CN105462861A - Process for efficiently producing rhizopus koji - Google Patents
Process for efficiently producing rhizopus koji Download PDFInfo
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- CN105462861A CN105462861A CN201610050328.3A CN201610050328A CN105462861A CN 105462861 A CN105462861 A CN 105462861A CN 201610050328 A CN201610050328 A CN 201610050328A CN 105462861 A CN105462861 A CN 105462861A
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Abstract
The invention discloses a process for efficiently producing rhizopus koji. The rhizopus koji is obtained through four-stage culture, and different culture media are selected for seeds of all the stages, wherein the potato dextrose agar culture medium is selected for the first-stage test tube seeds, the malt juice liquid culture medium is selected for the second-stage shake flask seeds, the potato-saccharose liquid culture medium is selected for the third-stage fermentation cylinder seeds, and the rice and wheat bran culture medium is selected for the fourth-stage production seeds. The production process technology with liquid-solid combined fermentation is adopted in the process, the liquid state fermentation technology is adopted for the first three stages of seeds, the solid state fermentation technology is adopted for the fourth-stage production seeds, the respective advantages of solid state fermentation and pure liquid state fermentation are combined, the prepared rhizopus koji is high in purity, high in activity, stable in quality, high in efficiency and short in period, the quality of bacteria of all the stages can be monitored online, and industrial production can be achieved.
Description
Technical field
The present invention relates to microorganism culturing field, particularly a kind of technique of High-efficient Production head mold song.
Background technology
The amylase activity of head mold is very strong, is conventional amylomyces in brewery industry.Most important effect is played in the brewing process of the bent making at fermented glutinous rice of head mold, yellow rice wine, Xiaoqu wine, the bent quality of head mold has directly been determined the yield of leavened prod and quality, the quality of head mold song mainly from its diastatic vigor, miscellaneous bacteria rate etc. decomposed because usually judging, and head mold song is always by each producer is paid attention to.The manufacture craft of tradition head mold song, adopt the method for solid medium progressively level enlarged culturing always, this method advantage is simple, convenient, practical, Gu weak point is that the performance of excellent species fails comparatively fast in solid-Secondary Culture process, easily cause into Qu Zhiliang unstable, activity is not high, preservation period is short, finally affects yield and the quality of downstream leavened prod; In addition in solid Secondary Culture process, can not on-line monitoring be carried out, contaminate a small amount of miscellaneous bacteria and also can't find, easily caused into Qu Buchun, the situation that productivity is not high; Also have the cycle of solid culture seed culture at different levels all to need 2-3 days, the time is longer, and seed, originally in drying process, controls tight also easy bacteria infection.It is bent that some R&D institution makes head mold with pure liquid state fermentation, Gu overcome some problems existed in solid-Secondary Culture production technique, the head mold mycelia that the method is produced is sturdy, active high, steady quality, can on-line Control, and level of automation is high, with short production cycle, miscellaneous bacteria infectivity also reduces greatly, but the finished product that the method is produced is mycelium, far away from the easy like that storage of spore that solid state fermentation is produced, use very inconvenience, practicality is low; Comparatively speaking, equipment investment is high in pure liquid state fermentation, and technical requirements is tight, and output is also much lower, and general factory is difficult to accomplish scale production.
Summary of the invention
For solving during existing head mold song is produced the problems referred to above existed, goal of the invention of the present invention is for providing a kind of head mold Qu Chundu produced high, active strong, steady quality, efficiency is high, cycle is short, can the quality of on-line monitoring bacterial classification at different levels, and can realize the technique of the High-efficient Production head mold song of suitability for industrialized production.
For achieving the above object, the invention provides a kind of technique of High-efficient Production head mold song, it is characterized in that, cultivated by level Four and obtain head mold song, the substratum that seed at different levels adopts is different: one-level test tube kind adopts potato dextrose agar; Second-level shake flask kind adopts malt juice liquid medium; Three grade fermemtation tank kind adopts potato-liquid sucrose substratum; Level Four produces kind of an employing rice wheat bran medium.
Preferably, in described rice wheat bran medium, the ratio of ground rice and wheat bran is 24:1, and described ground rice is rice abrasive dust, crosses 40 mesh sieves.
Preferably, described one-level test tube kind is adopted and is cultivated with the following method: preparation potato dextrose agar; In 115 DEG C of sterilizing 25min, divide and be filled in sterilized test tube, make test tube slant substratum; The rhizopus that picking has activated is inoculated in test tube slant, and 30 DEG C of constant temperature culture 48h obtain one-level test tube kind.
Preferably, the rhizopus original seed activation of the described rhizopus activated to be preserving number be HNCC6.197 obtains for 3-4 time.
Preferably, the compound method of described potato dextrose agar is: select fresh nothing to go mouldy potato, peeling, stripping and slicing, temperature fire boils rear filtration, moisturizing also adds glucose and agar powder, stirs, and final weight is than being potato 20%, glucose 2%, agar powder 2%.
Preferably, described second-level shake flask kind is adopted and is obtained with the following method: preparation maltose liquid nutrient medium, 115 DEG C of autoclaving 15min, after cooling, under gnotobasis, one-level test tube kind is inoculated in triangular flask, constant-temperature table cultivates 36h, and wherein temperature 30 DEG C, rotating speed 80rpm/min, obtain second-level shake flask kind.
Preferably, in described maltose liquid nutrient medium, maltose ratio is 13%.
Preferably, described three grade fermemtation tank kind is adopted and is obtained with the following method: preparation potato-liquid sucrose substratum adds in fermentor tank, 121 DEG C of sterilizing 20min, with sterilized stirrer, second-level shake flask kind is blended in an aseptic environment, the inoculum size access of 8% is pressed with in the fermentor tank of cooling by aseptic technique, add penicillin to 50 μ g/mL, 36h cultivated by 28-32 DEG C of bottom fermentation tank simultaneously, monitors pH value change in culturing process.
Preferably, described potato-liquid sucrose substratum is adopted and is prepared with the following method: select fresh nothing and to go mouldy potato, be cut into small pieces and pour in jacketed kettle after peeling, add the water of 5 times of volumes, after boiling 40-60min, with filter-cloth filtering, moisturizing to 5 times volume, stir to obtain fresh murphy juice; Fresh murphy juice is added in fermentor tank kind, add the sucrose of 2% and the defoamer of 0.5 ‰ simultaneously.
Preferably, the actual conditions of described fermentor cultivation is: fermentor tank mixing speed 200r/min, tank pressure 0.8-1.2kg/cm
2, ventilation 0.4V/Vmin.
Preferably, described level Four is produced to plant to adopt and is obtained with the following method: preparation rice wheat bran medium, takes out cooling, add the fresh pure water of 35% massfraction, stir when rice wheat bran medium is cooled to 50 DEG C after putting into Autoclave dry blowing 1h; Cultured three grades of kinds are released from fermentor tank, with sterilized stirrer, mycelium is blended, ratio in 10% is inoculated in above-mentioned rice wheat bran medium, after stirring, delivers to the fermenting house of having sterilized and cultivates 36h with the condition fixed temperature and humidity of temperature 30 DEG C, humidity 60%.
Preferably, also comprise into bent baking step, level Four produced to plant and put into drying room, 40-45 DEG C, hydrofuge dries 30-36h, obtains head mold song when moisture content is down to 8-10%.
The present invention has following beneficial effect: (1) this technique adopts the production technology of liquid-solid state combining with fermentation, first three grade of seed adopts liquid state fermentation technology, kind solid-state fermentation technology produced by the fourth stage, and combine solid state fermentation and pure liquid state fermentation advantage separately, the head mold Qu Chundu produced is high, active strong, steady quality, efficiency is high, and the cycle is short, can the quality of on-line monitoring bacterial classification at different levels, and can suitability for industrialized production be realized; (2) be with potato, rice, wheat bran for raw material, deposit number HNCC6.197's is original seed with mould, adopts mushy stage combining with fermentation to be trained the production and processing technology of quality rhizopus koji.The present invention adopts new production technology, can on-line monitoring seed quality at different levels in the production process of head mold song, the whole production cycle also obviously shortens, the head mold produced is bent, and saccharifying enzymic activity brings up to 1240U/g from 1172U/g, and miscellaneous bacteria number also greatly reduces, basic inspection does not measure, make kind of bent activity higher, quality is more stable, and security is better.
Embodiment
Below by specific embodiment, the present invention is further illustrated.
A technique for High-efficient Production head mold song, cultivated by level Four and obtain head mold song, the substratum that seed at different levels adopts is different: one-level test tube kind adopts potato dextrose agar; Second-level shake flask kind adopts malt juice liquid medium; Three grade fermemtation tank kind adopts potato-liquid sucrose substratum; Level Four produces kind of an employing rice wheat bran medium, and wherein, in rice wheat bran medium, the ratio of ground rice and wheat bran is 24:1, and described ground rice is rice abrasive dust, crosses 40 mesh sieves.
Wherein, the preparation process of one-level test tube kind is: select fresh nothing and to go mouldy potato, take 200g after peeling, be cut into small pieces, add after water temperature fire boils 40min, by 8 layers of filtered through gauze, moisturizing, to 1L, adds 20g glucose and 20g agar powder, stirs in 115 DEG C of sterilizing 25min, divide and be filled in 10 sterilized test tubes, make test tube slant substratum; Picking activated 3-4 time be inoculated in test tube slant with mould, 30 DEG C of constant temperature culture 48h obtain one-level test tube kind.
Wherein, the preparation process of second-level shake flask kind is: take malt extract powder (not containing agar) 130g, add distilled water or deionized water 1L, stirring heating is boiled to dissolving completely, dividing is filled in the triangular flask of two 1L capacity, the bottled 300ml of each triangle, 115 DEG C of autoclaving 15min, after cooling, under gnotobasis, picking 4 fritter one-level test tube kind is in each triangular flask, cultivate 36h as in constant-temperature table (temperature 30 DEG C, rotating speed 80rpm/min) again, obtain second-level shake flask kind.
Wherein, the preparation process of three grade fermemtation tank kind is: select fresh nothing and to go mouldy potato 1.2kg, be cut into small pieces after peeling and pour in jacketed kettle, add the water of 6L, after boiling 40-60min, with filter-cloth filtering, moisturizing is to 6L, and stir to obtain fresh murphy juice; Fresh murphy juice is all added in 10L fermentor tank kind, add the defoamer of 120g sucrose and 3ml simultaneously, 121 DEG C of sterilizing 20min; With sterilized stirrer, second-level shake flask kind is blended in an aseptic environment, by aseptic technique access 480ml bacterium liquid with in the fermentor tank of cooling, add quantitative penicillin simultaneously, make penicillin concn be 50 μ g/mL; Fermentor tank mixing speed 200r/min, tank pressure 0.8-1.2kg/cm
2, ventilation 0.4V/Vmin, 28-32 DEG C, in culturing process, the change of the change of on-line checkingi pH value, mycelium content and the limpid degree of nutrient solution, after cultivating 36h, obtains three grade fermemtation tank kind; In culturing process, pH value reduces gradually, and when being cultured to 33-36h, ph value is reduced to about 3.6 by initial value 7, if pH value is lower than 3 or all represent that seed is abnormal higher than 4 this moment; Along with the increase of incubation time, substratum becomes limpid transparent gradually, the bent distinctive sweet fragrance of head mold is more and more stronger, mycelium content also increases gradually, tank can be put when mycelium content is increased to 80-100g/L, now mycelium content reaches maximum, no longer increases with the prolongation of incubation time.
Wherein, the preparation process that level Four production is planted is: take 48kg rice, be crushed to 40 orders, mix after adding 2kg wheat bran, take out cooling after putting into Autoclave dry blowing 1h, add 17.5kg fresh sterile water when rice-wheat bran medium is cooled to 50 DEG C, stir; Cultured three grades of kinds are released from fermentor tank, with sterilized stirrer, mycelium is blended, take 3kg bacterium liquid and be inoculated in above-mentioned rice-wheat bran medium, after stirring, deliver to fermenting house fixed temperature and humidity and cultivate 36h, obtain level Four production kind;
Finally, become bent and dry, level Four produced to plant and put into drying room, 40-45 DEG C, hydrofuge dries 30-36h, must become bent.
The head mold Qu Chundu cultivated according to above-mentioned steps is high, active strong, steady quality, and efficiency is high, and the cycle is short, can the quality of on-line monitoring bacterial classification at different levels, and can realize suitability for industrialized production.The present invention adopts new production technology, can on-line monitoring seed quality at different levels in the production process of head mold song, the whole production cycle also obviously shortens, the head mold produced is bent, and saccharifying enzymic activity brings up to 1240U/g from 1172U/g, and miscellaneous bacteria number also greatly reduces, basic inspection does not measure, make kind of bent activity higher, quality is more stable, and security is better.
Apply specific embodiment herein to set forth principle of the present invention and embodiment, the explanation of above embodiment just understands method of the present invention and core concept thereof for helping.The above is only the preferred embodiment of the present invention, should be understood that, due to the finiteness of literal expression, and objectively there is unlimited concrete structure, for those skilled in the art, under the premise without departing from the principles of the invention, some improvement, retouching or change can also be made, also above-mentioned technical characteristic can be combined by rights; These improve retouching, change or combination, or the design of invention and technical scheme are directly applied to other occasion without improving, and all should be considered as protection scope of the present invention.
Claims (10)
1. a technique for High-efficient Production head mold song, is characterized in that, cultivated by level Four and obtain head mold song, the substratum that seed at different levels adopts is different: one-level test tube kind adopts potato dextrose agar; Second-level shake flask kind adopts malt juice liquid medium; Three grade fermemtation tank kind adopts potato-liquid sucrose substratum; Level Four produces kind of an employing rice wheat bran medium.
2. the technique of High-efficient Production head mold song according to claim 1, is characterized in that, in described rice wheat bran medium, the ratio of ground rice and wheat bran is 24:1, and described ground rice is rice abrasive dust, crosses 40 mesh sieves.
3. the technique of High-efficient Production head mold song according to claim 1, is characterized in that, described one-level test tube kind is adopted and cultivated with the following method: preparation potato dextrose agar; In 115 DEG C of sterilizing 25min, divide and be filled in sterilized test tube, make test tube slant substratum; The rhizopus that picking has activated is inoculated in test tube slant, and 30 DEG C of constant temperature culture 48h obtain one-level test tube kind.
4. the technique of High-efficient Production head mold song according to claim 3, it is characterized in that, the compound method of described potato dextrose agar is: select fresh nothing to go mouldy potato, peeling, stripping and slicing, temperature fire boils rear filtration, moisturizing also adds glucose and agar powder, stirs, and final weight is than being potato 20%, glucose 2%, agar powder 2%.
5. the technique of High-efficient Production head mold song according to claim 1, it is characterized in that, described second-level shake flask kind is adopted and is obtained with the following method: preparation maltose liquid nutrient medium, 115 DEG C of autoclaving 15min, after cooling, be inoculated in triangular flask under gnotobasis by one-level test tube kind, constant-temperature table cultivates 36h, wherein temperature 30 DEG C, rotating speed 80rpm/min, obtain second-level shake flask kind.
6. the technique of High-efficient Production head mold song according to claim 1, it is characterized in that, described three grade fermemtation tank kind is adopted and is obtained with the following method: preparation potato-liquid sucrose substratum adds in fermentor tank, 121 DEG C of sterilizing 20min, with sterilized stirrer, second-level shake flask kind is blended in an aseptic environment, the inoculum size access of 8% is pressed with in the fermentor tank of cooling by aseptic technique, add penicillin to 50 μ g/mL simultaneously, 36h cultivated by 28-32 DEG C of bottom fermentation tank, monitors pH value change in culturing process.
7. the technique of High-efficient Production head mold song according to claim 6, it is characterized in that, described potato-liquid sucrose substratum is adopted and is prepared with the following method: select fresh nothing and to go mouldy potato, be cut into small pieces after peeling and pour in jacketed kettle, add the water of 5 times of volumes, after boiling 40-60min, with filter-cloth filtering, moisturizing to 5 times volume, stir to obtain fresh murphy juice; Fresh murphy juice is added in fermentor tank kind, add the sucrose of 2% and the defoamer of 0.5 ‰ simultaneously.
8. the technique of High-efficient Production head mold song according to claim 7, is characterized in that, the actual conditions of described fermentor cultivation is: fermentor tank mixing speed 200r/min, tank pressure 0.8-1.2kg/cm
2, ventilation 0.4V/Vmin.
9. the technique of High-efficient Production head mold song according to claim 2, it is characterized in that, described level Four is produced to plant to adopt and is obtained with the following method: preparation rice wheat bran medium, cooling is taken out after putting into Autoclave dry blowing 1h, add the fresh pure water of 35% massfraction when rice wheat bran medium is cooled to 50 DEG C, stir; Cultured three grades of kinds are released from fermentor tank, with sterilized stirrer, mycelium is blended, ratio in 10% is inoculated in above-mentioned rice wheat bran medium, after stirring, delivers to the fermenting house of having sterilized and cultivates 36h with the condition fixed temperature and humidity of temperature 30 DEG C, humidity 60%.
10. the technique of High-efficient Production head mold song according to claim 9, is characterized in that, also comprise into bent baking step, and level Four produced to plant and put into drying room, 40-45 DEG C, hydrofuge dries 30-36h, obtains head mold song when moisture content is down to 8-10%.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106801018A (en) * | 2017-02-13 | 2017-06-06 | 湖南轻工研究院有限责任公司 | A kind of bent preparation method of vigor Mucor high |
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| 《中国优秀硕士学位论文全文数据库工程科技Ⅰ辑》 * |
| 《中国酿造》 * |
| 《酿酒科技》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106801018A (en) * | 2017-02-13 | 2017-06-06 | 湖南轻工研究院有限责任公司 | A kind of bent preparation method of vigor Mucor high |
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Application publication date: 20160406 |