CN1059463C - Violet monascus, monascus and their application. - Google Patents
Violet monascus, monascus and their application. Download PDFInfo
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- CN1059463C CN1059463C CN97100245A CN97100245A CN1059463C CN 1059463 C CN1059463 C CN 1059463C CN 97100245 A CN97100245 A CN 97100245A CN 97100245 A CN97100245 A CN 97100245A CN 1059463 C CN1059463 C CN 1059463C
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Abstract
The present invention relates to violet monascus and monascus which are new strains. The present invention has the advantages of very stable biochemical property and physicochemical property, high efficiency and stability of metabolites, wide requirement on culture mediums, no special requirement for environment, multiple function of products, high comprehensive utilization rate, no waste, low cost, simple process and manpower saving; monascus color and biologic nafenopin which are produced by the fermentation of the present invention have safe properties and multiple functions, can be used to develop various health foods (medicines), and can reduce triglyceride by 30 to 60%; in addition, the present invention has obvious effect, hypotensive effect, hypoglycemic effect, cancer prevention effect, etc., and is favourable for the heath of human bodies and the reduction of environmental pollution.
Description
The present invention relates to a kind of new bacterial strain and application thereof, belong to microbial technology field.
Monascorubin is the natural pigment of pure-blood ferment, and the past mostly obtains with the ethanol extraction, or directly red colouring agent for food, also used as a Chinese medicine is pulverized and used, the former cost height of these two kinds of methods, and it is few to obtain rate; The latter is extensive, and limitation is big, only makes soy cheese and so on usually.It also is like this that lipopenicillinase is loose.
Purpose of the present invention will overcome above-mentioned defective exactly, and the new bacterial strain purple Monascus anka Nakazawa et sato of extraction acquisition, Monascus anka Nakazawa et sato are produced monascorubin, lipopenicillinase is loose.
Technical solution of the present invention is:
New bacterial strain purple Monascus anka Nakazawa et sato N
4Monascus purpurus (AS3.4691), Monascus anka Nakazawa et sato N
14Monascus auka (AS3.4692) is characterized in that:
1-1-1. purple Monascus anka Nakazawa et sato N
4Monascus purpurus (AS3.4691) grows on malt extract medium and G25N substratum, the bacterium colony limitation, and quality is tight, and there are wrinkle and aerial hyphae in the surface, and mycelium is just orange, the orange red or purple grape in aging back, the bacterium colony quilt cover is red sauce.
1-1-2. bacterium colony presents radial wrinkle when growing on MEA and potato substratum, aerial hyphae is sparse and short and small, and quality is tight, and mycelium is just orange, and aging back is a red sauce, bacterium colony back side red sauce.
1-1-3. the microscopy form is: mycelia tool tabula, multinuclear, irregular branch, contain orange oil droplet, diameter 2-7 micron, conidium is accidental, Dan Sheng, pyriform, 810 microns, the quilt device is orange red, spherical, diameter 40-50 (70) micron, thecaspore is avette, smooth colourless, 6-7 * 4.5-5 micron.
1-2-1. Monascus anka Nakazawa et sato N
14Monascus auka (AS3.4692) grows on MEA and G25N substratum, bacterium colony topical type, many folds line likeness in form scar shape, no aerial hyphae, bacterium colony lava look or grape dark reddish purple look, the dark cologne earth look in the back side.
1-2-2. the microscopy form is: mycelia is separated branch, includes a large amount of orange red oil droplets, diameter 3-7 micron, conidium is many, pyriform 8-11 * 6-10 micron, quilt device sphere, orange red 30-50 micron, thecaspore oval, smooth, light red, 4-5 * 2-3 micron.
2, produce the method for monascorubin, it is characterized in that adopting new bacterial strain:
2-1. after the slant culture inoculation, temperature 30-32 ℃ cultivate down 13-16 days standby;
2-2. rice test tube or triangular flask are cultivated, the inoculation back under 30-34 ℃ of condition, cultivate 10-14 days standby;
Cultivate 2-3. the Quchi ventilation is cultivated or curtain is bent, after rice (bright rice) immersion steamed rice, the inoculation that cools, heap fermentation, went into pond (curtain) ventilating fermentation when treating the product temperature rise 4-5 days, bent low temperature (below 45 ℃) dry for standby to 50-51 ℃;
2-4. it is bent 25% usefulness by charging capacity of cryodrying is bent, and add the 75% warm boiling water (38-40 ℃) of total charging capacity, mix back fermentation 7-10 days about 28 ℃ thoroughly, with the wine hydraulic pressure after the fermentation press distill thick liquid (distillation back raffinate), again through essence filter after the filter residue and drying monascorubin.
3, fall the production method that ester looses, it is characterized in that adopting novel bacterial:
3-1. same 2-1,2-2,2-3 are described;
3-2. it is bent 25% usefulness by charging capacity of the described cryodrying of 3-1 is bent, and add the 75% warm boiling water (38-40 ℃) of total charging capacity, mix the back thoroughly about 28 ℃ fermentations of room temperature 7-10 days, with the wine hydraulic pressure after the fermentation press distill thick liquid (distillation back raffinate), be equipped with after the thick slag hyperthermia drying after its squeezing is pulverized that the correlation function material is refining to form.
New bacterial strain purple Monascus anka Nakazawa et sato of the present invention, its biochemistry of Monascus anka Nakazawa et sato, physicochemical property are very stable, and its meta-bolites efficient is high and stable, and is wide to the substratum requirement condition, and environment is not had particular requirement; The product function is many, comprehensive utilization ratio height, no waste material; Cost is low, and technology is simple, saves simple labour; The monascorubin of its fermentative production, lipopenicillinase diffusing (biological nafenopin), performance safety and function are many, can develop plurality kinds of health care food (medicine), can reach 30-60% to the triglyceride rate of falling, and instant effect, also have effects such as step-down, hypoglycemic and anti-cancer, help HUMAN HEALTH, reduce environmental pollution.
Fig. 1 is monascorubin of the present invention, lipopenicillinase day labor process flow figure.
Embodiment 1:
Purple Monascus anka Nakazawa et sato N
4Monascus purpureus (AS3.4691), Monascus anka Nakazawa et sato N
14Monascus auka (AS3.4692) is fungi, is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center on August 8th, 1996, and registering on the books is numbered: CGMCC NO.0269-1, CGMCC NO.0269-2.
The bacterial classification source: bacterium source of the present invention Jiangsu China prestige healthcare products factory red colouring agent for food, also used as a Chinese medicine under Yun Xi farm, the military region, Jiangsu Province is produced bacterium.Through separating with AS3.978 Monascus anka Nakazawa et sato preservation bacterial classification is reference, selects three group of 27 consistent relatively strain, i.e. purple Monascus anka Nakazawa et sato N
4Monascus purpureus (AS3.4691) and Monascus anka Nakazawa et sato N
14Monascus auka (AS3.4692).
Specimen preparation of bacterium source and purifying: gather 9 parts in bent sample from the red koji fermentation pond, 9 parts of bent samples are divided into three parts respectively, take out a little spoon with every part again, adding fills 10 milliliters of maltose liquid (10B that contain lactic acid 0.7%, alcohol 10%
e) test tube in, 30-32 ℃ leaves standstill and cultivated about 13 days, will float on the liquid level cenobium down in aseptic condition and choose in the triangular flask that fills 30 ml sterile waters, be placed with some sterilization granulated glass spherees in the bottle, shook 5 minutes, and smashed cenobium, filter with the sterilization funnel that is covered with the skim cotton, filtrate is diluted to proper concn, be coated with flat board,, select color depth, shallow, light in 35 ℃ of cultivations 6 days, the large, medium and small corresponding bacterium colony of bacterium colony moves into the maltose face and cultivates for screening usefulness.
Primary dcreening operation: with above strain isolated 27 strains, preservation strain 6 strains, carry out look valency, the bent diastatic fermentation simultaneous test of rice:
Will be under aseptic condition for examination bacterium slant strains, be transferred to triangular flask (500ml triangular flask, interior dress 50g rice medium, the postcooling 30-40 ℃ inoculation of sterilizing routinely) cultivates, under 40 ℃ ± 1 condition, cultivate after 48 hours, treat that the long adularescent spot of the grain of rice promptly transfers under 30-40 ℃ of condition in the triangular flask, cultivate after 168 hours, carry out look Jie, diastatic fermentation detection according to a conventional method.The result shows, the look valency is in 3 strains more than 1500,6 strains of look valency between 1000-1500, and other is all below 1000.Saccharogenic power is lower than 250 5 strains in 7 strains more than 700, and other are all about 500.
Multiple sieve: saccharogenic power is carried out producing behind the purifying at the bacterial strain more than 1500 again, sieved simultaneous test again 3 years at bacterial strain more than 700 and look valency, finally obtain new bacterial strain N
4Monascuspurpureus purple Monascus anka Nakazawa et sato (AS3.4691), look valency are stabilized in 1200-2000 (liquid state fermentation is higher than solid state fermentation), and saccharogenic power is at 600-800.N
14Monascusauka Monascus anka Nakazawa et sato (AS3.4691), its look valency is stabilized in 1000-1500 (liquid state fermentation is better than solid state fermentation), and saccharogenic power is at 700-850.Above-mentioned two strain bacterium are water-soluble more generally to be contrasted bacterium and has a clear superiority in, for food-processing provides convenience.
The composition of slant medium (%):
(1) MEA: malt extract 2%, peptone 0.1%, glucose 2%, agar 1.5%, pH value 5.1-5.4.
(2) G25N:N
aNO
30.3%, K
2HPO
40.1%, KCL0.05%, M
gSO
47H
2O 0.05%, F
eSO
47H
2O 0.01%, yeast extract 0.5%, agar 1.5%, glycerine 23%, pH value 6.5-6.7.
Triangular flask kind substratum: get the 1000g long-grained nonglutinous rice, soak and drain after 24 hours, steamed rice (ripe and sticking, interior no hard core) is sub-packed in after cooling in 20 500ml triangular flasks, and the 50g/ bottle can be inoculated under aseptic condition after sterilization.
Inoculation: draw 0.2% dilute acetic acid solution (0.3% glacial acetic acid solution) 5-10 milliliter with straight shape pipe, inject AS3.4691 or AS3.4692 in slant tube, under aseptic condition, stir lawn with glass stick or inoculating needle, use again through thick mouthful of suction pipe of sterilization and get 0.5-1 milliliter Monascus anka Nakazawa et sato liquid, shake up in the access rice triangular flask, be positioned in the 30-34 ℃ of thermostat container, should often shake during this time, rice is disperseed, does not make it to produce conglomeration and the irregular phenomenon of growing, cultivate 10-13 days standby.
Red colouring agent for food, also used as a Chinese medicine materials proportioning: get the 100kg long-grained nonglutinous rice and add cold boiling water 7kg, Glacial acetic acid 120-150kg, three bottles of Red kojic rice triangular flask kinds (150g), use is preceding with Red kojic rice kind porphyrize, and mixes with cold boiling water and Glacial acetic acid.
Immersion, steamed rice, inoculation: adopt first-class long-grained nonglutinous rice to be positioned in the vat, soak 120 fens kinds after, pick up and put into that the bamboo square-bottomed bamboo basket is few to be eluriated, the water in which rice has been washed flush away, drain, after all steaming around pouring in the steaming kier then, add a cover and steam three fens kinds again; The rice that steams after good moves in the treucher, rubs scattered caking, and heat radiation was cooled to 42-44 minute, inserts the porphyrize red colouring agent for food, also used as a Chinese medicine seed that has mixed Glacial acetic acid liquid, and the sterilization gunnysack of fully packing into behind the mix seals and enters deep closet and cultivate.
Cultivate: when entering deep closet, product temperature begins to drop to 34 ℃ by original 36 ℃, then rises to 39 ℃, needs 17-19 hour approximately; After approximately once per hour heat up, once heating up last half an hour, when treating that Qu Wen reaches 50-51 ℃, opens bag and move to the Quchi fermentation, cultivated 4-5 days under room temperature 25-30 ℃, 35 ℃ of conditions of bent temperature, ripe back cryodrying is standby.
The production method of monascorubin: rice or long-grained nonglutinous rice soaked drain after 24 hours, boiling, no sandwich, ripe and sticking getting final product, cool after 40 ℃, render in the altar, and press injected volume 25% and add red colouring agent for food, also used as a Chinese medicine, add 40 ℃ of warm boiling water that account for the total amount 75% that feeds intake again, stir evenly the back and add dry yeast, stir evenly back envelope altar fermentation 7-10 days, fermentation ends by 0.25%, open the altar squeezing, press liquid distill liquor and surplus liquid, with surplus liquid filter thick slag liquid and clear liquid, coarse filtration liquid is concentrated, oven dry, is the monascorubin elaboration after pulverizing.
The production method that lipopenicillinase is loose: will produce the squeezing vinasse in the monascorubin process, and oven dry, pulverize, refining to form lipopenicillinase diffusing with associated material compatibility again.
AS3.4691 or AS3.4692 are rich in Lovastain (C
24H
36O
5), a kind of HMG-COA reductase inhibitor (suppressing the synthetic of cholesterol) and monascin active substance Monacilink, has the blood fat reducing effect, wherein the Monas-corubin of red colouring agent for food, also used as a Chinese medicine citraurin and Rubuopunctation have active carbonyl, be easy to work with amino, therefore not only can treat amine mass formed by blood stasis (Ammoniemia), also can give protection against cancer effectively.
Embodiment 2:
Slant medium is with embodiment 1.
The monascorubin production method:
Get rice or long-grained nonglutinous rice 100Kg, after soaking, draining steamed rice, rendering to volume is in the big altar of 500Kg, and with meal before 24 hours, the Red kojic rice of getting the 25Kg cryodrying adds 40 ℃ of warm boiling water of 75Kg and goes into altar and soak song, after waiting to drop into rice, add the 0.25Kg dry yeast, seal fermentation after stirring evenly, stir once every day first three day, the fermentation room temperature should keep about 28 ℃, and fermentation in 7 to 10 days finishes.
Will fermented liquid after the squeezing wine liquid and vinasse, with wine liquid steam liquor and still residue, again still residue is filtered coarse filtration liquid and clear liquid, again coarse filtration liquid concentrated, oven dry, pulverize and be monascorubin pulvis elaboration, also can prepare the liquid monascorubin.
The scattered product method of lipopenicillinase:
After vinasse after the squeezing are carried out high temperature (100-120 ℃) oven dry, pulverize, make lipopenicillinase with relevant lipopenicillinase material compatibility again and loose or lipid-lowering pellet, capsule for descend of blood fat etc.Lipopenicillinase after refining is loose, and every gram contains Lovastain (C
24H
36O
5) more than 8 milligrams.
Attached: 1, strain identification book;
2, bacterial classification viability report.
Claims (6)
1, new bacterial strain purple Monascus anka Nakazawa et sato N
4(Monascus purpurus) now is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center, and its numbering of registering on the books is: CGMCCNO.0269-1.
2, new bacterial strain Monascus anka Nakazawa et sato N
14(Monascus anka) now is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center, and its numbering of registering on the books is: CGMCC NO.0269-2.
3, produce the method for monascorubin, it is characterized in that adopting claim 1 or 2 described new bacterial strains:
3-1. after the slant culture inoculation, temperature 30-32 ℃ cultivate down 13-16 days standby;
3-2. rice test tube or triangular flask are cultivated, the inoculation back under 30-34 ℃ of condition, cultivate 10-14 days standby;
Cultivate or the bent cultivation of curtain 3-3. Quchi ventilates, after rice (long-grained nonglutinous rice) immersion steamed rice, the inoculation that cools, heap fermentation, went into pond (curtain) ventilating fermentation when treating the product temperature rise 4-5 days, bent low temperature (below 45 ℃) dry for standby to 50-51 ℃;
3-4. it is bent 25% usefulness by charging capacity of cryodrying is bent, and add the 75% warm boiling water (38-40 ℃) of total charging capacity, mix back fermentation 7-10 days about 28 ℃ thoroughly, with the wine hydraulic pressure after the fermentation press distill thick liquid (distillation back raffinate), again through essence filter after the filter residue and drying monascorubin.
4, the production method of monascorubin according to claim 3 is characterized in that described slant medium comprises:
4-1.MEA (%): malt extract 2%, peptone 0.1%, glucose 2%, agar 1.5%, pH value 5.1-5.4.
4-2.G 25N (%): N
aNO
30.3%, K
2HPO
40.1%, KCL0.05%, M
gSO
47H
2O 0.05%, F
eSO
47H
2O 0.01%, yeast extract 0.5%, agar 1.5%, glycerine 23%, pH value 6.5-6.7.
5, fall the production method that ester looses, it is characterized in that adopting claim 1 or 2 described novel bacterials:
5-1. step 3-1,3-2,3-3 with claim 3 are described;
5-2. it is bent 25% usefulness by charging capacity of the described cryodrying of 5-1 is bent, and add the 75% warm boiling water (38-40 ℃) of total charging capacity, mix the back thoroughly about 28 ℃ fermentations of room temperature 7-10 days, with the wine hydraulic pressure after the fermentation press distill thick liquid (distillation back raffinate), be equipped with after the thick slag hyperthermia drying after its squeezing is pulverized that the correlation function material is refining to form.
6, the diffusing production method of lipopenicillinase according to claim 5 is characterized in that described slant medium comprises:
6-1.MEA (%): malt extract 2%, peptone 0.1%, glucose 2%, agar 1.5%, pH value 5.1-5.4.
6-2.G25N (%): N
aNO
30.3%, K
2HPO
40.1%, KCL0.05, M
gSO
47H
2O 0.05%, F
eSO
47H
2O 0.01%, yeast extract 0.5%, agar 1.5%, glycerine 23%, pH value 6.5-6.7.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN97100245A CN1059463C (en) | 1997-01-13 | 1997-01-13 | Violet monascus, monascus and their application. |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN97100245A CN1059463C (en) | 1997-01-13 | 1997-01-13 | Violet monascus, monascus and their application. |
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| Publication Number | Publication Date |
|---|---|
| CN1162640A CN1162640A (en) | 1997-10-22 |
| CN1059463C true CN1059463C (en) | 2000-12-13 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN97100245A Expired - Fee Related CN1059463C (en) | 1997-01-13 | 1997-01-13 | Violet monascus, monascus and their application. |
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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1908156B (en) * | 2005-08-02 | 2010-12-29 | 成都地奥九泓制药厂 | Monascus novel strain and traditional Chinese medicine monascus prepared by fermenting the same |
| CN100443583C (en) * | 2005-11-09 | 2008-12-17 | 朱兰琴 | Red monascus strain Danxi-1 |
| CN100443585C (en) * | 2005-11-09 | 2008-12-17 | 朱兰琴 | Monascus strain Danxi-3 |
| CN100443584C (en) * | 2005-11-09 | 2008-12-17 | 朱兰琴 | Purple monascus strain Danxi-2 |
| CN100382807C (en) * | 2005-11-09 | 2008-04-23 | 张帆 | Natural medicine composition for preventing and treating metabolic syndrome |
| CN101347476B (en) * | 2008-09-09 | 2011-01-19 | 北京东方红航天生物技术股份有限公司 | Health products capable of reducing blood pressure and preparation thereof |
| CN103966056B (en) * | 2014-06-03 | 2015-09-16 | 四川理工学院 | A kind of preparation method of white wine long-grained nonglutinous rice non-dramatic song |
| CN106591186A (en) * | 2016-12-13 | 2017-04-26 | 北京林业大学 | Preparation method of spirulina platensis protoplasts |
| CN110903983B (en) * | 2019-12-03 | 2021-04-13 | 武汉佳成生物制品有限公司 | Monascus monascus with high yield of saccharifying enzyme and esterifying enzyme and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1103891A (en) * | 1993-12-15 | 1995-06-21 | 广东江门生物技术开发中心 | Method for prepn. of water soluble monascin pigments |
-
1997
- 1997-01-13 CN CN97100245A patent/CN1059463C/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1103891A (en) * | 1993-12-15 | 1995-06-21 | 广东江门生物技术开发中心 | Method for prepn. of water soluble monascin pigments |
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