CN102010831A - Lovastatin-generating fungus and application thereof in production of Pu'er tea - Google Patents
Lovastatin-generating fungus and application thereof in production of Pu'er tea Download PDFInfo
- Publication number
- CN102010831A CN102010831A CN 201010182965 CN201010182965A CN102010831A CN 102010831 A CN102010831 A CN 102010831A CN 201010182965 CN201010182965 CN 201010182965 CN 201010182965 A CN201010182965 A CN 201010182965A CN 102010831 A CN102010831 A CN 102010831A
- Authority
- CN
- China
- Prior art keywords
- lovastatin
- tea
- monascus
- production
- purpurevs
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a lovastatin-generating fungus and the application thereof in the production of Pu'er tea, belonging to the biotechnology field. The production strain of the invention is obtained by separating, purifying and primarily filtering through thin layer chromatography and high efficiency liquid phase secondarily filtering. The production strain is lovastatin-generating Monascus purpurevs MPT13 with the preservation No. of CCTCC M 2010123, and is preserved on May 20, 2010, in the preservation department of China center for type culture collection located in Wuhan University, Wuhan. The lovastatin-generating fungus of the invention is applied to the production of Pu'er tea. The invention has the advantages that Monascus purpurevs is proportionally vaccinated and applied to the fermentation process of Pu'er tea to obtain the lovastatin-contained Pu'er tea with new characteristics in aspects of aroma and soup on a basis of keeping the flavor of traditional Pu'er tea, the processing time is shortened, and the invention plays an important role in the development of new Pu'er tea products and specific Pu'er tea.
Description
Technical field:
The present invention relates to a kind of produce lovastatin fungi and the application in Leaf of Assam Tea is produced thereof, belong to biological technical field.
Background technology:
In recent years, Leaf of Assam Tea is subjected to extensive concern owing to its good fat-reducing effect and unique quality, and there is the multidigit scholar to be reported in and found lovastatin in the Leaf of Assam Tea, his spit of fland medicine is hydroxyl first glutaryl coenzyme A (HMG-CoA) reductase inhibitor, it is the most classical and effective fat-reducing medicament, be widely used in the treatment of hyperlipidaemia, but at present also not relevant for the systematic study of microorganism in lovastatin and the Leaf of Assam Tea, also do not have as yet and produce the lovastatin bacterial strain and be applied in research in the fermentation of Pu'er.
Utilize monascus ruber to produce the production of Red kojic rice and monascorubin and the history of the existing more than one thousand years of application, its purposes also expands to field of food gradually by medical field.Monascus claims the secondary metabolite lovastatin that produces to become research and the focus of producing as good natural lipid lowerers at fermenting process, lovastatin in the natural red colouring agent for food, also used as a Chinese medicine mostly is acid, HMG-CoA is more approaching in its space structure and the body, need not hydrolysis, directly performance suppresses the effect of body inner cholesterol synthetic, its activity is high about one times than the lactone formula, and Fujian, Zhejiang one band has been developed protective foodss such as the multiple functional buckwheat red wine that contains lovastatin, biscuit.
In the Leaf of Assam Tea research field, because the research of Leaf of Assam Tea is especially carried out in the research of long-term lacking subject from the angle of microorganism.And how by the research to microorganism in the yunnan puer tea processing, the content of lovastatin is not seen disclosed bibliographical information as yet in the control Leaf of Assam Tea.
Summary of the invention:
The objective of the invention is Leaf of Assam Tea to be studied, produce the useful bacterium source that contains the lovastatin Leaf of Assam Tea for manufacturing enterprise provides from the angle of microorganism.Another object of the present invention is by the ratio of control probiotics in Leaf of Assam Tea is produced, and plays a role for promoting yunnan puer tea stay in grade and raising.
Product lovastatin fungi of the present invention is by the separation of producing bacterial strain, purifying, obtain after thin-layer chromatography primary dcreening operation, the multiple riddler's preface of high performance liquid phase.Produce bacterial strain for producing lovastatin purple Monascus anka Nakazawa et sato mould (Monascus purpurevs) MPT13, preservation date: on May 20th, 2010, preserving number: CCTCC M 2010123, depositary institution: Chinese typical culture collection center, address: Wuhan Wuhan University.
The morphological feature of monascus parpureus Went of the present invention (Monascus purpurevs) is:
Cultivate 7d, colony diameter reaches 2.9cm on the PDA substratum, and bacterium colony swells slightly, and radial rill is arranged, neat in edge, and the aerial hyphae rareness, the substrate mycelium prosperity, colony colour is based on orange, and the edge is a light salmon; There is radial rill at the bacterium colony back side, and neat in edge, color be based on dark orange, the edge orange.Colony diameter reaches 2.5cm on malt extract medium, and bacterium colony swells slightly, and radial zanjon line is arranged, neat in edge, aerial hyphae are very rare, the substrate mycelium prosperity, the colony edge depression, there is the epithelium shape on the bacterium colony surface, and colony colour becomes orange from ashen from inside to outside; There is radial rill at the bacterium colony back side, and the cracking of being shape is arranged, neat in edge, and color is ashen.A little less than the czapek's solution strain growth, bacterium colony is flat, and diameter is 1.2cm, and the edge is irregular, and boundary is unclear, and aerial hyphae is sparse, and a little less than the substrate mycelium growth, colony colour is light orange; Bacterium colony back side color is light orange, and the edge boundary is unclear.The present invention obtains after conventional processes such as known primary dcreening operation, multiple sieve, natural medium cultivation, the cultivation of head mold purifying.
Microscopic morphology be mycelia brokenly branch, barrier film is arranged, in oil droplet is arranged, with many particles, diameter is 2.9～9.2 μ m on the wall.Conidium is single to be given birth to or chaining, the sphere or the pyriform of falling, 6.6～9.2 * 7.9～13.1 μ m.Cleistothecium does not have the aperture, and is spherical or oval slightly, and diameter 20.4～71.4 μ m are orange or light red slightly.Thecaspore is avette or oval, 3.9～5.3 * 4.7～6.6 μ m.
A large amount of enzymes and pigment that monascus produced are being formed with important effect to the Leaf of Assam Tea quality, is produced from Pu'er of lovastatin bacterial strain (monascus parpureus Went Monascus purpurevs) MPT13 fermentation in inoculation, on the basis that keeps traditional Leaf of Assam Tea product local flavor, fragrance and Tang Se aspect also have new characteristics.
The fungi of product lovastatin of the present invention is applied in the production of Leaf of Assam Tea.
The invention has the advantages that: monascus parpureus Went (Monascus purpurevs) is inoculated according to a certain percentage be applied in the Leaf of Assam Tea fermenting process, obtained containing the Leaf of Assam Tea of lovastatin and on the basis that keeps traditional puer tea flavor, fragrance and Tang Se aspect also have new characteristics, and shortened process period, the exploitation of Leaf of Assam Tea product innovation and characteristic Leaf of Assam Tea has been played a significant role.
Embodiment:
Embodiment 1: the separation and purification of monascus parpureus Went (Monascus purpurevs) bacterial classification
From the sample that collects, (the 200g potato is cleaned the peeling stripping and slicing by the PDA substratum, add water 1000mL and boil half an hour, filtered through gauze, add 20g glucose and 18g agar again, 121 ℃ of sterilizations are fallen dull and stereotyped after 30 minutes) and malt extract medium (wort 750～800ml adds water to 1000mL, add 15～18g agar, 121 ℃ of sterilizations are fallen dull and stereotyped after 30 minutes) separation and purification.
Separation purification method is as follows:
(1) with the solid medium fusing, be cooled to 45 ℃, inject sterile petri dish, about 15～20 milliliters of every ware, rocking-turn is a little left standstill, and makes dull and stereotyped standby.
(2) get and want isolating pulverizing sample 1g, be added in the test tube that the 9ml sterilized water is housed in that flame is other, what obtained this moment is 10
-2Bacteria suspension.
(3) dilution: bacteria suspension is diluted to 10 by decimal dilution method
-8, the bacteria suspension that ten times of methods are meant next pipe is than rare ten times of a last pipe.
(4) inoculation: dull and stereotyped 9 covers of previously prepd, label is 10 respectively
-6, 10
-7, 10
-8, each extent of dilution is write three cover plates, so that relatively.Inject the corresponding bacteria suspension of 1ml in the plate of label at each, be coated with evenly, put into 25～28 ℃ and cultivate observation in 24 hours with spreading rod.
(5) cultivate after 2～5 days, the single bacterium colony of picking, the purifying of repeatedly ruling obtains the purifying bacterial strain.
Embodiment 2: the screening of producing lovastatin monascus parpureus Went (Monascus purpurevs)
(1) liquid state fermentation
The bacterial strain that separation and purification obtains, inoculation wort inclined-plane, 28 ℃ cultivate 6d after, wash with the 10mL sterilized water in the little triangular flask of band granulated glass sphere and make spore suspension.Get the 3mL spore suspension and be inoculated in (the bottled liquid 50ml of every 250ml triangle) 30 ℃ in the 50mL liquid seed culture medium, 150r/m, it is little red to liquid color to cultivate 2～3d.
Inoculum size by 5% inserts (250mL liquid amount 50mL) 28 ℃ in the liquid fermentation medium with liquid seed culture medium, and 150r/m cultivates about 12～14d.
(2) thin-layer chromatography primary dcreening operation
Get fermented liquid 1mL, add ethyl acetate 5mL → 30 ℃ following 150r/min jolting 3 hours → centrifugal, collect supernatant liquor → precipitation again with 5mL ethyl acetate extracting 2 times → merging supernatant liquor, after evaporate to dryness → with a little benzene dissolves dry thing → treat that benzole soln can be sent out, add the 0.2mL dissolve with methanol.Liquid to be measured is point sample 10 μ L on thin layer chromatography board, at hexanaphthene: chloroform: launch in the developping agent of Virahol=6: 3: 1, after drying up, develop the color with 10% phospho-molybdic acid.
(3) high performance liquid phase sieves again
Get the 1mL fermented liquid, add 10mL methyl alcohol, ultrasonic extraction 1h uses earlier filter paper filtering, filters with mocromembrane again, collects filtrate, carries out the high performance liquid phase check and analysis, and analysis condition is: ultraviolet detection wavelength 238nm; Chromatographic column: EclipseXDB-C18 (5 μ m, 4.4 * 150mm); Moving phase:: A methyl alcohol; B 60% methyl alcohol; 30 ℃ of column temperatures; Elution program: 0-12min (A liquid, 100%-0; B liquid, 0-100%), 12-15min (100%B liquid), 15-20min (100%A liquid); Flow velocity: 1mL/min; Sample size: 10 μ l.
Embodiment 3: the application of monascus parpureus Went (Monascus purpurevs) in the Leaf of Assam Tea fermentation
Undertaken damp and hot by traditional method gross tea, the solid state fermentation agent of monascus parpureus Went (Monascus purpurevs) of inoculation weight ratio 5% allows its fermentation in gross tea subsequently, turning regularly, when heap temperature rise to 25 ± 3 ℃, Monascus purpurevs begins to carry out metabolism, lovastatin is the secondary metabolite of monascus, begins to produce after the 3-5 behind the inoculation fermentation days, and output reached the highest in 12-15 days.
The Leaf of Assam Tea of producing behind inoculation monascus parpureus Went (Monascus purpurevs), lovastatin content is up to 0.263mg/mL, and it is dense bright to have formed the soup look red, the sweet cunning of flavour, the fragrant outstanding flavor characteristic of fragrance ester.
Claims (2)
1. one kind is produced the lovastatin fungi, by the separation of producing bacterial strain, purifying, after thin-layer chromatography primary dcreening operation, the multiple riddler's preface of high performance liquid phase, obtain, it is characterized in that producing bacterial strain for producing lovastatin monascus parpureus Went (Monascus purpurevs) MPT13, preservation date: on May 20th, 2010, preserving number CCTCC M 2010123, depositary institution: Chinese typical culture collection center, address: Wuhan Wuhan University.
2. right 1 described product lovastatin fungi is characterized in that producing the lovastatin fungi and is applied in the production of Leaf of Assam Tea.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101829659A CN102010831B (en) | 2010-05-26 | 2010-05-26 | Lovastatin-generating fungus and application thereof in production of Pu'er tea |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101829659A CN102010831B (en) | 2010-05-26 | 2010-05-26 | Lovastatin-generating fungus and application thereof in production of Pu'er tea |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102010831A true CN102010831A (en) | 2011-04-13 |
| CN102010831B CN102010831B (en) | 2012-04-18 |
Family
ID=43841163
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010101829659A Active CN102010831B (en) | 2010-05-26 | 2010-05-26 | Lovastatin-generating fungus and application thereof in production of Pu'er tea |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102010831B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586119A (en) * | 2012-01-03 | 2012-07-18 | 西藏月王生物技术有限公司 | Functional monascus strain, and preparation method and application of functional highland barley monascus prepared by using functional monascus strain |
| CN103060203A (en) * | 2012-12-21 | 2013-04-24 | 楚雄师范学院 | Fungus and application thereof |
| CN103349099A (en) * | 2012-10-12 | 2013-10-16 | 谢春生 | Simvastatin-containing Pu'er tea, and fermenting method and application thereof |
| CN104770487A (en) * | 2015-04-21 | 2015-07-15 | 张淑芹 | Production method for monascus pine needle tea |
| CN105685284A (en) * | 2016-03-03 | 2016-06-22 | 云南农业大学 | Processing method for Lovastatin Pu'er tea |
| CN109055235A (en) * | 2018-08-20 | 2018-12-21 | 云南农业大学 | A kind of preparation of the inoculating microbes strain for Pu'er tea pile fermentation and its application method |
| CN110373332A (en) * | 2019-07-19 | 2019-10-25 | 江南大学 | A kind of purple Monascus and its total ferment produce the method and application of Lovastatin |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1908156A (en) * | 2005-08-02 | 2007-02-07 | 成都地奥九泓制药厂 | Monascus novel strain and traditional Chinese medicine monascus prepared by fermenting the same |
| CN101028023A (en) * | 2007-04-06 | 2007-09-05 | 云南农业大学 | Green trichodermin and its use in production of Pu'er tea |
| CN101490271A (en) * | 2006-05-24 | 2009-07-22 | 加利福尼亚大学董事会 | Methods and materials for making simvastatin and related compounds |
-
2010
- 2010-05-26 CN CN2010101829659A patent/CN102010831B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1908156A (en) * | 2005-08-02 | 2007-02-07 | 成都地奥九泓制药厂 | Monascus novel strain and traditional Chinese medicine monascus prepared by fermenting the same |
| CN101490271A (en) * | 2006-05-24 | 2009-07-22 | 加利福尼亚大学董事会 | Methods and materials for making simvastatin and related compounds |
| CN101028023A (en) * | 2007-04-06 | 2007-09-05 | 云南农业大学 | Green trichodermin and its use in production of Pu'er tea |
Non-Patent Citations (1)
| Title |
|---|
| 《中国抗生素杂志》 19990430 王立新等 红曲霉固态发酵生产洛伐他汀的研究 , 第02期 2 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586119A (en) * | 2012-01-03 | 2012-07-18 | 西藏月王生物技术有限公司 | Functional monascus strain, and preparation method and application of functional highland barley monascus prepared by using functional monascus strain |
| CN102586119B (en) * | 2012-01-03 | 2014-07-16 | 西藏月王生物技术有限公司 | Functional monascus strain, and preparation method and application of functional highland barley monascus prepared by using functional monascus strain |
| CN103349099A (en) * | 2012-10-12 | 2013-10-16 | 谢春生 | Simvastatin-containing Pu'er tea, and fermenting method and application thereof |
| CN103060203A (en) * | 2012-12-21 | 2013-04-24 | 楚雄师范学院 | Fungus and application thereof |
| CN104770487A (en) * | 2015-04-21 | 2015-07-15 | 张淑芹 | Production method for monascus pine needle tea |
| CN105685284A (en) * | 2016-03-03 | 2016-06-22 | 云南农业大学 | Processing method for Lovastatin Pu'er tea |
| CN109055235A (en) * | 2018-08-20 | 2018-12-21 | 云南农业大学 | A kind of preparation of the inoculating microbes strain for Pu'er tea pile fermentation and its application method |
| CN110373332A (en) * | 2019-07-19 | 2019-10-25 | 江南大学 | A kind of purple Monascus and its total ferment produce the method and application of Lovastatin |
| CN110373332B (en) * | 2019-07-19 | 2021-11-02 | 江南大学 | Monascus purpureus and method for co-fermenting monascus purpureus to produce lovastatin and application of monascus purpureus |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102010831B (en) | 2012-04-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102010831B (en) | Lovastatin-generating fungus and application thereof in production of Pu'er tea | |
| CN103224890A (en) | Monascus ruber for high generation of lovastatin and gamma-aminobutyric acid and low generation of pigment, use thereof, and functional Monascus sp. prepared through using Monascus ruber | |
| CN106635820B (en) | A kind of Aspergillus niger strain of high yield theabrownin and its application | |
| CN106978350A (en) | One plant of aspergillus niger and its application in the preparation of Pu'er tea chlorins compound | |
| CN108018218A (en) | One plant height produces ethyl acetate yeast strain and its cultural method and application | |
| CN106754438B (en) | A kind of Christian Breton silk spore Pichia pastoris and application thereof | |
| CN104630076A (en) | Monascus purpureus (Monascus purpureus) Mp-42 strain capable of producing amylase at high yield and application of monascus purpureus Mp-42 strain | |
| CN107189949A (en) | Rhizopus oryzae LJH3 and the application in bioconversion Sophoricoside prepares genistein | |
| CN109468232B (en) | Feruloyl esterase-producing eurotium schlegelii and application thereof in white spirit Daqu | |
| CN108315265B (en) | Aspergillus versicolor Av-2 strain and application thereof | |
| CN103073527B (en) | Diterpene Libertellenone G and preparation method and use thereof | |
| CN110551636B (en) | Monascus purpureus MY-21 strain and application thereof | |
| CN104611236A (en) | Cumminghamella echinulata(Thaxter) thaxter FAR3 and method for fermentation preparation of Gamma-linolenic acid grease with Cumminghamella echinulata(Thaxter) thaxter FAR3 | |
| CN102373166B (en) | Bacillus thuringiensis PX-95 strain capable of producing poly-beta-hydroxybutyric acid at high yield and application thereof | |
| CN102154123B (en) | Dioscorea nipponica Makino Fusarium sp. and application thereof | |
| CN101603011B (en) | Method for preparing epipodophyllotoxin diglucoside and special strain used thereby | |
| CN106754430A (en) | Dendritic bacterial strains of cladosporium Cc 106 of high yield monascus purpureus and application thereof | |
| CN105802896A (en) | Komagataeibacter rhaeticus for producing saccharic acid-1,4-lactone | |
| CN110128392A (en) | It is a kind of by compound isolated in aspergillus flavus secondary metabolite | |
| CN115634241A (en) | Preparation method of Shandong ganoderma lucidum extract and application of Shandong ganoderma lucidum extract in preparation of hypoglycemic drugs | |
| CN106479900B (en) | High yield monascus purpureus penicillium oxalicum Po-25 bacterial strain and application thereof | |
| CN101575576A (en) | Aspergillus fumigatus and application thereof | |
| CN106282032B (en) | Penicillium citrinum LB and the application in phillygenol is prepared in bioconversion forsythin | |
| CN103865804B (en) | Beta-glucosidase Producing Strain and the application in resveratrol is prepared in conversion thereof | |
| TWI486452B (en) | Method for producing resveratrol through biotransformation by microorganism |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190715 Address after: 650000 No. 305, 8 Blocks, Fenghuajunyuan, Chunrong Street, Chenggong District, Kunming City, Yunnan Province Patentee after: Yunnan Zhifang Industrial Co., Ltd. Address before: 650201 Tea College of Yunnan Agricultural University, Heilongtan, Kunming City, Yunnan Province Patentee before: Yunnan Agricultural University |
|
| TR01 | Transfer of patent right |