CN102409067B - Culture medium for producing kojic acid by fermenting aspergillus oryzae, and fermentation method thereof - Google Patents
Culture medium for producing kojic acid by fermenting aspergillus oryzae, and fermentation method thereof Download PDFInfo
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- CN102409067B CN102409067B CN2011104134158A CN201110413415A CN102409067B CN 102409067 B CN102409067 B CN 102409067B CN 2011104134158 A CN2011104134158 A CN 2011104134158A CN 201110413415 A CN201110413415 A CN 201110413415A CN 102409067 B CN102409067 B CN 102409067B
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Abstract
The invention provides a culture medium for producing kojic acid by standing fermentation of aspergillus oryzae. Every 1.5 liters of culture medium comprises the following components in weight: 10-30g of mannitol, 1-5g of yeast extract, 10-30g of maltose, 1-5g of monosodium glutamate, 3-15g of glucose, 1-10g of peptone, 50-200g of mashed potato and the balance of distilled water. The invention also provides a new standing fermentation method. The invention has the advantages of easy availability of raw materials, low energy consumption and high yield, and provides a new clue and a new method for research on fermentation of aspergillus oryzae and medical research on kojic acid.
Description
Technical field
The present invention relates to the microorganism culturing technical field, more specifically relate to a kind of substratum and fermentation process that produces kojic acid with the aspergillus oryzae fermentation.
Background technology
Kojic acid (5-hydroxyl-2-methylol-1, pyrokomane, English name Kojic acid), it is a kind of melanochrome specificity inhibitor, it enter after skin cells can with cell in the cupric ion complexing, change the three-dimensional arrangement of tyrosine oxidase, stop the activation of tyrosine oxidase, thus the formation of check melanin.Kojic acid class whitening promoting agent has better tyrosine oxidase inhibition than other whitening promoting agent.It does not act on other biological enzyme in cell, and cell is not had to toxic action, and it can also enter in intercellular substance simultaneously, forms the intercellular colloid, plays the effect of water conservation and increase skin elasticity.Allocated in various makeup at present, made the skin-lightening cosmetic for freckle, senile plaque, pigmentation and acne.
Kojic acid is the organic acid that utilizes fermentative Production, China's kojic acid producing process that begins one's study from the eighties, but take in the past import in 97 years as main always.Calendar year 2001, Shaanxi Institute of Microbiology adopted microorganism compound mutation breeding technology, selected flavus mutant strain UCN7-17, for China's kojic acid production is laid a good foundation.At present, the kojic acid production technology of China is also more backward, and production unit is many, complicated operation, and energy consumption is large, and production technique need to improve.
Summary of the invention
Substratum and the fermentation process thereof for the aspergillus oryzae fermentation product kojic acid that the object of the present invention is to provide a kind of raw material to be easy to get, the method energy consumption is low, cultural method is simple.
Technical scheme of the present invention is as follows:
The invention provides a kind of substratum that produces kojic acid for the aspergillus oryzae fermentation, contain in every 1.5 liters of substratum: N.F,USP MANNITOL 10～30g, yeast extract paste 1～5g, maltose 10～30g, monosodium glutamate 1～5g, glucose 3～15g, peptone 1～10g, mashed potato 50～200g, all the other are water.
More preferably, in every 1.5 liters of substratum, contain: N.F,USP MANNITOL 15g, yeast extract paste 2.75g, maltose 15g, monosodium glutamate 3.75g, glucose 7.5g, peptone 3.75g, mashed potato 150g, all the other are water.
Described water is first water.
The present invention also provides a kind of fermentation process that utilizes above-mentioned culture medium culturing aspergillus oryzae to produce kojic acid, comprises the following steps:
(1) female enlarged culturing of planting;
(2) inoculation medium standing for fermentation;
(3) condition of described standing for fermentation is: anaerobically fermenting, and leavening temperature is 26-28 ℃, the time of standing for fermentation is 32-36 days.
The condition optimization of described standing for fermentation is: anaerobically fermenting, and leavening temperature is 28 ℃, the time of standing for fermentation is 34 days.
Remarkable advantage of the present invention:
Culture medium raw material of the present invention is easy to get, easy for the production technique of producing kojic acid, energy consumption is low, output is large.
Embodiment
The mensuration of kojic acid output
Get 1,50ml volumetric flask, add 1ml developer (1%FeCl
3-HCL), add the clarified broth 9.8ml after fermentation, with first water, be settled to 50ml, with ultraviolet spectrophotometer, measure its OD
500Value, compare with typical curve, obtains substratum kojic acid output.
Content for a better understanding of the present invention, be described further below in conjunction with specific embodiment, but the present invention is not limited only to this.
Embodiment 1
1.1 the configuration of substratum:
Test group: take N.F,USP MANNITOL 15g, yeast extract paste 2.75g, maltose 15g, monosodium glutamate 3.75g, glucose 7.5g, peptone 3.75g, mashed potato 150g, first with appropriate warm water, yeast extract paste is dissolved, after progressively add other composition, be settled to 1500ml with first water after all dissolving.
Control group 1: take yeast extract paste 2.25g, N.F,USP MANNITOL 15g, monosodium glutamate 7.5g, glucose 7.5g, maltose 15g, sal epsom (MgSO
47H
2O) 0.225g, potassium hydrogen phosphate 0.375g, sodium-chlor 11.25g, first dissolve yeast extract paste with appropriate warm water, after progressively add other composition, be settled to 1500ml with first water after all dissolving.
Control group 2: take SODIUMNITRATE 4.5g, potassium hydrogen phosphate 1.5g, sal epsom (MgSO
47H
2O) 0.75g, Repone K 0.75g, ferric sulfate 0.015g, sucrose 45g, be settled to 1500ml with first water.
Control group 3: take peptone 7.5g, yeast extract paste 3.75g, glucose 15g, first dissolve yeast extract paste with appropriate warm water, after progressively add other composition, be settled to 1500ml with first water after all dissolving.
Control group 4: take glucose 30g, peptone 7.5g, sodium-chlor 3.75g, add first and with appropriate warm water, yeast extract paste dissolved, after progressively add other composition, be settled to 1500ml with first water after all dissolving.
1.2 the fermentation aspergillus oryzae is produced kojic acid
By the aspergillus oryzae of stored frozen (aspergillus oryzae be aspergillus oryzae (
Aspergillus oryzae) IBPT-1, be preserved in Chinese Typical Representative culture collection center (CCTCC) on December 1st, 2011, address: Wuhan Wuhan University, deposit number CCTCC NO:M2011437) inclined-plane thaws, and spreads cultivation to plate culture medium 3-5 days, then spreads cultivation to test tube, after 2-3 days, spread cultivation to above-mentioned substratum (tool plug Erlenmeyer flask), at 28 ℃ of temperature, standing cultivation 34 days, stop cultivating.
1.3 the mensuration of kojic acid output
Get 1,50ml volumetric flask, add 1ml developer (1%FeCl
3-HCL), add clarified broth 9.8ml after 1.2 fermentations, with first water, be settled to 50ml, with ultraviolet spectrophotometer, measure its OD
500Value and typical curve are compared, and obtain substratum kojic acid output.
The substratum kojic acid output that test group and control group 1-4 obtain is respectively 968mg/L, 173.4mg/L, 55.8mg/L, 167.6mg/L, 133.8mg/L.
By the comparison of above 5 kinds of different culture medias, find the output maximum of substratum of the present invention.
Claims (3)
1. the aspergillus oryzae for generation of kojic acid is characterized in that: described aspergillus oryzae be aspergillus oryzae (
Aspergillus oryzae) IBPT-1, be preserved in Chinese Typical Representative culture collection center C CTCC on December 1st, 2011, address: Wuhan Wuhan University, deposit number CCTCC NO:M2011437; Adopt the described aspergillus oryzae of following culture medium culturing to produce kojic acid:
In every 1.5 liters of substratum, contain: N.F,USP MANNITOL 10～30g, yeast extract paste 1～5g, maltose 10～30g, monosodium glutamate 1～5g, glucose 3～15g, peptone 1～10g, mashed potato 50～200g, all the other are water.
2. a fermentation process that utilizes aspergillus oryzae claimed in claim 1 to produce kojic acid, is characterized in that, comprises the following steps:
(1) female enlarged culturing of planting;
(2) inoculation medium, standing for fermentation; The condition of described standing for fermentation is: anaerobically fermenting, and leavening temperature is 26-28 ℃, the time of standing for fermentation is 32-36 days.
3. fermentation process according to claim 2, it is characterized in that: the condition of described standing for fermentation is: anaerobically fermenting, leavening temperature is 28 ℃, the time of standing for fermentation is 34 days.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105018352A (en) * | 2015-07-23 | 2015-11-04 | 云南大学 | Fungal strain capable of producing kojic acid and preparation method of fungal strain |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106755164A (en) * | 2016-12-23 | 2017-05-31 | 江南大学 | A kind of method for improving aspergillus oryzae kojic acid yield |
Citations (3)
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|---|---|---|---|---|
| CN1263153A (en) * | 1999-09-24 | 2000-08-16 | 崔英波 | Method for preparing amino acylase by means of immobilization of Aspergillus oryzae and equipment for resolution and suspension of amino acid |
| CN100338226C (en) * | 2005-07-29 | 2007-09-19 | 天津科技大学 | Method for preparing 11 alpha bydroxy canrenone through conversion method of batch fermentation of microbe |
| CN102187947A (en) * | 2011-07-01 | 2011-09-21 | 湖南农业大学 | Feed additive and feed containing same |
-
2011
- 2011-12-10 CN CN2011104134158A patent/CN102409067B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1263153A (en) * | 1999-09-24 | 2000-08-16 | 崔英波 | Method for preparing amino acylase by means of immobilization of Aspergillus oryzae and equipment for resolution and suspension of amino acid |
| CN100338226C (en) * | 2005-07-29 | 2007-09-19 | 天津科技大学 | Method for preparing 11 alpha bydroxy canrenone through conversion method of batch fermentation of microbe |
| CN102187947A (en) * | 2011-07-01 | 2011-09-21 | 湖南农业大学 | Feed additive and feed containing same |
Non-Patent Citations (4)
| Title |
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| 曲酸菌的选育及发酵工艺条件研究;李凤林等;《发酵科技通讯》;20090731;第38卷(第3期);15-18 * |
| 曲酸高产菌株的诱变选育;赵鑫等;《激光生物学报》;20100831;第19卷(第4期);542-545 * |
| 李凤林等.曲酸菌的选育及发酵工艺条件研究.《发酵科技通讯》.2009,第38卷(第3期),15-18. |
| 赵鑫等.曲酸高产菌株的诱变选育.《激光生物学报》.2010,第19卷(第4期),542-545. |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105018352A (en) * | 2015-07-23 | 2015-11-04 | 云南大学 | Fungal strain capable of producing kojic acid and preparation method of fungal strain |
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