CN107805574B - Method for reducing citrinin content in pigment produced by monascus and preparing health-care monascus wine - Google Patents
Method for reducing citrinin content in pigment produced by monascus and preparing health-care monascus wine Download PDFInfo
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- 241000228347 Monascus <ascomycete fungus> Species 0.000 title claims abstract description 58
- CQIUKKVOEOPUDV-IYSWYEEDSA-N antimycin Chemical compound OC1=C(C(O)=O)C(=O)C(C)=C2[C@H](C)[C@@H](C)OC=C21 CQIUKKVOEOPUDV-IYSWYEEDSA-N 0.000 title claims abstract description 32
- CQIUKKVOEOPUDV-UHFFFAOYSA-N citrinine Natural products OC1=C(C(O)=O)C(=O)C(C)=C2C(C)C(C)OC=C21 CQIUKKVOEOPUDV-UHFFFAOYSA-N 0.000 title claims abstract description 32
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- 229940026314 red yeast rice Drugs 0.000 description 5
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- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 4
- 238000007865 diluting Methods 0.000 description 4
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- BWWAFUZQSLIIIH-UHFFFAOYSA-N 2-phenyl-3H-chromen-3-id-4-one Chemical compound O1C(=[C-]C(=O)C2=CC=CC=C12)C1=CC=CC=C1 BWWAFUZQSLIIIH-UHFFFAOYSA-N 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
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- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
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Abstract
The invention discloses a method for reducing the content of citrinin in pigment produced by monascus and preparing health-care monascus wine, belonging to the field of brewing engineering. The method for reducing the content of citrinin in pigment produced by monascus comprises the following steps: inoculating the activated monascus seed solution into a culture solution, adding the kudzu root powder subjected to steam explosion treatment and silkworm cocoons coated with silkworm chrysalis powder, and fermenting for 2-2.5 days; adding ultrasonic-crushing-treated fen-flavor medium-temperature distiller's yeast and activated carbon for adsorbing quinoa hydrolysis (amylase hydrolysis) filtrate, and fermenting for 6-7d to obtain Monascus fermentation liquid with reduced citrinin content. Filtering the fermentation liquor, and blending with the fen-flavor base liquor to obtain the health red koji wine. The health red koji wine produced by utilizing the kudzu roots has good health care effect; the metabolism of monascus is controlled by the slow release of the nitrogen source, so that the output of citrinin in the monascus is reduced, and the high-quality health-care monascus wine is obtained.
Description
Technical Field
The invention belongs to the field of brewing engineering, and particularly relates to a method for reducing the content of citrinin in pigment produced by monascus and preparing health-care monascus wine.
Background
The monascus pigment is a natural pigment produced by monascus in the growth and metabolism process, and is widely used for coloring food in China due to the advantages of bright red color, strong tinting strength, good stability, natural taste and the like. The monascus pigment also has various biological activities, such as obvious inhibition effect on carcinogenesis induced by some conditions. In 1995, French scholars BLANC P J et al demonstrated that Monascus purpureus CBS 109.07 (Monascus ruber) and Monascus ruber van Tieghem produced Monascidin A while the pigment was metabolized, and that this substance was the mycotoxin-citrinin, thereby drawing attention to the safety of Monascus pigment. Citrinin, also known as citrinin, was first discovered to be produced by penicillium in the 30 th 20 th century and was highly regarded as an antibiotic, and was later discovered to be nephrotoxic, precluding its use as a therapeutic antibiotic and identified as a mycotoxin. The target organ of citrinin is the kidney, and administration of citrinin causes kidney toxicity in various experimental animals, which is characterized by enlargement of the kidney, ultimately leading to renal failure. Research also found that citrinin has teratogenic, hepatic metabolism impairing, carcinogenic and mutagenic effects and may inhibit the central nervous system.
The production and application history of red yeast rice in China is long, and the scale is continuously enlarged, but the citrinin can not be effectively controlled in the red yeast rice product, so that the potential safety problem of the red yeast rice product exists, and the export trade of the red yeast rice in China is greatly limited. Therefore, the key to solving the problems is to seek a production process for reducing the content of citrinin in the red yeast rice product.
Disclosure of Invention
The invention aims to overcome the problems in the prior art and provide a method for reducing the content of citrinin in pigment produced by monascus. The invention also aims to provide a method for preparing the health-care red koji wine.
The purpose of the invention is realized by the following technical scheme:
a method for reducing the content of citrinin in pigment produced by monascus comprises the following steps:
(1) pretreatment of raw materials
Pretreatment of distiller's yeast: and (3) carrying out ultrasonic crushing on the fen-flavor medium-temperature distiller's yeast to obtain the pretreated fen-flavor medium-temperature distiller's yeast.
Silkworm cocoon wrapping silkworm chrysalis powder: loading silkworm pupa powder into silkworm cocoons, and sealing the silkworm cocoon openings with threads to obtain silkworm pupa powder-coated silkworm cocoons; each silkworm cocoon is preferably wrapped with 2-3g silkworm pupa powder.
Pretreating kudzu roots: and (4) performing steam explosion on the kudzu root powder to obtain the pretreated kudzu root powder.
Hydrolyzing quinoa: and adding quinoa powder and amylase into water for hydrolysis to obtain quinoa hydrolysate, and further filtering to obtain quinoa hydrolysis filtrate. Wherein, the amount of water is preferably 2-2.2 times of the weight of the quinoa wheat flour, the amount of amylase is preferably 2-3% of the weight of the quinoa wheat flour, and the hydrolysis conditions are preferably as follows: hydrolyzing at 45-50 deg.C for 3-4 h.
Adsorption of quinoa hydrolysis filtrate by activated carbon: and adding activated carbon into the quinoa hydrolyzed filtrate to enable the activated carbon to fully adsorb the quinoa hydrolyzed filtrate, and then filtering to obtain the activated carbon adsorbing the quinoa hydrolyzed filtrate. Wherein the amount of activated carbon is preferably 1/5-1/3 of the weight of quinoa hydrolyzed filtrate.
(2) Fermentation of
Inoculating the monascus seed liquid into a culture solution, adding pretreated pueraria powder and silkworm cocoon wrapping the silkworm chrysalis powder, wherein the addition amount of the pretreated pueraria powder is 200-225g/L of the culture solution, and the addition amount of the silkworm cocoon wrapping the silkworm chrysalis powder is 50-100g/L of the culture solution, and fermenting for 2-2.5 days; adding pretreated fen-flavor medium-temperature distiller's yeast with the mass of 0.5-1% of the fermented mash and 5-8% of activated carbon for adsorbing quinoa hydrolyzed filtrate, and fermenting for 6-7d to obtain Monascus fermentation liquor with reduced citrinin content.
The formula of the culture solution in the step (2) is preferably as follows: glucose 0.1g, peptone 0.2g, K2HPO40.5g, 1000mL of water.
In the step (2), the fermentation temperature is preferably 35 ℃, and the rotation speed is preferably 130-160 r/min.
A method for preparing health red koji wine comprises the following steps: and (3) filtering the monascus fermentation liquor obtained by the method to obtain filtrate, and blending the filtrate with the fen-flavor base liquor to obtain the monascus liquor. Preferably, the degree of the fen-flavor base wine is 60 degrees; the blending volume ratio of the filtrate obtained by filtering the monascus fermentation liquor to the fen-flavor base wine is 2: 7.
The invention has the following advantages and beneficial effects:
(1) the kudzu root contains components with excellent health care effects such as flavone and the like, and the health red koji wine produced by the kudzu root has good health care effect.
(2) According to the invention, the metabolism of monascus is effectively controlled through the slow release of the nitrogen source, so that the output of citrinin in the monascus is reduced, and the high-quality health-care monascus wine is obtained.
Detailed Description
The following examples are intended to further illustrate the invention but should not be construed as limiting it. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
Example 1
1. Materials and apparatus
Monascus: monascus purpureus strain CCTCC AF 93208. Fresh scent type medium temperature yeast: longshan permanent biological koji Co. Quinoa: beijing tianci Chenopodium quinoa SpA. BF 7658-alpha amylase (solid): 1000u/g, from the company Schchen Wanda bioengineering, Ltd. Coconut shell activated carbon: consolidates the new sharp water purification materials ltd.
An ultrasonic crusher LC-N-4.3: ningbo Licheng instruments Ltd. Steam explosion machine QB-200: crane wall city is just in the way of heavy machinery factories.
2. Method of producing a composite material
(1) Pretreatment of raw materials
Pretreatment of distiller's yeast: taking the fen-flavor medium-temperature Daqu, and carrying out ultrasonic crushing for 20min by using an ultrasonic crusher to obtain the pretreated fen-flavor medium-temperature Daqu.
Silkworm cocoon wrapping silkworm chrysalis powder: the silkworm chrysalis is crushed by a crusher, the crushed silkworm chrysalis is sieved by a 20-mesh sieve, the sieved silkworm chrysalis passes through the 20-mesh sieve and is sieved by a 50-mesh sieve, and the crushed silkworm chrysalis powder is arranged on the 50-mesh sieve. Adding pulverized silkworm pupa powder into silkworm cocoon, adding silkworm pupa powder 2g into each silkworm cocoon, sewing the silkworm cocoon opening with natural silk, sewing two lines of sealing lines with sewing needles, and controlling the distance between the needle hole and the needle hole within 0.3 mm.
Pretreating kudzu roots: and (3) putting the crushed kudzu root powder which is sieved by a 20-mesh sieve into a steam explosion machine for treatment to obtain the pretreated kudzu root powder. The conditions of steam explosion are as follows: the release is carried out instantly after 2min of heat preservation under 2.0 MPa.
Hydrolyzing quinoa: crushing quinoa by a crusher, and sieving by a 20-mesh sieve to obtain quinoa powder. Adding quinoa powder into tap water which is 2 times of quinoa powder in mass and is sterilized and cooled to room temperature, adding amylase according to the amount of 2g of amylase added into every 100g of quinoa powder, and hydrolyzing for 3h at 50 ℃ to obtain quinoa hydrolysate. Filtering the quinoa hydrolysate with filter paper to obtain quinoa hydrolyzed filtrate.
Adsorption of quinoa hydrolysis filtrate by activated carbon: sterilizing the hydrolyzed filtrate at 121 deg.C for 30min, and cooling to room temperature in a fume hood. Transferring the cooled quinoa hydrolyzed filtrate into a 250mL triangular flask, adding 100mL of activated carbon with the mass of 1/4 of the quinoa hydrolyzed filtrate, oscillating at 180r/min for 30min, and filtering with filter paper in a fume hood to obtain the activated carbon for adsorbing the quinoa hydrolyzed filtrate.
(2) Preparation of culture medium
Monascus seed culture solution: 20g of malt meal, 1g of peptone, 4g of glucose and K2HPO40.5g, 1000mL of water, pH5.8, and sterilizing at 121 ℃ for 20 min.
Fermentation culture solution of monascus: glucose 0.1g, peptone 0.2g, K2HPO40.5g, 1000mL water, pH5.8, 121 ℃ sterilization for 20 min.
(3) Fermentation of
Activating monascus: adding 5mL of sterile water into Monascus purpureus CCTCC AF93208 test tube slant strains, stirring by using an inoculating needle, inoculating into a 250mL triangular flask filled with 50mL of seed culture solution, and culturing at 35 ℃ at 160r/min for 24h to obtain the Monascus purpureus seed solution.
Liquid state fermentation of monascus: inoculating 2mL of monascus seed liquid into a 500mL triangular flask filled with 100mL of fermentation culture liquid, adding 22.5g of pretreated radix puerariae powder and 10g of silkworm cocoon wrapping silkworm chrysalis powder, fermenting at 35 ℃ and 160r/min for 2 days, then adding pretreated fen-flavor medium-temperature distiller's yeast with the mass of 0.5% of fermented mash, adding activated carbon with the mass of 5% of fermented mash and adsorbing quinoa hydrolysis filtrate, and fermenting at 35 ℃ and 160r/min for 6 days again to obtain monascus fermentation liquid.
(4) Determination of color value, citrinin content and total flavone content of monascus fermentation liquor
1) Determination of color number of pigment
The method for measuring the color value of the alcohol-soluble pigment comprises the following steps: centrifuging Monascus fermentation liquid at 5000r/min for 10min, collecting supernatant, diluting with 70% ethanol by a certain multiple, water bathing at 60 deg.C for 1h, and filtering; diluting the filtrate with 70% ethanol by a certain multiple, and respectively measuring OD505、OD465、OD410The color value (OD) is calculated based on the volume of the fermentation liquid obtainedThe color value of alcohol-soluble red, orange and yellow pigments is obtained.
The water-soluble pigment color value determination method comprises the following steps: centrifuging Monascus fermentation liquid at 5000r/min for 10min, collecting supernatant, diluting with distilled water by a certain multiple, water bathing at 60 deg.C for 1h, and filtering; diluting the filtrate with distilled water by a certain multiple, and respectively measuring OD505、OD465、OD410The color number of the water-soluble red, orange and yellow pigments was calculated from the color number OD × dilution factor/volume of the obtained fermentation broth.
The total color value is the sum of the color value of the alcohol-soluble pigment and the color value of the water-soluble pigment.
2) Determination of citrinin content
10mL of monascus fermentation liquid is taken to be centrifuged at 5000r/min for 10min, 20mL of 70% ethanol solution is added into the supernatant to be mixed uniformly, and the mixture is filtered by a 0.22-micron microporous membrane and then is subjected to high performance liquid chromatography detection according to the national standard GB/T5009.222-2008.
3) Determination of Total Flavonoids content
Taking Monascus fermentation liquid, centrifuging at 5000r/min for 10min, filtering supernatant with filter paper, and measuring total flavone content in filtrate. The total flavone content was measured according to the method described in the literature (Wang Yongming et al, research on the conditions of the extraction process of pueraria flavonid, Xuzhou institute of engineering, 2007.).
The measurement results are as follows: the concentration of pigment in the monascus fermentation liquid is 2.34U/mL, the content of citrinin is 0.0175mg/mL, and the content of total flavonoids is 12.5 mg/L.
(5) Preparation of red yeast wine
The monascus fermentation liquor is centrifuged for 15min at 5000r/min, then filtered by filter paper, and the filtrate is blended with fen-flavor base liquor (60 degrees, Sichuan Baishiyuan liquor Co., Ltd.) according to the volume ratio of 2:7 to obtain the monascus wine with red color and soft and full taste.
Comparative experiment 1
Referring to example 1, the procedure of example 1 was otherwise the same except that 10g of silkworm chrysalis powder-coated silkworm cocoon was not added during the fermentation, and the pigment concentration and the citrinin content of the monascus fermentation broth were measured to be 1.98U/mL and 0.0233mg/mL, respectively.
Comparative experiment 2
Referring to example 1, the procedure of example 1 was otherwise the same as that of example 1 except that 10g of silkworm chrysalis powder-coated silkworm cocoons were added during fermentation and 10g of the pulverized silkworm chrysalis powder was added, and the pigment concentration and the citrinin content of the monascus fermentation broth were measured to be 2.50U/mL and 0.0221mg/mL, respectively.
Comparative experiment 3
Referring to example 1, 10g of silkworm cocoon coated with silkworm pupa powder was added during fermentation, and the other operations were the same as example 1 except that the pigment concentration in the monascus fermentation broth was 1.98U/mL and the citrinin content was 0.0212 mg/mL.
Comparative experiment 4
Referring to example 1, after 2 days of fermentation, the same procedure as in example 1 was repeated except that no pretreated medium-temperature koji having a fen-flavor with a fermented mash mass of 0.5% was added, and the pigment concentration of the Monascus fermentation broth was measured to be 2.32U/mL and the citrinin content was measured to be 0.0221 mg/mL.
Comparative experiment 5
Referring to example 1, after 2 days of fermentation, the fermentation was continued for 6 days without adding the pre-treated fen-flavor medium temperature distiller's yeast and the activated carbon for adsorbing the quinoa hydrolyzed filtrate, and the other operations were the same as example 1, except that the pigment concentration in the monascus fermentation broth was measured to be 2.02U/mL and the citrinin content was measured to be 0.0243 mg/mL.
Comparative test 6
Referring to example 1, after 2 days of fermentation, activated carbon adsorbing 5% of the mass of the fermented mash and adsorbing the quinoa hydrolyzed filtrate was added instead of activated carbon adsorbing 5% of the mass of the fermented mash, and the pigment concentration of the monascus fermentation broth was measured to be 2.31U/mL and the citrinin content was measured to be 0.0259mg/mL in the same manner as in example 1.
Comparative experiment 7
Referring to example 1, after 2 days of fermentation, the procedure of adding 5% by mass of activated carbon adsorbing quinoa hydrolyzed filtrate of fermented mash into 0.5% by mass of quinoa hydrolyzed filtrate of fermented mash was changed, and the pigment concentration in the monascus fermentation broth was measured to be 2.08U/mL and the citrinin content was measured to be 0.0199mg/mL in the same manner as in example 1.
Comparative experiment 8
Referring to example 1, after 2 days of fermentation, the procedure of adding quinoa hydrolysate having a mass of 0.5% of the mass of the fermented mash instead of adding quinoa activated carbon adsorbing quinoa hydrolyzed filtrate having a mass of 5% of the mass of the fermented mash was otherwise the same as in example 1, and the pigment concentration in the monascus fermentation broth was measured to be 2.02U/mL, and the citrinin content was measured to be 0.0207 mg/mL.
Comparative test 9
Referring to example 1, 10g of silkworm cocoon coated with silkworm chrysalis powder is not added during fermentation, after 2d of fermentation, activated carbon which is 5% of the mass of fermented liquor and adsorbs quinoa hydrolysis filtrate is added instead of yeast solution which is 1.6% of the mass of fermented liquor, and the other operations are the same as example 1, and the pigment concentration of the monascus fermentation liquor is measured to be 2.18U/mL, and the citrinin content is measured to be 0.0215 mg/mL.
Wherein the yeast liquid is obtained by the following treatment: 50 μ L of Saccharomyces cerevisiae ATCC 36859 seed solution was inoculated into a large test tube containing 5mL of YEPD (peptone 20g/L, yeast extract 10g/L, glucose 20g/L, pH 6.0, sterilized at 121 ℃ for 20min), and cultured at 30 ℃ for 8h at 160 r/min.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (9)
1. A method for reducing the content of citrinin in pigment produced by monascus is characterized in that: the method comprises the following steps:
(1) pretreatment of raw materials
Pretreatment of distiller's yeast: carrying out ultrasonic crushing on the fen-flavor medium-temperature distiller's yeast to obtain pretreated fen-flavor medium-temperature distiller's yeast;
silkworm cocoon wrapping silkworm chrysalis powder: loading silkworm pupa powder into silkworm cocoons, and sealing the silkworm cocoon openings with threads to obtain silkworm pupa powder-coated silkworm cocoons;
pretreating kudzu roots: the kudzu root powder is subjected to steam explosion to obtain pretreated kudzu root powder;
hydrolyzing quinoa: adding quinoa powder and amylase into water, hydrolyzing to obtain quinoa hydrolysate, and further filtering to obtain quinoa hydrolysis filtrate;
adsorption of quinoa hydrolysis filtrate by activated carbon: adding activated carbon into the quinoa hydrolyzed filtrate to enable the activated carbon to fully adsorb the quinoa hydrolyzed filtrate, and then filtering to obtain activated carbon adsorbing the quinoa hydrolyzed filtrate;
(2) fermentation of
Inoculating the monascus seed liquid into a culture solution, adding pretreated pueraria powder and silkworm cocoon wrapping the silkworm chrysalis powder, wherein the addition amount of the pretreated pueraria powder is 200-225g/L of the culture solution, and the addition amount of the silkworm cocoon wrapping the silkworm chrysalis powder is 50-100g/L of the culture solution, and fermenting for 2-2.5 days; adding pretreated fen-flavor medium-temperature distiller's yeast with the mass of 0.5-1% of the fermented mash and 5-8% of activated carbon for adsorbing quinoa hydrolyzed filtrate, and fermenting for 6-7d to obtain Monascus fermentation liquor with reduced citrinin content.
2. The method of claim 1, wherein: and (2) coating silkworm cocoons in the step (1) with silkworm chrysalis powder, wherein each silkworm cocoon is coated with 2-3g of silkworm chrysalis powder.
3. The method of claim 1, wherein: hydrolyzing quinoa in the step (1): the amount of water is 2-2.2 times of the quinoa powder; the amylase accounts for 2-3% of the quinoa powder; the hydrolysis conditions were: hydrolyzing at 45-50 deg.C for 3-4 h.
4. The method of claim 1, wherein: adsorbing the quinoa hydrolyzed filtrate by using the activated carbon in the step (1), wherein the amount of the activated carbon is 1/5-1/3 of the quinoa hydrolyzed filtrate.
5. The method of claim 1, wherein: the formula of the culture solution in the step (2) is as follows: glucose 0.1g, peptone 0.2g, K2HPO40.5g, 1000mL of water.
6. The method of claim 1, wherein: in the step (2), the fermentation temperature is 35 ℃.
7. A method for preparing health-care red koji wine is characterized by comprising the following steps: the method comprises the following steps: filtering Monascus fermentation liquid obtained by the method of any one of claims 1-6, and blending the obtained filtrate with fen-flavor base liquor to obtain health Monascus liquor.
8. The method for preparing a health red koji wine according to claim 7, wherein: the degree of the fen-flavor base wine is 60 degrees.
9. A method of preparing red koji wine according to claim 7 or 8, characterized in that: the blending volume ratio of the filtrate obtained by filtering the monascus fermentation liquor to the fen-flavor base wine is 2: 7.
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