains spore suspension, through 50-65 DEG C of water-bath 5-20 minute, then cooling rapidly, be placed in 1-4 hour on 35-37 DEG C of shaking table, shaking speed 200-240rpm.
Wherein the described cooling rear readjustment pH6.0-7.0 employing mass ratio of step (1) is that 30%NaOH solution regulates pH.
It is that 30%NaOH solution regulates pH that the described tune of step (2) pH7.2～7.5 adopt mass ratio.
Operation is simple in the present invention, and gordian technique is preparation, water-bath hot activation, the spore germination of spore germination substratum, makes the spore germination rate of micromonospora echinospora bring up to 30-50% by 10-20%, has improved the synchronism of spore germination and mycelial growth simultaneously; The present invention has reduced cost, has simplified technique, makes original level Four fermentation be improved to three grade fermemtation, has improved the utilization ratio of equipment, and production efficiency is greatly improved, and has increased the predictability of fermenting process.
Embodiment
Embodiment 1:
1. the preparation of spore germination substratum: soybean cake powder 2g, Semen Maydis powder 3g, is settled to 100ml, adjusts pH3.0 with 6N HCl, boils and maintains 1 hour, adds the moisture evaporating in process, cooling rear with 30%NaOH solution readjustment pH6.0, filters out that to soak juice for subsequent use;
2. in above-mentioned diffusion juice, add Zulkovsky starch 1g, glucose 0.1g, saltpetre 0.05g, peptone 0.05g, Tryptones 0.1g, cobalt chloride 200ug, potassium primary phosphate 0.005g, dipotassium hydrogen phosphate 0.03g, 30%NaOH solution regulates pH7.5, makes spore germination substratum;
3. micromonospora echinospora, after slant activation, adds 15ml spore germination substratum to inclined-plane scraping spore and prepares spore suspension, is placed in 55 DEG C of water-bath hot activations 10 minutes, then cooling rapidly, be placed on shaking table 35 DEG C of temperature, shaking speed 220rpm, cultivates 2 hours.
Present method makes spore germination rate bring up to 30% by 10%, has improved the synchronism of spore germination and mycelial growth simultaneously; The present invention has reduced cost, simplify technique, make the fermentation of original level Four be improved to three grade fermemtation (existing technical process: spore is sprouted in advance---shaking flask mycelium culture---first class seed pot---secondary breeding tank---fermentor tank), improve the utilization ratio of equipment, production efficiency is greatly improved, has increased the predictability of fermenting process.
Embodiment 2:
(1) preparation of spore germination substratum: soybean cake powder 4g, Semen Maydis powder 1.5g, is settled to 100ml, with 6N HCl tune pH2.0, boil and maintain 1.5 hours, in process, add the moisture evaporating, cooling rear with 30%NaOH solution readjustment pH7.0, filter out that to soak juice for subsequent use;
(2) in above-mentioned diffusion juice, add Zulkovsky starch 2g, glucose 0.1g, saltpetre 0.1g, peptone 0.1g, Tryptones 0.05g, cobalt chloride 250ug, potassium primary phosphate 0.007g, dipotassium hydrogen phosphate 0.02g, adjusts pH7.4, makes spore germination substratum;
(3) micromonospora echinospora is after slant activation, add 20ml spore germination substratum to inclined-plane scraping spore and prepare spore suspension, be placed in 60 DEG C of water-bath hot activations 5 minutes, then cooling rapidly, be placed on shaking table 36 DEG C of temperature, shaking speed 240rpm, pre-sprouting 1 hour, finally accesses in seed culture medium.
Present method makes spore germination rate bring up to 50% by 20%, has improved the synchronism of spore germination and mycelial growth simultaneously; The present invention has reduced cost, has simplified technique, makes original level Four fermentation be improved to three grade fermemtation, has improved the utilization ratio of equipment, and production efficiency is greatly improved, and has increased the predictability of fermenting process.
Embodiment 3:
(1) preparation of spore germination substratum: soybean cake powder 3g, Semen Maydis powder 2.5g, is settled to 100ml, with 6N HCl tune pH3.0, boil and maintain 0.5 hour, in process, add the moisture evaporating, cooling rear with 30%NaOH solution readjustment pH6.5, filter out that to soak juice for subsequent use;
(2) in above-mentioned diffusion juice, add Zulkovsky starch 1.5g, glucose 0.05g, saltpetre 0.07g, peptone 0.075g, Tryptones 0.075g, cobalt chloride 100ug, potassium primary phosphate 0.01g, dipotassium hydrogen phosphate 0.03g, adjusts pH7.2, makes spore germination substratum;
(3) micromonospora echinospora is after slant activation, adds 30ml spore germination substratum to inclined-plane scraping spore and prepares spore suspension (the whole solubility of spore is 5-6 × 10
8/ ml), be placed in 65 DEG C of water-bath hot activations 5 minutes, then cooling rapidly, be placed on shaking table, 37 DEG C of temperature, shaking speed 200rpm, sprouts 4 hours in advance, finally accesses in seed culture medium.
Present method makes spore germination rate bring up to 30% by 20%, has improved the synchronism of spore germination and mycelial growth simultaneously; The present invention has reduced cost, has simplified technique, makes original level Four fermentation be improved to three grade fermemtation, has improved the utilization ratio of equipment, and production efficiency is greatly improved, and has increased the predictability of fermenting process.
Claims (3)
1. a method that improves micromonospora echinospora spore germination rate, is characterized in that:
(1) preparation of spore germination substratum: get soybean cake powder 2-4% and Semen Maydis powder 1.5-3%, the constant volume that adds water, with 6N HCl tune pH2.0-3.0, boil and maintain 0.5-1.5 hour, in process, add the moisture evaporating, cooling rear readjustment pH6.0-7.0, filters out that to soak juice for subsequent use;
(2) add Zulkovsky starch 1-2% above-mentioned in soaking juice, glucose 0.05-0.1%, saltpetre 0.05-0.1%, peptone 0.05-0.1%, Tryptones 0.05-0.1%, cobalt chloride 0.0001-0.0003%, potassium primary phosphate 0.005-0.01%, dipotassium hydrogen phosphate 0.01-0.03%, adjusts pH7.2～7.5, makes spore germination substratum;
(3) micromonospora echinospora, after slant activation, adds spore germination substratum to inclined-plane scraping spore, obtains spore suspension, through 50-65 DEG C of water-bath 5-20 minute, then cooling rapidly, be placed in 1-4 hour on 35-37 DEG C of shaking table, shaking speed 200-240rpm;
Described in step (3), the whole solubility of spore suspension miospore is 5～6 × 10
8/ ml.
2. the method for raising micromonospora echinospora spore germination rate according to claim 1, is characterized in that: it is that 30%NaOH solution regulates pH that the described cooling rear readjustment pH6.0-7.0 of step (1) adopts mass ratio.
3. the method for raising micromonospora echinospora spore germination rate according to claim 1, is characterized in that: it is that 30%NaOH solution regulates pH that the described tune of step (2) pH7.2～7.5 adopt mass ratio.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210424789.4A CN102899273B (en) | 2012-10-31 | 2012-10-31 | Method for improving spore germination rate of micromonospora echinospora |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210424789.4A CN102899273B (en) | 2012-10-31 | 2012-10-31 | Method for improving spore germination rate of micromonospora echinospora |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102899273A CN102899273A (en) | 2013-01-30 |
| CN102899273B true CN102899273B (en) | 2014-09-17 |
Family
ID=47571763
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210424789.4A Expired - Fee Related CN102899273B (en) | 2012-10-31 | 2012-10-31 | Method for improving spore germination rate of micromonospora echinospora |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102899273B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106498026A (en) * | 2016-11-07 | 2017-03-15 | 河南省农业科学院植物保护研究所 | A kind of method for promoting sweet potato black rot pathogen conidium stably to sprout and obtain optimal viewing effect |
| US20200288723A1 (en) * | 2017-09-20 | 2020-09-17 | Bayer Cropscience Biologics Gmbh | Method of improving storage stability and fitness of fungal spores |
| CN108456706A (en) * | 2018-03-30 | 2018-08-28 | 丽珠集团福州福兴医药有限公司 | A kind of micromonospora cultural method improving output of gentamycin |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101979647B (en) * | 2010-11-09 | 2012-09-19 | 福州大学 | Method for increasing yield of gentamicin |
| CN102732580A (en) * | 2011-04-09 | 2012-10-17 | 浙江海正药业股份有限公司 | Preparation method of efficient antitumor antibiotic calicheamicin |
| CN102747115B (en) * | 2011-04-19 | 2015-05-27 | 上海医药工业研究院 | Fermentation medium for fermentation production of antibiotic calicheamicin gamma1 I, fermentation method suitable for the fermentation medium and use of the fermentation medium |
-
2012
- 2012-10-31 CN CN201210424789.4A patent/CN102899273B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN102899273A (en) | 2013-01-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102172174B (en) | Antrodia camphorate quick liquid fermentation process based on asexual spores | |
| CN102258059B (en) | Method for preparing ralstonia solanacearum-resistant bacillus subtilis microbial inoculum | |
| CN108085354A (en) | A kind of culture medium and its method of fermenting and producing FR901379 | |
| CN102899273B (en) | Method for improving spore germination rate of micromonospora echinospora | |
| CN103740790A (en) | Production method capable of increasing yield of neomycin | |
| CN109593807A (en) | A kind of method of high level fermenting and producing apramycin | |
| CN107828702A (en) | A kind of kasugarnycin fermentation medium and fermentation process | |
| CN104232709A (en) | Method for preparing apramycin through fermentation | |
| CN109486881A (en) | A kind of fermentation medium and zymotechnique of kasugarnycin | |
| CN110468080B (en) | Microbial agent for promoting rice growth and reducing cadmium as well as preparation method and application method thereof | |
| CN103898181A (en) | Method for producing nosiheptide by virtue of fermentation | |
| CN105420148A (en) | Preparation method for kitasamycin industrial production strains | |
| CN103103135B (en) | Aspergillus oryzae strain and method for producing fungal alpha-amylase by liquid-state fermentation of aspergillus oryzae strain | |
| CN103087967B (en) | Bacillus subtilis for efficiently producing beta glucanase as well as application thereof | |
| CN102864082B (en) | Method for culturing citric acid fermenting seeds and method for preparing citric acid by fermenting | |
| CN102634553B (en) | Hainanmycin fermentation method | |
| CN101880703B (en) | Method for fermenting daptomycin by adding caprate | |
| CN102154419A (en) | Fermentation medium for improving enramycin yield | |
| CN104388501A (en) | Preparation method of erythromycin by using bioenzyme | |
| CN109722408B (en) | Method for promoting bacillus subtilis to produce spores | |
| CN109258305A (en) | A kind of method for promoting sclerotium of morchella esculenta to be formed and its fluid nutrient medium used | |
| RU2413707C2 (en) | STRAIN OF Sinorhizobium fredii KB-11 TYPE LEGUME BACTERIA FOR PRODUCING BACTERIAL FERTILISER FOR SOYA BEAN | |
| CN105586374B (en) | A method of sugar production doractin is mended based on metabolizing parameters reduced sugar | |
| CN106282045B (en) | Fermentation medium and fermentation method of Bacillus licheniformis | |
| CN108277243A (en) | A kind of preparation method of Pseudomonic Acid A |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140917 Termination date: 20191031 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |