CN1483810A - Method for culturing funcional red aspergillus quick solid - Google Patents
Method for culturing funcional red aspergillus quick solid Download PDFInfo
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Mushroom Cultivation (AREA)
Abstract
The present invention relates to a functional monoscaceae quick solid culture method. Said invention utilizes the regulation of air intake and air exhaustion of culture chamber or plastic film greenhouse or opened type ventilation system, and increases the air intake in the logarithmic phase of mycelium growth to change temp. and regulate humidity, and regulates and controls the change of water content of culture medium material so as to make culture medium material retain pultaceous state and accelerate mycelium growth rate and synthesis and accumulation of functional metabolic product monacolin K. In said obtained product the jiemycin is not detected.
Description
Technical field
The present invention relates to a kind of functional monoscaceae quick solid culture method.
Background technology
Red kojic rice is the traditional zymotic product of China, is divided into the storehouse song, and light bent and look bent three major types is used for food color or wine brewing for a long time.Red colouring agent for food, also used as a Chinese medicine is called red bent in ancient times, and record early can trace back to " clear different record " " cooking meat with red colouring agent for food, also used as a Chinese medicine " that later stage Tang Dynasty Tao Gu writes; To unit, the Ming Dynasty, relevant red colouring agent for food, also used as a Chinese medicine preparation and the record of using day by day increase, and Compendium of Material Medica, " new compilation of materia medica " are thought, red colouring agent for food, also used as a Chinese medicine " sweet temperature is nontoxic ", " warm in nature, flavor is sweet, has effect promoting blood circulation and removing blood stasis, strengthening the spleen to promote digestion, curing mainly diseases such as the stomachache of being retarded by silt, dyspepsia are glutted, wound ".What deserves to be mentioned is that the successive dynasties then record for wine of rice fermented with red yeast: " wine brewing person, then hot, slightly poisonous ", in Qing Dynasty's document, think wine of rice fermented with red yeast " hot in nature, as should not to drink " more.As for the alcoholic strength heat that red colouring agent for food, also used as a Chinese medicine led to, slightly poisonous saying is unsolved riddle for many years always.
1979, Japan's rattan chapter far away was found to be the active substance of the synthetic key enzyme inhibitor of body's cholesterol from the nutrient solution of red Monascus anka Nakazawa et sato (Monascus ruber), and (Monacolin K is also made Lovastatin, Mevinolin) to called after Monacolin K; People such as Hong Kong Wang Hin-Chung in 1981 find that the monascorubin of thick extraction has anti-microbial activity, at that time with its called after Monascidin A, but discover that people such as nineteen ninety-five France Blanc it has identical feature with Citrinin that Penicillium Citrinum produces (mycotoxins Citrinin) aspect analyses such as absorption spectrum, NMR, MS, is same substance by identification with Citrinin, be a kind of neurovirulent secondary metabolite that has, it is a kind of nephrotoxin material.After this, the research of the functional meta-bolites of Monascus anka Nakazawa et sato is very active, France, Japan, the U.S., Holland and China have successively carried out research, have affirmed the existence of Citrinin in red colouring agent for food, also used as a Chinese medicine and the related products thereof, and have inquired into the measuring method and the some effects factor of Citrinin.In recent years monascus product is subjected to strict check in countries such as Japan, France, clearly from Chinese import red colouring agent for food, also used as a Chinese medicine, must have safety in production bacterial classification and product not to contain Citrinin.Therefore, the formation that how to make Monascus anka Nakazawa et sato suppress Monascidin A (Citrinin) in chromogenesis and functional meta-bolites Monacolin K has become an important topic, screening does not produce Citrinin or the extremely micro-bacterial strain of Citrinin output, and the bacterial strain and the related process that improve red colouring agent for food, also used as a Chinese medicine look valency and functional metabolite content, become the research focus of Chinese scholars.
Summary of the invention
The purpose of this invention is to provide a kind of functional monoscaceae quick solid culture method, make Monascus anka Nakazawa et sato in chromogenesis and functional meta-bolites Monacolin K, suppress the formation of Monascidin A (Citrinin), product does not contain Citrinin.
Method of the present invention is to set up a kind of alternating temperature, the Monascus anka Nakazawa et sato mycelium quick solid cultural method of damping and regulation and control culture base-material water content, in spawn culture bag or vial filling culture base-material, insert little ventpipe, by regulating culturing room or plastics hothouse air output exhaust air rate or open type ventilation system, at the air flow of mycelial growth logarithmic phase increasing to strain bag or bottle, reach alternating temperature, the variation of damping and regulation and control culture base-material water content, the ability that suppresses mycelial growth logarithmic phase secreting amylase, reduction is to starchiness culture base-material liquefying-saccharifying speed, make culture base-material keep loose, quicken the synthetic and accumulation of mycelial growth speed and meta-bolites Monacolin K.Detect according to one's analysis, the mycelium culture cycle is when 7d, 14d, 21d and 28d, and Monascus anka Nakazawa et sato meta-bolites Monacolin K content reaches 0.3%, 0.9%, 1.25% and 1.6% respectively, and living contaminants does not appear in whole mycelium culture cycle; The functional Monascus that is obtained does not detect Citrinin, and Citrinin is detected extremely trace (less than 0.2ppm) of content in the utmost point individual samples.
The concrete steps of the inventive method are as follows:
1, screening does not produce Citrinin or the extremely micro-Monascus anka Nakazawa et sato bacterial strain of Citrinin output, comprise red Monascus anka Nakazawa et sato Monascus rubber vanTieghem, purple Monascus anka Nakazawa et sato Monascus Purpureus Went, the Monascus anka Nakazawa et sato Monascus albidus Sato that turns white, pale Monascus anka Nakazawa et sato Monascus pallens Cannon, Abdullah and Abbas, Florida Monascus anka Nakazawa et sato Monascus floridanus Cannon andBarnard, feathering Monascus anka Nakazawa et sato Monascus pilosus Sato ex Hawksworth ﹠amp; Pitt, Monascus rubber van Tieghem Monascus fuliginosusSato;
2, starchiness culture base-material main raw material is rice (comprising brown rice), wheat (comprising wheat, barley), the culture base-material prescription is: rice 10-96% or wheat 10-96%, also can add wheat bran 4-30%, soybean cake powder 0.1-2%, inorganic salt 0.01-0.5%, acetic acid or acetate 0.01-0.8%, culture base-material can add one or more; Culture base-material control moisture content is 25-95%, is generally 35-45%;
3, can be soaked in water or directly to add poach earlier ripe or cook for rice, wheat, adds wheat bran, soybean cake powder, inorganic salt, acetic acid or acetate (water-soluble back adds), stirs; Adopt high temperature resistant strain bag or vial filling culture base-material, behind the filling culture base-material, insert 1 to 10 of little ventpipe, auxiliary and strengthen culture base-material ventilation during the mycelial growth; Cover plug special double-deck ventilation tampon in culture base-material filling back places 121 ℃ of sterilization 30min or 128 ℃ of sterilization 20min then, is cooled to below 45 ℃;
4, little ventpipe adopts recyclable nonexpondable bamboo trunk or stainless steel tube or high-temperature resistance plastice pipe or vitrified pipe; Little ventpipe diameter is 4-12mm, is generally 6-8mm; Length is 100-350mm, is generally 150-250mm; Tube wall is arranged and is drilled with countless apertures in order or disorderly, and the size in hole is 0.5-2mm, is generally 1-1.2mm;
5, special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with 40% above white lime water or saturated white lime water logging bubble, drying treatment;
6, inoculation solid spawn or liquid spawn can be adopted and be disseminated the method inoculation, impel inoculation back bacterial classification multiple spot to sprout, and make culture base-material be covered with the Monascus anka Nakazawa et sato mycelium in the utmost point short period of time, form growth vigor.Inoculum size is 1-15%, is generally 3-6%; 16-34 ℃ of mycelium culture temperature is generally 19-28 ℃;
7, remove the upper strata tampon of special ventilation tampon in the vigorous growth logarithmic phase of Monascus anka Nakazawa et sato mycelial growth, and tighten three layers of gauze that stay and cover in bottleneck; By the temperature difference between regulation and control culturing room or plastics hothouse air output exhaust air rate and open type ventilation system widen round the clock between round the clock, the mycelium culture temperature is 20-34 ℃ between regulation and control daytimes, is generally 24-27 ℃; Relative humidity 65-95% is generally 75-90%; Night, the mycelium culture temperature was 14-26 ℃, was generally 16-23 ℃; Relative humidity 40-70% is generally 45-60%; The adaptation Monascus anka Nakazawa et sato growth of culture environment temperature difference alternating temperature and damping between realizing round the clock and the good culture environment that functional meta-bolites synthesizes and accumulates.
The inorganic salt that the present invention relates to are one or more of sal epsom, lime carbonate, potassium primary phosphate or dipotassium hydrogen phosphate.
Studies show that the screening of bacterial classification is the primary factor that improves functional Monascus look valency, meta-bolites Monacolin K content and reduce Citrinin output.The accumulation of Monascus anka Nakazawa et sato meta-bolites Monacolin K generally reaches the climax in the mycelium late stage of culture, because Monascus anka Nakazawa et sato late stage of culture starchiness culture base-material is dissolved existing culture base-material ventilation environmental degradation by the vigorous amylase liquid of Monascus anka Nakazawa et sato secretion in the traditional technology, assorted bacterium follows and gives birth to, and the formation of Citrinin is unavoidable.Experiment is found, Monascus anka Nakazawa et sato soaks in the be liquefied high density of saccharification, the glucose solution of high osmotic pressure of growth logarithmic phase mycelial cell wall, transformation reactions appears in mycelial growth metabolism, and meta-bolites Monacolin K content reduces, and secondary metabolite Citrinin output significantly increases.By regulating culturing room or plastics hothouse air output exhaust air rate or open type ventilation system, at the air flow of mycelial growth logarithmic phase increasing to the mycelium production environment, reach alternating temperature, damping, the variation of regulation and control culture base-material water content, make culture base-material keep loose, quicken the synthetic and accumulation of mycelial growth speed and functional meta-bolites Monacolin K.Living contaminants does not appear in whole mycelium culture cycle, and the functional Monascus that is obtained does not detect Citrinin.
Embodiment
Below in conjunction with embodiment the present invention is described in further detail.
Embodiment one:
1, screening does not produce the red Monascus anka Nakazawa et sato Monascus rubber van Tieghem bacterial strain of Citrinin.
2, starchiness culture base-material composition of raw materials: rice 90%, wheat bran 9.8%, soybean cake powder 0.1%, sal epsom 0.01%, lime carbonate 0.07%, acetic acid 0.02%; Culture base-material control moisture content is 32-35%.
3, the rice 2h that is soaked in water earlier cooks then, adds wheat bran, soybean cake powder, inorganic salt and acetic acid (water-soluble back adds), regulates moisture content, stirs.Adopt high temperature resistant strain bag filling culture base-material, behind the filling culture base-material, insert 10 of little ventpipes; Cover plug special double-deck ventilation tampon in culture base-material filling back places 121 ℃ of sterilization 30min then, is cooled to below 45 ℃.
4, little ventpipe adopts bamboo trunk; Little ventpipe diameter is 10mm, and length is 100mm, and tube wall is arranged and is drilled with countless apertures in order, and the size in hole is 0.5mm.
5, special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with 40% white lime water logging bubble, drying treatment.
6, inoculation solid spawn is adopted and disseminates the method inoculation, and the bacterial classification grain is scattered in the culture base-material with loop-carrier, and inoculum size is 10%.24 ℃ of mycelium culture temperature.
7, the 3rd day vigorous growth logarithmic phase of Monascus anka Nakazawa et sato mycelial growth, remove the upper strata tampon of special ventilation tampon, and tighten three layers of gauze that stay and cover in bottleneck.By regulation and control plastics hothouse air output exhaust air rate ventilation system, widen round the clock between the temperature difference, the mycelium culture temperature is 26 ℃ between daytime, relative humidity 85%; Night, the mycelium culture temperature was 20 ℃, relative humidity 45%.Regulation and control culture base-material moisture content is 30-32%.
8, Monascus anka Nakazawa et sato mycelium culture cycle is when 7d, and functional ingredient Monacolin K content is 0.3% in the Monascus anka Nakazawa et sato, and after testing, the functional Monascus bacterium that is obtained does not detect Citrinin, living contaminants do not occur.
Embodiment two:
1, the extremely micro-purple Monascus anka Nakazawa et sato Monascus Purpureus Went bacterial strain of screening Citrinin output.
2, starchiness culture base-material composition of raw materials: rice 80%, wheat bran 19%, soybean cake powder 1%, lime carbonate 0.03%, potassium primary phosphate 0.1%, acetic acid 0.08%; Culture base-material control moisture content is 40-45%.
3, the rice 3h that is soaked in water earlier cooks then, adds wheat bran, soybean cake powder, inorganic salt and acetic acid (water-soluble back adds), regulates moisture content, stirs.Adopt high temperature resistant strain bag filling culture base-material, behind the filling culture base-material, insert 3 of little ventpipes; Cover plug special double-deck ventilation tampon in culture base-material filling back places 128 ℃ of sterilization 20min then, is cooled to below 45 ℃.
4, little ventpipe adopts stainless steel tube; Little ventpipe diameter is 5mm, and length is 150mm, and tube wall is arranged and is drilled with countless apertures in order, and the size in hole is 1mm.
5, special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with saturated white lime water logging bubble, drying treatment.
6, inoculation solid spawn is adopted and disseminates the method inoculation, and the bacterial classification grain is scattered in the culture base-material with loop-carrier, and inoculum size is 3%, and the mycelium culture temperature is 28 ℃.
7, the 4th day vigorous growth logarithmic phase of Monascus anka Nakazawa et sato mycelial growth, remove the upper strata tampon of special ventilation tampon, and tighten three layers of gauze that stay and cover in bottleneck.By regulation and control culturing room open type ventilation system, widen round the clock between the temperature difference, the mycelium culture temperature is 28 ℃ between daytime, relative humidity 90%; Night, the mycelium culture temperature was 18 ℃, relative humidity 50%.Regulation and control culture base-material moisture content is 35-40%.
8, Monascus anka Nakazawa et sato mycelium culture cycle is when 14d, and functional ingredient Monacolin K content is 0.9% in the Monascus anka Nakazawa et sato, and after testing, the functional Monascus bacterium that is obtained does not detect Citrinin, living contaminants do not occur.
Embodiment three:
1, the extremely micro-Monascus anka Nakazawa et sato Monascus albidus Sato bacterial strain that turns white of screening Citrinin output.
2, starchiness culture base-material composition of raw materials: rice 50%, wheat 50%, potassium primary phosphate 0.05%, dipotassium hydrogen phosphate 0.057%, sodium-acetate 0.1%, culture base-material control moisture content is 40-43%.
3, rice, wheat directly boil, and add inorganic salt and acetate (water-soluble back adds), regulate moisture content, stir.Adopt vial filling culture base-material, behind the filling culture base-material, insert 8 of little ventpipes; Cover plug special double-deck ventilation tampon in culture base-material filling back places 121 ℃ of sterilization 30min then, is cooled to below 45 ℃.
4, little ventpipe adopts vitrified pipe; Little ventpipe diameter is 12mm, and length is 200mm, and tube wall is arranged the unordered countless apertures that are drilled with, and the size in hole is 1.2mm.
5, special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with 50% white lime water logging bubble, drying treatment.
6, inoculation liquid spawn, inoculum size is 3%, the mycelium culture temperature is 20 ℃.
7, the 3rd day vigorous growth logarithmic phase of Monascus anka Nakazawa et sato mycelial growth, remove the upper strata tampon of special ventilation tampon, and tighten three layers of gauze that stay and cover in bottleneck.By regulation and control plastics hothouse open type ventilation system, widen round the clock between the temperature difference, the mycelium culture temperature is 30 ℃ between daytime, relative humidity 95%; Night, the mycelium culture temperature was 18 ℃, relative humidity 53%.Regulation and control culture base-material moisture content is 36-42%.
8, Monascus anka Nakazawa et sato mycelium culture cycle is when 28d, and functional ingredient Monacolin K content is 1.6% in the Monascus anka Nakazawa et sato, and after testing, the functional Monascus Citrinin that is obtained is detected content less than 0.1ppm, living contaminants do not occur.
Embodiment four:
1, screening does not produce the Florida Monascus anka Nakazawa et sato Monascus flridanus Cannon and Barnard bacterial strain of Citrinin.
2, starchiness culture base-material composition of raw materials: barley 90%, wheat bran 9.8%, soybean cake powder 0.2%, sal epsom 0.05%, lime carbonate 0.02%, acetic acid 0.2%, culture base-material control moisture content is 38-40%.
3, barley directly cooks earlier, adds wheat bran, soybean cake powder, inorganic salt and acetic acid (water-soluble back adds), regulates moisture content, stirs.Adopt high temperature resistant strain bag filling culture base-material, behind the filling culture base-material, insert 6 of little ventpipes; Cover plug special double-deck ventilation tampon in culture base-material filling back places 128 ℃ of sterilization 20min then, is cooled to below 45 ℃.
4, little ventpipe adopts bamboo trunk; Little ventpipe diameter is 8mm, and length is 200mm, and tube wall is arranged and is drilled with countless apertures in order, and the size in hole is 1.5mm.
5, special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with 40% white lime water logging bubble, drying treatment.
6, inoculation solid spawn is adopted and disseminates the method inoculation, and the bacterial classification grain is scattered in the culture base-material with loop-carrier, and inoculum size is 5%, and the mycelium culture temperature is 23 ℃.
7, the 3.5th day vigorous growth logarithmic phase of Monascus anka Nakazawa et sato mycelial growth, remove the upper strata tampon of special ventilation tampon, and tighten three layers of gauze that stay and cover in bottleneck.By regulation and control culturing room's air output and exhaust air rate ventilation system, widen round the clock between the temperature difference, the mycelium culture temperature is 29 ℃ between daytime, relative humidity 95%; Night, the mycelium culture temperature was 18 ℃, relative humidity 50%.Regulation and control culture base-material moisture content is 35-38%.
8, Monascus anka Nakazawa et sato mycelium culture cycle is when 21d, and functional ingredient Monacolin K content is 1.25% in the Monascus anka Nakazawa et sato, and after testing, the functional Monascus bacterium that is obtained does not detect Citrinin, living contaminants do not occur.
Embodiment five:
1, screening does not produce the red Monascus anka Nakazawa et sato Monascus rubber van Tieghem bacterial strain of Citrinin.
2, starchiness culture base-material composition of raw materials: rice 98%, wheat bran 2%, soybean cake powder 0.1%, sal epsom 0.1%, acetic acid 0.08%; Culture base-material control moisture content is 42-43%.
3, rice directly boils, and adds wheat bran, soybean cake powder, inorganic salt and acetic acid (water-soluble back adds), regulates moisture content, stirs.Adopt vial filling culture base-material, behind the filling culture base-material, insert 5 of little ventpipes; Cover plug special double-deck ventilation tampon in culture base-material filling back places 121 ℃ of sterilization 30min then, is cooled to below 45 ℃.
4, little ventpipe adopts vitrified pipe; Little ventpipe diameter is 12mm, and length is 230mm, and tube wall is arranged the unordered countless apertures that are drilled with, and the size in hole is 0.8mm.
5, special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with 40% white lime water logging bubble, drying treatment.
6, inoculation solid spawn is adopted and disseminates the method inoculation, and the bacterial classification grain is scattered in the culture base-material with loop-carrier, and inoculum size is 1%, and the mycelium culture temperature is 22 ℃.
7, the 5th day vigorous growth logarithmic phase of Monascus anka Nakazawa et sato mycelial growth, remove the upper strata tampon of special ventilation tampon, and tighten three layers of gauze that stay and cover in bottleneck.By regulation and control plastics hothouse open type ventilation system, widen round the clock between the temperature difference, the mycelium culture temperature is 30 ℃ between daytime, relative humidity 88%; Night, the mycelium culture temperature was 20 ℃, relative humidity 52%.Regulation and control culture base-material moisture content is 28-31%.
8, Monascus anka Nakazawa et sato mycelium culture cycle is when 28d, and functional ingredient Monacolin K content is 1.61% in the Monascus anka Nakazawa et sato, and after testing, the functional Monascus bacterium that is obtained does not detect Citrinin, living contaminants do not occur.
Embodiment six:
1, screening does not produce the purple Monascus anka Nakazawa et sato Monascus Purpureus Went bacterial strain of Citrinin.
2, starchiness culture base-material composition of raw materials: brown rice 45%, barley 47%, wheat bran 8%, acetic acid 0.02%; Culture base-material control moisture content is 38-48%.
3, brown rice, the barley 2h that is soaked in water earlier cooks then, adds wheat bran, acetic acid (water-soluble back adds), regulates moisture content, stirs.Adopt high temperature resistant strain bag filling culture base-material, behind the filling culture base-material, insert 8 of little ventpipes; Cover plug special double-deck ventilation tampon in culture base-material filling back places 121 ℃ of sterilization 30min then, is cooled to below 45 ℃.
4, little ventpipe adopts bamboo trunk; Little ventpipe diameter is 10mm, and length is 100mm, and tube wall is arranged and is drilled with countless apertures in order, and the size in hole is 0.5mm.
5, special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with 60% white lime water logging bubble, drying treatment.
6, inoculation solid spawn is adopted and disseminates the method inoculation, and the bacterial classification grain is scattered in the culture base-material with loop-carrier, and inoculum size is 12%, and the mycelium culture temperature is 27 ℃.
7, the 2.5th day vigorous growth logarithmic phase of Monascus anka Nakazawa et sato mycelial growth, remove the upper strata tampon of special ventilation tampon, and tighten three layers of gauze that stay and cover in bottleneck.By regulation and control plastics hothouse open type ventilation system, widen round the clock between the temperature difference, the mycelium culture temperature is 26 ℃ between daytime, relative humidity 93%; Night, the mycelium culture temperature was 20 ℃, relative humidity 52%.Regulation and control culture base-material moisture content is 39-42%.
8, Monascus anka Nakazawa et sato mycelium culture cycle is when 7d, and functional ingredient Monacolin K content is 0.41% in the Monascus anka Nakazawa et sato, and after testing, the functional Monascus bacterium that is obtained does not detect Citrinin, living contaminants do not occur.
Embodiment seven:
1, screening does not produce the red Monascus anka Nakazawa et sato Monascus rubber van Tieghem bacterial strain of Citrinin.
2, starchiness culture base-material composition of raw materials: rice 61%, wheat 39%, soybean cake powder 0.1%, lime carbonate 0.1%, culture base-material control moisture content is 55-60%.
3, rice, wheat directly boil, and add soybean cake powder, inorganic salt and acetate (water-soluble back adds), regulate water-content, stir.Adopt vial filling culture base-material, behind the filling culture base-material, insert 2 of little ventpipes; Cover plug special double-deck ventilation tampon in culture base-material filling back places 121 ℃ of sterilization 30min then, is cooled to below 45 ℃.
4, little ventpipe adopts the high-temperature resistance plastice pipe; Little ventpipe diameter is 6mm, and length is 180mm, and tube wall is arranged and is drilled with countless apertures in order, and the size in hole is 0.6mm.
5, special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with 40% white lime water logging bubble, drying treatment.
6, inoculation solid spawn is adopted and disseminates the method inoculation, and the bacterial classification grain is scattered in the culture base-material with loop-carrier, and inoculum size is 8%, and the mycelium culture temperature is 25 ℃.
7, the 3rd day vigorous growth logarithmic phase of Monascus anka Nakazawa et sato mycelial growth, remove the upper strata tampon of special ventilation tampon, and tighten three layers of gauze that stay and cover in bottleneck.By regulation and control plastics hothouse open type ventilation system, widen round the clock between the temperature difference, the mycelium culture temperature is 31 ℃ between daytime, relative temperature 92%; Night, the mycelium culture temperature was 25 ℃, relative humidity 60%.Regulation and control culture base-material moisture content is 60-70%.
8, Monascus anka Nakazawa et sato mycelium culture cycle is when 14d, and functional ingredient Monacolin K content is 0.9% in the Monascus anka Nakazawa et sato, and after testing, the functional Monascus bacterium that is obtained does not detect Citrinin, living contaminants do not occur.
Embodiment eight:
1, screening does not produce the purple Monascus anka Nakazawa et sato Monascus Purpureus Went bacterial strain of Citrinin.
2, starchiness culture base-material composition of raw materials: rice 96%, wheat bran 4%, lime carbonate 0.07%, potassium primary phosphate 0.15%, acetic acid 0.02%; Culture base-material control moisture content is 27-28%.
3, the rice 1h that is soaked in water earlier cooks then, adds wheat bran, inorganic salt and acetic acid (water-soluble back adds), regulates moisture content, stirs.Adopt high temperature resistant strain bag filling culture base-material, behind the filling culture base-material, insert 7 of little ventpipes; Cover plug special double-deck ventilation tampon in culture base-material filling back places 121 ℃ of sterilization 30min then, is cooled to below 45 ℃.
4, little ventpipe adopts vitrified pipe; Little ventpipe diameter is 12mm, and length is 250mm, and tube wall is arranged and is drilled with countless apertures in order, and the size in hole is 1.6mm.
5, special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with 40% white lime water logging bubble, drying treatment.
6, inoculation solid spawn is adopted and disseminates the method inoculation, and the bacterial classification grain is scattered in the culture base-material with loop-carrier, and inoculum size is 2%, and the mycelium culture temperature is 28 ℃.
7, the 4th day vigorous growth logarithmic phase of Monascus anka Nakazawa et sato mycelial growth, remove the upper strata tampon of special ventilation tampon, and tighten three layers of gauze that stay and cover in bottleneck.By regulation and control plastics hothouse open type ventilation system, draw back round the clock between the temperature difference, the mycelium culture temperature is 25 ℃ between daytime, relative humidity 95%; Night, the mycelium culture temperature was 18 ℃, relative humidity 48%.Regulation and control culture base-material moisture content is 37-40%.
8, Monascus anka Nakazawa et sato mycelium culture cycle is when 21d, and functional ingredient Monacolin K content is 1.25% in the Monascus anka Nakazawa et sato, and after testing, the functional Monascus bacterium that is obtained does not detect Citrinin, living contaminants do not occur.
Embodiment nine:
1, screening does not produce the red Monascus anka Nakazawa et sato Monascus rubber van Tieghem bacterial strain of Citrinin.
2, starchiness culture base-material composition of raw materials: wheat 96%, wheat bran 2%, soybean cake powder 2%, sal epsom 0.35%, sodium-acetate 0.01%, culture base-material control moisture content is 44-45%.
3, wheat directly boils, and adds wheat bran, soybean cake powder, inorganic salt and acetate (water-soluble back adds), regulates moisture content, stirs.Adopt vial filling culture base-material, behind the filling culture base-material, insert 4 of little ventpipes; Cover plug special double-deck ventilation tampon in culture base-material filling back places 121 ℃ of sterilization 30min then, is cooled to below 45 ℃.
4, little ventpipe adopts stainless steel tube; Little ventpipe diameter is 8mm, and length is 180mm, and tube wall is arranged the unordered countless apertures that are drilled with, and the size in hole is 0.8mm.
5, special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with saturated white lime water logging bubble, drying treatment.
6, inoculation liquid spawn, inoculum size is 3%, the mycelium culture temperature is 26 ℃.
7, the 3rd day vigorous growth logarithmic phase of Monascus anka Nakazawa et sato mycelial growth, remove the upper strata tampon of special ventilation tampon, and tighten three layers of gauze that stay and cover in bottleneck.By regulation and control culturing room open type ventilation system, widen round the clock between the temperature difference, cultivation filament culture temperature is 28 ℃ between daytime, relative temperature 95%; Night, the mycelium culture temperature was 19 ℃, relative temperature 60%.Regulation and control culture base-material moisture content is 40-50%.
8, Monascus anka Nakazawa et sato mycelium culture cycle is when 28d, and functional ingredient Monacolin K content is 1.62% in the Monascus anka Nakazawa et sato, and after testing, the functional Monascus bacterium that is obtained does not detect Citrinin, living contaminants do not occur.
Embodiment ten:
1, screening does not produce the red Monascus anka Nakazawa et sato Monascus rubber van Tieghem bacterial strain of Citrinin.
2, starchiness culture base-material composition of raw materials: rice 90%, wheat bran 8.8%, soybean cake powder 1.2%, potassium primary phosphate 0.07%, dipotassium hydrogen phosphate 0.06%, acetic acid 0.01%, culture base-material control moisture content is 80-88%.
3, the rice 1h that is soaked in water earlier cooks then, adds wheat bran, soybean cake powder, inorganic salt and acetic acid (water-soluble back adds), stirs.Adopt high temperature resistant strain bag filling culture base-material, behind the filling culture base-material, insert 10 of little ventpipes; Cover plug special double-deck ventilation tampon in culture base-material filling back places 121 ℃ of sterilization 30min then, is cooled to below 45 ℃.
4, little ventpipe adopts bamboo trunk; Little ventpipe diameter is 8mm, and length is 200mm, and tube wall is arranged and is drilled with countless apertures in order, and the size in hole is 1mm.
5, special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with saturated white lime water logging bubble, drying treatment.
6, inoculation solid spawn is adopted and disseminates the method inoculation, and the bacterial classification grain is scattered in the culture base-material with loop-carrier, and inoculum size is 5%, and the mycelium culture temperature is 22 ℃.
7, the 4th day vigorous growth logarithmic phase of Monascus anka Nakazawa et sato mycelial growth, remove the upper strata tampon of special ventilation tampon, and tighten three layers of gauze that stay and cover in bottleneck.By regulation and control plastics hothouse open type ventilation system, widen round the clock between the temperature difference, bacterium mycelium culture temperature is 27 ℃ between daytime, relative humidity 95%; Night, the mycelium culture temperature was 20 ℃, relative humidity 55%.Regulation and control culture base-material moisture content is 68-69%
8, Monascus anka Nakazawa et sato mycelium culture cycle is when 14d, and functional ingredient Monacolin K content is 0.9% in the Monascus anka Nakazawa et sato, and after testing, the functional Monascus bacterium that is obtained does not detect Citrinin, living contaminants do not occur.
Claims (7)
1. functional monoscaceae quick solid culture method, it is characterized in that setting up a kind of alternating temperature, the Monascus anka Nakazawa et sato mycelium quick solid cultural method of damping and regulation and control culture base-material water content, in spawn culture bag or vial filling culture base-material, insert little ventpipe, by regulating culturing room or plastics hothouse air output exhaust air rate or open type ventilation system, at the air flow of mycelial growth logarithmic phase increasing to strain bag or bottle, reach alternating temperature, damping, the variation of regulation and control culture base-material water content, make culture base-material keep loose, quicken the synthetic and accumulation of mycelial growth speed and functional meta-bolites Monacolin K.
2. the method for claim 1 is characterized in that concrete steps are:
(1) screening does not produce Citrinin or the extremely micro-Monascus anka Nakazawa et sato bacterial strain of Citrinin output, comprise red Monascus anka Nakazawa et sato Monascus rubbervan Tieghem, purple Monascus anka Nakazawa et sato Monascus Purpureus Went, the Monascus anka Nakazawa et sato Monascus albidus Sato that turns white, pale Monascus anka Nakazawa et sato Monascus pallens Cannon, Abdullah and Abbas, Florida Monascus anka Nakazawa et sato Monascus flridanus Cannon andBarnard, feathering Monascus anka Nakazawa et sato Monascus pilosus Sato ex Hawksworth ﹠amp; Pitt, Monascus rubber van Tieghem Monascus fuliginosusSato;
(2) starchiness culture base-material main raw material is rice, wheat, and the culture base-material prescription is: rice 10-96% or wheat 10-96%, also can add wheat bran 4-30%, soybean cake powder 0.1-2%, inorganic salt 0.01-0.5%, acetic acid or acetate 0.01-0.8% can add one or more; Culture base-material control moisture content is 25-95%;
(3) can be soaked in water or directly to add poach earlier ripe or cook for rice, wheat, adds wheat bran, soybean cake powder, inorganic salt, acetic acid or acetate, stirs.Adopt high temperature resistant strain bag or vial filling culture base-material, behind the filling culture base-material, insert 1 to 10 of little ventpipe, cover plug special double-deck ventilation tampon in culture base-material filling back places 121 ℃ of sterilization 30min or 128 ℃ of sterilization 20min then, is cooled to below 45 ℃;
(4) little ventpipe adopts recyclable nonexpondable bamboo trunk or stainless steel tube or high-temperature resistance plastice pipe or vitrified pipe; Little ventpipe diameter is 4-12mm, and length is 100-350mm, and tube wall is arranged and is drilled with countless apertures in order or disorderly, and the size in hole is 0.5-2mm;
(5) special ventilation tampon is made up of three layers of gauze cover and tampon, and gauze is standby with 40% above white lime water or saturated white lime water logging bubble, drying treatment;
(6) inoculation solid spawn or liquid spawn can be adopted and be disseminated the method inoculation, and inoculum size is 1-15%, 19-34 ℃ of mycelium culture temperature;
(7) by the temperature difference between regulation and control culturing room or plastics hothouse air output exhaust air rate and open type ventilation system widen round the clock between round the clock, the mycelium culture temperature is 20-34 ℃ between regulation and control daytimes; Relative humidity 65-95%, night, the mycelium culture temperature was 14-26 ℃; Relative humidity 40-70%.
3. method as claimed in claim 2 is characterized in that said inorganic salt are one or more of sal epsom, lime carbonate, potassium primary phosphate or dipotassium hydrogen phosphate.
4. method as claimed in claim 2 is characterized in that culture base-material control moisture content is 35-45% in the concrete steps (2).
5. method as claimed in claim 2 is characterized in that the medium and small ventpipe diameter of concrete steps (4) is 6-8mm, and length is 150-250mm, and the size that tube wall is arranged aperture is 1-1.2mm.
6. method as claimed in claim 2 is characterized in that the inoculum size of concrete steps (6) middle inoculation solid spawn or liquid spawn is 3-6%, and the mycelium culture temperature is 19-28 ℃.
7. method as claimed in claim 2 is characterized in that in the concrete steps (7) that the mycelium culture temperature is 24-27 ℃ between regulation and control daytimes, relative humidity 75-90%, and night, the mycelium culture temperature was 16-23 ℃, relative humidity 45-60%.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101659923B (en) * | 2008-08-26 | 2011-10-26 | 北京北大维信生物科技有限公司 | Method for preparing open-loop lovastatin |
| CN103952317A (en) * | 2013-04-11 | 2014-07-30 | 武汉佳成生物制品有限公司 | Esterification monascus strain and production technology thereof |
| CN109055242A (en) * | 2018-09-10 | 2018-12-21 | 李芳� | A kind of monascus purpureus bacterial strain and its zymotechnique and application |
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| CN100386425C (en) * | 2006-01-20 | 2008-05-07 | 中山大学 | Culture method of functional Monascus color mycelium not containing citrinin |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101659923B (en) * | 2008-08-26 | 2011-10-26 | 北京北大维信生物科技有限公司 | Method for preparing open-loop lovastatin |
| CN103952317A (en) * | 2013-04-11 | 2014-07-30 | 武汉佳成生物制品有限公司 | Esterification monascus strain and production technology thereof |
| CN103952317B (en) * | 2013-04-11 | 2019-08-20 | 武汉佳成生物制品有限公司 | Esterified red yeast strain and its production technology |
| CN109055242A (en) * | 2018-09-10 | 2018-12-21 | 李芳� | A kind of monascus purpureus bacterial strain and its zymotechnique and application |
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