CN1193048A - Method for producing beta-carotene by fermentation - Google Patents
Method for producing beta-carotene by fermentation Download PDFInfo
- Publication number
- CN1193048A CN1193048A CN98111199A CN98111199A CN1193048A CN 1193048 A CN1193048 A CN 1193048A CN 98111199 A CN98111199 A CN 98111199A CN 98111199 A CN98111199 A CN 98111199A CN 1193048 A CN1193048 A CN 1193048A
- Authority
- CN
- China
- Prior art keywords
- spore
- fermentation
- cultivated
- culture
- tank
- 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.)
- Pending
Links
Abstract
The present invention relates to a technological process for preparing beta-carotin by using spore inoculation in air-lift type fermentation tank and culturing filamentous fungi to make fermentation, and includes the procedures of primary inoculum culture, secondary inoculum culture and fermentation, and is characterized by that under a certain condition, culturing microbe inoculum capable of metabolizing and producing beta-carotin to form a lot of spores, using air-lift fermentation tank as bioreactor for culturing inoculum and making fermentation. In this type of tank, the spore is cultured to obtain a lot of hyphae, then these hyphae as seeds inoculated into fermentation culture medium, and fermented to produce beta-carotin.
Description
The present invention is the technology for preparing β-Hu Luobusu with spore inoculating fermentation culture filamentous fungus in airlift fermentor.β-Hu Luobusu is a vitamin medicaments, and it is the precursor of vitamin A, by the oxidasic effect two molecule vitamin A that dissociate, plays vitamin A in human body.It has scavenging(action) to the peroxidation base that the sun exposure protoporphyrin is produced, and therefore has been widely used at medicine and grocery trade.
The fermentative preparation β-Hu Luobusu seeding tank methods of advancing of shaking bottle mycelia that adopt are cultivated seed more before this invention, be bacterial classification by slant culture or shake-flask culture become to insert again behind the mycelia seeding tank breeding produce a large amount of mycelia do fermentation with kind, this seed preparation technology operation steps is many, cycle is long, increases the chance of living contaminants.In addition, the general form fermentor tank that has mechanical stirring device is adopted in the fermentation of seed culture before the present invention.Mechanical stirring easily interrupts the mycelia of filamentous fungus, causes mycelia damage, and biomass reduces, and the output of fermentation β-Hu Luobusu is reduced, and fermentation requires the ventilation height, and energy consumption is big, the cost height.
The object of the present invention is to provide a kind of technology simple, fermentation beta carotene output height, the method for the producing beta-carotene by fermentation of low cost of manufacture.
The present invention realizes by following scheme.
This method comprises: a, one-level kind are cultivated; B, secondary kind are cultivated; C, fermentation; Wherein the bio-reactor that the secondary kind is cultivated and fermentation procedure adopted is airlift fermentor, and the structure of this airlift fermentor is internal-circulation type or outer circulation type.The one-level kind cultivates in the operation, the energy metabolism of preservation is produced the slant strains of β-Hu Luobusu or the bacterial classification spore of sand tube preservation inserts on the PDA substratum, under temperature 24-30 ℃ condition, cultivated 48-72 hour, become suspension with stroke-physiological saline solution wash-out spore after treating to grow in a large number spore, the amount that contains spore in the suspension should be 7-9 * 10
5Individual/milliliter.The PDA substratum is potato, glucose and nutrient agar or potato, dextrose culture-medium, cultivates the vessel of spore and can use glass triangle flask or eggplant bottle and culture dish.The bacterial classification that produces β-Hu Luobusu is a kind of filamentous fungus, and Blakeslea frispora divides "+", "-" two kinds of bacterial strains, is preserved in Jiangsu Prov. Inst. of Microbiology culture presevation place, and bacterium number is 9203 (positive strains), 9204 (negative strains).In the secondary kind was cultivated operation, fresh spore spore was in suspension sprouted in seeding tank and is bred into a large amount of mycelium to use seed, the content of spore suspension miospore as fermenting be 7-9 * 10
5Individual/milliliter.The secondary kind is cultivated and fermentation procedure adopts airlift fermentor, the aspect ratio of tank body is 10: 1-20: 1, tank body is 1 with the diameter ratio of circulation tube: 0.3-1: 0.5, pressurized air sprays at a high speed from the nozzle of device the nutrient solution circulation tube and is the gas-liquid mixed state in the nutrient solution, cause nutrient solution severe to reduce, and obtain the power of high velocity air and follow endless tube and rise and enter tank body, produce continual circulation.Cultivate in the operation in the secondary kind, substratum is glucose 1-2%, starch 2-4%, corn steep liquor 3-7%, potassium primary phosphate 0.1-0.3%, sal epsom 0.01-0.05%, vitamin 0.001-0.003, PH6.0-6.5, culture temperature 24-30 ℃, ventilation 1: 0.8-1: 1.5, incubation time 24-48 hour.In zymotechnique, substratum is starch 2-4%; Dregs of rice cake powder 3-5%; Vegetables oil 3-5%; Corn steep liquor 3-5%; Dipotassium hydrogen phosphate 0.1-0.3%; Sal epsom 0.01-0.05%; Vitamin 0.001-0.003%, PH7.0-8.0, culture temperature 24-30 ℃, ventilation 1: 1.0-1: 20, tank pressure 0.04-0.08MPa, incubation time 90-120 hour.
Advantage of the present invention is 1, because profit has airlift fermentor, thereby when having avoided original use common fermentation jar, because of paddle causes bigger shearing force, make mycelium injured, biomass descends, thereby reduces the problem of the output of β-Hu Luobusu, concrete experimental result such as table one.
Table one:
2, the common fermentation jar satisfies the high problem of fermenting process oxygen-consumption by improving ventilation.Thereby energy consumption is big, and the utilization ratio of oxygen is also low.And adopt the airlift fermentor fermentation just can address this problem.Experimental result sees Table two.
Table two:
| Blade diameter length ratio | Ventilation | |
| Common jar | ????1∶2.5 | ????1∶5 |
| The gas lift jar | ????1∶10 | ????1∶1 |
3, the present invention replaces the technology of shake-flask seed as one-level kind seed inoculation jar with spore suspension, can shorten a jar seed culture time, improves the vigor of seed, and β-Hu Luobusu output is improved greatly.Experimental data sees Table three.
Table three:
| Shake bottle and plant the jar seed that connects | The cylinder seed that spore suspension connects | ||||
| ????03 | ????04 | ????03 | ????04 | ||
| The secondary seed cultivation results | ???PH | ??5.8 | ????＜5.4 | ????6.7 | ????5.8 |
| Biomass | ??0.67％ | ????0.55％ | ????0.91％ | ????0.8％ | |
| Time | 42-44 hour | 30-33 hour | |||
| β-Hu Luobusu output | 1053mg/L | ????1500mg/L | |||
Example 1:
1, the one-level kind is cultivated: the PDA substratum is formulated as follows: 150 gram peeling potatos are cut into fragment, place 100 ml waters to boil 30 minutes, filter through gauze, add 20 gram glucose in the filtrate, 20 gram agar, flame enrichment is settled to 1000ml, the PH nature, pack in 1000 milliliters the Erlenmeyer flask every bottled 80 milliliters of substratum, 120 ℃ of sterilizations 20 minutes into.
To be preserved in Jiangsu Prov. Inst. of Microbiology culture presevation place, bacterium number is that BLakeslea frispora "+", "-" bacterial strain sand tube spore of 9203 (positive strains), 9204 (negative strains) is seeded to respectively in 4 triangular flasks, and each bacterial strain respectively connects two.Cultivated 48 hours for 26 ℃, treat that media surface grows behind a large amount of black spores, add stroke-physiological saline solution 200ml in every triangular flask, contain 7 * 10 during the spore wash-out is become every milliliter
5The spore suspension of individual spore.
2, the secondary kind is cultivated, and seed culture medium is pressed following formulation: starch 2%; Glucose 2%; Corn steep liquor 5%; Sal epsom 0.01%; Potassium primary phosphate 0.1%; Vitamin 0.001% is adjusted to PH6.2 with sodium hydroxide solution.
Seeding tank is 2 of the internal circulation gas-lift type glass fermentor tanks of volume 6 liters, 2 liters of every dress seed culture mediums.Substratum was through 120 ℃ of sterilizations 30 minutes.Insert above-mentioned "+", the spore suspension of "-" bacterium pearl of cultivating base unit weight 1% respectively, cultivated air quantity 1: 1, tank pressure 0.05MPa 32 hours for 26 ℃.
3, fermentation
The prescription of fermention medium is a starch 4%; Cottonseed meal 4%; Corn steep liquor 3%: soya-bean oil 3%; Potassium primary phosphate 0.1%; Sal epsom 0.01%, vitamin 0.001% is transferred PH to 8.0 with sodium hydroxide solution.
Fermentor tank is the outer circulation air lift type glass fermentor tank of 30 milliliters of volumes, aspect ratio 15: 1, circulation tube is 0.4: 1 with jar diameter ratio, and dress fermention medium 18 liters were sterilized 40 minutes for 120 ℃, all insert the seed of cultured "+", "-" two bacterial strains in the fermention medium, inoculum size is 20%, "+", "-" bacterial strain each 10%, 27 ℃ ± 1 ℃ of leavening temperature, ventilation 1: 2, tank pressure 0.08MPa.The 48 hours stream that ferments adds alpha, beta-lonone 0.1%, ferments 90 hours, gets the fermented liquid of 1751.24 milligrams/liter of content beta-carotene.With the centrifugal collection thalline of fermented liquid, oven dry is pulverized to such an extent that dry bacterial powder 606.2 restrains, and every gram dry bacterial powder contains 52.0 milligrams of β-Hu Luobusus.Dry bacterial powder extracts with industrial hexane, and extracting solution adds the dehydrated alcohol low temperature crystallization behind concentrating under reduced pressure, a small amount of absolute ethanol washing of crystallization, and vacuum-drying gets β-Hu Luobusu crystal 19.3g, crystal purity is 95.6%.
Example 2
1, the one-level kind is cultivated: except that not adding the agar, PDA substratum and culture condition be with the triangular flask of 1,3000 milliliter of example, 150 milliliters of every bottled substratum.Two bacterial strains of aforementioned "+", "-" respectively connect 8 bottles, contain 9 * 10 in every milliliter
5Individual spore.
2, the secondary kind is cultivated, and seed culture medium is with example 1, and seeding tank adopts two of the stainless cylinder of steels of 300 liter outer circulation air lift types.Jar aspect ratio be 20: 1, circulation tube is 0.36: 1 with the diameter ratio of jar, two jars are respectively adorned substratum 240 liters, insert to cultivate "+", "-" bacterium pearl spore suspension of base unit weight 0.5% after the sterilization respectively, cultivate 30 hours, ventilation 1: 0.8 for 27 ℃.
3, fermentation: fermention medium removes 3% soya-bean oil and makes 3% Oleum Gossypii semen into, cottonseed meal changes into outside the soybean-cake flour, all the other are with example 1, fermentor tank is the stainless steel fermentor tank of the outer circulation air lift type of volume 3000 liters, the tank body aspect ratio is 20: 1, and circulation tube is 0.44: 1 with the diameter ratio of jar, dress substratum 2000 liters in jar, sterilized 1 hour, and after the cooling cultured "+", "-" two bacterial strains were inserted in the substratum together for 120 ℃.27 ° ± 1 ℃ of leavening temperature.Ventilation 1: 1.4.Tank pressure 0.05MPa.Fermented 48 hours, stream adds alpha, beta-lonone 0.1%, the end in 114 hours of fermenting, and β-Hu Luobusu output is 1328 milligrams/liter in the fermented liquid.After extracting, make 29.5 kilograms of the oily products of content beta-carotene 8%.
Claims (8)
1, a kind of method of producing beta-carotene by fermentation comprises a, the cultivation of one-level kind; B, secondary kind are cultivated; C, fermentation; It is characterized in that the bio-reactor that the secondary kind is cultivated and fermentation procedure adopted is airlift fermentor, the structure of this airlift fermentor is internal-circulation type or outer circulation type.
2, method according to claim 1, it is characterized in that cultivating in the operation in the one-level kind, the energy metabolism of preservation is produced the slant strains of β-Hu Luobusu or the bacterial classification spore of sand tube preservation inserts on the PDA substratum, under temperature 24-30 ℃ condition, cultivated 48-72 hour, become suspension with stroke-physiological saline solution wash-out spore after treating to grow in a large number spore, the amount that contains spore in the suspension should be 7-9 * 10
5Individual/milliliter.
3,, it is characterized in that the PDA substratum is potato, glucose and nutrient agar or potato, dextrose culture-medium, cultivates the vessel of spore and can use glass triangle flask or eggplant bottle and culture dish according to claim 1,2 described methods.
4, according to claim 1,2 described methods, the bacterial classification that it is characterized in that producing β-Hu Luobusu is a kind of filamentous fungus, Blakeslea frispora, divide "+", "-" two kinds of bacterial strains, be preserved in Jiangsu Prov. Inst. of Microbiology culture presevation place, bacterium number is 9203 (positive strains), 9204 (negative strains).
5, method according to claim 1 is characterized in that in the secondary kind is cultivated operation, and the spore in the fresh spore suspension is sprouted in seeding tank and bred into a large amount of mycelium to use seed, the content of spore suspension miospore as fermenting be 7-9 * 10
5Individual milliliter.
6, method according to claim 1, it is characterized in that the secondary kind is cultivated and fermentation procedure adopts airlift fermentor, the aspect ratio of tank body is 10: 1-20: 1, tank body is 1 with the diameter ratio of circulation tube: 0.3-1: 0.5, pressurized air sprays at a high speed from the nozzle of device the nutrient solution circulation tube and is the gas-liquid mixed state in the nutrient solution, cause nutrient solution severe to reduce, and obtain the power of high velocity air and follow endless tube and rise and enter tank body, produce continual circulation.
7, method according to claim 1, it is characterized in that cultivating in the operation in the secondary kind, substratum is glucose 1-2%, starch 2-4%, corn steep liquor 3-7%, potassium primary phosphate 0.1-0.3%, sal epsom 0.01-0.05%, vitamin 0.001-0.003,24-30 ℃ of PH6.0-6.5. culture temperature, ventilation 1: 0.8-1: 1.5, incubation time 24-48 hour.
8, method according to claim 1 is characterized in that in zymotechnique, and substratum is starch 2-4%; Dregs of rice cake powder 3-5%; Vegetables oil 3-5%; Corn steep liquor 3-5%; Dipotassium hydrogen phosphate 0.1-0.3%; Sal epsom 0.01-0.05%; Vitamin 0.001-0.003%, PH7.0-8.0, culture temperature 24-30 ℃, ventilation 1: 1.0-1: 20, tank pressure 0.04-0.0 8MPa, incubation time 90-120 hour.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN98111199A CN1193048A (en) | 1998-03-23 | 1998-03-23 | Method for producing beta-carotene by fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN98111199A CN1193048A (en) | 1998-03-23 | 1998-03-23 | Method for producing beta-carotene by fermentation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1193048A true CN1193048A (en) | 1998-09-16 |
Family
ID=5221196
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN98111199A Pending CN1193048A (en) | 1998-03-23 | 1998-03-23 | Method for producing beta-carotene by fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1193048A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102757995A (en) * | 2012-06-25 | 2012-10-31 | 湖北工业大学 | Method for preparing beta-carotene through fermentation of Blakeslea trispora |
| WO2012159446A1 (en) | 2011-05-20 | 2012-11-29 | 浙江医药股份有限公司新昌制药厂 | METHOD FOR PRODUCING NATURAL β-CAROTENE BY FERMENTATION AND USE THEREOF |
| CN104531538A (en) * | 2014-11-17 | 2015-04-22 | 嘉必优生物工程（武汉）有限公司 | Blakeslea trispora mutant strain and application thereof |
-
1998
- 1998-03-23 CN CN98111199A patent/CN1193048A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012159446A1 (en) | 2011-05-20 | 2012-11-29 | 浙江医药股份有限公司新昌制药厂 | METHOD FOR PRODUCING NATURAL β-CAROTENE BY FERMENTATION AND USE THEREOF |
| CN102757995A (en) * | 2012-06-25 | 2012-10-31 | 湖北工业大学 | Method for preparing beta-carotene through fermentation of Blakeslea trispora |
| CN104531538A (en) * | 2014-11-17 | 2015-04-22 | 嘉必优生物工程（武汉）有限公司 | Blakeslea trispora mutant strain and application thereof |
| CN105861323A (en) * | 2014-11-17 | 2016-08-17 | 嘉必优生物技术（武汉）股份有限公司 | Beta-carotene-containing bacterial powder, microbial oil and oil suspension |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1313498C (en) | Process for producing pullulan | |
| CN102057836B (en) | Method for quickly producing edible fungus liquid strain by utilizing primary-secondary type culture tank | |
| CN104893983B (en) | Liquid state fermentation low citrinin, the preparation method of High color values monascorubin and product | |
| CN101513161A (en) | Formula for culture medium of cordyceps militaris liquid strains and method for culturing same | |
| CN103250568A (en) | Method for preparing hericium erinaceus rich in selenium | |
| CN1232632C (en) | New strain APC-20 of Paecilomyces cicadae and fementation process for artificial culture | |
| CN102845225A (en) | Hypsizygus marmoreus liquid strain fermenting technique | |
| CN110317734A (en) | A kind of monascus and its isolated culture method and the application of high-yield glucoamylase, Esterified Enzyme and protease | |
| CN101768541A (en) | Preparation method and system of beta-glucan | |
| CN109504725A (en) | A kind of method and fermentation medium of fermentation Hericium erinaceus preparation high-purity Hericium erinaceus Polysaccharides | |
| CN1063788C (en) | Fermenting production process of cordyceps fungus | |
| CN111763636A (en) | Method for producing coenzyme Q10 by industrial fermentation | |
| CN1193048A (en) | Method for producing beta-carotene by fermentation | |
| CN106035985A (en) | Method for producing single cell proteins by using processed waste from mixed bacteria liquid fermentation of yellow wine | |
| CN100412186C (en) | Tinder fungus and process for deep liquid fermentation preparation of tinder fungus | |
| CN101861796B (en) | Method for culturing amanita pantherina by using waste distillage after fermentation of coloured rice | |
| CN112625914B (en) | Method for repeatedly fermenting ganoderma lucidum in batches by using airlift fermentation tank | |
| CN1456673A (en) | Production of alcohol by fermenting by yeast tolerant to high concentrated sugar and alcohol | |
| CN106754401A (en) | A kind of production method of hirsutella sinensis fungal | |
| CN114164122A (en) | Schizochytrium limacinum for high yield of EPA and application thereof | |
| CN101974500A (en) | Production method of high-purity and intermediate-temperate alpha-amylase | |
| CN115039633A (en) | Artificial culture method for sporocarp of Isaria japonica | |
| CN110903983A (en) | Monascus monascus with high yield of saccharifying enzyme and esterifying enzyme and application thereof | |
| CN105018410A (en) | Method for inducing Blakeslea trispora aging strain to rapidly produce a large number of spores | |
| CN113136339A (en) | Method for mixotrophic-autotrophic continuous culture of photosynthetic microorganisms, culture system and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C01 | Deemed withdrawal of patent application (patent law 1993) | ||
| WD01 | Invention patent application deemed withdrawn after publication |