CN104561137A - Method for producing polyphenol by fermenting lachnum - Google Patents
Method for producing polyphenol by fermenting lachnum Download PDFInfo
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- CN104561137A CN104561137A CN201510020451.6A CN201510020451A CN104561137A CN 104561137 A CN104561137 A CN 104561137A CN 201510020451 A CN201510020451 A CN 201510020451A CN 104561137 A CN104561137 A CN 104561137A
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
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Abstract
The invention belongs to the technical field of microbial culture and discloses a method for producing polyphenol by fermenting lachnum. The method particularly comprises the following steps: taking hyphae from a bevel of preserved lachnum CCTCC No:M 209193, inoculating the hyphae to a potato glucose solid culture medium, activating, and culturing at 28-30 DEG C for 72-96 hours; inoculating a 1cm*1cm fungus block into a triangular flask containing a seed liquid culture medium, and carrying out shaking culturing at 28-30 DEG C for 48-72 hours, so as to obtain a seed liquid; inoculating 2%-5% of seed liquid by volume of a fermentation culture medium into a 5L mechanical stirring fermenting tank, and fermenting, wherein the fermentation culture medium contains the following components in percentage by weight: 1.5%-3% of glycerin, 0.1%-0.5% of yeast powder and the balance of water. The method has the beneficial effects that the constitution of the fermentation culture medium is simple, few bubbles are generated in the fermentation process, the fermentation time is short, and the content of polyphenol in a fermentation liquid is high, so that the production efficiency is improved, and the production cost is lowered.
Description
Technical field
The invention belongs to field of microbial culture technology, relate to the Technology that a kind of Lachnum liquid fermenting produces polyphenol.
Background technology
Polyphenol is the general name containing multiple phenolic hydroxyl-compounds, and it can be divided into flavonoid, tannins, phenolic acids and anthocyanin class etc.Polyphenol substance is because having anti-oxidant, radioprotective, the physiologically active such as antitumor, hypotensive, hypoglycemic and in widespread attention.Current active polyphenol class material mainly by extracting and utilizing fungal metabolite to produce from plant, extractive technique comparative maturity from plant, but cost is higher when producing in batches, extraction yield is lower, extracts polyphenol as Chinese Patent Application No. 201310504273.5 and 201410179149.0 discloses in a kind of bamboo shoots respectively and extract polyphenol from pomelo peel.
It is less that fungal metabolite produces polyphenol report, and as Zhu Jinwei utilizes Phaeopoms obliquus liquid submerged fermentation to produce polyphenol, its substratum weight percentage consists of: Semen Maydis powder 3.5, maize straw 3.0, peptone 0.3, KH
2pO
40.1, ZnSO
4.2H
2o 0.001, K
2hPO
40.04, FeSO
4.7H
2o 0.005, MgSO
4.7H
2o 0.05, CuSO
4.5H
2o 0.002, CoCl
20.001, MnSO
4.H2O 0.008, pH 6.0, in fermented liquid, polyphenol content reaches maximum constantly fermentation 216 is little, for 0.1357mg/mL (Zhu Jinwei, the liquid submerged fermentation preparation of Phaeopoms obliquus polyphenol and anti-oxidant activity [D], Hangzhou: Institutes Of Technology Of Zhejiang's master thesis, 2010).The production cycle that this method produces polyphenol is longer, and in fermented liquid, polyphenol content is low.Phaeopoms obliquus belongs to white-rot fungi, can lignocellulose degradation and Lachnum can not utilize lignocellulose.Qian Meishuan utilizes Lachnum fermentative production polyphenol, its substratum weight percentage consists of: sucrose 2.0, peptone 0.5, magnesium chloride 0.081, tyrosine 0.001, Sodium Nitroprusside 0.003, pH6, the outer polyphenol output of born of the same parents reaches maximum constantly fermentation 192 is little, for 0.148mg/ml (Qian Meishuan, Lachnum polyphenol fermentation condition, physicochemical characteristics and bioactivity research [D], Hefei: HeFei University of Technology's master thesis, 2013).The shortcoming of this substratum is that polyphenol output is lower.The invention provides a kind of new technical scheme.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, provide one to improve the quality of products, the Lachnum liquid fermenting reducing production cost produces the method for polyphenol.
Object of the present invention can be achieved through the following technical solutions:
Lachnum liquid fermenting produces a method for polyphenol, and the method concrete steps are as follows:
(1), mycelium inoculation is got on potato glucose solid medium from the inclined-plane of preservation Lachnum CCTCC No:M 209193;
(2), getting 1 centimetre * 1 centimetre bacterium block is seeded in the triangular flask that seed liquor substratum is housed;
(3), seed liquor is inoculated in by the 2-5% of fermention medium volume the 5L mechanical agitating fermentation tank that fermention medium is housed;
In described fermention medium, each composition weight percentage composition is: glycerine 1.5 ~ 3%, yeast powder 0.1 ~ 0.5%, and surplus is water.
Preferably, the mycelia described in step (1) on potato glucose solid medium, activation culture 72 ~ 96 hours at 28 ~ 30 DEG C.
Preferably, the bacterium block described in step (2) in seed liquor substratum, 28 ~ 30 DEG C of shaking culture 48 ~ 72 hours.
Preferably, the fermentation condition of the fermentation cylinder for fermentation substratum described in step (3) is: at 28 ~ 30 DEG C, and rotating speed is 180 ~ 250r/min, and air flow is 2 ~ 3L/min, cultivates 120 ~ 144 hours.
Preferably, in the seed liquor substratum described in step (2), each composition weight percentage composition is: glycerine 2 ~ 4%, yeast powder 0.5 ~ 1%, and surplus is water.
Beneficial effect of the present invention: substratum composition of the present invention is simple, and in fermenting process, foam is few, and polyphenol content is high, and fermentation time is short, thus improves production efficiency and ratio defective product, reduces production cost.
Embodiment
Be described in more detail the present invention below by embodiment, these embodiments are only the descriptions to best mode for carrying out the invention, not restricted to protection scope of the present invention.
Embodiment 1
Shovel mycelium inoculation on potato glucose solid medium from the inclined-plane of preservation Lachnum CCTCC No:M 209193 with inoculation, 30 DEG C of constant temperature culture 72 hours, get 1 centimetre * 1 centimetre bacterium block to be seeded to and 100mL seed liquor substratum is housed (consists of glycerine 2%, yeast powder 0.6%) 250mL triangular flask in, 30 DEG C, shaking culture 48 hours under 150r/min.(fermention medium forms: glycerine 1.5% 100mL seed liquor to be inoculated in the 5L mechanical agitating fermentation tank that 4L fermention medium is housed, yeast powder 0.2%, surplus is water, pH nature), 28 ~ 30 DEG C, 200r/min, air flow 2.5L/min, cultivate 132 hours, in fermented liquid, polyphenol content is 0.7963mg/ml (Forint phenol method, with gallic acid conversion).
Embodiment 2
Use from the inclined-plane of preservation Lachnum CCTCC No:M 209193 transfering loop picking one ring mycelium inoculation on potato glucose solid medium, cultivate 96 hours at 30 DEG C.Getting 1 centimetre * 1 centimetre bacterium block is seeded in the 500mL triangular flask that 200mL seed liquor substratum (consisting of glycerine 2%, yeast powder 0.8%) is housed, 30 DEG C, shaking culture 72 hours under 150r/min.(fermention medium forms: glycerine 2% 200mL seed liquor to be inoculated in the 5L mechanical agitating fermentation tank that 4L fermention medium is housed, yeast powder 0.5%, surplus is water, pH nature), 28 ~ 30 DEG C, 230r/min, air flow 3.0L/min, cultivate 144 hours, in fermented liquid, polyphenol content is 0.7663mg/ml (Forint phenol method, with gallic acid conversion).
Embodiment 3
Shovel mycelium inoculation on potato glucose solid medium from the inclined-plane of preservation Lachnum CCTCC No:M 209193 with inoculation, cultivate 72 hours at 28 DEG C, get 1 centimetre * 1 centimetre bacterium block to be seeded to and 150mL seed liquor substratum is housed (consists of glycerine 2%, yeast powder 0.5%) 500mL triangular flask in, 30 DEG C, shaking culture 48 hours under 150r/min.(fermention medium forms: glycerine 3% 150mL seed liquor to be inoculated in the 5L mechanical agitating fermentation tank that 4L fermention medium is housed, yeast powder 0.1%, 4L water, pH nature), 28 ~ 30 DEG C, 180r/min, air flow 2.2L/min, cultivate 120 hours, in fermented liquid, polyphenol content is 0.8573mg/ml (Forint phenol method, with gallic acid conversion).
Comparative example
Shovel mycelium inoculation on potato glucose solid medium from the inclined-plane of preservation Lachnum CCTCC No:M 209193 with inoculation, cultivate 72 hours at 30 DEG C, get 1 centimetre * 1 centimetre bacterium block to be seeded to and 150mL seed liquor substratum is housed (consists of glycerine 2%, yeast powder 0.5%) 500mL triangular flask in, 30 DEG C, shaking culture 48 hours under 150r/min.(fermention medium forms: glycerine 3% 150mL seed liquor to be inoculated in the 5L mechanical agitating fermentation tank that 4L fermention medium is housed, yeast powder 1%, 4L water, pH nature), 28 ~ 30 DEG C, 180r/min, air flow 2.2L/min, cultivate 120 hours, in fermented liquid, polyphenol content is 0.5265mg/ml (Forint phenol method, with gallic acid conversion).
Above-mentioned Lachnum CCTCC No:M 209193, be preserved in China typical culture collection center, address: Luo Jia Shan, wuchang, wuhan Wuhan University Life Science College, preservation date on September 3rd, 2009, deposit number CCTCC No:M 209193, Classification And Nomenclature: Lachnum (Lachnum sp.).
Found by research, the somatomedin of adding in conventional medium, as tyrosine, inorganic salt etc., is cultivated by changing and is formed and do not need to add somatomedin in addition.Lachnum CCTCC No:M 209193 fermention medium of the present invention consists of glycerine, yeast powder, does not need to add other somatomedins.
Glycerine is as the by product of production of biodiesel process, cheap.We study and find what the outer polyphenol of Lachnum born of the same parents was mainly synthesized by shikimic acid pathway and polyketone approach, and glycerine can promote the metabolic flux of shikimic acid pathway and polyketone approach in Lachnum, facilitates the synthesis of polyphenol; On the other hand, glycerine can also promote that Lachnum metabolism produces kojic acid, and kojic acid can suppress the activity of polyphenoloxidase, thus decreases polyphenol to melanic conversion, improves the output of polyphenol.In fermenting process, foam is less.
The polyphenol of Lachnum synthesis is secondary metabolite, just starts synthesis when nutritive substance approach exhaustion.We study and find that culture media nitrogen source is conducive to the synthesis of polyphenol when being yeast powder, and add a small amount of yeast powder and Lachnum can be made to enter the secondary metabolism stage in advance, fermentation time reduction was by 120 ~ 144 hours; A small amount of yeast powder makes biomass reduce, but to be conducive to more transformation of glycerol be polyphenol; The yeast powder of low levels reduces amino acid concentration in fermented liquid, is conducive to being further purified of polyphenol, reduces production cost.
Medium pH of the present invention is about 6.5 (after sterilizings), and Lachnum is well-grown in the scope of pH4.5 ~ 8.0, and therefore basal culture medium is before sterilization without the need to acid-alkali accommodation pH, simplifies operation.
Carbon-nitrogen ratio in substratum of the present invention is relatively more reasonable, and the potential of hydrogen change of prior fermentation process is little, and the fermentation later stage makes pH value reduce due to the generation of polyphenol and kojic acid, is about 4.0 ~ 4.5, is conducive to the formation of polyphenol to pH during fermentation ends.
The object of seed liquor substratum of the present invention is long thalline, and it is little that seed culture medium changes the impact of other composition tunning amounts into, but the possibility cycle can slightly long point.
Adopt substratum of the present invention, make fermentation time reduction by 120 ~ 144 hours, in 5L fermentor tank, polyphenol content is 0.75 ~ 0.90mg/ml (forint ~ phenol method, converts as gallic acid), improves quality product, reduces production cost.
Leavening temperature of the present invention is higher, is 28 ~ 30 DEG C, and in this temperature range, thalli growth rapidly and be beneficial to the synthesis of polyphenol, and shorten fermentation period.
Above content is only citing made for the present invention and explanation; affiliated those skilled in the art make various amendment to described specific embodiment or supplement or adopt similar mode to substitute; only otherwise depart from invention or surmount this scope as defined in the claims, protection scope of the present invention all should be belonged to.
Claims (5)
1. Lachnum liquid fermenting produces a method for polyphenol, and the method concrete steps are as follows:
(1) mycelium inoculation is got on potato glucose solid medium from the inclined-plane of preservation Lachnum CCTCC No:M 209193;
(2) getting 1 centimetre * 1 centimetre bacterium block is seeded in the triangular flask that seed liquor substratum is housed;
(3) seed liquor is inoculated in by 2 ~ 5% of fermention medium volume the 5L mechanical agitating fermentation tank that fermention medium is housed;
It is characterized in that, in the fermention medium described in step (3), each composition weight percentage composition is: glycerine 1.5 ~ 3%, yeast powder 0.1 ~ 0.5%, and surplus is water.
2. a kind of Lachnum liquid fermenting according to claim 1 produces the method for polyphenol, it is characterized in that, the mycelia described in step (1) on potato glucose solid medium, activation culture 72 ~ 96 hours at 28 ~ 30 DEG C.
3. a kind of Lachnum liquid fermenting according to claim 1 produces the method for polyphenol, it is characterized in that, the bacterium block described in step (2) in seed liquor substratum, 28 ~ 30 DEG C of shaking culture 48 ~ 72 hours.
4. a kind of Lachnum liquid fermenting according to claim 1 produces the method for polyphenol, it is characterized in that, the fermentation condition of the fermentation cylinder for fermentation substratum described in step (3) is: at 28 ~ 30 DEG C, mixing speed is 180 ~ 250r/min, air flow is 2 ~ 3L/min, cultivates 120 ~ 144 hours.
5. a kind of Lachnum liquid fermenting according to claim 1 produces the method for polyphenol, and it is characterized in that, in the seed liquor substratum described in step (2), each composition weight percentage composition is: glycerine 2 ~ 4%, yeast powder 0.5 ~ 1%, and surplus is water.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108715880A (en) * | 2018-05-29 | 2018-10-30 | 合肥工业大学 | A kind of preparation method of the fat-soluble uranidin of Lachnum |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009101959A1 (en) * | 2008-02-13 | 2009-08-20 | Daiichi Sankyo Company, Limited | Novel compound, lachnochromonin compound |
| JP2012051897A (en) * | 2011-09-27 | 2012-03-15 | Forestry & Forest Products Research Institute | Antibacterial agent, oral cavity composition and food and drink containing the same |
| CN102746705A (en) * | 2012-06-28 | 2012-10-24 | 合肥工业大学 | Method for modifying lachnum extracellular melanin into water-soluble melanin |
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2015
- 2015-01-15 CN CN201510020451.6A patent/CN104561137B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009101959A1 (en) * | 2008-02-13 | 2009-08-20 | Daiichi Sankyo Company, Limited | Novel compound, lachnochromonin compound |
| JP2012051897A (en) * | 2011-09-27 | 2012-03-15 | Forestry & Forest Products Research Institute | Antibacterial agent, oral cavity composition and food and drink containing the same |
| CN102746705A (en) * | 2012-06-28 | 2012-10-24 | 合肥工业大学 | Method for modifying lachnum extracellular melanin into water-soluble melanin |
Non-Patent Citations (2)
| Title |
|---|
| 叶明等: "辛格粒毛盘菌胞内多酚提取及其抗氧化性研究", 《食品科学》 * |
| 钱梅双等: "粒毛盘菌YＭＵ50多酚发酵与纯化", 《合肥工业大学学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108715880A (en) * | 2018-05-29 | 2018-10-30 | 合肥工业大学 | A kind of preparation method of the fat-soluble uranidin of Lachnum |
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