CN104195178A - High tone monascus red pigment preparation method - Google Patents
High tone monascus red pigment preparation method Download PDFInfo
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- CN104195178A CN104195178A CN201410323842.0A CN201410323842A CN104195178A CN 104195178 A CN104195178 A CN 104195178A CN 201410323842 A CN201410323842 A CN 201410323842A CN 104195178 A CN104195178 A CN 104195178A
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Abstract
The invention provides a high tone monascus red pigment preparation method which comprises the following steps: 1, culturing monascus in a seed culture medium to obtain a seed liquid; 2, inoculating the seed liquid into a fermentation culture liquid for fermentation culture, regulating the pH value of the fermentation culture liquid, centrifuging to obtain monascus wet cells, and extracting intracellular monascus pigment; 3, putting the extracted intracellular monascus pigment into a surfactant micellar solution, and adding sodium glutamate to realize micellar catalysis for converting monascus orange pigment into monascus red pigment; or inoculating the seed liquid obtained by the step 1 into a fermentation culture medium containing a nonionic surfactant, adding the sodium glutamate, regulating the pH value of the fermentation culture medium for culturing to obtain the monascus red pigment. According to the method, the surfactant and the sodium glutamate are added into the fermentation culture medium for preparation of the high tone and high color value monascus red pigment with low content of the monascus orange pigment.
Description
Technical field
The present invention relates to a kind of fermentation process of technical field of bioengineering, particularly, relate to a kind of application surface promoting agent secretion intracellular product and further born of the same parents, realize the method that the monascorubin of high tone (component is relatively single) is prepared in nicellar catalysis outward.
Background technology
The Red kojic rice that monascus ruber (Monascus sp) solid state fermentation is produced is the traditional food color additive of China, is also a kind of Chinese medicine, in China, has had the use of more than 1,000 year historical.The modern microbic liquid deep layer fermentation technology of current employing is produced monascorubin, and this pigment is widely used in East Asian countries as food color additive, as China, Japan, Korea S, Taiwan, Thailand, Vietnam etc.Usually, the monascorubin that monascus ruber fermentation produces comprises the amino acid derivative of two kinds of monascus yellow pigment components, two kinds of red colouring agent for food, also used as a Chinese medicine citraurin components, two kinds of monascorubin components and monascorubin.Commodity monascorubin is generally the mixture of above-mentioned various colour components, and two parameters of look valency and tone are commonly used to characterize concentration and the relative content of each component in monascorubin.Wherein look valency represents the pigment concentration of monascorubin, as the absorbancy at 510nm place represents the look valency of monascorubin.Tone represents the relative content of each component in monascorubin, as the tone of monascorubin with respect to red colouring agent for food, also used as a Chinese medicine citraurin can represent with monascorubin and the look valency ratio of red colouring agent for food, also used as a Chinese medicine citraurin, this illness that has not attacked the vital organs of the human body the relative content of monascorubin and red colouring agent for food, also used as a Chinese medicine citraurin.
The experiment of chicken embryo shows that red colouring agent for food, also used as a Chinese medicine citraurin has certain teratogenesis [Martinkova L, Patakova-Juzlova P, Kren V, Kucerova Z, Havlicek V, Olsovsky P, Hovorka O, Rihova B, Vesely D, Vesela D, Ulrichova J, Prikrylova V.Biological activities of oligoketide pigments of Monascus purpureus.Food Add Contamin.1999,16 (1): the biological activity of 15-24. monascus ruber polyketone pigment.< < food additive agent pollution > >, 1999,16 (1): 15-24].Therefore, still monascorubin is not as the important technology index of pigment additive for the tone of raising monascorubin, and the while is also the important quality index that improves monascorubin safety about food additives.During the fermentation, utilize modern biotechnology means transformation microorganism strains to there is theoretically feasibility to control minute rate of red colouring agent for food, also used as a Chinese medicine citraurin, but the leavened prod that genetic engineering modified Institute of Micro-biology obtains belong to Genetic engineering food.Due to the singularity of food safety, it is very loaded down with trivial details and difficult that Genetic engineering food obtains in a lot of countries the license of producing and selling.Adopt the secondary metabolism of fermentation engineering regulation and control monascus ruber to become important means.
In order to meet the demand of market to natural monascorubin, ensure food safety, in the urgent need to developing the monascorubin fermentation technique that in high tone, monascorubin, colour component is relatively single simultaneously.Find by prior art documents, Zhiqiang Hu, Xuehong Zhang, Zhenqiang Wu, Hanshi Qi, Zhilong Wang.Export of intracellular Monascus pigments by two-stage microbial fermentation in nonionic surfactant micelle aqueous solution.Journal of biotechnology, 2012, 162:202-209. (Hu Zhiqiang, Zhang Xuehong, Wu Zhenqiang, Qi Hanshi, Wang Zhilong. " in nonionogenic tenside micellar solution, apply two sections of fermentation techniques and discharge monascorubin in born of the same parents ". < < biotechnology journal > >, 2012, 162:202-209), introduce, monascus ruber in nonionogenic tenside micellar solution fermentation energy by born of the same parents in monascorubin be secreted into extracellular environment.But different fermentation conditions and mode form the impact of minute rate on monascorubin, especially, to eliminating the effect of red colouring agent for food, also used as a Chinese medicine citraurin, also not studies have reported that.In further retrieving, about apply nicellar catalysis red colouring agent for food, also used as a Chinese medicine citraurin in Free Energy of Surfactant Micelle Solution, be converted into the relevant report of monascorubin.
The patent of invention of application before applicant: 201310123517.5, name is called extractive fermentation and regulates and controls the method that pH prepares monascorubin, this patented technology combines with regulation and control pH technology by extractive fermentation technology, prepares the monascorubin that citrinin content is low and monascus pigment component is relatively single.
Existing fermentation technique, no matter be solid fermentation or liquid state fermentation, gained monascorubin is all the mixture of monascorubin, red colouring agent for food, also used as a Chinese medicine citraurin, monascus yellow pigment substantially, due to the potential potential safety hazard of red colouring agent for food, also used as a Chinese medicine citraurin.The present invention is chemically converted to monascorubin by red colouring agent for food, also used as a Chinese medicine citraurin, thereby eliminates the potential safety hazard of red colouring agent for food, also used as a Chinese medicine citraurin, prepares the monascorubin product of high tone.
In prior art, be generally in organic solvent, to realize red colouring agent for food, also used as a Chinese medicine citraurin to react with Sodium Glutamate, can not realize during the fermentation, otherwise microorganism is difficult to survival.The above-mentioned first method of the present invention in Free Energy of Surfactant Micelle Solution outside born of the same parents nicellar catalysis red colouring agent for food, also used as a Chinese medicine citraurin be converted into monascorubin, in extractive fermentation and nicellar catalysis, next step realizes red colouring agent for food, also used as a Chinese medicine citraurin and reacts with Sodium Glutamate second method, therefore, get rid of the degraded during the fermentation of red colouring agent for food, also used as a Chinese medicine citraurin, improved production concentration.Before being different from, patent application is extractive fermentation at 201310123517.5: 201310123517.5, does not add the Sodium Glutamate of high density, and what therefore obtain is the outer hybrid pigments of born of the same parents.After the present invention adds Sodium Glutamate, citraurin has been removed, and controls the formation that higher pH has suppressed monascus yellow pigment, and what therefore obtain is mainly monascorubin.
Sodium Glutamate of the present invention is the reaction reagent that red colouring agent for food, also used as a Chinese medicine citraurin is converted into monascorubin, and nonionogenic tenside is reaction medium in this reaction, has Micellar catalysis, otherwise very slow in reactant aqueous solution speed.Nonionic surfactant solution can be realized extractive fermentation in step 22 simultaneously, and red colouring agent for food, also used as a Chinese medicine citraurin in born of the same parents is transferred to extracellular.
The present invention has used the micellar solution of nonionogenic tenside (as Triton X-114, Tween 80, Triton X-100) to realize the secretion of monascorubin in born of the same parents and the nicellar catalysis that the outer red colouring agent for food, also used as a Chinese medicine citraurin of born of the same parents is converted into monascorubin.Can obtain the monascorubin product of the high tone that red colouring agent for food, also used as a Chinese medicine citraurin content is low.
The present invention adopts the object of intracellular product secretion and nicellar catalysis micella to be to realize during the fermentation the release of monascorubin in born of the same parents and the process integration that the outer nicellar catalysis red colouring agent for food, also used as a Chinese medicine citraurin of born of the same parents is chemically converted to monascorubin, obtains the high tone monascorubin product that component is relatively single, red colouring agent for food, also used as a Chinese medicine citraurin content is low.
The present invention adopts the object of tensio-active agent to be parachrome to be secreted into extracellular.
The present invention adopts the object of nicellar catalysis technology to be further outside born of the same parents, in micellar solution, to realize by being secreted into the outer red colouring agent for food, also used as a Chinese medicine citraurin of born of the same parents the nicellar catalysis that red colouring agent for food, also used as a Chinese medicine citraurin is converted into monascorubin.
Compared with prior art, the present invention has following beneficial effect: the present invention adopts intracellular product secretion and nicellar catalysis is integrated has controlled minute rate of red colouring agent for food, also used as a Chinese medicine citraurin in monascorubin.The requirement of the market that the monascorubin that in monascorubin, component is relatively single has met food color additive on the one hand to product specific aim and sense organ, has eliminated red colouring agent for food, also used as a Chinese medicine citraurin component, the security that has improved foodstuff additive on the other hand.Improved the look valency of monascorubin in fermenting process simultaneously.The inventive method is simple, easily realizes, and productive rate is high, and cost is low, and repeatability is strong, and environmental protection, has good application prospect.
Summary of the invention
For defect of the prior art, the object of this invention is to provide a kind of combine method of monascorubin of the high tone of preparation of the outer nicellar catalysis chemical reaction of intracellular product and born of the same parents of secreting, that the method application surface promoting agent micellar solution is secreted into parachrome born of the same parents is during the fermentation outer and further born of the same parents, realize the nicellar catalysis that red colouring agent for food, also used as a Chinese medicine citraurin is converted into monascorubin outward, has prepared the high tone that red colouring agent for food, also used as a Chinese medicine citraurin content is low, the monascorubin product of high look valency.
For achieving the above object, the present invention can adopt in following two kinds of technical schemes any one:
A preparation method for high tone monascorubin, described method comprises the steps:
Step 11, monascus ruber is cultivated in seed culture medium, obtains seed liquor; Described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 12, in order to protect the outer nicellar catalysis of intracellular product secretion and born of the same parents, prepare the monascorubin of high tone, described seed liquor is inoculated in the fermentation culture of carbonaceous sources, carry out fermentation culture, regulate the pH value of fermentation culture, centrifugal, obtain monascus ruber wet cell, extract monascus ruber wet cell and obtain take the monascorubin that red colouring agent for food, also used as a Chinese medicine citraurin is main ingredient; Resulting monascorubin is the high intracellular product of red colouring agent for food, also used as a Chinese medicine citraurin content.
Step 13; in order to protect nicellar catalysis can further control red colouring agent for food, also used as a Chinese medicine citraurin, be converted into monascorubin, the monascorubin of described extraction added to the nicellar catalysis reaction that is converted into monascorubin containing carrying out red colouring agent for food, also used as a Chinese medicine citraurin in the solution of nonionogenic tenside and Sodium Glutamate.
Preferably, in step 13, the scope of concentration of sodium glutamate is at 5-30g/L.The concentration of nonionogenic tenside is less than or equal to 100g/L.
Aforesaid method of the present invention, by adding excessive Sodium Glutamate, in Free Energy of Surfactant Micelle Solution, red colouring agent for food, also used as a Chinese medicine citraurin reacts in Free Energy of Surfactant Micelle Solution with Sodium Glutamate, realizes red colouring agent for food, also used as a Chinese medicine citraurin and is converted into monascorubin.
A preparation method for high tone monascorubin, described method comprises the steps:
Step 21, monascus ruber is cultivated in seed culture medium, obtains seed liquor; Described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 22; in order to protect intracellular product secretion, can further control the formation of high tone monascorubin; control the content of red colouring agent for food, also used as a Chinese medicine citraurin simultaneously; resulting seed liquor in step 21 is inoculated into containing fermentation culture in the substratum of tensio-active agent and Sodium Glutamate to the centrifugal outer monascorubin of born of the same parents that obtains.This step has realized the fermentation of the high tone monascorubin that the component of high look valency is relatively single, the monascorubin of minute high tone that rate is low, component is relatively single of red colouring agent for food, also used as a Chinese medicine citraurin in gained monascorubin.
Preferably, in step 22, surfactant concentration remains on the scope of 50-100g/l.The concentration of Sodium Glutamate remains on the scope of 5-30g/l.
Preferably, in step 22, described fermention medium is comprised of each component of following content: water 100ml, glucose 5g, KH
2pO
40.4g, MgSO
40.1g, CaCl
20.005g, nonionogenic tenside 1～10g/100ml, aminoglutaric acid concentration is 5g/100～30g/1000ml.
Aforesaid method of the present invention, by adding excessive Sodium Glutamate, fermentation in surfactant soln can be secreted into extracellular by monascorubin in born of the same parents, and under Micellar catalysis, red colouring agent for food, also used as a Chinese medicine citraurin reacts with Sodium Glutamate and has realized red colouring agent for food, also used as a Chinese medicine citraurin and be converted into monascorubin in born of the same parents' external surfactants micellar solution.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.Following examples will contribute to those skilled in the art further to understand the present invention, but not limit in any form the present invention.It should be pointed out that to those skilled in the art, without departing from the inventive concept of the premise, can also make some distortion and improvement.These all belong to protection scope of the present invention.Method based on described in summary of the invention, some details that relate in following examples are as follows:
Look valency detection method is specially: cell free fermentation liquid or intracellular product release liquid are suitably diluted with ethanol (pH=2) solution of 70% (V/V), adopt the light absorption value at spectrophotometer measure of spread 410,470,510nm place.Above-mentioned light absorption value is multiplied by the look valency that absorbance unit that extension rate obtains represents respectively yellow pigment, citraurin and haematochrome.Accordingly, the hue table of monascorubin is shown the ratio of the look valency of monascus yellow pigment and red colouring agent for food, also used as a Chinese medicine citraurin.
The detection method of determining the basal component of monascorubin is specially: cell free fermentation liquid or intracellular product release liquid are suitably absorbed with ethanol (pH=2) solution of 70% (V/V), adopt thin-layer chromatography (the silica gel F254 of Merck & Co., Inc. plate), solvent systems is acetic acid: methyl alcohol, chloroform=9:21:285, under natural light, at Rf, be respectively 0.8,0.68 and 0 place and red colouring agent for food, also used as a Chinese medicine citraurin can be detected, monascus yellow pigment and monascorubin.
In step 11 and step 21, described seed culture medium is comprised of each component of following content: water 100ml, peptone 2g, yeast powder 2g, glucose 2g.
In step 11 and step 21, described cultivation is 30 ℃ in temperature, in the shaking table of 200rpm, carries out, and incubation time is 32 hours.
In step 12, described fermentation culture is 30 ℃ in temperature, in the shaking table of 200rpm, carries out, and incubation time is 6-8 days.The initial pH value of described fermentation culture is 4.5.Carbon source in described fermention medium can use the natural agricultural-food such as rice meal, Semen Maydis powder, analysis for soybean powder to substitute.Carbon source is the microbial metabolism necessary matter and energy source of growing, and can make glucose.But the cheap agricultural-food of industrial normal employing, or even agriculture residue, the present invention considers food safety, adopts the good natural agricultural-food of quality.
In step 13, described reaction refers to that in temperature be 30 ℃, in the shaking table of 200rpm, carries out, and the reaction times is 6～12 hours; Reaction pH is 2-7.
In step 22, described cultivation refers to that in temperature be 30 ℃, in the shaking table of 200rpm, carries out, and incubation time is 6～8 days; Described separation refers in the water-bath of 70 ℃ standing 2 hours.The initial pH value of described fermention medium is 4～6.5.
embodiment 1
The present embodiment relates to a kind of nicellar catalysis method that red colouring agent for food, also used as a Chinese medicine citraurin is converted into monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4.5 that described seed liquor is inoculated into initial pH value, in the fermentation culture that the Semen Maydis powder of take is nitrogenous source, in temperature, is 30 ℃, carries out the fermentation culture of 7 days in the shaking table of 200rpm, centrifugal, obtains monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 2,1 and 1.8 absorbance units; Fermented liquid final pH is 6.8.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin; 410,470 and the absorbancy at 510nm place be respectively 25,38 and 20 absorbance units.The tone of monascorubin is 20/38.
Step 3, after concentrated, in the tensio-active agent Triton-100 aqueous solution that the concentrated pigment that takes a morsel adds to, regulates pH=2 by the extract of monascorubin in above-mentioned born of the same parents; Surfactant concentration is 5%, and concentration of sodium glutamate is 30g/L, and the look valency of controlling red colouring agent for food, also used as a Chinese medicine citraurin is 0.8; In temperature, be 30 ℃, in the shaking table of 200rpm, carry out, the reaction times is 8 hours;
The absorption spectrum of analytical reaction liquid, 410,470 and the absorbancy at 510nm place be respectively 0.3,0.7 and 0.4 absorbance unit.The tone of monascorubin is 0.4/0.7; Adopt thin layer chromatography analysis to show that a minute rate for each component of monascorubin does not have considerable change.
embodiment 2
The present embodiment relates to a kind of method that red colouring agent for food, also used as a Chinese medicine citraurin is converted into the nicellar catalysis of monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4.5 that described seed liquor is inoculated into initial pH value, in the fermentation culture that the Semen Maydis powder of take is nitrogenous source, in temperature, is 30 ℃, carries out the fermentation culture of 7 days in the shaking table of 200rpm, centrifugal, obtains monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 2,1 and 1.8 absorbance units; Fermented liquid final pH is 6.8.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin; 410,470 and the absorbancy at 510nm place be respectively 25,38 and 20 absorbance units.The tone of monascorubin is 20/38.
Step 3, after concentrated, in the tensio-active agent Trition-100 aqueous solution that the concentrated pigment that takes a morsel adds to, regulates pH=7 by the extract of monascorubin in above-mentioned born of the same parents; Surfactant concentration is 5%, and concentration of sodium glutamate is 30g/L, and the look valency of controlling red colouring agent for food, also used as a Chinese medicine citraurin is 0.8; In temperature, be 30 ℃, in the shaking table of 200rpm, carry out, the reaction times is 8 hours;
The absorption spectrum of analytical reaction liquid, 410,470 and the absorbancy at 510nm place be respectively 0.6,0.4 and 0.5 absorbance unit.The tone of monascorubin is 0.5/0.4; Adopt thin layer chromatography analysis to show that monascorubin red colouring agent for food, also used as a Chinese medicine citraurin minute rate obviously reduces and a monascorubin minute rate obviously increases.
embodiment 3
The present embodiment relates to a kind of method that red colouring agent for food, also used as a Chinese medicine citraurin is converted into the nicellar catalysis of monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4.5 that described seed liquor is inoculated into initial pH value, in the fermentation culture that the Semen Maydis powder of take is nitrogenous source, in temperature, is 30 ℃, carries out the fermentation culture of 7 days in the shaking table of 200rpm, centrifugal, obtains monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 2,1 and 1.8 absorbance units; Fermented liquid final pH is 6.8.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin; 410,470 and the absorbancy at 510nm place be respectively 25,38 and 20 absorbance units.The tone of monascorubin is 20/38.
Step 3, after concentrated, in the tensio-active agent Trition-100 aqueous solution that the concentrated pigment that takes a morsel adds to, regulates pH=4 by the extract of monascorubin in above-mentioned born of the same parents; Surfactant concentration is 5%, and concentration of sodium glutamate is 30g/L, and the look valency of controlling red colouring agent for food, also used as a Chinese medicine citraurin is 0.8; In temperature, be 30 ℃, in the shaking table of 200rpm, carry out, the reaction times is 8 hours;
The absorption spectrum of analytical reaction liquid, 410,470 and the absorbancy at 510nm place be respectively 0.3,0.5 and 0.4 absorbance unit.The tone of monascorubin is 0.4/0.5; Adopt thin layer chromatography analysis to show that monascorubin red colouring agent for food, also used as a Chinese medicine citraurin minute rate reduces and monascorubin minute rate increase.
embodiment 4
The present embodiment relates to a kind of method that red colouring agent for food, also used as a Chinese medicine citraurin is converted into the nicellar catalysis of monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4.5 that described seed liquor is inoculated into initial pH value, in the fermentation culture that the Semen Maydis powder of take is nitrogenous source, in temperature, is 30 ℃, carries out the fermentation culture of 7 days in the shaking table of 200rpm, centrifugal, obtains monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 2,1 and 1.8 absorbance units; Fermented liquid final pH is 6.8.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin; 410,470 and the absorbancy at 510nm place be respectively 25,38 and 20 absorbance units.The tone of monascorubin is 20/38.
Step 3, after concentrated, in the tensio-active agent Trition-100 aqueous solution that the concentrated pigment that takes a morsel adds to, regulates pH=7 by the extract of monascorubin in above-mentioned born of the same parents; Surfactant concentration is 10%, and concentration of sodium glutamate is 30g/L, and the look valency of controlling red colouring agent for food, also used as a Chinese medicine citraurin is 0.8; In temperature, be 30 ℃, in the shaking table of 200rpm, carry out, the reaction times is 8 hours;
The absorption spectrum of analytical reaction liquid, 410,470 and the absorbancy at 510nm place be respectively 0.65,0.4 and 0.55 absorbance unit.The tone of monascorubin is 0.55/0.4; Adopt thin layer chromatography analysis to show that monascorubin red colouring agent for food, also used as a Chinese medicine citraurin spot almost disappears and a monascorubin minute rate obviously increases.
embodiment 5
The present embodiment relates to a kind of method that red colouring agent for food, also used as a Chinese medicine citraurin is converted into the nicellar catalysis of monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4.5 that described seed liquor is inoculated into initial pH value, in the fermentation culture that the Semen Maydis powder of take is nitrogenous source, in temperature, is 30 ℃, carries out the fermentation culture of 7 days in the shaking table of 200rpm, centrifugal, obtains monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 2,1 and 1.8 absorbance units; Fermented liquid final pH is 6.8.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin; 410,470 and the absorbancy at 510nm place be respectively 25,38 and 20 absorbance units.The tone of monascorubin is 20/38.
Step 3, after concentrated, in the tensio-active agent Trition-100 aqueous solution that the concentrated pigment that takes a morsel adds to, regulates pH=7 by the extract of monascorubin in above-mentioned born of the same parents; Surfactant concentration is 0.5%, and concentration of sodium glutamate is 30g/L, and the look valency of controlling red colouring agent for food, also used as a Chinese medicine citraurin is 0.8; In temperature, be 30 ℃, in the shaking table of 200rpm, carry out, the reaction times is 8 hours;
The absorption spectrum of analytical reaction liquid, 410,470 and the absorbancy at 510nm place be respectively 0.5,0.5 and 0.4 absorbance unit.The tone of monascorubin is 0.4/0.5; Adopt thin layer chromatography analysis to show that monascorubin red colouring agent for food, also used as a Chinese medicine citraurin spot exists and a monascorubin minute rate obviously increases.
embodiment 6
The present embodiment relates to a kind of method that red colouring agent for food, also used as a Chinese medicine citraurin is converted into the nicellar catalysis of monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4.5 that described seed liquor is inoculated into initial pH value, in the fermentation culture that the Semen Maydis powder of take is nitrogenous source, in temperature, is 30 ℃, carries out the fermentation culture of 7 days in the shaking table of 200rpm, centrifugal, obtains monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 2,1 and 1.8 absorbance units; Fermented liquid final pH is 6.8.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin; 410,470 and the absorbancy at 510nm place be respectively 25,38 and 20 absorbance units.The tone of monascorubin is 20/38.
Step 3, after concentrated, in the tensio-active agent Trition-100 aqueous solution that the concentrated pigment that takes a morsel adds to, regulates pH=7 by the extract of monascorubin in above-mentioned born of the same parents; Surfactant concentration is 10%, and concentration of sodium glutamate is 30g/L, and the look valency of controlling red colouring agent for food, also used as a Chinese medicine citraurin is 0.8; In temperature, be 30 ℃, in the shaking table of 200rpm, carry out, the reaction times is 8 hours;
The absorption spectrum of analytical reaction liquid, 410,470 and the absorbancy at 510nm place be respectively 0.7,0.4 and 0.6 absorbance unit.The tone of monascorubin is 0.6/0.5; Adopt thin layer chromatography analysis to show that monascorubin red colouring agent for food, also used as a Chinese medicine citraurin spot disappears and a monascorubin minute rate obviously increases.
embodiment 7
The present embodiment relates to a kind of method that red colouring agent for food, also used as a Chinese medicine citraurin is converted into the nicellar catalysis of monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4.5 that described seed liquor is inoculated into initial pH value, in the fermentation culture that the Semen Maydis powder of take is nitrogenous source, in temperature, is 30 ℃, carries out the fermentation culture of 7 days in the shaking table of 200rpm, centrifugal, obtains monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 2,1 and 1.8 absorbance units; Fermented liquid final pH is 6.8.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin; 410,470 and the absorbancy at 510nm place be respectively 25,38 and 20 absorbance units.The tone of monascorubin is 20/38.
Step 3, after concentrated, in the tensio-active agent Trition-100 aqueous solution that the concentrated pigment that takes a morsel adds to, regulates pH=7 by the extract of monascorubin in above-mentioned born of the same parents; Surfactant concentration is 10%, and concentration of sodium glutamate is 5g/L, and the look valency of controlling red colouring agent for food, also used as a Chinese medicine citraurin is 0.8; In temperature, be 30 ℃, in the shaking table of 200rpm, carry out, the reaction times is 8 hours;
The absorption spectrum of analytical reaction liquid, 410,470 and the absorbancy at 510nm place be respectively 0.4,0.6 and 0.3 absorbance unit.The tone of monascorubin is 0.3/0.6; Adopt thin layer chromatography analysis to show each component of monascorubin.
embodiment 8
The present embodiment relates to and a kind ofly at fermenting process, parachrome is secreted into outside born of the same parents and further red colouring agent for food, also used as a Chinese medicine citraurin is converted into the method for the nicellar catalysis of monascorubin outside born of the same parents, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4 that described seed liquor is inoculated into initial pH value, the fermentation culture that the Semen Maydis powder of take is nitrogenous source, in substratum, adding surfactant concentration is that 5g/L, concentration of sodium glutamate are 30g/L.In temperature, be 30 ℃, in the shaking table of 200rpm, carry out the fermentation culture of 7 days, centrifugal, obtain monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
Analyzing fermented liquid final pH is 6.5; The absorbancy of cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 25,15 and 28 absorbance units; The tone of monascorubin is 28/15; Thin-layer chromatography shows that monascorubin main component is monascorubin.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is also monascorubin; 410,470 and the absorbancy at 510nm place be respectively 7,3 and 4 absorbance units; The tone of monascorubin is 4/3.
embodiment 9
The present embodiment relates to and a kind ofly at fermenting process, parachrome is secreted into outside born of the same parents and further red colouring agent for food, also used as a Chinese medicine citraurin is converted into the method for the nicellar catalysis of monascorubin outside born of the same parents, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4 that described seed liquor is inoculated into initial pH value, take and mixes the fermentation culture that Semen Maydis powder and analysis for soybean powder (1:1) are nitrogenous source, in substratum, adding surfactant concentration is that 5g/L, concentration of sodium glutamate are 20g/L.In temperature, be 30 ℃, in the shaking table of 200rpm, carry out the fermentation culture of 6 days, centrifugal, obtain monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
Analyzing fermented liquid final pH is 6.8; The absorbancy of cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 23,12 and 25 absorbance units; The tone of monascorubin is 25/12; Thin-layer chromatography shows that monascorubin main component is monascorubin.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is also monascorubin; 410,470 and the absorbancy at 510nm place be respectively 6,3 and 5 absorbance units, the tone of monascorubin is 5/3.
embodiment 10
The present embodiment relates to and a kind ofly at fermenting process, parachrome is secreted into outside born of the same parents and further red colouring agent for food, also used as a Chinese medicine citraurin is converted into the method for the nicellar catalysis of monascorubin outside born of the same parents, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4 that described seed liquor is inoculated into initial pH value, take the fermentation culture that mixing rice powder and analysis for soybean powder (1:1) they are nitrogenous source, in substratum, adding surfactant concentration is that 5g/L, concentration of sodium glutamate are 20g/L.In temperature, be 30 ℃, in the shaking table of 200rpm, carry out the fermentation culture of 7 days, centrifugal, obtain monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
Analyzing fermented liquid final pH is 6.7; The absorbancy of cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 22,11 and 24 absorbance units; The tone of monascorubin is 24/11; Thin-layer chromatography shows that monascorubin main component is monascorubin.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is also monascorubin; 410,470 and the absorbancy at 510nm place be respectively 7,2 and 3 absorbance units, the tone of monascorubin is 3/2.
embodiment 11
The present embodiment relates to and a kind ofly at fermenting process, parachrome is secreted into outside born of the same parents and further red colouring agent for food, also used as a Chinese medicine citraurin is converted into the method for the nicellar catalysis of monascorubin outside born of the same parents, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4 that described seed liquor is inoculated into initial pH value, take glucose as the fermentation culture that carbon source, L-glutamic acid are nitrogenous source, in substratum, adding surfactant concentration is that 5g/L, concentration of sodium glutamate are 5g/L.In temperature, be 30 ℃, in the shaking table of 200rpm, carry out the fermentation culture of 8 days, centrifugal, obtain monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
Analyzing fermented liquid final pH is 6.2; The absorbancy of cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 15,30 and 17 absorbance units; The tone of monascorubin is 17/30; Thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is also red colouring agent for food, also used as a Chinese medicine citraurin; 410,470 and the absorbancy at 510nm place be respectively 7,3 and 2 absorbance units, the tone of monascorubin is 2/3.
embodiment 12
The present embodiment relates to and a kind ofly at fermenting process, parachrome is secreted into outside born of the same parents and further red colouring agent for food, also used as a Chinese medicine citraurin is converted into the method for the nicellar catalysis of monascorubin outside born of the same parents, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4 that described seed liquor is inoculated into initial pH value, take glucose as the fermentation culture that carbon source, L-glutamic acid are nitrogenous source, in substratum, adding surfactant concentration is that 5g/L, concentration of sodium glutamate are 20g/L.In temperature, be 30 ℃, in the shaking table of 200rpm, carry out the fermentation culture of 7 days, centrifugal, obtain monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
Analyzing fermented liquid final pH is 6.5; The absorbancy of cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 23,13 and 21 absorbance units; The tone of monascorubin is 21/13; Thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is also monascorubin; 410,470 and the absorbancy at 510nm place be respectively 5,3 and 4 absorbance units, the tone of monascorubin is 4/3.
embodiment 13
The present embodiment relates to and a kind ofly at fermenting process, parachrome is secreted into outside born of the same parents and further red colouring agent for food, also used as a Chinese medicine citraurin is converted into the method for the nicellar catalysis of monascorubin outside born of the same parents, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4.5 that described seed liquor is inoculated into initial pH value, take glucose as the fermentation culture that carbon source, L-glutamic acid are nitrogenous source, in substratum, adding surfactant concentration is that 5g/L, concentration of sodium glutamate are 35g/L.In temperature, be 30 ℃, in the shaking table of 200rpm, carry out the fermentation culture of 8 days, centrifugal, obtain monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
Analyzing fermented liquid final pH is 6.8; The absorbancy of cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 31,17 and 33 absorbance units; The tone of monascorubin is 33/17; Thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is also monascorubin; 410,470 and the absorbancy at 510nm place be respectively 7,3 and 5 absorbance units, the tone of monascorubin is 5/3.
embodiment 14
The present embodiment relates to and a kind ofly at fermenting process, parachrome is secreted into outside born of the same parents and further red colouring agent for food, also used as a Chinese medicine citraurin is converted into the method for the nicellar catalysis of monascorubin outside born of the same parents, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 6.5 that described seed liquor is inoculated into initial pH value, take glucose as the fermentation culture that carbon source, L-glutamic acid are nitrogenous source, in substratum, adding surfactant concentration is that 5g/L, concentration of sodium glutamate are 35g/L.In temperature, be 30 ℃, in the shaking table of 200rpm, carry out the fermentation culture of 7 days, centrifugal, obtain monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
Analyzing fermented liquid final pH is 6.8; The absorbancy of cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 33,17 and 35 absorbance units; The tone of monascorubin is 35/17; Thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is also monascorubin; 410,470 and the absorbancy at 510nm place be respectively 7,3 and 5 absorbance units, the tone of monascorubin is 5/3.
embodiment 15
The present embodiment relates to and a kind ofly at fermenting process, parachrome is secreted into outside born of the same parents and further red colouring agent for food, also used as a Chinese medicine citraurin is converted into the method for the nicellar catalysis of monascorubin outside born of the same parents, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium water (100ml, peptone 2g, yeast powder 2g, glucose 2g) in, cultivate 32 hours, obtain seed liquor, described monascus ruber provides (Monascus anka, CICC 5013) for Chinese industrial microorganism strains preservation center;
Step 2, it is 4.5 that described seed liquor is inoculated into initial pH value, take glucose as the fermentation culture that carbon source, SODIUMNITRATE are nitrogenous source, in substratum, adding surfactant concentration is that 5g/L, concentration of sodium glutamate are 25g/L.In temperature, be 30 ℃, in the shaking table of 200rpm, carry out the fermentation culture of 7 days, centrifugal, obtain monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, the look valency of the monascorubin inside and outside detection fermented liquid pH, cell; Wherein, described pre-treatment is specially: get 0.5g wet cell and soak 1 hour in ethanol (pH=2) solution of isopyknic fermented liquid 70% (V/V);
Analyzing fermented liquid final pH is 6.9; The absorbancy of cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 23,11 and 21 absorbance units; The tone of monascorubin is 21/11; Thin-layer chromatography shows that monascorubin main component is red colouring agent for food, also used as a Chinese medicine citraurin.Parachrome extracts through ethanolic soln, and thin-layer chromatography shows that monascorubin main component is also monascorubin; 410,470 and the absorbancy at 510nm place be respectively 7,4 and 5 absorbance units, the tone of monascorubin is 5/4.
Above specific embodiments of the invention are described.It will be appreciated that, the present invention is not limited to above-mentioned specific implementations, and those skilled in the art can make various distortion or modification within the scope of the claims, and this does not affect flesh and blood of the present invention.
Claims (10)
1. a preparation method for high tone monascorubin, is characterized in that, described method comprises the steps:
Step 11, monascus ruber is cultivated in seed culture medium, obtains seed liquor;
Step 12, is inoculated into described seed liquor in the fermentation culture of carbonaceous sources, carries out fermentation culture, regulates the pH value of fermentation culture, centrifugal, obtains monascus ruber wet cell, and extraction monascus ruber wet cell obtains take the monascorubin that red colouring agent for food, also used as a Chinese medicine citraurin is main ingredient;
Step 13, adds the monascorubin of described extraction to the nicellar catalysis reaction that is converted into monascorubin containing carrying out red colouring agent for food, also used as a Chinese medicine citraurin in the solution of nonionogenic tenside and Sodium Glutamate.
2. the preparation method of high tone monascorubin according to claim 1, is characterized in that, in step 13, the scope of concentration of sodium glutamate is at 5-30g/L.
3. the preparation method of high tone monascorubin according to claim 1, is characterized in that, in step 13, the concentration of nonionogenic tenside is less than or equal to 100g/L.
4. according to the preparation method of the high tone monascorubin described in claim 1-3 any one, it is characterized in that, in step 13, described reaction refers to that in temperature be 30 ℃, in the shaking table of 200rpm, carries out, and the reaction times is 6～12 hours; Reaction pH is 2-7.
5. according to the preparation method of the high tone monascorubin described in claim 1-3 any one, it is characterized in that, in step 12, described fermentation culture is 30 ℃ in temperature, in the shaking table of 200rpm, carries out, and incubation time is 6-8 days; The initial pH value of described fermentation culture is 4.5.
6. a preparation method for high tone monascorubin, is characterized in that, described method comprises the steps:
Step 21, monascus ruber is cultivated in seed culture medium, obtains seed liquor;
Step 22, is inoculated into resulting seed liquor in step 1 containing fermentation culture in the substratum of nonionogenic tenside and Sodium Glutamate, the centrifugal outer monascorubin of born of the same parents that obtains high look valency, high tone.
7. the preparation method of high tone monascorubin according to claim 6, is characterized in that, in step 22, nonionogenic tenside concentration remains on the scope of 50-100g/l.
8. the preparation method of high tone monascorubin according to claim 6, is characterized in that, in step 22, the concentration of Sodium Glutamate remains on the scope of 5-30g/l.
9. according to the preparation method of the high tone monascorubin described in claim 6-8 any one, it is characterized in that, in step 22, the initial pH value of described substratum is 4～6.5.
10. according to the preparation method of the high tone monascorubin described in claim 6-8 any one, it is characterized in that, in step 22, described substratum is comprised of each component of following content: water 100ml, glucose 5g, KH
2pO
40.4g, MgSO
40.1g, CaCl
20.005g, nonionogenic tenside 1～10g/100ml, aminoglutaric acid concentration is 5g/100～30g/1000ml.
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Application publication date: 20141210 |