CN102321700A - Kojic acid fermentation technique and apparatus - Google Patents
Kojic acid fermentation technique and apparatus Download PDFInfo
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- CN102321700A CN102321700A CN201110221006A CN201110221006A CN102321700A CN 102321700 A CN102321700 A CN 102321700A CN 201110221006 A CN201110221006 A CN 201110221006A CN 201110221006 A CN201110221006 A CN 201110221006A CN 102321700 A CN102321700 A CN 102321700A
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Abstract
The invention discloses a kojic acid fermentation technique and apparatus. In the invention, fed-batch fermentation is utilized to control the concentration of the fermentation liquid, and the product concentration is controlled by continually separating the product, thereby improving the fermentation environment, greatly enhancing the fermentation yield of the kojic acid, enhancing the apparatus utilization ratio and lowering the energy consumption. The invention capable of effectively enhancing the fermentation level and efficiency is based on the following principles: every microbe has the respective optimal conditions for growth and product conversion, the fed-batch fermentation is utilized to control the concentration of the fermentation liquid, and the microbe is fed according to the metabolic level, so that the concentration of the fermentation liquid is kept on an optimal level; and the product accumulated to a certain concentration can inhibit the microbial growth and metabolism, a separation device is utilized to continually separate the product kojic acid by using a fermentation tank external circulation separation technique, so that the product concentration in the fermentation liquid is always kept on a low level, and the whole fermentation process is carried out under the optimal conditions.
Description
Technical field
The invention belongs to technical field of biological fermentation, relate to a kind of kojic acid fermentative prodn technology, specifically is a kind of through control fermented liquid concentration and production concentration, the kojic acid zymotechnique machine fermentation equipment that improve the product level, improves plant factor and cut down the consumption of energy.
Background technology
Kojic acid has another name called kojic acid, Qu acid, is a kind of melanochrome specificity suppressor factor, its get into behind skin cells can with the cupric ion complexing in the cell, change the three-dimensional arrangement of tyrosine oxidase, stop the activation of tyrosine oxidase, thereby suppress melanic formation.It does not act on other enzyme in the cell, and pair cell does not have toxic action, and it can also get in the intercellular substance simultaneously, forms the intercellular colloid, plays water conservation and the effect that increases skin elasticity.Kojic acid also have remove radical, strengthen cell viability, food fresh keeping is protected effects such as look, is widely used in medicine and field of food.As can do the raw material of producing food flavoring agent voitol and veltol plus; Be used to produce the midbody of cephalosporin analog antibiotic; , insecticide without environmental pollution nontoxic to people and animals with doing is used for agricultural chemicals; Impose on farm crop as foliage fertilizer; As iron analytical reagent, film spotting agent etc.The most popular and the most promising purposes is aspect makeup and food two at present.
Kojic acid is the product of obtaining through fermentation under carbon source, the nitrogenous source situation having, and fermentation industry is a high energy consumption industry, how effectively to improve single jar of level and output, is the direction of studying in the industry.Having in the kojic acid production technology of report at present; As be called " kojic acid producing process " in name, the patent No. is " CN01135102.0 ", and publication number discloses a kind of kojic acid production technology in the patent document of " CN1417341 "; It directly drops into fermentor tank liquefaction and saccharification with starch; Liquid glucose inoculation fermentation after sterilizing is cultivated kojic acid, and the repeatedly condensing crystal product of kojic acid clear liquid is through after dissolving, decolouring, and decrease temperature crystalline is purified and obtained the kojic acid crystal of finished product.Fermentation and acid acidity＞6% in this production technology is that present kojic acid ferments normal one and produces acid number.But up to the present, owing to lack effective process means, in commercial scale prodn, kojic acid product acidity can only be about 6%, the zymotechnique that not have appearance can kojic acid product acidity be increased substantially all the time.
Summary of the invention
The objective of the invention is to overcome the deficiency that exists in the prior art, provide a kind of, the kojic acid zymotechnique and the fermentation equipment that improve the product level, improve plant factor and cut down the consumption of energy through control fermented liquid concentration and production concentration.
According to technical scheme provided by the invention: a kind of kojic acid zymotechnique is characterized in that comprising the steps:
(1), with glucose carbon source preparation feeding medium during fermentation, through sterilization and subsequent use after being cooled to the suitable growth temperature scope of production bacterium;
(2), be carbon source preparation fermentation basestocks with glucose, after sterilization and being cooled to the suitable growth temperature scope of producing bacterium, insert the production bacterium, the march acid-fermentation;
(3) when the remaining sugar concentration in the fermented liquid is reduced to 30～40g/L, begin to mend feeding medium during fermentation, in whole fermentation process, make the remaining sugar concentration in the fermented liquid be controlled at 30～60g/L, finish until feed supplement, produce bacteria growing, metabolic needs to satisfy;
(4) when the kojic acid acidity in the fermented liquid during, begin to carry out kojic acid and separate, until fermentation ends at 30～40g/L.
As the further improvement of said zymotechnique, said production bacterium is adopted aspergillus oryzae (Aspergillus oryzae) ACCC 30163; Above-mentioned aspergillus oryzae is in " foodstuffs industry science and technology " 24 (4): 17～19,2003, " China brewages " (6): 42～44,2006 publications such as grade are open.
As the further improvement of said zymotechnique, the prescription of said fermentation basestocks consists of: glucose 50～60g/L, and yeast extract 7.5～9g/L, sal epsom 0.5g/L, potassium hydrogenphosphate 0.5g/L, skimmer 0.2g/L transfers pH3.0～3.2 with phosphoric acid.
As the further improvement of said zymotechnique, the prescription of said feeding medium during fermentation consists of: glucose 350～370g/L, and sal epsom 0.5g/L, potassium hydrogenphosphate 0.5g/L, skimmer 0.2g/L transfers pH3.0～3.2 with phosphoric acid.
As the further improvement of said zymotechnique, said fermentation basestocks and feeding medium during fermentation all are to adopt high pressure steam sterilization, are cooled to 29～31 ℃ then.
As the further improvement of said zymotechnique, the inoculum size of said production bacterium is 3～7% of fermentation basestocks volume.
As the further improvement of said zymotechnique, the initial add-on of said fermentation basestocks is 50～60% of a fermentor tank volume capacity.
As the further improvement of said zymotechnique, in the said step (3), feeding medium during fermentation be adopt the computer control continuous and quantitative mend into, the time that begins to mend into feeding medium during fermentation is at fermentation 24～36h; In the said step (4), beginning to carry out the kojic acid separation time is at fermentation 48～60h.
A kind of kojic acid fermentation equipment; Comprise fermentor tank; Fermentor tank is used for splendid attire fermentation basestocks, is provided with whisking appliance and thermostatically heating pipe in the fermentor tank, and it is characterized in that: said fermentation equipment also comprises feed supplement jar and tripping device; Said feed supplement jar is used for splendid attire fermentation fluid infusion, and the feed supplement jar links to each other with material-feeding port on the fermentor tank; Said tripping device is used for separating during the fermentation kojic acid; Tripping device comprises water cooler, cone-shaped body and scoop; The fluid inlet of water cooler links to each other with the liquid outlet that separates of fermentor tank through liquid-outlet valve, and the liquid outlet of water cooler links to each other with the osculum of cone-shaped body upper end, big mouthful of connection scoop of cone-shaped body lower end; Scoop sidewall top is provided with liquid outlet, and the liquid outlet of scoop links to each other with the liquid return hole that separates of fermentor tank through returning the liquid valve.
The method that said tripping device separates kojic acid is: before beginning to separate kojic acid work, open the water coolant of tripping device earlier, tripping device is carried out precooling, make chiller temperature remain on 10～15 ℃; When beginning to separate, open earlier slowly liquid-outlet valve, make fermented liquid get into water cooler and fully cooled off, and guarantee that the fermented liquid temperature of water cooler afterbody is lower than 20 ℃; After being full of fermented liquid in the whole tripping device, open back the liquid valve again, utilize temperature official post fermented liquid in tripping device, from up to down to flow automatically, thereby make the fluid power that ferments in fermented liquid and the fermentor tank in the tripping device exchange of in the regular hour, flowing; Fermented liquid in the tripping device after being cooled to the kojic acid supersolubility temperature, kojic acid crystallization and separating out; Kojic acid that crystallization goes out and fermented liquid descend in tripping device simultaneously; When dropping to cone-shaped body; Because the sectional area of cone-shaped body increases gradually; The fermented liquid flow velocity slows down, and in scoop, the lower concentration fermented liquid of removing the part kojic acid is back in the fermentor tank through returning the liquid valve crystal in the action of gravity settle.
The present invention compared with prior art, advantage is: the present invention has that fermentation level height, fermentation period are short, energy consumption is low relatively, fermentation efficiency is high, produce advantages such as acidacidity＞10%; The present invention adds feeding medium during fermentation through stream and controls fermented liquid concentration, controls production concentration through continuous separated product, thereby can improve yeasting, significantly improves the kojic acid fermentation yield, improves plant factor, cuts down the consumption of energy.The present invention can effectively improve fermentation level and efficient is based on principle: because the condition that any mikrobe all has the righttest growth separately and product to transform; The present invention adds feeding medium during fermentation control fermented liquid concentration through stream; How many metabolism what mends, and makes fermented liquid concentration remain on an optimum level; And, also can suppress product microorganism growth and metabolism when running up to finite concentration; The present invention utilizes tripping device to adopt fermentor tank outer circulation stripping technique that the product kojic acid is constantly separated; Make the production concentration in the fermented liquid remain on lower level always, whole fermentation process is all carried out under optimum.
Description of drawings
Fig. 1 is a kojic acid fermentation equipment synoptic diagram of the present invention.
Embodiment
Below in conjunction with concrete accompanying drawing and embodiment the present invention is described further, but content of the present invention is not limited to following embodiment.
Embodiment 1
Produce bacterium: adopt aspergillus oryzae (Aspergillus oryzae) ACCC 30163.Above-mentioned aspergillus oryzae is in " foodstuffs industry science and technology " 24 (4): 17～19,2003, " China brewages " (6): 42～44,2006 publications such as grade are open.
(1), take by weighing glucose 2450kg, sal epsom 3.5kg, potassium hydrogenphosphate 3.5kg, skimmer 1.4kg, the adding volume capacity is 25m
3Feed supplement jar 4 in, add water to 7m then
3, using phosphoric acid to regulate pH is 3.0, high pressure steam sterilization (pressure is 0.1MPa, and temperature is that 121 ℃, time are 30min) postcooling to 29 ℃ is subsequent use as feeding medium during fermentation;
(2), take by weighing glucose 700kg, yeast extract 105kg, sal epsom 7kg, potassium hydrogenphosphate 7kg, skimmer 2.8kg, the adding volume capacity is 25m
3Fermentor tank 1 in, add water to 14m then
3, stir, using phosphoric acid to regulate pH is 3.0, high pressure steam sterilization (pressure is 0.1MPa, and temperature is that 121 ℃, time are 30min) postcooling to 29 ℃ is as the fermentation basestocks; Insert bacterial classification, march acid-fermentation under 29 ℃ of conditions by 3% of fermentation basestocks volume;
(3), when fermenting, record remaining sugar concentration 42g/L in the fermented liquid, begin to mend feeding medium during fermentation, PM is mended into 2.3L, and the remaining sugar concentration in the control fermented liquid is controlled at 30～60g/L, finishes until feed supplement, produces bacteria growing, metabolic needs to satisfy to 36h;
When (4) fermenting, record kojic acid acidity in the fermented liquid, start the tripping device 6 in the fermentation equipment at 33g/L to 56h; Begin to carry out the product kojic acid and separate, until fermentation ends, fermentation period adds up to 98 hours; After the fermentation ends, the remaining sugar concentration in the fermented liquid is 9g/L.
The isolating method of said kojic acid is specific as follows: before beginning to separate kojic acid work, open the water coolant of tripping device 6 earlier, tripping device 6 is carried out precooling, make water cooler 6.3 temperature remain on 10 ℃; When beginning to separate, open earlier slowly liquid-outlet valve 7, make fermented liquid get into water cooler 6.3 and fully cooled off, the fermented liquid temperature of controlled chilling device 6.3 afterbodys is 16 ℃; After being full of fermented liquid in the whole tripping device 6, open back liquid valve 5 again; Utilize temperature official post fermented liquid in tripping device 6, from up to down to flow automatically, thereby make the fluid power that ferments in fermented liquid and the fermentor tank 1 in the tripping device 6 exchange of in the regular hour, flowing; Fermented liquid in the tripping device 6 after being cooled to the kojic acid supersolubility temperature, kojic acid crystallization and separating out; Kojic acid that crystallization goes out and fermented liquid descend in tripping device 6 simultaneously; When dropping to cone-shaped body 6.2; Because the sectional area of cone-shaped body 6.2 increases gradually; The fermented liquid flow velocity slows down, and in scoop 6.1, the lower concentration fermented liquid of removing the part kojic acid is back in the fermentor tank 1 through returning liquid valve 5 crystal in the action of gravity settle.Fermentation and acid level among the embodiment 1 is 106g/L (the kojic acid acidity of promptly finally separating is 10.6%), and the remaining sugar concentration in the fermented liquid is 9g/L.
Embodiment 2
Produce bacterium: adopt aspergillus oryzae (Aspergillus oryzae) ACCC 30163.Above-mentioned aspergillus oryzae is in " foodstuffs industry science and technology " 24 (4): 17～19,2003, " China brewages " (6): 42～44,2006 publications such as grade are open.
(1), take by weighing glucose 2520kg, sal epsom 3.5kg, potassium hydrogenphosphate 3.5kg, skimmer 1.4kg, the adding volume capacity is 25m
3Feed supplement jar 4 in, add water to 7m then
3, using phosphoric acid to regulate pH is 3.1, high pressure steam sterilization (pressure is 0.1MPa, and temperature is that 121 ℃, time are 30min) postcooling to 30 ℃ is subsequent use as feeding medium during fermentation;
(2), take by weighing glucose 770kg, yeast extract 115kg, sal epsom 7kg, potassium hydrogenphosphate 7kg, skimmer 2.8kg, the adding volume capacity is 25m
3Fermentor tank 1 in, add water to 14m then
3, stir, using phosphoric acid to regulate pH is 3.1, high pressure steam sterilization (pressure is 0.1MPa, and temperature is that 121 ℃, time are 30min) postcooling to 30 ℃ is as the fermentation basestocks; Insert bacterial classification, march acid-fermentation under 30 ℃ of conditions by 5% of fermentation basestocks volume;
(3), when fermenting, record remaining sugar concentration 45g/L in the fermented liquid, begin to mend feeding medium during fermentation, PM is mended into 2.1L, and the remaining sugar concentration in the control fermented liquid is controlled at 30～60g/L, finishes until feed supplement, produces bacteria growing, metabolic needs to satisfy to 30h;
When (4) fermenting, record kojic acid acidity in the fermented liquid, start the tripping device 6 in the fermentation equipment at 35g/L to 50h; Begin to carry out the product kojic acid and separate, until fermentation ends, fermentation period adds up to 96 hours; After the fermentation ends, the remaining sugar concentration in the fermented liquid is 7g/L.
The isolating method of said kojic acid is identical with embodiment 1, and difference is: tripping device 6 precoolings, and water cooler 6.3 temperature remain on 12 ℃; The fermented liquid temperature of controlled chilling device 6.3 afterbodys is 18 ℃.Fermentation and acid level among the embodiment 1 is 118g/L (the kojic acid acidity of promptly finally separating is 11.8%), and the remaining sugar concentration in the fermented liquid is 7g/L.
Embodiment 3
(1), take by weighing glucose 2590kg, sal epsom 3.5kg, potassium hydrogenphosphate 3.5kg, skimmer 1.4kg, the adding volume capacity is 25m
3Feed supplement jar 4 in, add water to 7m then
3, using phosphoric acid to regulate pH is 3.2, high pressure steam sterilization (pressure is 0.1MPa, and temperature is that 121 ℃, time are 30min) postcooling to 31 ℃ is subsequent use as feeding medium during fermentation;
(2), take by weighing glucose 840kg, yeast extract 126kg, sal epsom 7kg, potassium hydrogenphosphate 7kg, skimmer 2.8kg, the adding volume capacity is 25m
3Fermentor tank 1 in, add water to 14m then
3, stir, using phosphoric acid to regulate pH is 3.2, high pressure steam sterilization (pressure is 0.1MPa, and temperature is that 121 ℃, time are 30min) postcooling to 31 ℃ is as the fermentation basestocks; Insert bacterial classification, march acid-fermentation under 30 ℃ of conditions by 7% of fermentation basestocks volume;
(3), when fermenting, record remaining sugar concentration 44g/L in the fermented liquid, begin to mend feeding medium during fermentation, PM is mended into 1.9L, and the remaining sugar concentration in the control fermented liquid is controlled at 30～60g/L, finishes until feed supplement, produces bacteria growing, metabolic needs to satisfy to 24h;
When (4) fermenting, record kojic acid acidity in the fermented liquid, start the tripping device 6 in the fermentation equipment at 31g/L to 44h; Begin to carry out the product kojic acid and separate, until fermentation ends, fermentation period adds up to 93 hours; After the fermentation ends, the remaining sugar concentration in the fermented liquid is 11g/L.
The isolating method of said kojic acid is identical with embodiment 1, and difference is: tripping device 6 precoolings, and water cooler 6.3 temperature remain on 15 ℃; The fermented liquid temperature of controlled chilling device 6.3 afterbodys is 20 ℃.Fermentation and acid level among the embodiment 1 is 109g/L (the kojic acid acidity of promptly finally separating is 10.9%), and the remaining sugar concentration in the fermented liquid is 11g/L.
Claims (10)
1. a kojic acid zymotechnique is characterized in that comprising the steps:
(1), with glucose carbon source preparation feeding medium during fermentation, through sterilization and subsequent use after being cooled to the suitable growth temperature scope of production bacterium;
(2), be carbon source preparation fermentation basestocks with glucose, after sterilization and being cooled to the suitable growth temperature scope of producing bacterium, insert the production bacterium, the march acid-fermentation;
(3) when the remaining sugar concentration in the fermented liquid is reduced to 30 ~ 40g/L, begin to mend feeding medium during fermentation, in whole fermentation process, make the remaining sugar concentration in the fermented liquid be controlled at 30 ~ 60g/L, finish until feed supplement;
(4) when the kojic acid acidity in the fermented liquid during, begin to carry out kojic acid and separate, until fermentation ends at 30 ~ 40g/L.
2. kojic acid zymotechnique as claimed in claim 1 is characterized in that: said production bacterium is adopted aspergillus oryzae (Aspergillus oryzae) ACCC 30163.
3. kojic acid zymotechnique as claimed in claim 1 is characterized in that: the prescription of said fermentation basestocks consists of: glucose 50 ~ 60g/L, yeast extract 7.5 ~ 9g/L; Sal epsom 0.5g/L; Potassium hydrogenphosphate 0.5g/L, skimmer 0.2g/L transfers pH3.0 ~ 3.2 with phosphoric acid.
4. kojic acid zymotechnique as claimed in claim 1 is characterized in that: the prescription of said feeding medium during fermentation consists of: glucose 350 ~ 370g/L, and sal epsom 0.5g/L, potassium hydrogenphosphate 0.5g/L, skimmer 0.2g/L transfers pH3.0 ~ 3.2 with phosphoric acid.
5. kojic acid zymotechnique as claimed in claim 1 is characterized in that: said fermentation basestocks and feeding medium during fermentation all are to adopt high pressure steam sterilization, are cooled to 29～31 ℃ then.
6. kojic acid zymotechnique as claimed in claim 1 is characterized in that: the inoculum size of said production bacterium is 3～7% of fermentation basestocks volume.
7. kojic acid zymotechnique as claimed in claim 1 is characterized in that: the initial add-on of said fermentation basestocks is 50～60% of fermentor tank (a 1) volume capacity.
8. kojic acid zymotechnique as claimed in claim 1 is characterized in that: in the said step (3), feeding medium during fermentation be adopt the computer control continuous and quantitative mend into, the time that begins to mend into feeding medium during fermentation is at fermentation 24 ~ 36h; In the said step (4), beginning to carry out the kojic acid separation time is at fermentation 48 ~ 60h.
9. kojic acid fermentation equipment; Comprise fermentor tank (1); Fermentor tank (1) is used for splendid attire fermentation basestocks, is provided with whisking appliance (2) and thermostatically heating pipe (3) in the fermentor tank (1), and it is characterized in that: said fermentation equipment also comprises feed supplement jar (4) and tripping device (6); Said feed supplement jar (4) is used for splendid attire fermentation fluid infusion, and feed supplement jar (4) links to each other with material-feeding port on the fermentor tank (1); Said tripping device (6) is used for separating during the fermentation kojic acid; Tripping device (6) comprises water cooler (6.3), cone-shaped body (6.2) and scoop (6.1); The fluid inlet of water cooler (6.3) links to each other with the liquid outlet that separates of fermentor tank (1) through liquid-outlet valve (7); The liquid outlet of water cooler (6.3) links to each other with the osculum of cone-shaped body (6.2) upper end; Big mouthful of connection scoop (6.1) of cone-shaped body (6.2) lower end, scoop (6.1) side wall upper part is provided with liquid outlet, and the liquid outlet of scoop (6.1) links to each other with the liquid return hole that separates of fermentor tank (1) through returning liquid valve (5).
10. kojic acid fermentation equipment as claimed in claim 9; It is characterized in that: the method that said tripping device (6) separates kojic acid is: before beginning to separate kojic acid work; Open the water coolant of tripping device (6) earlier; Tripping device (6) is carried out precooling, make water cooler (6.3) temperature remain on 10 ~ 15 ℃; When beginning to separate, open earlier slowly liquid-outlet valve (7), make fermented liquid get into water cooler (6.3) and fully cooled off, and guarantee that the fermented liquid temperature of water cooler (6.3) afterbody is lower than 20 ℃; After being full of fermented liquid in the whole tripping device (6), open back liquid valve (5) again; Utilize temperature official post fermented liquid in tripping device (6), from up to down to flow automatically, thereby make the fluid power that ferments in fermented liquid and the fermentor tank (1) in the tripping device (6) exchange of in the regular hour, flowing; Fermented liquid in the tripping device (6) after being cooled to the kojic acid supersolubility temperature, kojic acid crystallization and separating out; Kojic acid that crystallization goes out and fermented liquid descend in tripping device (6) simultaneously; When dropping to cone-shaped body (6.2); Because the sectional area of cone-shaped body (6.2) increases gradually; The fermented liquid flow velocity slows down, and in scoop (6.1), the lower concentration fermented liquid of removing the part kojic acid is back in the fermentor tank (1) through returning liquid valve (5) crystal in the action of gravity settle.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107099451A (en) * | 2017-06-12 | 2017-08-29 | 成都金开生物工程有限公司 | A kind of kojic acid production system of energy-conserving and environment-protective |
| CN114908132A (en) * | 2022-05-06 | 2022-08-16 | 成都金开生物工程有限公司 | Method for preparing kojic acid by using biosynthesis method |
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| 李学如 等: "复合诱变对米曲霉产曲酸的影响", 《激光生物学报》 * |
| 陆正清 等: "曲酸的发酵法生产及其在食品加工中的应用", 《中国调味品》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107099451A (en) * | 2017-06-12 | 2017-08-29 | 成都金开生物工程有限公司 | A kind of kojic acid production system of energy-conserving and environment-protective |
| CN114908132A (en) * | 2022-05-06 | 2022-08-16 | 成都金开生物工程有限公司 | Method for preparing kojic acid by using biosynthesis method |
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