CN101775384A - Method for producing crude enzyme preparations of esterifying enzymes by using monascus - Google Patents
Method for producing crude enzyme preparations of esterifying enzymes by using monascus Download PDFInfo
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- CN101775384A CN101775384A CN 201010115946 CN201010115946A CN101775384A CN 101775384 A CN101775384 A CN 101775384A CN 201010115946 CN201010115946 CN 201010115946 CN 201010115946 A CN201010115946 A CN 201010115946A CN 101775384 A CN101775384 A CN 101775384A
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Abstract
The invention discloses a method for producing crude enzyme preparations of esterifying enzymes by using monascus strains. In the method, the crude enzyme preparations of the esterifying enzymes are produced by using the monascus strains which can produce the esterifying enzymes with high yield as production strains, using rice flour, soy flour, lactic acid or yellow water as main materials, using rice husks as auxiliary materials and adopting a solid state fermentation process. The method has the advantages of simple process, difficult pollution, easy operation and short fermentation time (five days); and the esterifying power of the produced crude enzyme preparations of the esterifying enzymes can reach 80 to 160mg per 100ml in 7 days.
Description
Affiliated technical field
The present invention relates to a kind of preparation method of crude zyme preparation, especially utilize high yield esterification enzyme monascus strain to adopt solid-state fermentation process to produce the method for crude enzyme preparations of esterifying enzymes.Affiliated field is a microbial fermentation technology.
Background technology
Aromatic Chinese spirit is an output maximum in China's liquor 12 big odor types, the branch cloth cover is the widest, manufacturer maximum, market covers a widest class liquor, occupies an important position in national economy and people's daily life.At present, also there is long, problem such as high-grade-goods rate is low of production cycle in the production of high-quality aromatic Chinese spirit, has seriously restricted the development of aromatic Chinese spirit enterprise.One of the main reasons is that the slow and final content of the formation speed of the main fragrance matter ethyl hexanoate of aromatic Chinese spirit is low.Producing at present and going up the method that improves ethyl hexanoate content mainly is that caproic acid bacteria is irritated the cellar for storing things method, and this method has little effect, and is to produce caproic acid because caproic acid bacteria mainly acts on, and a little less than the esterification, caproic acid is changed into the ability of ethyl hexanoate.Therefore using catalyst caproic acid and ethanol to generate ethyl hexanoate is to improve the quality of aromatic Chinese spirit and shorten one of major measure of production cycle.Discover that a lot of bacterial strains in bacterium, yeast, the mould can both synthesize the lipase of energy catalysis short chain fatty acid and ethyl esterification, this quasi-microorganism is referred to as esterifying bacteria by liquor industry, but the esterifying bacteria that is fit to wine brewing production is also few.
Monascus is the traditional medicinal fungi of China, and it is inoculated in the red colouring agent for food, also used as a Chinese medicine that rice forms through fermentative preparation is the traditional Chinese medicine that a kind of dietotherapy is had both, and has effects such as strengthening the spleen to promote digestion promoting blood circulation and removing blood stasis.Studies show that red colouring agent for food, also used as a Chinese medicine also has effects such as lipopenicillinase, step-down, hypoglycemic and inhibition tumor growth simultaneously.Early 1960s, some bacterial strain of the biological monascus of finding in Chinese Academy of Sciences Chengdu can make short chain fatty acid and alcohol generate ester, has opened up new way for the application of red colouring agent for food, also used as a Chinese medicine in liquor production, is a great innovation of China's liquor industry.Because monascus can be bred growth in wine brewing places such as Daqu, koji workshop, wine brewing mash, poor unstrained spirits, thereby becomes the focus of wine brewing circle research.The research of domestic in recent years wine bound pair monascus has obtained certain effect, as the biological Lu type esterifying bacteria M101 bacterial strain that is separated in Chinese Academy of Sciences Chengdu, utilizes this strain culturing " to strengthen the bacterium song ", is used for fermentation and produces wine, has stronger living Xiang Gong energy.The two foreign wine factories in Jiangsu are separated to the monascus strain that a strain has extremely strong product ester ability, and this bacterial strain is used for the production of aromatic Chinese spirit, and to improving output, the quality of yeast wine, the ratio of adjusting ethyl hexanoate/ethyl lactate all has positive effect.
Though liquor industry is being obtained important achievement aspect the seed selection of monascus specie in recent years, monascus utilization aspect also exists technological deficiency with not enough.The main technique of utilizing monascus to produce crude enzyme preparations of esterifying enzymes at present is raw material with the wheat bran, and the tray solid state fermentation prepares wheat bran.Nutritious because of wheat bran, numerous microorganism growth that suit, and monascus is compared poor growth with other mould, cause this technology contaminated easily, to place and operational requirement strictness, the large-scale production Pollution risk is big.Rice is the natural medium of monascus growth, monascus is grown on rice matrix, competitive edge is obvious, be difficult for polluting, but employing is the traditional technology production red colouring agent for food, also used as a Chinese medicine crude enzyme preparations of esterifying enzymes of the red colouring agent for food, also used as a Chinese medicine production of raw material with the rice also exists defective, and mainly show as: traditional technology is a raw material with whole grain rice, and monascus and matrix contact surface are little, mycelia is slow toward the interior growth of matrix seepage velocity, causes fermentation period long (generally more than 10 days); Complex technical process is loaded down with trivial details, consuming time (will drain through soaking, cook, cooling such as breaks up at pretreatment procedure); The control of technology key point requires high to workman's experience, implement difficulty big (soak as rice, cook etc. all just right).
Summary of the invention
Easily pollute in order to overcome existing red colouring agent for food, also used as a Chinese medicine crude enzyme preparations of esterifying enzymes production technique, fermentation period is long, complex technical process is loaded down with trivial details, difficult defective and the deficiency of key point control, the invention provides a kind of method of utilizing monascus to produce crude enzyme preparations of esterifying enzymes, this method serves as to produce bacterial strain with high yield esterification enzyme monascus strain, with rice meal, analysis for soybean powder, lactic acid or yellow water is major ingredient, with the rice husk is auxiliary material, adopt solid-state fermentation process to produce the wine crude enzyme preparations of esterifying enzymes, the esterification power of the crude enzyme preparations of esterifying enzymes that this method is produced reaches (80-160) mg/100ml7day.This method technology is simple, easy to operate, is difficult for polluting, and is not high to site requirements, and can significantly shorten fermentation period (can shorten to 5 days).
Technical scheme: rice meal is broken into powdery makes carbon source, analysis for soybean powder is replenished nitrogenous source, and consumption is the 3%-7% of rice meal quality, regulates pH value with lactic acid or yellow water, the lactic acid consumption is the 0.5%-1% of rice meal quality, yellow water is the 19%--27% of rice meal quality, with the rice husk is to support auxiliary material, plays fluffy ventilative oxygen supply, it is agglomerating to remedy rice meal chance water, the ventilation property variation, oxygen supply is not enough and the defective that causes monascus not grow, and its consumption is the 40%-50% of rice meal quality.Can adopt triangular flask solid state fermentation pattern or tray solid state fermentation pattern to cultivate.When adopting the triangular flask solid state fermentation, 80% of adding rice meal quality water in the described in the above material, fully mix thoroughly, divide and install to (the bottling amount is 90g/500ml-100g/500ml triangular flask or 200g/1000ml-220g/1000ml triangular flask) in the triangular flask, autoclaving (121 ℃) 30 minutes, being heat breaks up, when being cooled to 40 ℃ of left and right sides, insert 10ml monascus ruber liquid, shake up, at 34 ℃ of following constant temperature culture 115-125 hours, treat that whole culture medium covers with mycelia, slightly redly, arrive and cultivate terminal point, receive bent the oven dry down and be red colouring agent for food, also used as a Chinese medicine crude enzyme preparations of esterifying enzymes finished product in 45 ℃; When adopting the tray solid state fermentation, the water that adds the 120%-150% of rice meal quality, fully mix thoroughly, with gauze or calico bag by matrix, autoclaving (121 ℃) 30 minutes is heat and breaks up, when being cooled to 40 ℃ of left and right sides, insert the solid-state seed of triangular flask secondary of the 8%-10% of quality of material, mix thoroughly and shakeout in the tray, making material thickness is 4cm, covers 4 layers of wet gauze or 2 layers of wet calico in the above, in 34 ℃ of following constant temperature culture 48 hours, cool to 32 ℃, mend 10% water of quality of material, gauze or the calico that covered this moment have become dry, be replaced with wet gauze or wet calico, continue to cultivate 52h-62h, treat that whole culture medium covers with mycelia, slightly redly, to cultivating terminal point, receive song oven dry under 45 ℃ and be red colouring agent for food, also used as a Chinese medicine crude enzyme preparations of esterifying enzymes finished product.
The invention has the beneficial effects as follows: a kind of method of utilizing monascus to produce crude enzyme preparations of esterifying enzymes is provided, the shortcoming of easily having polluted when this method has overcome existing technology and is the raw material production crude enzyme preparations of esterifying enzymes with the wheat bran, and improved the esterification power (bringing up to 80-160mg/100ml7day) of crude enzyme preparations of esterifying enzymes from 40-80mg/100ml7day; It is to moisten grain in the production technique of raw material (generally to need 24 hours with the rice that this method has been saved, make suction between 60%-70%), drain, cook and (require just to care, ripe and oiliness, difficult control), need loaded down with trivial details step consuming time such as repeatedly moisturizing in the culturing process, and significantly shortened fermentation period (from shortening to about 5 days more than 10 days).Production technique provided by the invention is simple, and is easy to operate, and Pollution risk is little, easily large-scale production.
Embodiment
The present invention is further described below in conjunction with embodiment.
Implementation column 1
(1) slant culture:
Slant medium: 10
0The BX wort, agar 3%, PH nature; Monascus specie is inoculated on the slant medium, cultivated 7 days down, treat that mycelia is equipped with next step use when covering with test tube and producing spore at 34 ℃.
(2) preparation of bacterium liquid:
3 (1) described test tube seed is washed in the 500ml triangular flask that fills the 100ml sterilized water, add a certain amount of granulated glass sphere and 1g glucose, at 34 ℃, isothermal vibration was cultivated 24 hours under the rotating speed of 180r/min, got monascus ruber liquid.
(3) triangular flask solid state fermentation:
Solid-state fermentation culture medium: rice meal 400g, analysis for soybean powder 20g, lactic acid 4ml, rice husk 200g, distilled water 320ml; Mixing rice meal and analysis for soybean powder thoroughly back adds rice husk and mixes thoroughly.With distilled water, add behind the abundant mixing of lactic acid in the above-mentioned solid substrate, rub even (can not with energetically) gently with the hands, allow rice meal fully be distributed on the rice husk and guarantee that material is fluffy.Above-mentioned material average mark is installed in 10 500ml triangular flasks, and tampon is wrapped tampon with kraft paper beyond the Great Wall, high pressure (121 ℃) sterilization 30 minutes.Being heat after the taking-up shakes agglomerating matrix diffusing, to be cooled during to 40 ℃ of left and right sides, each triangular flask inserts the described bacterium liquid of 10ml (2), after shaking up, detains bottle after 72 hours 34 ℃ of following constant temperature culture, continue to cultivate 48 hours, this moment, fermented substrate covered with mycelia, slightly redly, and to cultivating terminal point, receive bent under 45 ℃, dry red colouring agent for food, also used as a Chinese medicine crude enzyme preparations of esterifying enzymes finished product 390g, esterification power is 110mg/100ml7day.
Implementation column 2
(1) slant culture:
With implementation column 1.
(2) preparation of bacterium liquid:
With implementation column 1.
(3) the solid-state seed of triangular flask:
Early stage, operation was with implementation column 1, about 72 hours of 34 ℃ of following constant temperature culture, the concrete time decided according to the matrix growing state, treats that matrix covers with white hypha, when microscopy had a large amount of generation of conidium, button shook and makes matrix disperse to be equipped with tray cultivation use as seed as far as possible at this moment.
(4) tray solid state fermentation:
Solid-state fermentation culture medium: rice meal 750g, analysis for soybean powder 37.5g, yellow water 180ml, rice husk 375g, distilled water 900ml; Mixing rice meal and analysis for soybean powder thoroughly back adds rice husk and mixes thoroughly.With distilled water, add behind the abundant mixing of yellow water in the above-mentioned solid substrate, rub even (can not with energetically) gently with the hands, allow rice meal fully be distributed on the rice husk and guarantee that material is fluffy.With gauze or calico bag by matrix, autoclaving (121 ℃) 30 minutes, being heat breaks up, to be cooled during to 40 ℃ of left and right sides, inserting the solid-state seed of the described triangular flask of 100g (3) (about 2 bottles of 500ml triangular flask cultures) mixes thoroughly, it is 4cm that material is shakeout the thickness that makes it in the tray, cover 4 layers of wet gauze or 2 layers of wet calico in the above, under 34 ℃, constant temperature culture is 48 hours under the condition of humidity 〉=80%, temperature is dropped to 32 ℃, mend the water of weight of material 10%, gauze or the calico that covered this moment have become dry, are replaced with wet gauze or wet calico, continue to cultivate 58h under the condition of maintenance humidity 〉=90%, treat that whole culture medium covers with mycelia, slightly redly, to cultivating terminal point, receive bent under 45 ℃, dry red colouring agent for food, also used as a Chinese medicine crude enzyme preparations of esterifying enzymes 730g, esterification power is 100mg/100ml7day.
Claims (6)
1. utilize monascus strain to produce the method for crude enzyme preparations of esterifying enzymes, it is characterized in that: with rice meal, analysis for soybean powder, lactic acid or yellow water is major ingredient, rice husk is an auxiliary material, and the monascus strain of high yield esterification enzyme is a fermentation strain, adopts solid-state fermentation process to produce the wine crude enzyme preparations of esterifying enzymes.
2. the method for utilizing monascus strain to produce crude enzyme preparations of esterifying enzymes according to claim 1, it is characterized in that: be carbon source with the rice meal, soyflour is a nitrogenous source, is stopping composition with the rice husk, regulates pH value with lactic acid or yellow water.
3. according to claim 1, the 2 described methods of utilizing monascus strain to produce crude enzyme preparations of esterifying enzymes, it is characterized in that: soyflour: the ratio of rice meal (w/w) is 3/100-7/100; Rice husk: rice meal (w/w) is 40/100-50/100; Lactic acid: rice meal (v/m) is 0.5/100-1/100 or yellow water: rice meal (v/m) is 19/100-27/100.
4. according to the described method of utilizing monascus strain to produce crude enzyme preparations of esterifying enzymes of claim 1-3, it is characterized in that: during the triangular flask solid state fermentation, water content is 80% (rice meal quality 80%), inoculum size is a 10ml monascus ruber liquid, the bottling amount is (90-100) g/500ml triangular flask or (200g-220) g/1000ml triangular flask, culture temperature is 34 ℃, and incubation time is 115h-125h.
5. according to the described method of utilizing monascus strain to produce crude enzyme preparations of esterifying enzymes of claim 1-3, it is characterized in that: during the tray solid state fermentation, water content is 120%-150% (percentage ratio of rice meal quality), inoculum size is the solid-state seed of triangular flask of 8%-10% (percentage ratio of quality of material), material thickness is 4cm, culture temperature was 34 ℃ in preceding 48 hours, and the later stage is 32 ℃, and incubation time is 100h-110h.
6. the method for utilizing monascus strain to produce crude enzyme preparations of esterifying enzymes according to claim 5, it is characterized in that: during the tray solid state fermentation, material is during fermentation covered by the wet calico of 4 layers of wet gauze or 2 layers, mends 10% moisture (percentage ratio of quality of material) and change the wet gauze or the wet calico of having sterilized after cultivating 48h.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101993825A (en) * | 2010-07-27 | 2011-03-30 | 湖北工业大学 | Monascus aurantiacus |
| CN103045558A (en) * | 2012-12-18 | 2013-04-17 | 山东中惠食品有限公司 | Method for producing esterifying enzyme preparation from monascus |
| CN103173316A (en) * | 2013-04-02 | 2013-06-26 | 四川省食品发酵工业研究设计院 | Method for reducing ethyl lactate content in luzhou-flavor liquor |
| CN103502429A (en) * | 2010-10-28 | 2014-01-08 | 道达尔研究技术弗吕股份有限公司 | Process for polylactic acid production using monascus |
| CN110283806A (en) * | 2019-07-15 | 2019-09-27 | 北京工商大学 | A kind of monascus parpureus Went Lipase absobed enzyme LIP05-50, encoding gene and its application |
| CN110283805A (en) * | 2019-07-15 | 2019-09-27 | 北京工商大学 | A kind of monascus parpureus Went Lipase absobed enzyme LIP05, encoding gene and its application |
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| CN1297056A (en) * | 1999-11-18 | 2001-05-30 | 张军 | Process for preparing monascorubin with higher yield |
| CN101240301A (en) * | 2008-01-22 | 2008-08-13 | 贵州大学 | Solid-state fermentation preparation method for gamma-aminobutyric acid |
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Patent Citations (2)
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| CN1297056A (en) * | 1999-11-18 | 2001-05-30 | 张军 | Process for preparing monascorubin with higher yield |
| CN101240301A (en) * | 2008-01-22 | 2008-08-13 | 贵州大学 | Solid-state fermentation preparation method for gamma-aminobutyric acid |
Non-Patent Citations (1)
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| 《中国酿造》 20041231 周立平 红曲霉在中国的生产及其应用（下） 第5-7页 1-6 , 第10期 2 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101993825A (en) * | 2010-07-27 | 2011-03-30 | 湖北工业大学 | Monascus aurantiacus |
| CN101993825B (en) * | 2010-07-27 | 2012-09-05 | 湖北工业大学 | Monascus aurantiacus |
| CN103502429A (en) * | 2010-10-28 | 2014-01-08 | 道达尔研究技术弗吕股份有限公司 | Process for polylactic acid production using monascus |
| CN103502429B (en) * | 2010-10-28 | 2018-07-17 | 道达尔研究技术弗吕股份有限公司 | The method for producing polylactic acid using monascus |
| CN103045558A (en) * | 2012-12-18 | 2013-04-17 | 山东中惠食品有限公司 | Method for producing esterifying enzyme preparation from monascus |
| CN103173316A (en) * | 2013-04-02 | 2013-06-26 | 四川省食品发酵工业研究设计院 | Method for reducing ethyl lactate content in luzhou-flavor liquor |
| CN103173316B (en) * | 2013-04-02 | 2014-05-07 | 四川省食品发酵工业研究设计院 | Method for reducing ethyl lactate content in luzhou-flavor liquor |
| CN110283806A (en) * | 2019-07-15 | 2019-09-27 | 北京工商大学 | A kind of monascus parpureus Went Lipase absobed enzyme LIP05-50, encoding gene and its application |
| CN110283805A (en) * | 2019-07-15 | 2019-09-27 | 北京工商大学 | A kind of monascus parpureus Went Lipase absobed enzyme LIP05, encoding gene and its application |
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Application publication date: 20100714 |