CN107190026B - Method for improving secondary metabolite of monascus - Google Patents

Method for improving secondary metabolite of monascus Download PDF

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CN107190026B
CN107190026B CN201710607599.9A CN201710607599A CN107190026B CN 107190026 B CN107190026 B CN 107190026B CN 201710607599 A CN201710607599 A CN 201710607599A CN 107190026 B CN107190026 B CN 107190026B
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张婵
王成涛
孙宝国
杨乐
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Beijing Technology and Business University
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Abstract

The invention provides a method for improving monascus secondary metabolite, which is characterized in that 5-azacytidine is added in the initial liquid fermentation or fermentation process of monascus. The 5-azacytidine is added within 48 hours of liquid fermentation culture of monascus, and the addition amount is 10‑6g/L to 1 g/L. The invention can improve most secondary metabolites of monascus, is simple, convenient and quick, and does not produce other harmful byproducts.

Description

Method for improving secondary metabolite of monascus
Technical Field
The invention belongs to the field of microbial culture, and relates to a method for improving a monascus secondary metabolite.
Background
Monascus purpureus Went, Chinese alias Monascus, red vinasse and red rice exist in trees, soil, deposits and the like. Is an important microbial resource in China, can produce wide natural products with biological activity, and has high commercial value. The temperature range suitable for the growth of monascus is wide, the general temperature is 15-42 ℃, the optimal temperature is 25-30 ℃, the optimal pH value for the growth is 3.5-5, and the monascus can resist ethanol with the concentration of 10%. The bacterial colony grows well on a malt extract agar culture medium, is initially white, becomes light pink, purple or gray black after aging, and is red in color. The application of red yeast rice has been in the history for thousands of years, the medical efficacy of the red yeast rice is recorded in ancient books of China, and the red yeast rice can be used for brewing wine, making vinegar, making a coloring agent and a flavoring agent of fermented bean curd, and can also be used as a traditional Chinese medicine. The existing research shows that the red yeast rice has multiple effects of reducing blood fat, reducing blood pressure, resisting oxidation, resisting cancer, resisting bacteria and fatigue, preventing senile dementia and the like.
The monascus produces secondary metabolites mainly including: monascus pigment, Monacolins compounds, citrinin and other metabolites. The monascus pigment is a mixed pigment and mainly comprises a red pigment, a yellow pigment and an orange pigment. With respect to the biosynthesis of monascin, it is generally believed that the possible processes are: polyketide synthase firstly catalyzes acetyl CoA and malonyl CoA to generate a hexanone chromophore, the hexanone chromophore and medium-chain fatty acid generated in a fatty acid synthesis way generate orange pigment through transesterification, and the orange pigment respectively forms red pigment and yellow pigment through ammoniation reaction and reduction action. The current research shows that amino acid is an important influence factor in the synthesis process of monascus pigment, and different types and contents of amino acid can obviously influence the composition and yield of monascus pigment. The Monacolins compounds have lipid-lowering and anticancer activities. With the increase of hypertension patients, the annual demand of MonacolinK is also increasing year by year. Therefore, the method is simple and quick, the yield is improved, and the practical significance is achieved.
At present, the aim of improving the secondary metabolite of monascus is achieved mainly by regulating and controlling the fermentation process of monascus. For example, application No. 201710071293.6 entitled Monascus liquid fermentation method for producing monascus aurantiol is disclosed, wherein after inoculating monascus seed culture liquid into liquid culture medium, fermenting at 30 deg.C for 6-8 days, and adding surfactant (Span series, Tween series, TritonX-100, PEG series or Chinese honeylocust) into the fermentation culture liquid within 24h after fermentation begins to promote monascus to synthesize the active substance monascus aurantiol during growth process. Application No. 201410117001.4 discloses a coupled in-situ extraction and fermentation method for preparing monascus orange pigment by a two-liquid fermentation method, which is characterized in that the synthetic monascus orange pigment is rapidly enriched into a fat-soluble solvent by designing reasonable carbon and nitrogen source components, controlling the content of free amino acid in a culture medium and a two-liquid fermentation system, so that the production efficiency of the monascus orange pigment is improved. Application No. 201610475354.0 discloses a fermentation method for increasing the yield of haematochrome and yellow pigment in monascus purpureus, wherein histidine or tyrosine is additionally added into an amino nitrogen source-free culture medium, so that the synthesis amount of the haematochrome or the yellow pigment can be remarkably increased.
However, the prior art and the method can only improve partial secondary metabolites of the monascus and have limited application range. At present, no relevant report for simultaneously improving most monascus secondary metabolites exists.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a simple, convenient and quick method for improving most secondary metabolites of monascus.
In order to achieve the above object, the present invention provides a method for increasing a secondary metabolite of monascus, which comprises adding 5-azacytidine at the beginning of or during liquid fermentation of monascus.
The 5-azacytidine is added within 48h of liquid fermentation culture of monascus, preferably, the 5-azacytidine is added within 24h of liquid fermentation culture of monascus.
The addition amount of the 5-azacytidine is 10-6g/L-1 g/L, the best addition amount is 10-2g/L。
The fermentation culture conditions are air bath, 33 ℃, 200r/min culture for 18 days.
The monascus is monascus M1 or monascus RP 2.
In the liquid fermentation process of red yeast rice RP2 and M1, the inventor of the application unexpectedly finds that after the 5-azacytidine is added, secondary metabolites of the two strains are improved (namely, the yields of monascus haematochrome, orange pigment, yellow pigment and MonacolinK are improved), and the wrinkles and filaments of the strains added with the 5-azacytidine are increased as shown by electron microscope observation. Presumably, 5-azacytidine contributes to the generation of DNA methylation, which promotes the increase of metabolites of filamentous fungi; the folds and filaments increase the surface area of the cells, so that secondary metabolites in the cells are easier to discharge out of the body, and the accumulation of the secondary metabolites in the body is reduced, thereby increasing the amount of the secondary metabolites.
The invention has the following beneficial effects:
(1) can greatly improve the yield of the secondary metabolites of the monascus.
The invention is characterized in that the purpose of simultaneously improving the yield of the monascus red pigment, the orange pigment, the yellow pigment and the MonacolinK is realized by adding a substance. The research of the invention shows that the thallus dry weights of two strains of monascus M1 and monascus RP2 added with 5-azacytidine are respectively improved by 7.7 percent and 5.1 percent compared with the strains without the 5-azacytidine original culture medium, the red pigment in the color value of the monascus pigment is respectively improved by 19.6 percent and 15.6 percent, the orange pigment is respectively improved by 16.3 percent and 20.6 percent, and the yellow pigment is respectively improved by 16.4 percent and 15 percent; the MonacolinK yield of the monascus M1 is improved by 58.6%.
(2) Is economical and efficient.
The method can achieve the aim of simultaneously improving various secondary metabolites by only adding a proper amount of 5-azacytidine in a proper culture time, and is more economic and efficient compared with the method in the prior art.
(3) The applicability is wider.
In the prior art, different substances are usually added for certain secondary metabolites to achieve the purpose of improving the yield. The method has strong pertinence but limited application range, and if high-yield secondary metabolites are required to be obtained, separate fermentation production is required. The method can achieve the aim of improving various metabolites simultaneously in the process of one-time fermentation.
(4) No by-product.
The method of the invention does not produce other harmful byproducts in the fermentation process and does not increase the yield of the citrinin.
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FIG. 1 shows the result of a single-factor experiment of adding 5-azacytidine to monascus purpureus M1.
FIG. 2 shows the result of a single-factor experiment of adding 5-azacytidine to monascus purpureus RP 2.
FIG. 3 shows the results of a single-factor experiment of the concentration of 5-azacytidine in monascus purpureus RP 2.
FIG. 4 shows the result of a single-factor experiment of adding 5-azacytidine to monascus purpureus M1.
FIG. 5 is a comparison of the dry cell weights of the experimental group and the control group of Monascus purpureus M1 at the optimum amount and time of addition of 5-azacytidine.
FIG. 6 is a comparison of the cell dry weights of the experimental group and the control group of monascus purpureus RP2 at the optimum amount and time of addition of 5-azacytidine.
FIG. 7 is a comparison of monascus red pigment (A), orange pigment (B) and yellow pigment (C) in the experimental group and the control group of monascus purpureus M1 at the optimum amount and time of addition of 5-azacytidine.
FIG. 8 is a graph showing the comparison of monascus red pigment (A), orange pigment (B) and yellow pigment (C) in the experimental group and the control group of monascus purpureus RP2 at the optimum amount and time of addition of 5-azacytidine.
FIG. 9 is a graph showing a comparison of MonacolinK against the control group of the test group of MonacolinK of Monascus purpureus M1 at the optimum amount and time of addition of 5-azacytidine.
FIG. 10 is a high performance liquid phase diagram of the test group (a) and the control group (b) of Monascus purpureus M1 at the optimum amount and time of addition of 5-azacytidine.
FIG. 11 is an electron micrograph of the experimental group and the control group after culturing for 8 days at the optimum addition amount and addition time of 5-azacytidine. Wherein (a, c and e) are graphs of RP2 control strains, and (b, d and f) are graphs of RP2 experimental strains added with 5-azacytidine; (g, i, k) is a map of M1 control strain, and (h, j, l) is a map of M1 experimental strain supplemented with 5-azacytidine.
Detailed Description
The technical solution and the technical effects thereof are further illustrated by the following specific test methods and the accompanying drawings, and the following description is only for explaining the present invention but not limiting the present invention in any way, and any modifications or alterations based on the teaching of the present invention are within the protection scope of the present invention. The methods of the present invention are conventional in the art unless otherwise specified. The reagents used are commercially available, unless otherwise specified.
Example 1
1. Strains and materials
The strain of the monascus purpureus M1 (provided by China center for Collection of microbial resources) and the strain of the monascus purpureus RP2 (collected in the laboratory of Beijing university of Industrial and commercial sciences) is stored in a refrigerator at the temperature of 80 ℃ below zero in a laboratory.
Plate culture medium: PDA culture medium
Seed liquid culture medium of RP2 strain: 40g/L of long-shaped rice flour, 8g/L of peptone and 5g/L, KH of soybean meal2PO4 2g/L、NaNO3 2g/L、MgSO4Shaking in air bath at 33 deg.C and 200r/min for 48h at 1g/L, and transferring the fermentation broth.
RP2 strain fermentation broth medium: 77g/L of indica rice flour, 75g/L of glucose and 2g/L, KH of soybean meal2PO4 0.5g/L、NaNO31.8g/L、MgSO41g/L of corn steep liquor, 3.5g/L of corn steep liquor and an air bath shaker under the conditions of 33 ℃ and 200 r/min.
M1 strain seed liquid medium: 30g/L glucose and 15g/L, MgSO soybean meal4 1g/L KH 2PO42g/L, glycerol 70g/L, peptone 10g/L, NaNO32g/L, shaking in an air bath for 48h at 30 ℃ and 200r/min, and transferring the fermentation liquor.
M1 strain fermentation liquid culture medium: 90g/L of glycerin, 20g/L of long-shaped rice flour and 10g/L, NaNO of peptone3 5g/L、MgSO41g/L、ZnSO4 2g/L、KH 2PO 4Shaking in air bath at 30 deg.C and 150r/min for 48h at 2.5g/L, and adjusting shaking table conditions to 25 deg.C and 150 r/min.
2. Time point and addition amount of 5-azacytidine
5-azacytidine mainly acts on the G2 phase of cell mitosis, and the specific time period of the G2 phase of red yeast rice is not reported at present, so the optimal addition time is determined. After fermenting the red yeast rice M1 and the red yeast rice RP2 for two days, the logarithmic phase begins, so the G2 phase of the red yeast rice is necessarily within 48 hours of the seed liquid fermentation liquid. In the experiment, 4 hours are taken as an adding time point, 5-azacytidine is respectively added into 50ml of fermentation liquor, so that the concentration of the 5-azacytidine reaches 10-3g/L to determine the optimal addition timing.
After the optimal time point was obtained, by adding different concentrations of 5-azacytidine to 50ml of fermentation broth. The concentration gradient of 5-azacytidine in the fermentation broth was: 1g/L, 10-1g/L、10-2g/L、10-3g/L、10-4g/L、10-5g/L、10-6g/L. To investigate the optimum amount of 5-azacytidine added.
The method comprises the following steps:
(1) the monascus strain and the culture medium are adopted. Adding 5-azacytidine into 50ml fermentation liquid within 48 hr of fermentation liquid of Monascus ruber seed liquid, respectively, at a time point of 4 hr, to allow fermentationThe concentration of 5-azacytidine in the fermentation liquid reaches 10-3g/L. The monascus seed liquid culture conditions are as follows: culturing in air bath at 33 deg.C and 200r/min for 48 hr to obtain fermentation solution; fermentation culture conditions: inoculating the seed liquid into a fermentation culture medium according to the inoculation amount of 10% (V/V), and culturing at the temperature of 33 ℃ and 200r/min in an air bath. The results are shown in FIGS. 1 and 2.
(2) Adding 5-azacytidine with different concentrations into 50ml of fermentation liquor within 24 hours after the seed liquor is inoculated to the fermentation liquor; the concentration gradient of the 5-azacytidine in the fermentation liquor is as follows: 1g/L, 10-1g/L、10-2g/L、10-3g/L、10-4g/L、10-5g/L、10-6g/L. As shown in FIGS. 3 and 4, when 5-azacytidine was at a low concentration, the yield of secondary metabolites increased with the increase in concentration, and reached 10 at the concentration of 5-azacytidine-2When the yield reaches the maximum g/L, the concentration is continuously increased, the side effect of the 5-azacytidine begins to appear, and the yield of the secondary metabolite is gradually reduced.
3. Comparative experiment of 5-azacytidine-added experimental group and 5-azacytidine-not-added control group
In the test group, 5-azacytidine is added within 48 hours after the liquid fermentation culture is started, and the addition amount is 10-2g/L. The control group was supplemented with 5-azacytidine in the same amount as sterile water. Comparing the dry weight of the two groups of thalli, the pH value, the pigment color value of the monacolin, the yield of the MonacolinK and the shape of the mycelia observed by an endoscope on the 8 th day.
3.1 comparison of the Dry weight of Red Rice cells
Adding 5-azacytidine experimental monascus M1 and RP2 and control monascus M1 and RP2 which are not added with 5-azacytidine and added with the same amount of sterilized water into 2, 5, 8, 10, 12, 14, 16 and 18 days of fermentation, respectively taking 5ml of fermentation liquor, filtering the fermentation liquor by using 4 layers of gauze, washing the fermentation liquor on the gauze by using sterile water until the liquid flowing out of the gauze becomes colorless, drying the gauze to constant weight at the temperature of 40 ℃, and weighing the gauze. As shown in FIGS. 5 and 6, 5-azacytidine had an effect on the growth of cells at the beginning, so that the dry weight of the experimental group was significantly lower than that of the control group. However, from the general trend, 5-azacytidine has the effect of promoting cell growth. As can be seen from FIGS. 5 and 6, the experimental group to which 5-azacytidine was added from day 8 showed a significant increase in dry weight of the cells as compared to the control group to which 5-azacytidine was not added, and reached a maximum value when the fermentation was continued for 12 days. Following this time, the cells enter the decline phase and the dry weight of the bacteria begins to gradually decrease.
3.2 comparison of pH value of Monascus purpureus
The pH values of the monascus M1 and RP2 in the experimental group and the control group were measured by pH meter at 2, 5, 8, 10, 12, 14, 16, 18 days, respectively, and the results are shown in Table 1.
TABLE 1 pH values of the test group and the control group at different incubation times
Figure BDA0001358745630000061
As can be seen from Table 1, 5-azacytidine had a slight effect on the pH of Red Rice M1 with little effect on Red Rice RP 2.
3.3 comparison of color values of Monascus purpureus pigment
Fermenting for 2, 5, 8, 10, 12, 14, 16 and 18 days, respectively taking 3ml of fermentation liquor of monascus M1 and RP2 of an experimental group and a control group, then adding 6ml of 70% ethanol, carrying out water bath for 1h at 60 ℃, centrifuging for 15min at 4000r, diluting by corresponding times (diluting by 70% ethanol), and respectively measuring the color values of monascus red, orange and yellow pigments at 505nm, 465nm and 410 nm. Measuring the OD value of the red pigment by a spectrophotometer under the wavelength of 505nm, and multiplying the value by the dilution factor to obtain the color value of the red pigment; measuring the OD value at the wavelength of 465nm, and multiplying the value by the dilution factor to obtain the color value of the orange pigment; the OD value was measured at a wavelength of 410nm and multiplied by the dilution to obtain the yellow pigment color number. Color number units are expressed in U/mL.
The results are shown in FIGS. 7-8, and the 5-azacytidine has significant effects on the improvement of the color values of red, orange and yellow pigments in the red yeast M1 and red yeast RP2 pigments.
3.4 comparison of MonacolinK yields
Taking 5ml of fermentation liquor of monascus of experimental groups and control groups fermented for 2, 5, 8, 10, 12, 14, 16 and 18 days respectively, adding 15ml of 75% methanol, carrying out ultrasonic disruption for 30min at 30 ℃ and 300W, standing for 6 hours in a dark place, filtering by using an organic filter membrane with the diameter of 0.45 mu m, putting the filtrate into a liquid phase vial, and measuring the content of MonacolinK.
Detection conditions for MonacolinK: shimadzu liquid phase, column C18Column (150mm × 4.6mm), mobile phase methanol: 0.1% strength phosphoric acid 75:25, flow rate: 1ml/min, detector: ultraviolet detector, wavelength: 237nm, temperature; at 30 ℃.
The liquid phase detection results are shown in fig. 9 and 10, and compared with the control group without 5-azacytidine, the experimental group with 5-azacytidine added has a remarkable effect of improving the yield of MonacolinK.
3.5 cultivation day 8 endoscope comparison
(1) Culturing red yeast M1 and red yeast RP2 for 8 days, centrifuging at 12000r/min for 5min at low temperature of 4 ℃, collecting thalli cells, and suspending the cells in 2.5% glutaraldehyde solution for fixing for 12 hours.
(2) Cells were rinsed twice with 0.1M PBS and the supernatant was discarded.
(3) Sequentially dehydrating the cells with ethanol solutions (30%, 50%, 70%, 80%, 90%, 100%) of different concentrations, standing for 10min at each concentration, centrifuging at 12000r/min at 4 deg.C for 5min, and discarding the supernatant.
(4) The cells are respectively replaced by isoamyl acetate and ethanol (v: v ═ 1:1) solution and isoamyl acetate solution, the cells are suspended in each solvent and kept still for 10min, and then the cells are centrifuged for 5min at 12000r/min under the low temperature condition of 4 ℃, and the supernatant is discarded.
(5) Adding hexamethyldisilazane, plugging the centrifugal tube with absorbent cotton, placing in a 60 ℃ oven, drying until the sample becomes powder, and observing by an electron microscope.
As shown in FIG. 11, a-e are electron micrographs of Red Yeast RP 2. Wherein, a (2000X), c (5000X), and e (10000X) are photographs of a Monascus purpureus RP2 mycelium obtained by adding an equal amount of sterile water to a control group in place of 5-azacytidine. b (2000X), d (5000X), and f (10000X) are photographs of the filament of Monascus purpureus RP2 to which an equal amount of 5-azacytidine was added. The data indicate that the red yeast RP2 control group had full spores, smooth surfaces, and fewer mycelial filaments. The experimental group has rough spore surface, more wrinkles and bulges, more folds of mycelium and bad plumpness. g-l is an electron micrograph of Monascus purpureus M1. g (2000X), i (5000X), k (10000X) are photographs of a control group to which an equal amount of sterile water was added in place of the mycelium of 5-azacytidine monascus M1, and h (2000X), j (5000X), l (10000X) are photographs to which the mycelium of 5-azacytidine experimental monascus M1 was added. The results showed that the mycelium surface of the control group of monascus M1 was smooth, did not have as many filaments and had plump spore particles with no noticeable wrinkles. The surface of the mycelium of the experimental group of monascus M1 is rough, a plurality of filaments are formed, spore particles are not full, and wrinkles are formed.
The experimental group has more thalli folds and filaments, increases the surface area of cells, enables secondary metabolites in cells to be easily discharged out of the body, reduces the accumulation of the secondary metabolites in the body, and increases the amount of the secondary metabolites. It is explained and confirmed in principle that the addition of 5-azacytidine according to the present invention can increase the amount of secondary metabolites.

Claims (3)

1. A method for improving monascus secondary metabolite is characterized in that 5-azacytidine is added in the liquid fermentation initial stage or fermentation process of monascus;
the addition amount of the 5-azacytidine is 10-2g/L；
The secondary metabolite is monascus red pigment, orange pigment, yellow pigment and MonacolinK;
the 5-azacytidine is added within 48 hours of liquid fermentation culture of monascus;
the Monascus purpureus is Monascus purpureus.
2. The method for increasing the secondary metabolite content of monascus according to claim 1, wherein the fermentation culture conditions are air bath, 33 ℃ and 200r/min for 18 days.
3. The method for increasing secondary metabolites of monascus according to claim 1, wherein the method can simultaneously increase the yields of monascus red pigment, orange pigment, yellow pigment and MonacolinK.
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105969673A (en) * 2016-06-27 2016-09-28 北京工商大学 Fermentation method for increasing yield of haematochrome and uranidin in monascus purpureus

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105969673A (en) * 2016-06-27 2016-09-28 北京工商大学 Fermentation method for increasing yield of haematochrome and uranidin in monascus purpureus

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
5-杂氮胞苷对牛蒡内生真菌黑曲霉(NY-1)次级代谢产物杀虫活性的影响;李昉等;《青岛科技大学学报（自然科学版）》;20160430;第37卷(第2期);第127-130页，尤其是摘要，第128页左栏第1、5-6段，第129页左栏第3段 *

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