CN103404785B - Preparation method of high-color-value antimicrobial water soluble functional red yeast rice - Google Patents
Preparation method of high-color-value antimicrobial water soluble functional red yeast rice Download PDFInfo
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Abstract
The invention discloses a preparation method of high-color-value antimicrobial water soluble functional red yeast rice. The preparation method is capable of solving a problem that the color value of water soluble functional red yeast rice products is low, and the introduction of carboxylation chitosan is capable of endowing the high-color-value water soluble functional red yeast rice with bacteriostatic activity. The preparation method comprises key steps such as early stage fermentation of the functional red yeast rice, processing of materials after fermentation, a second time of fermentation of the functional red yeast rice, processing of fermentation broth and acquisition of the high-color-value antimicrobial water soluble functional red yeast rice. Drying efficiency is high, product quality is stable, the color value and solubility are high, the high-color-value antimicrobial water soluble functional red yeast rice possesses bacteriostatic activity, and application value of the high-color-value antimicrobial water soluble functional red yeast rice is excellent.
Description
Technical field
The present invention relates to a kind of preparation method of red colouring agent for food, also used as a Chinese medicine, particularly a kind of preparation method of high look valency bacteriostatic water dissolubility functional red yeast rice.
Background technology
Red colouring agent for food, also used as a Chinese medicine is as fermented food, have a long history in China, 20 century 70s, day wood and the researcher of the U.S. isolate a kind of active material-Monacolin K (Monacolin K) that can powerful inhibition cholesterol biosynthesis in succession from monascus nutrient solution, thereby accelerate the research of people to functional Monascus, the traditional mode of production of red colouring agent for food, also used as a Chinese medicine is improved, also some products taking functional Monascus as raw material have been developed, because it regulates cardiovascular and cerebrovascular disease determined curative effect, obtain the accreditation of extensive patients.
Antilipemic monascus product be in the market mainly taking process for solid state fermentation produce functional red yeast rice as raw material, except the Monacolin K that contains fat-reducing effect, also contain a large amount of remaining starch, due to its poorly water-soluble, color range is low, often with red colouring agent for food, also used as a Chinese medicine particle, the forms such as red colouring agent for food, also used as a Chinese medicine capsule, end product as health products or medicine is sold, cannot directly use at the field of food such as liquid beverage or cake, limit to a great extent the scope of application of functional Monascus, therefore on functional Monascus basis, be developed to directly water-soluble water-soluble functional red yeast rice, be applied to food, the industries such as medicine, there is good application prospect.
At present, for the exploitation of water-soluble monascus, there is bibliographical information, as application number: 200810234815.0 Chinese invention patent, explain a kind of preparation method of water-soluble monascus, adopt solid state fermentation to produce functional red yeast rice, then taking functional red yeast rice as raw material, adopt the method for solvent extraction to obtain extract, the spray-dried water-soluble monascus powder that makes of extract, the look valency that the method exists water-soluble monascus is low, there is no the problems such as antibacterial characteristic; Application number is 200910095494.5 Chinese invention patent, adopt enzymolysis to send out extraction water-soluble monascus, adopt spray drying process to make finished product, but do not disclose spray-dired concrete grammar and condition, and the water-soluble monascus of this explained hereafter, there is the problems such as painted valency is low.
Can be found out by above-mentioned documents and materials, though water-soluble monascus has had many research, also imperfection of current technology, product quality has much room for improvement, and has no the application report using using carboxyl chitosan as composition Adsorption Concentration agent such as monascorubins simultaneously.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of high look valency bacteriostatic water dissolubility functional red yeast rice, have that drying efficiency is high, product with stable quality, look valency is high, solubility is high, have the features such as biocidal property, has good using value.
The technical solution adopted for the present invention to solve the technical problems is:
A preparation method for high look valency bacteriostatic water dissolubility functional red yeast rice, comprises the steps:
(1) prior fermentation of functional Monascus
High temperature and cold fermentation: by functional Monascus strain transfer in fermentation materials, inoculum concentration is the 1-5% of fermentation materials weight, then cultivate in hot fermentation chamber, cultivation temperature is 30-35 DEG C, incubation time is 2-5 days, after having cultivated, transfers to cold fermentation chamber and cultivates, cultivation temperature is 16-25 DEG C, and incubation time is 10-25 days.Functional Monascus is the red colouring agent for food, also used as a Chinese medicine that mainly can produce Monacolin K, but the pigment color value of its product is very low.
Adopt the advantage of high temperature and low temperature two-step fermentation to be: to adopt higher cultivation temperature in early stage, be conducive to monascus ruber and grow, allow mycelium be covered with whole culture medium material; Later stage adopts low temperature to cultivate, and is conducive to the secondary metabolism of Monascus, produces secondary metabolite Lovastatin, improves the content of active ingredient.
(2) processing of material after fermentation
Material after step (1) fermentation is carried out to sterilizing, sterilising conditions is 121 ± 1 DEG C, and then sterilizing 30-45min mixes with sterilized water material according to the weight ratio of 1:2-4, then process with colloid mill making beating, through making beating emulsifying materials fineness 10-50 micron after treatment.
By material pull an oar process advantage be: on the one hand, can pulverize large granular materials, so that monascus making full use of nutrition composition; On the other hand, making beating is processed and can be made material be evenly distributed in fermentation tank, is beneficial to stirring; And in the process of next step liquid fermentation, if there is large particulate matter, easily form caking phenomenon, be unfavorable for fermentation.
By the first sterilized material benefit of processing of pulling an oar be again, the gelatinization that can avoid making beating to cause, that is to say, first making beating, then sterilizing, and high temperature causes the starch gelatinization in material, is unfavorable for fermentation.
Sterilized water is running water or drinking water, adopts moist hear heat test, under 121 DEG C of conditions, and sterilizing 20-25min and get final product.
(3) look song ferments again
Pack the material after step (2) making beating into full-automatic liquid fermentation tank, access look aspergillus kind is fermented again, inoculum concentration is the 1-5% of the weight of material after making beating, fermentation condition is again: temperature 28-30 DEG C, mixing speed 60-200r/min, throughput 2.0-5.0L/min, zymotic fluid pH value is 5.0-6.0, fermented incubation time is 10-20 days.Look song is the Monascus of high yield monascus pigment.
(4) fermentation liquor treatment
Get step (3) the gained zymotic fluid that ferments again, add the edible alcohol of 0.5-2 times of weight, regulate pH value to regulate the effect of pH value 6.5-7.5 to 6.5-7.5(, on the one hand can be by active ingredient Lovastatin from alkali formula thaumatropy acid structure, acid structure does not need to enter liver metabolism, reduce burden of liver, be beneficial to absorption; On the other hand, can increase the dissolution rate of monascorubin, improve product look valency), stir, leave standstill 1-2h, then use 300-350 object filter-cloth filtering, gained decompression filtrate recycling ethanol obtains concentrate.
The common solid-state Hongqu powder (red colouring agent) product that obtains high-quality after filtration gained filter residue and drying, makes full use of the antibacterial water-soluble functional red yeast rice powder of the high look valency product of getting back to fermentation materials.
Add edible alcohol (60%-95wt%) for extracting solvent, can improve on the one hand the recovery rate of Lovastatin and pigment, be conducive on the other hand the precipitation of the materials such as starch, the clarification of guarantee extract, accelerate the rate of filtration, be beneficial to filtration, the ethanol of suitable concentration also helps the stripping of Monascus polysaccharides simultaneously.
(5) acquisition of high look valency bacteriostatic water dissolubility functional red yeast rice
Account for to adding in step (4) gained concentrate the using carboxyl chitosan that concentrate quality is 0.1%-10%, then regulate pH value to 2.0-5, stir, leave standstill 1-2h, then, regulate pH to 5.5-7.5, now can produce a large amount of red precipitates, migrate out part supernatant, the part by weight that makes to remain supernatant and precipitation is 0.5-2.0:1, stir and make suspension, in whipping process, add and have malt sugar group-beta-cyclodextrin and hydroxy propyl-Beta cyclodextrin to suspension, in every kilogram of suspension, add malt sugar group-beta-cyclodextrin 10-30g, hydroxy propyl-Beta cyclodextrin 10-50g, after suspension spraying is dry, obtain high look valency bacteriostatic water dissolubility functional red yeast rice.
Using carboxyl chitosan is a kind of positive biological polyoses, under suitable pH, its molecular conformation difference, in the time of pH2.0-5, it has dissolubility, can have suction-operated to the little molecule of feminine gender (as Lovastatin, monascorubin etc.), under the condition of pH5.5-7.5, can form a large amount of precipitations, using carboxyl chitosan has stronger bacteriostatic activity to common fungus and bacterium simultaneously.Using carboxyl chitosan is applied to the preparation of water-soluble monascus, can brings into play on the one hand the ability of its absorption Lovastatin and monascorubin, reach the concentrated of Lovastatin and monascorubin; By the introducing of using carboxyl chitosan, can give monascorubin product bacteriostatic activity on the other hand, improve its anti-living contaminants ability in food or pharmaceutical applications.
Produce after a large amount of red precipitates, precipitation can directly be dried or dry, make high look valency bacteriostatic water dissolubility functional red yeast rice, but such red colouring agent for food, also used as a Chinese medicine rate of drying is slow, efficiency is low, and water-soluble is not very high, dissolve by precipitating again, add malt sugar group-beta-cyclodextrin and hydroxy propyl-Beta cyclodextrin, and spraying dry after, rate of drying is fast, water-soluble height.Stay part supernatant to mix and be dried with precipitation, can utilize so active ingredient and pigment remaining in supernatant, improve product quality.
Add malt sugar group-beta-cyclodextrin and hydroxy propyl-Beta cyclodextrin, by different proportionings, regulate the character of water-soluble monascus product, cyclodextrin structure is more special, and molecule outside is hydrophilic group, and intramolecule is lipophilic structure, there is clathration, the solubility that can improve the materials such as Monacolin K, monascorubin, this inclusion compound can also improve the stability of water-soluble monascus, avoids the destruction producing because baking temperature is high.
As preferably, described functional Monascus bacterial classification cultural method is:
First class inoculum is cultivated: plant after activation female functional Monascus, be transferred on test tube slant culture medium, in constant temperature bacterial classification incubator, 25-35 DEG C of cultivation obtains first class inoculum for 2-5 days;
Second class inoculum is cultivated: first class inoculum is transferred in liquid culture medium at desinfection chamber, after inoculating, at 30-32 DEG C, in the isothermal vibration shaking table of rotating speed 150-200r/min, cultivates and within 3-7 days, obtain second class inoculum, second class inoculum is functional Monascus bacterial classification.
As preferably, slant medium is prepared as: taking the water of 100 weight portions as standard, add maltose modulation 5-8 Baume degrees, then add soy peptone 3-5 weight portion, glucose 2-5 weight portion, agar 2-3 weight portion; After all components is dissolved, pack test tube into, every test tube 8-12ml, then, with cotton plug sealing, carries out moist heat sterilization, and sterilising conditions is 121 DEG C ± 1 DEG C, 15-25min; After sterilizing, while hot test tube is tiltedly put, to be cooled.
As preferably, liquid culture medium is prepared as: taking the water of 100 weight portions as standard, add glucose 3-6 weight portion, soy peptone 3-5 weight portion, white sugar 1-3 weight portion, ZnSO
40.1-0.5 weight portion, NaNO
30.1-0.5 weight portion, KH
2pO
40.1-0.3 weight portion, ground rice 1-2 weight portion, analysis for soybean powder 1-3 weight portion, mealy potato 0.5-1 weight portion, glycerine 1-3 weight portion; After mixing dissolving, pack in 500ml triangular flask, every bottle of 150-250ml, with after gauze sealing, uses moist hear heat test sterilizing, and sterilising conditions is: 121 DEG C ± 1 DEG C, and 20-30 minute.
As preferably, in step (1) high temperature and cold fermentation process, fermentation materials is broken up to processing, within during hot fermentation 2-3 days, break up once, during cold fermentation, broke up once every 5-7 days.
As preferably, the described fermentation materials preparation method of step (1) is: formula: brown rice 52-57wt%, and wheat bran 4-6wt %, soy meal 4-6 wt %, soy peptone 2-3 wt %, glucose 2-3 wt %, glycerine 1-2 wt %, surplus is water; Brown rice is pulverized, added other each component, and then add water, stir, packing volume into is plastics fermentation flask or the glass fermentation flask of 1L, and every bottled amount is 250-350g, with gauze sealing, adopts moist hear heat test sterilizing, 121 ± 1 DEG C, and 30-45min.
As preferably, the spray-dired setting parameter of step (5): air inlet actual temperature is at 102-115 DEG C, air outlet actual temperature 60-80 DEG C, sample size is 2.0-5.0ml/min, intake is 100-400NL/h.Intake air temperature is set in 102-115 DEG C, is owing to containing polysaccharose substance in water-soluble monascus, excess Temperature, and easily gelatinization, coking, temperature is too low, the shortage of heat that can provide, material atomization fast, affects output.
As preferably, described look aspergillus kind cultural method is:
First class inoculum cultivation: by look bent female kind after activation, be transferred on test tube slant culture medium, 25-30 DEG C of cultivation obtains first class inoculum for 2-5 days in constant temperature bacterial classification incubator;
Second class inoculum is cultivated: first class inoculum is transferred in liquid culture medium at desinfection chamber, after inoculating, at 25-30 DEG C, cultivates and within 4-7 days, obtain second class inoculum in the isothermal vibration shaking table of rotating speed 145-180r/min, second class inoculum is look aspergillus kind.
As preferably, slant medium is prepared as: taking the water of 100 weight portions as standard, add maltose modulation 5-8 Baume degrees, then add soy peptone 3-5 weight portion, glucose 2-5 weight portion, agar 2-3 weight portion; After all components is dissolved, pack test tube into, every test tube 8-12ml, then, with cotton plug sealing, carries out moist heat sterilization, and sterilising conditions is 121 DEG C ± 1 DEG C, 15-25min; After sterilizing, while hot test tube is tiltedly put, to be cooled.
As preferably, liquid culture medium is prepared as: formula: rice meal 0.1-0.5wt%, and soy peptone is 2.0-5.0 wt %, glucose 1.0-3.0 wt %, sucrose 1.5-2.5 wt %, surplus is water; After mixing, pack in 500ml triangular flask, every bottle of 180-250ml, with after gauze sealing, uses moist hear heat test sterilizing, and sterilising conditions is: 121 DEG C ± 1 DEG C, and 20-30 minute.
The invention has the beneficial effects as follows:
1, solved the not high problem of the soluble red rice red product look of activated water valency, look valency is by the 500-1000u/g of general water-soluble monascus, more than bringing up to 5000u/g.
2, the solubility of product is high, under 25 DEG C of conditions, can reach 50g/100ml water.
3, the antibacterial water-soluble functional red yeast rice of high look valency that this method obtains, the content of Monacolin K is high, and spray drying process to split closed loop proportional impact less, product Open-closed-loop ratio is more than 95%.
4,, in spray-drying process, the malt sugar group-beta-cyclodextrin adding and hydroxy propyl-Beta cyclodextrin, in improving product stability, change the character of material, utilized and be dried, can improve the yield and quality, but also can increase the solubility of product, there is multi-efficiency.
5, add using carboxyl chitosan, can bring into play on the one hand the ability of its absorption Lovastatin and monascorubin, reach the concentrated of Lovastatin and monascorubin; By the introducing of using carboxyl chitosan, can give monascorubin product bacteriostatic activity on the other hand, improve its anti-living contaminants ability in food or pharmaceutical applications.
Detailed description of the invention
Below by specific embodiment, technical scheme of the present invention is described in further detail.
In the present invention, if not refer in particular to, raw material and the equipment etc. adopting all can be buied from market or this area is conventional, wherein using carboxyl chitosan can be CMC or carboxyetbyl chitosan, percent grafting can be 10 ~ 50%, molecular weight can be 50kDa ~ 2000kDa, in following examples, only describes as an example of CMC example.Separately, the related test strain of bacteriostatic test can be Escherichia coli (
e.coli), staphylococcus aureus (
s.aureus), salmonella (
salmonella), pseudomonas aeruginosa (
p.aeruginosa), Friedlander's bacillus (
k.peneumoniae) or single increasing listeria spp (
l.monocytogenes) in any, in following examples, only describe as an example of Escherichia coli example.Method in following embodiment, if no special instructions, is the conventional method of this area.
Functional Monascus bacterial classification cultural method is:
First class inoculum is cultivated: by (commercially available female functional Monascus kind, purchased from Hangzhou Qingzheng Bio-Tech Co., Ltd., be numbered: GM02, for producing the red colouring agent for food, also used as a Chinese medicine bacterial classification of Monacolin k) activate at 30 DEG C, soak time is 24-36h, be transferred on test tube slant culture medium, in constant temperature bacterial classification incubator, 25-35 DEG C of cultivation obtains first class inoculum for 2-5 days.
Slant medium is prepared as: taking the water of 100 weight portions as standard, add maltose modulation 5-8 Baume degrees, then add soy peptone 3-5 weight portion, glucose 2-5 weight portion, agar 2-3 weight portion; After all components is dissolved, pack test tube into, every test tube 8-12ml, then, with cotton plug sealing, carries out moist heat sterilization, and sterilising conditions is 121 DEG C ± 1 DEG C, 15-25min; After sterilizing, while hot test tube is tiltedly put, to be cooled.
Second class inoculum is cultivated: first class inoculum is transferred in liquid culture medium at desinfection chamber, after inoculating (inoculum concentration is the 1-2% of liquid culture medium weight), at 30-32 DEG C, in the isothermal vibration shaking table of rotating speed 150-200r/min, cultivate and within 3-7 days, obtain second class inoculum, second class inoculum is functional Monascus bacterial classification.
Liquid culture medium is prepared as: taking the water of 100 weight portions as standard, add glucose 3-6 weight portion, soy peptone 3-5 weight portion, white sugar 1-3 weight portion, ZnSO
40.1-0.5 weight portion, NaNO
30.1-0.5 weight portion, KH
2pO
40.1-0.3 weight portion, ground rice 1-2 weight portion, analysis for soybean powder 1-3 weight portion, mealy potato 0.5-1 weight portion, glycerine 1-3 weight portion; After mixing dissolving, pack in 500ml triangular flask, every bottle of 150-250ml, with after gauze sealing, uses moist hear heat test sterilizing, and sterilising conditions is: 121 DEG C ± 1 DEG C, and 20-30 minute.
Look aspergillus kind cultural method is:
First class inoculum is cultivated: by (commercially available look bent female kind, purchased from Hangzhou Qingzheng Bio-Tech Co., Ltd., be numbered: FM4000, for the Monascus of high yield monascus pigment) under 25-30 DEG C condition, activation 24-36h, be transferred on test tube slant culture medium, in constant temperature bacterial classification incubator, 25-30 DEG C of cultivation obtains first class inoculum for 2-5 days.Slant medium is the same.
Second class inoculum is cultivated: first class inoculum is transferred in liquid culture medium at desinfection chamber, after inoculating (inoculum concentration is the 1-2% of liquid culture medium weight), at 25-30 DEG C, in the isothermal vibration shaking table of rotating speed 145-180r/min, to cultivate and within 4-7 days, obtain second class inoculum, second class inoculum is look aspergillus kind.Liquid culture medium is prepared as: formula: rice meal 0.1-0.5wt%, and soy peptone is 2.0-5.0 wt %, glucose 1.0-3.0 wt %, sucrose 1.5-2.5 wt %, surplus is water; After mixing, pack in 500ml triangular flask, every bottle of 180-250ml, with after gauze sealing, uses moist hear heat test sterilizing, and sterilising conditions is: 121 DEG C ± 1 DEG C, and 20-30 minute.
Embodiment 1:
A preparation method for high look valency bacteriostatic water dissolubility functional red yeast rice, comprises the steps:
(1) prior fermentation of functional Monascus
High temperature and cold fermentation: by functional Monascus strain transfer in fermentation materials, inoculum concentration is 1% of fermentation materials weight, then cultivate in hot fermentation chamber, cultivation temperature is 30 DEG C, incubation time is 5 days, after having cultivated, transfers to cold fermentation chamber and cultivates, cultivation temperature is 16 DEG C, and incubation time is 25 days.
In high temperature and cold fermentation process, fermentation materials is broken up to processing, within during hot fermentation 3 days, break up once, during cold fermentation, broke up once every 5 days.
Fermentation materials preparation method is: formula: brown rice 52wt%, and wheat bran 6wt %, soy meal 6 wt %, soy peptone 3 wt %, glucose 3 wt %, glycerine 1 wt %, surplus is water; Brown rice is pulverized, added other each component, and then add water, stir, packing volume into is the plastics fermentation flask of 1L, and every bottled amount is 250g, with gauze sealing, adopts moist hear heat test sterilizing, 121 ± 1 DEG C, and 30min.
(2) processing of material after fermentation
Material after step (1) fermentation is carried out to sterilizing, sterilising conditions is 121 ± 1 DEG C, and then sterilizing 30min mixes with sterilized water material according to the weight ratio of 1:2, then process with colloid mill making beating, through making beating emulsifying materials fineness 10-50 micron after treatment.
(3) look song ferments again
Pack the material after step (2) making beating into full-automatic liquid fermentation tank, access look aspergillus kind is fermented again, inoculum concentration be making beating after weight of material 1%, fermentation condition is again: 28 DEG C of temperature, mixing speed 60r/min, throughput 2.0L/min, zymotic fluid pH value is 5.0-6.0, fermented incubation time is 20 days.
(4) fermentation liquor treatment
Get step (3) the gained zymotic fluid that ferments again, add the 95wt% edible alcohol of 0.5 times of weight, regulate pH value to 6.5, stir, standing 2h, then uses 300 object filter-cloth filterings, and gained decompression filtrate recycling ethanol obtains concentrate.After gained filter residue and drying, obtain the common solid-state Hongqu powder (red colouring agent) product of high-quality.
(5) acquisition of high look valency bacteriostatic water dissolubility functional red yeast rice
To adding in step (4) gained concentrate, to account for concentrate quality be 0.1% CMC (percent grafting 10%, molecular weight 50kDa), then with acid for adjusting pH value to 2.0, stir, leave standstill 1h, then, with alkali adjusting pH to 5.5, now can produce a large amount of red precipitates, migrate out part supernatant, the part by weight that makes to remain supernatant and precipitation is 0.5:1, stir and make suspension, in whipping process, add and have malt sugar group-beta-cyclodextrin and hydroxy propyl-Beta cyclodextrin to suspension, in every kilogram of suspension, add malt sugar group-beta-cyclodextrin 10g, hydroxy propyl-Beta cyclodextrin 10g, after suspension spraying is dry, obtain high look valency bacteriostatic water dissolubility functional red yeast rice, spray-dired setting parameter: air inlet actual temperature is at 102 DEG C, 60 DEG C of air outlet actual temperatures, sample size is 2.0ml/min, intake is 400NL/h.
(6) bacteriostatic activity of high look valency bacteriostatic water dissolubility functional red yeast rice
The made Hongqu powder (red colouring agent) of step (5) is dissolved in distilled water, is mixed with mass percent concentration and is 1% the red colouring agent for food, also used as a Chinese medicine aqueous solution.Get this red colouring agent for food, also used as a Chinese medicine aqueous solution 1mL, add to and carry that in colibacillary LB culture medium, (e. coli concentration is 10
4cFU/mL), and at 37 DEG C, after 200 turn/min shaken cultivation 24h, absorption bacterium liquid is measured it, and under 600nm wavelength, light absorption value is lower than 10% of control group, and Escherichia coli Growth is significantly inhibited.
Embodiment 2:
A preparation method for high look valency bacteriostatic water dissolubility functional red yeast rice, comprises the steps:
(1) prior fermentation of functional Monascus
High temperature and cold fermentation: by functional Monascus strain transfer in fermentation materials, inoculum concentration is 5% of fermentation materials weight, then cultivate in hot fermentation chamber, cultivation temperature is 35 DEG C, incubation time is 2 days, after having cultivated, transfers to cold fermentation chamber and cultivates, cultivation temperature is 25 DEG C, and incubation time is 10 days.
In high temperature and cold fermentation process, fermentation materials is broken up to processing, within during hot fermentation 2 days, break up once, during cold fermentation, broke up once every 7 days.
Fermentation materials preparation method is: formula: brown rice 57wt%, and wheat bran 4wt %, soy meal 4wt %, soy peptone 2 wt %, glucose 2 wt %, glycerine 2 wt %, surplus is water; Brown rice is pulverized, added other each component, and then add water, stir, packing volume into is the glass fermentation flask of 1L, and every bottled amount is 350g, with gauze sealing, adopts moist hear heat test sterilizing, 121 ± 1 DEG C, and 45min.
(2) processing of material after fermentation
Material after step (1) fermentation is carried out to sterilizing, sterilising conditions is 121 ± 1 DEG C, and then sterilizing 45min mixes with sterilized water material according to the weight ratio of 1:4, then process with colloid mill making beating, through making beating emulsifying materials fineness 10-50 micron after treatment.
(3) look song ferments again
Pack the material after step (2) making beating into full-automatic liquid fermentation tank, access look aspergillus kind is fermented again, inoculum concentration be making beating after weight of material 5%, fermentation condition is again: 30 DEG C of temperature, mixing speed 200r/min, throughput 5.0L/min, zymotic fluid pH value is 5.0-6.0, fermented incubation time is 10 days.
(4) fermentation liquor treatment
Get step (3) the gained zymotic fluid that ferments again, add the 60wt% edible alcohol of 2 times of weight, regulate pH value to 7.5, stir, standing 2h, then uses 350 object filter-cloth filterings, and gained decompression filtrate recycling ethanol obtains concentrate.After gained filter residue and drying, obtain the common solid-state Hongqu powder (red colouring agent) product of high-quality.
(5) acquisition of high look valency bacteriostatic water dissolubility functional red yeast rice
To adding in step (4) gained concentrate, to account for concentrate quality be 10% CMC (percent grafting 25%, molecular weight 1000kDa), then with acid for adjusting pH value to 5, stir, leave standstill 2h, then, with alkali adjusting pH to 7.5, now can produce a large amount of red precipitates, migrate out part supernatant, the part by weight that makes to remain supernatant and precipitation is 2:1, stir and make suspension, in whipping process, add and have malt sugar group-beta-cyclodextrin and hydroxy propyl-Beta cyclodextrin to suspension, in every kilogram of suspension, add malt sugar group-beta-cyclodextrin 30g, hydroxy propyl-Beta cyclodextrin 50g, after suspension spraying is dry, obtain high look valency bacteriostatic water dissolubility functional red yeast rice, spray-dired setting parameter: air inlet actual temperature is at 115 DEG C, 80 DEG C of air outlet actual temperatures, sample size is 5.0ml/min, intake is 100NL/h.
(6) bacteriostatic activity of high look valency bacteriostatic water dissolubility functional red yeast rice
The made Hongqu powder (red colouring agent) of step (5) is dissolved in distilled water, is mixed with mass percent concentration and is 5% the red colouring agent for food, also used as a Chinese medicine aqueous solution.Get this red colouring agent for food, also used as a Chinese medicine aqueous solution 1mL, add to and carry that in colibacillary LB culture medium, (e. coli concentration is 10
4cFU/mL), and at 37 DEG C, after 200 turn/min shaken cultivation 24h, absorption bacterium liquid is measured it, and under 600nm wavelength, light absorption value is lower than 10% of control group, and Escherichia coli Growth is significantly inhibited.
Embodiment 3:
A preparation method for high look valency bacteriostatic water dissolubility functional red yeast rice, comprises the steps:
(1) prior fermentation of functional Monascus
High temperature and cold fermentation: by functional Monascus strain transfer in fermentation materials, inoculum concentration is 2% of fermentation materials weight, then cultivate in hot fermentation chamber, cultivation temperature is 32 DEG C, incubation time is 3 days, after having cultivated, transfers to cold fermentation chamber and cultivates, cultivation temperature is 20 DEG C, and incubation time is 15 days.
In high temperature and cold fermentation process, fermentation materials is broken up to processing, within during hot fermentation 2 days, break up once, during cold fermentation, broke up once every 6 days.
Fermentation materials preparation method is: formula: brown rice 55wt%, and wheat bran 5wt %, soy meal 5 wt %, soy peptone 3 wt %, glucose 3 wt %, glycerine 1 wt %, surplus is water; Brown rice is pulverized, added other each component, and then add water, stir, packing volume into is the glass fermentation flask of 1L, and every bottled amount is 300g, with gauze sealing, adopts moist hear heat test sterilizing, 121 ± 1 DEG C, and 40min.
(2) processing of material after fermentation
Material after step (1) fermentation is carried out to sterilizing, sterilising conditions is 121 ± 1 DEG C, and then sterilizing 40min mixes with sterilized water material according to the weight ratio of 1:3, then process with colloid mill making beating, through making beating emulsifying materials fineness 10-50 micron after treatment.
(3) look song ferments again
Pack the material after step (2) making beating into full-automatic liquid fermentation tank, access look aspergillus kind is fermented again, inoculum concentration be making beating after weight of material 2%, fermentation condition is again: 30 DEG C of temperature, mixing speed 100r/min, throughput 3.0L/min, zymotic fluid pH value is 5.0-6.0, fermented incubation time is 15 days.
(4) fermentation liquor treatment
Get step (3) the gained zymotic fluid that ferments again, add the 75wt% edible alcohol of 1 times of weight, regulate pH value to 7.0, stir, standing 1.5h, then uses 300 object filter-cloth filterings, and gained decompression filtrate recycling ethanol obtains concentrate.After gained filter residue and drying, obtain the common solid-state Hongqu powder (red colouring agent) product of high-quality.
(5) acquisition of high look valency bacteriostatic water dissolubility functional red yeast rice
To adding in step (4) gained concentrate, to account for concentrate quality be 5% CMC (percent grafting 50%, molecular weight 2000kDa), then regulate pH value to 3, stir, leave standstill 1.5h, then, with alkali adjusting pH to 7, now can produce a large amount of red precipitates, migrate out part supernatant, the part by weight that makes to remain supernatant and precipitation is 1:1, stir and make suspension, in whipping process, add and have malt sugar group-beta-cyclodextrin and hydroxy propyl-Beta cyclodextrin to suspension, in every kilogram of suspension, add malt sugar group-beta-cyclodextrin 20g, hydroxy propyl-Beta cyclodextrin 30g, after suspension spraying is dry, obtain high look valency bacteriostatic water dissolubility functional red yeast rice, spray-dired setting parameter: air inlet actual temperature is at 105 DEG C, 70 DEG C of air outlet actual temperatures, sample size is 3.0ml/min, intake is 200NL/h.
(6) bacteriostatic activity of high look valency bacteriostatic water dissolubility functional red yeast rice
The made Hongqu powder (red colouring agent) of step (5) is dissolved in distilled water, is mixed with mass percent concentration and is 10% the red colouring agent for food, also used as a Chinese medicine aqueous solution.Get this red colouring agent for food, also used as a Chinese medicine aqueous solution 1mL, add to and carry that in colibacillary LB culture medium, (e. coli concentration is 10
4cFU/mL), and at 37 DEG C, after 200 turn/min shaken cultivation 24h, absorption bacterium liquid is measured it, and under 600nm wavelength, light absorption value is lower than 10% of control group, and Escherichia coli Growth is significantly inhibited.
Product look valency of the present invention is by the 500-1000u/g of general water-soluble monascus, more than bringing up to 5000u/g.The solubility of product is high, under 25 DEG C of conditions, can reach 50g/100ml water.The antibacterial water-soluble functional red yeast rice of high look valency that the present invention obtains, the content of Monacolin K is high, and spray drying process to split closed loop proportional impact less, product Open-closed-loop ratio is more than 95%.In spray-drying process, the malt sugar group-beta-cyclodextrin adding and hydroxy propyl-Beta cyclodextrin, in improving product stability, change the character of material, utilized and be dried, can improve the yield and quality, but also can increase the solubility of product, there is multi-efficiency.Add using carboxyl chitosan, can bring into play on the one hand the ability of its absorption Lovastatin and monascorubin, reach the concentrated of Lovastatin and monascorubin; By the introducing of using carboxyl chitosan, can give monascorubin product bacteriostatic activity on the other hand, improve its anti-living contaminants ability in food or pharmaceutical applications.
Product configuration of the present invention becomes the aqueous solution of 0.01% ~ 1% mass percent concentration, to Escherichia coli (
e.coli), staphylococcus aureus (
s.aureus), salmonella (
salmonella), pseudomonas aeruginosa (
p.aeruginosa), Friedlander's bacillus (
k.peneumoniae) and single increasing listeria spp (
l.monocytogenes) etc. there is remarkable bacteriostatic activity.
Above-described embodiment is preferably scheme of one of the present invention, not the present invention is done to any pro forma restriction, also has other variant and remodeling under the prerequisite that does not exceed the technical scheme that claim records.
Claims (10)
1. a preparation method for high look valency bacteriostatic water dissolubility functional red yeast rice, is characterized in that: comprise the steps:
(1) prior fermentation of functional Monascus
High temperature and cold fermentation: by functional Monascus strain transfer in fermentation materials, inoculum concentration is the 1-5% of fermentation materials weight, then cultivate in hot fermentation chamber, cultivation temperature is 30-35 DEG C, incubation time is 2-5 days, after having cultivated, transfers to cold fermentation chamber and cultivates, cultivation temperature is 16-25 DEG C, and incubation time is 10-25 days;
(2) processing of material after fermentation
Material after step (1) fermentation is carried out to sterilizing, sterilising conditions is 121 ± 1 DEG C, and then sterilizing 30-45min mixes with sterilized water material according to the weight ratio of 1:2-4, then process with colloid mill making beating, through making beating emulsifying materials fineness 10-50 micron after treatment;
(3) look song ferments again
Pack the material after step (2) making beating into full-automatic liquid fermentation tank, access look aspergillus kind is fermented again, inoculum concentration is the 1-5% of the weight of material after making beating, fermentation condition is again: temperature 28-30 DEG C, mixing speed 60-200r/min, throughput 2.0-5.0L/min, zymotic fluid pH value is 5.0-6.0, fermented incubation time is 10-20 days;
(4) fermentation liquor treatment
Get step (3) the gained zymotic fluid that ferments again, add the edible alcohol of 0.5-2 times of weight, regulate pH value to 6.5-7.5, stir, leave standstill 1-2h, then use 300-350 object filter-cloth filtering, gained decompression filtrate recycling ethanol obtains concentrate;
(5) acquisition of high look valency bacteriostatic water dissolubility functional red yeast rice
Account for to adding in step (4) gained concentrate the using carboxyl chitosan that concentrate quality is 0.1%-10%, then regulate pH value to 2.0-5, stir, leave standstill 1-2h, then, regulate pH to 5.5-7.5, now can produce a large amount of red precipitates, migrate out part supernatant, the part by weight that makes to remain supernatant and precipitation is 0.5-2.0:1, stir and make suspension, in whipping process, add and have malt sugar group-beta-cyclodextrin and hydroxy propyl-Beta cyclodextrin to suspension, in every kilogram of suspension, add malt sugar group-beta-cyclodextrin 10-30g, hydroxy propyl-Beta cyclodextrin 10-50g, after suspension spraying is dry, obtain high look valency bacteriostatic water dissolubility functional red yeast rice.
2. preparation method according to claim 1, is characterized in that: described functional Monascus bacterial classification cultural method is:
First class inoculum is cultivated: plant after activation female functional Monascus, be transferred on test tube slant culture medium, in constant temperature bacterial classification incubator, 25-35 DEG C of cultivation obtains first class inoculum for 2-5 days;
Second class inoculum is cultivated: first class inoculum is transferred in liquid culture medium at desinfection chamber, after inoculating, at 30-32 DEG C, in the isothermal vibration shaking table of rotating speed 150-200r/min, cultivates and within 3-7 days, obtain second class inoculum, second class inoculum is functional Monascus bacterial classification.
3. preparation method according to claim 2, it is characterized in that: slant medium is prepared as: taking the water of 100 weight portions as standard, add maltose modulation 5-8 Baume degrees, then add soy peptone 3-5 weight portion, glucose 2-5 weight portion, agar 2-3 weight portion; After all components is dissolved, pack test tube into, every test tube 8-12ml, then, with cotton plug sealing, carries out moist heat sterilization, and sterilising conditions is 121 DEG C ± 1 DEG C, 15-25min; After sterilizing, while hot test tube is tiltedly put, to be cooled.
4. preparation method according to claim 2, is characterized in that: liquid culture medium is prepared as: taking the water of 100 weight portions as standard, add glucose 3-6 weight portion, soy peptone 3-5 weight portion, white sugar 1-3 weight portion, ZnSO
40.1-0.5 weight portion, NaNO
30.1-0.5 weight portion, KH
2pO
40.1-0.3 weight portion, ground rice 1-2 weight portion, analysis for soybean powder 1-3 weight portion, mealy potato 0.5-1 weight portion, glycerine 1-3 weight portion; After mixing dissolving, pack in 500ml triangular flask, every bottle of 150-250ml, with after gauze sealing, uses moist hear heat test sterilizing, and sterilising conditions is: 121 DEG C ± 1 DEG C, and 20-30 minute.
5. according to the preparation method described in claim 1 or 2 or 3 or 4, it is characterized in that: in step (1) high temperature and cold fermentation process, fermentation materials is broken up to processing, within during hot fermentation 2-3 days, break up once, during cold fermentation, broke up once every 5-7 days.
6. according to the preparation method described in claim 1 or 2 or 3 or 4, it is characterized in that: the described fermentation materials preparation method of step (1) is: formula: brown rice 52-57wt%, wheat bran 4-6wt %, soy meal 4-6 wt %, soy peptone 2-3 wt %, glucose 2-3 wt %, glycerine 1-2 wt %, surplus is water; Brown rice is pulverized, added other each component, and then add water, stir, packing volume into is plastics fermentation flask or the glass fermentation flask of 1L, and every bottled amount is 250-350g, with gauze sealing, adopts moist hear heat test sterilizing, 121 ± 1 DEG C, and 30-45min.
7. according to the preparation method described in claim 1 or 2 or 3 or 4, it is characterized in that: the spray-dired setting parameter of step (5): air inlet actual temperature is at 102-115 DEG C, air outlet actual temperature 60-80 DEG C, sample size is 2.0-5.0ml/min, intake is 100-400NL/h.
8. preparation method according to claim 1, is characterized in that: described look aspergillus kind cultural method is:
First class inoculum cultivation: by look bent female kind after activation, be transferred on test tube slant culture medium, 25-30 DEG C of cultivation obtains first class inoculum for 2-5 days in constant temperature bacterial classification incubator;
Second class inoculum is cultivated: first class inoculum is transferred in liquid culture medium at desinfection chamber, after inoculating, at 25-30 DEG C, cultivates and within 4-7 days, obtain second class inoculum in the isothermal vibration shaking table of rotating speed 145-180r/min, second class inoculum is look aspergillus kind.
9. preparation method according to claim 8, it is characterized in that: slant medium is prepared as: taking the water of 100 weight portions as standard, add maltose modulation 5-8 Baume degrees, then add soy peptone 3-5 weight portion, glucose 2-5 weight portion, agar 2-3 weight portion; After all components is dissolved, pack test tube into, every test tube 8-12ml, then, with cotton plug sealing, carries out moist heat sterilization, and sterilising conditions is 121 DEG C ± 1 DEG C, 15-25min; After sterilizing, while hot test tube is tiltedly put, to be cooled.
10. preparation method according to claim 8, is characterized in that: liquid culture medium is prepared as: formula: rice meal 0.1-0.5wt%, and soy peptone is 2.0-5.0 wt %, glucose 1.0-3.0 wt %, sucrose 1.5-2.5 wt %, surplus is water; After mixing, pack in 500ml triangular flask, every bottle of 180-250ml, with after gauze sealing, uses moist hear heat test sterilizing, and sterilising conditions is: 121 DEG C ± 1 DEG C, and 20-30 minute.
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| CN107156638B (en) * | 2017-06-16 | 2021-05-25 | 江苏神华药业有限公司 | Preparation method of lipid-lowering red yeast powder |
| CN107212268A (en) * | 2017-07-24 | 2017-09-29 | 中国医药集团总公司四川抗菌素工业研究所 | A kind of preparation method of red yeast rice |
| CN109627804B (en) * | 2019-01-18 | 2020-03-27 | 福州隆利信生物制品有限公司 | A kind of method for extracting Monascus pigment |
| CN110623205A (en) * | 2019-09-20 | 2019-12-31 | 湖南微著生物科技有限公司 | Preparation method of red yeast rice with high color value |
| CN111139268B (en) * | 2020-01-22 | 2023-07-04 | 安诺康(福建)生物科技有限公司 | Preparation method of water-soluble functional red yeast rice powder |
| CN114395489A (en) * | 2022-01-27 | 2022-04-26 | 中南林业科技大学 | Preparation method and application of monascus koji |
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