CN103224958A - Method for preparing monascus pigment through extraction fermentation and pH value regulation - Google Patents
Method for preparing monascus pigment through extraction fermentation and pH value regulation Download PDFInfo
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- monascorubin
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- monascus
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- 230000004151 fermentation Effects 0.000 title claims abstract description 179
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- 239000000049 pigment Substances 0.000 title claims abstract description 29
- 238000000605 extraction Methods 0.000 title abstract 3
- CQIUKKVOEOPUDV-IYSWYEEDSA-N antimycin Chemical compound OC1=C(C(O)=O)C(=O)C(C)=C2[C@H](C)[C@@H](C)OC=C21 CQIUKKVOEOPUDV-IYSWYEEDSA-N 0.000 claims abstract description 109
- CQIUKKVOEOPUDV-UHFFFAOYSA-N citrinine Natural products OC1=C(C(O)=O)C(=O)C(C)=C2C(C)C(C)OC=C21 CQIUKKVOEOPUDV-UHFFFAOYSA-N 0.000 claims abstract description 109
- 238000000136 cloud-point extraction Methods 0.000 claims abstract description 27
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- 235000013305 food Nutrition 0.000 claims description 25
- 239000001052 yellow pigment Substances 0.000 claims description 25
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 23
- 239000008103 glucose Substances 0.000 claims description 23
- 239000002609 medium Substances 0.000 claims description 21
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 21
- SPBPMWXNKPPVSX-KXOLNMLNSA-N Citraurin Natural products CC(=C/C=C/C(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1C(=CC(O)CC1(C)C)C)/C)C=O SPBPMWXNKPPVSX-KXOLNMLNSA-N 0.000 claims description 18
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- 229920004890 Triton X-100 Polymers 0.000 claims description 14
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 claims description 14
- 238000005516 engineering process Methods 0.000 claims description 14
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 240000007594 Oryza sativa Species 0.000 claims description 12
- 235000007164 Oryza sativa Nutrition 0.000 claims description 12
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 12
- 230000000813 microbial effect Effects 0.000 claims description 9
- 235000009566 rice Nutrition 0.000 claims description 7
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 6
- 238000000926 separation method Methods 0.000 claims description 6
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- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 5
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- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The present invention provides a method for preparing a monascus pigment through extraction fermentation and pH value regulation. The method comprises the following steps: 1, culturing monascus sp in a seed culture medium to obtain a seed liquid; 2, inoculating the seed liquid into a nitrogen source-containing fermentation culture medium to carry out fermentation culture, adjusting the pH value of the fermentation culture solution, and centrifuging to obtain wet cells of the monascus sp; and 3, inoculating the obtained wet cells of the monascus sp into a non-ionic surfactant-containing fermentation culture medium to culture, adjusting the pH value of the fermentation culture medium, carrying out cloud point extraction, and separating to obtain the finished product. According to the present invention, the pH value of the fermented solution is adjusted to control formation of the monascus pigment and citrinin, and the extraction fermentation and the cloud point extraction are adopted to adjust the pigment composition so as to remove the byproduct citrinin to prepare the low citrinin content monascus pigment. The method has advantages of simpleness, easy achievement, high yield, low cost, strong repeatability, environmental protection, and good application prospect.
Description
Technical field
The present invention relates to a kind of fermentation process of technical field of bioengineering, particularly, relate to the method that a kind of extractive fermentation and regulation and control pH prepare monascorubin.
Background technology
The Red kojic rice of monascus ruber (Monascus sp) solid state fermentation production is the traditional food color additive of China, also is a kind of traditional Chinese medicine, and the use history in more than 1,000 year has been arranged in China.The modern microbic liquid deep layer fermentation technology of current employing is produced monascorubin, and this pigment is extensive use of in East Asian countries as the food color additive, as China, Japan, Korea S, Taiwan, Thailand, Vietnam etc.Since having found that monascus ruber fermentative production monascorubin also produces the by product Citrinin with neurotoxicity and renal toxicity, monascorubin is produced the America and Europe and has been forbidden production and selling.Especially, the monascorubin product is the mixture of monascorubin, monascorubin, red colouring agent for food, also used as a Chinese medicine citraurin composition.The national standard of having formulated monascorubin except that some countries of East Asia allows its production and selling, and monascus yellow pigment and red colouring agent for food, also used as a Chinese medicine citraurin are not also put into effect corresponding national standards.
In order to ensure food safety, control content of by-products in the monascorubin product, particularly have the content of the Citrinin of toxic side effect, be the important research content of food microorganisms always.During the fermentation, utilize the modern biotechnology means to transform microorganism strains and have feasibility technically, but the leavened prod that genetic engineering modified Institute of Micro-biology obtains belongs to Genetic engineering food with the formation that suppresses Citrinin.Because the singularity of food safety, Genetic engineering food is very loaded down with trivial details and difficult in the license that a lot of countries obtain to produce and sell.Adopt fermentation engineering and become important means, as regulating operating parameters such as dissolved oxygen in conjunction with the content that the downstream separation technology reduces Citrinin.
The formation of microbial secondary meta-bolites depends on fermentation condition consumingly.In the fermenting process of monascus ruber, the pH that regulates fermentation media can change the branch rate of each colour component in the monascorubin.Under nearly condition of neutral pH, the branch rate of monascorubin is higher, and this adopts the rice fermentation Red kojic rice consistent with traditional technology.The pH that reduces fermented liquid makes the corresponding increase of branch rate of red colouring agent for food, also used as a Chinese medicine citraurin and monascus yellow pigment, causes the pigmentary colours flavescence.Guangdong “ salt baked chicken in 2012 " the food disturbance is exactly because added monascus yellow pigment.Though the security of monascus yellow pigment has obtained the confirmation of domestic and international research, pH changes the influence that the monascus ruber metabolic by-prods is formed and but lacks research.Part scholar infers the monascus yellow pigment security of products from the security of monascus yellow pigment itself, because of by product (cut down as Citrinin, Lip river he is left alone without help etc.) content and toxicity are difficult to obtain conclusive evidence, can not allow the human consumer convince, also be difficult to obtain the approval of state food administration.
In order to satisfy the demand of market to monascus yellow pigment, ensure food safety simultaneously, press for the single relatively monascus ruber fermentation technique of colour component in low, the monascorubin of exploitation citrinin content.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, use two sections fermentation techniques and discharge monascorubin in the born of the same parents ". " biotechnology journal ", 2012, introduce 162:202-209), monascus ruber in nonionogenic tenside micellar solution fermentation energy with born of the same parents in monascorubin be secreted into extracellular environment, realized the intracellular product extractive fermentation to born of the same parents' external surfactants micellar solution.But different fermention mediums and fermentation mode are formed the influence that divides rate to monascorubin, the especially influence that the by product Citrinin is formed, and also research is not reported.In further retrieving, also do not combine with regulation and control pH and realize the bibliographical information of control pigment composition and citrinin content about extractive fermentation in surfactant micelle solution.
Summary of the invention
At defective of the prior art, the purpose of this invention is to provide the method that a kind of extractive fermentation and regulation and control pH prepare monascorubin, this method is used serial monascorubin products such as the extractive fermentation water-soluble/hydrophobicity monascorubin low with regulating and control pH technological development citrinin content, hydrophobicity red colouring agent for food, also used as a Chinese medicine citraurin, hydrophobicity monascus yellow pigment.
The invention provides the method that a kind of extractive fermentation and regulation and control pH prepare monascorubin, described method combines the low and single relatively monascorubin of monascus pigment component of preparation citrinin content by the extractive fermentation technology with regulation and control pH technology; Described method comprises the steps:
Step 1, monascus ruber is cultivated in seed culture medium, seed liquor, described monascus ruber for Chinese industrial microorganism strains preservation center provide (Monascus anka, CICC5013);
Step 2 is regulated the composition that pH can control monascorubin in order to protect, and can control the formation of Citrinin simultaneously under low pH condition, described seed liquor is inoculated in the fermentation culture that contains nitrogenous source, carries out fermentation culture, regulate the pH value of fermentation culture, centrifugal, get the monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, detect fermented liquid pH, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell;
Wherein, described pre-treatment is specially: get the 0.5g wet cell at isopyknic fermented liquid 70%(V/V) ethanol (pH=2) solution in soaked 1 hour;
Step 3 can further be controlled the formation of monascus yellow pigment in order to protect extractive fermentation, controls the content of Citrinin simultaneously, described monascus ruber wet cell is inoculated in the fermention medium that contains nonionogenic tenside, cultivates, regulate the pH value of fermention medium, cloud point extraction separates; Get the pre-treatment of part cell, discharge intracellular product, detect fermented liquid pH, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell.
Preferably, described pigment concentration, its detection method is specially: cell free fermentation liquid or intracellular product are discharged liquid 70%(V/V) ethanol (pH=2) solution suitably absorb the light absorption value at employing spectrophotometer measure of spread 410,470,510nm place.Above-mentioned light absorption value is multiplied by and absorbs the concentration that absorbance unit that multiple gets is represented yellow pigment, citraurin and haematochrome respectively.Accordingly, working sample is at visible wavelength region (390～540nm) absorbancy, absorption spectrum that can the check sample correspondence.
Described citrinin content, its detection method is specially: cell free fermentation liquid or intracellular product are discharged liquid 70%(V/V) ethanol (pH=2) solution suitably absorb, adopt thin-layer chromatography (the silica gel F254 of Merck ﹠ Co., Inc. plate), solvent systems is an acetic acid: methyl alcohol, chloroform=9:21:285, the detection wavelength is 365nm, is the fluorescence power that 0.56 place can detect Citrinin at Rf.Simultaneously, under natural light, be respectively 0.8,0.68 and 0 place at Rf and can detect the red colouring agent for food, also used as a Chinese medicine citraurin, monascus yellow pigment and monascorubin.
Preferably, in the step 1, described seed culture medium is made up of each component of following content: water 100ml, peptone 2g, yeast powder 2g, glucose 2g.
Preferably, in the step 1, described cultivation is 30 ℃ in temperature, carries out in the shaking table of 200rpm, and incubation time is 32 hours.
Preferably, in the step 2, described fermentation culture is 30 ℃ in temperature, carries out in the shaking table of 200rpm, and incubation time is 5-7 days.
Preferably, in the step 2, the initial pH value of described fermentation culture is 2.5～6, and the final pH value is 2.3～7.
Preferably, in the step 3, described cultivation is meant that in temperature be 30 ℃, carries out in the shaking table of 200rpm, and incubation time is 3～4 days; Described separation is meant left standstill in 70 ℃ water-bath 2 hours.
Preferably, in the step 3, the initial pH value of described fermention medium is 3～5, and the final pH value is 2.2～6.2.
Preferably, in the step 3, described fermention medium is made up of each component of following content: water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, nonionogenic tenside 1～4g/100ml, nitrogenous source 5g/100～15g/100ml.
Preferably, described nitrogenous source has comprised inorganic nitrogen-sourced and organic natural nitrogenous source commonly used.Described nitrogenous source is rice meal, Semen Maydis powder, analysis for soybean powder, Sodium Glutamate, ammonium sulfate, ammonium chloride, SODIUMNITRATE.
Preferably, the present invention has used the micellar solution of nonionogenic tenside Triton X-100 to realize the extractive fermentation and the cloud point extraction of parachrome.With ammonium sulfate, ammonium chloride etc. is the nitrogenous source of fermention medium, and extractive fermentation has been realized intracellular product is extracted into cell free fermentation liquid, and pigment is a monascus yellow pigment outside the born of the same parents, and citrinin content is lower.Further realized the cloud point extraction of cell free fermentation liquid, Citrinin mainly is distributed in the tensio-active agent dilute phase and monascus yellow pigment is distributed in the tensio-active agent concentrated phase, thereby further reduced the content of Citrinin in the tensio-active agent concentrated phase at separation phase, can obtain the monascus yellow pigment product of low citrinin content by the tensio-active agent concentrated phase.
Preferably, the present invention has used the micellar solution of nonionogenic tenside Triton X-100 to realize the extractive fermentation and the cloud point extraction of parachrome.With the nitrogenous source of fermention mediums such as SODIUMNITRATE, extractive fermentation has been realized intracellular product is extracted into cell free fermentation liquid, and the outer pigment of born of the same parents is a monascorubin, and citrinin content is higher.Further realized the cloud point extraction of cell free fermentation liquid, Citrinin mainly is distributed in the tensio-active agent dilute phase and monascorubin is distributed in the tensio-active agent concentrated phase, thereby further reduced the content of Citrinin in the tensio-active agent concentrated phase at separation phase, can obtain the monascorubin product of low citrinin content by the tensio-active agent concentrated phase.
Preferably, the present invention has used the micellar solution of nonionogenic tenside Triton X-100 to realize the extractive fermentation and the cloud point extraction of parachrome.With Sodium Glutamate etc. is the nitrogenous source of fermention medium, and extractive fermentation has been realized intracellular product is extracted into cell free fermentation liquid, and the outer pigment of born of the same parents is the water-soluble red pigment of red rice derivative, and citrinin content is higher.Further cloud point extraction cell free fermentation liquid, Citrinin mainly is distributed in the tensio-active agent dilute phase, the water-soluble red pigment of red rice derivative has certain distribution in two-phase, but mainly be distributed in the tensio-active agent concentrated phase, thereby further reduced the content of Citrinin in the tensio-active agent concentrated phase at separation phase, can obtain the water-soluble red pigment of red rice derivative product (wetting ability monascorubin derivative) of low citrinin content by the tensio-active agent concentrated phase.
The present invention adopts the purpose of regulation and control pH and extractive fermentation to be to obtain relative single, the serial monascorubin product that citrinin content is low of component, as water-soluble red pigment of red rice, fat-soluble red colouring agent for food, also used as a Chinese medicine citraurin, fat-soluble monascus yellow pigment and fat-soluble monascorubin etc.
The present invention adopts the purpose of regulation and control pH to be to regulate the branch rate of colour component in the monascorubin and the formation of control metabolic by-prods Citrinin.
The present invention adopts the purpose of extractive fermentation technology to be to change the branch rate of monascus pigment component, and parachrome is secreted into the extracellular.
The present invention adopts the extractive fermentation technology that intracellular product is secreted into the cloud point extraction that the outer purpose of born of the same parents is further to realize cell free fermentation liquid.
The present invention adopts the purpose of cloud point extraction cell free fermentation liquid to be to realize the inhomogeneous distribution in biphasic system of monascorubin and Citrinin, further reduces the content of Citrinin.
Compared with prior art, the present invention has following beneficial effect: the present invention adopts the pH value of fermentation and control fermentation, and then controlled the formation of monascorubin and Citrinin, the present invention uses the extractive fermentation technology simultaneously and cloud point extraction is regulated the pigment composition and removed the by product Citrinin, has realized hydrophobicity monascorubin, hydrophobicity red colouring agent for food, also used as a Chinese medicine citraurin, hydrophobicity monascus yellow pigment and water-soluble red pigment of red rice (glutamic acid derivatives of the monascorubin) series product development of low citrinin content.The inventive method is simple, realizes easily, and the productive rate height, cost is low, and repeatability is strong, and environmental protection has application promise in clinical practice.
Embodiment
The present invention is described in detail below in conjunction with specific embodiment.Following examples will help those skilled in the art further to understand the present invention, but not limit the present invention in any form.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.
Embodiment 1
Present embodiment relates to a kind of extractive fermentation and regulates and control the method that pH prepares 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) cultivated 32 hours in, seed liquor, described monascus ruber for Chinese industrial microorganism strains preservation center provide (Monascus anka, CICC5013);
Step 2, it is 4 that described seed liquor is inoculated into initial pH value, is in the fermentation culture of nitrogenous source with the SODIUMNITRATE, is 30 ℃ in temperature, carries out 5 days fermentation culture in the shaking table of 200rpm, regulates the pH value of fermentation culture, centrifugal, gets the monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, detect fermented liquid pH, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell; Wherein, described pre-treatment is specially: get the 0.5g wet cell at isopyknic fermented liquid 70%(V/V) ethanol (pH=2) solution in soaked 1 hour;
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 2,1 and 1.8 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 35,25 and 32 absorbance units.
The fermented liquid final pH is 6.8, and the inside and outside citrinin content of born of the same parents shows the content height through thin layer chromatography analysis, and obvious distribution is all arranged inside and outside born of the same parents, is more prone to be distributed in cell free fermentation liquid.Thin-layer chromatography shows that the monascorubin main component is a monascorubin.
Step 3 with the cell 0.5g of fermentation culture, adds initial pH and is 4 fermention medium (water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, ammonium sulfate 5g/100ml, Triton X-100 4g/100ml) and middle the cultivation after 5 days, the absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 65,45 and 22 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 25,20 and 15 absorbance units.The fermentation final pH is 2.2, and the inside and outside citrinin content of born of the same parents shows through thin layer chromatography analysis and do not contain Citrinin substantially.Thin-layer chromatography shows that the monascorubin main component is a monascus yellow pigment.Further cloud point extraction cell free fermentation liquid in 70 degree water-baths, in the tensio-active agent concentrated phase 410,470 and the absorbancy of 510nm place monascorubin be respectively 120,60 and 20 absorbance units; In the tensio-active agent dilute phase 410,470 and the absorbancy at 510nm place be respectively 15,8 and 7 absorbance units.
Implementation result: the present embodiment product can get through the thin layer chromatography analysis result: can detect the existence of Citrinin in the tensio-active agent dilute phase, the tensio-active agent concentrated phase detect less than.
Embodiment 2
Present embodiment relates to a kind of extractive fermentation and regulates and control the method that pH prepares monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium (100ml, peptone 2g, yeast powder 2g, glucose 2g) cultivated 32 hours in, seed liquor, described monascus ruber for Chinese industrial microorganism strains preservation center provide (Monascus anka, CICC5013);
Step 2, it is 4 that described seed liquor is inoculated into initial pH value, is in the fermentation culture of nitrogenous source with the Sodium Glutamate, carries out fermentation culture after 7 days, regulates the pH value of fermentation culture, centrifugal, gets the monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, detect fermented liquid pH value, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell; Wherein, described pre-treatment is specially: get the 0.5g wet cell at isopyknic fermented liquid 70%(V/V) ethanol (pH=2) solution in soaked 1 hour;
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 10,8 and 12 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 30,22 and 32 absorbance units.
The fermented liquid final pH is 6.5, and the inside and outside citrinin content of born of the same parents shows the content height through thin layer chromatography analysis, and obvious distribution is all arranged inside and outside born of the same parents, is more prone to be distributed in cell free fermentation liquid.Thin-layer chromatography shows that the monascorubin main component is the monascorubin derivative.
Step 3 with the cell 0.5g of fermentation culture, adds initial pH and is 4 fermention medium (water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, ammonium sulfate 15g/100ml, Triton X-100 3g/100ml) and middle the cultivation after 3 days, the absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 45,25 and 20 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 35,30 and 18 absorbance units.The fermentation final pH is 2.2, and the inside and outside citrinin content of born of the same parents shows through thin layer chromatography analysis and do not contain Citrinin substantially.Thin-layer chromatography shows that the monascorubin main component is a monascus yellow pigment.Further cloud point extraction cell free fermentation liquid in 70 degree water-baths, in the tensio-active agent concentrated phase 410,470 and the absorbancy of 510nm place monascorubin be respectively 110,60 and 30 absorbance units; In the tensio-active agent dilute phase 410,470 and the absorbancy at 510nm place be respectively 15,8 and 7 absorbance units.
Implementation result: the present embodiment product can get through the thin layer chromatography analysis result: thin layer chromatography analysis can detect the existence of Citrinin in the tensio-active agent dilute phase, the tensio-active agent concentrated phase detect less than.
Embodiment 3
Present embodiment relates to a kind of extractive fermentation and regulates and control the method that pH prepares monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium (100ml, peptone 2g, yeast powder 2g, glucose 2g) cultivated 32 hours in, seed liquor, described monascus ruber for Chinese industrial microorganism strains preservation center provide (Monascus anka, CICC5013);
Step 2, it is 6 that described seed liquor is inoculated into initial pH value, is in the fermentation culture of nitrogenous source with the Sodium Glutamate, carries out fermentation culture after 7 days, regulates the pH value of fermentation culture, centrifugal, gets the monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, detect fermented liquid pH value, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell; Wherein, described pre-treatment is specially: get the 0.5g wet cell at isopyknic fermented liquid 70%(V/V) ethanol (pH=2) solution in soaked 1 hour;
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 5,3 and 5 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 20,12 and 22 absorbance units.
The fermented liquid final pH is 7, and the inside and outside citrinin content of born of the same parents shows the content height through thin layer chromatography analysis, and obvious distribution is all arranged inside and outside born of the same parents, is more prone to be distributed in cell free fermentation liquid.Thin-layer chromatography shows that the monascorubin main component is the monascorubin derivative.
Step 3 with the cell 0.5g of fermentation culture, adds initial pH and is 5.5 fermention medium (water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, SODIUMNITRATE 15g/100ml, Triton X-100 3g/100ml) and middle the cultivation after 4 days, the absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 55,40 and 53 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 15,8 and 12 absorbance units.The fermentation final pH is 6.2, and obviously there is Citrinin in the inside and outside citrinin content of born of the same parents through thin layer chromatography analysis inside and outside cell, and Citrinin is more prone to be distributed in outside the born of the same parents.Thin-layer chromatography shows that the monascorubin main component is a monascorubin.Further cloud point extraction cell free fermentation liquid in 70 degree water-baths, in the tensio-active agent concentrated phase 410,470 and the absorbancy of 510nm place monascorubin be respectively 108,80 and 110 absorbance units; In the tensio-active agent dilute phase 410,470 and the absorbancy at 510nm place be respectively 5,2 and 5 absorbance units.
Implementation result: the present embodiment product can get through the thin layer chromatography analysis result: thin layer chromatography analysis can obviously detect the existence of Citrinin in the tensio-active agent dilute phase, the tensio-active agent concentrated phase almost detect less than.
Embodiment 4
Present embodiment relates to a kind of extractive fermentation and regulates and control the method that pH prepares monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium (100ml, peptone 2g, yeast powder 2g, glucose 2g) cultivated 32 hours in, seed liquor, described monascus ruber for Chinese industrial microorganism strains preservation center provide (Monascus anka, CICC5013);
Step 2, it is 2.5 that described seed liquor is inoculated into initial pH value, is in the fermentation culture of nitrogenous source with the Sodium Glutamate, carries out fermentation culture after 5 days, regulates the pH value of fermentation culture, centrifugal, gets the monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, detect fermented liquid pH value, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell; Wherein, described pre-treatment is specially: get the 0.5g wet cell at isopyknic fermented liquid 70%(V/V) ethanol (pH=2) solution in soaked 1 hour;
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 1,0.3 and 0.8 absorbance unit; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 30,45 and 12 absorbance units.
The fermented liquid final pH is 2.5, and the inside and outside citrinin content of born of the same parents shows through thin layer chromatography analysis and do not contain Citrinin substantially.Thin-layer chromatography shows that the monascorubin main component is the red colouring agent for food, also used as a Chinese medicine citraurin.
Step 3 with the cell 0.5g of fermentation culture, adds initial pH and is 3.5 fermention medium (water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, SODIUMNITRATE 15g/100ml, Triton X-100 3g/100ml) and middle the cultivation after 4 days, the absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 53,40 and 50 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 14,9 and 12 absorbance units.The fermentation final pH is 6.2, and obviously there is Citrinin in the inside and outside citrinin content of born of the same parents through thin layer chromatography analysis inside and outside cell, and Citrinin is more prone to be distributed in outside the born of the same parents.Thin-layer chromatography shows that the monascorubin main component is a monascorubin.Further cloud point extraction cell free fermentation liquid in 70 degree water-baths, in the tensio-active agent concentrated phase 410,470 and the absorbancy of 510nm place monascorubin be respectively 100,80 and 108 absorbance units; In the tensio-active agent dilute phase 410,470 and the absorbancy at 510nm place be respectively 5,2 and 5 absorbance units.
Implementation result: the present embodiment product can get through the thin layer chromatography analysis result: thin layer chromatography analysis can obviously detect the existence of Citrinin in the tensio-active agent dilute phase, the tensio-active agent concentrated phase almost detect less than.
Embodiment 5
Present embodiment relates to a kind of extractive fermentation and regulates and control the method that pH prepares monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium (100ml, peptone 2g, yeast powder 2g, glucose 2g) cultivated 32 hours in, seed liquor, described monascus ruber for Chinese industrial microorganism strains preservation center provide (Monascus anka, CICC5013);
Step 2, it is 6 that described seed liquor is inoculated into initial pH value, is in the fermentation culture of nitrogenous source with the Sodium Glutamate, carries out fermentation culture after 5 days, regulates the pH value of fermentation culture, centrifugal, gets the monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, detect fermented liquid pH value, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell; Wherein, described pre-treatment is specially: get the 0.5g wet cell at isopyknic fermented liquid 70%(V/V) ethanol (pH=2) solution in soaked 1 hour;
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 5,3 and 5 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 20,32 and 18 absorbance units.
The fermented liquid final pH is 4, and the inside and outside citrinin content of born of the same parents shows the content height through thin layer chromatography analysis, and obvious distribution is all arranged inside and outside born of the same parents, is more prone to be distributed in cell free fermentation liquid.Thin-layer chromatography shows that the monascorubin main component is the red colouring agent for food, also used as a Chinese medicine citraurin.
Step 3 with the cell 0.5g of fermentation culture, adds initial pH and is 5 fermention medium (water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, Sodium Glutamate 15g/100ml, Triton X-100 3g/100ml) and middle the cultivation after 3 days, the absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 50,35 and 50 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 15,10 and 14 absorbance units.The fermentation final pH is 6.2, and obviously there is Citrinin in the inside and outside citrinin content of born of the same parents through thin layer chromatography analysis inside and outside cell, and Citrinin is more prone to be distributed in outside the born of the same parents.Thin-layer chromatography shows that the monascorubin main component is the monascorubin derivative.Further cloud point extraction cell free fermentation liquid in 70 degree water-baths, in the tensio-active agent concentrated phase 410,470 and the absorbancy of 510nm place monascorubin be respectively 90,75 and 98 absorbance units; In the tensio-active agent dilute phase 410,470 and the absorbancy at 510nm place be respectively 15,12 and 13 absorbance units.
Implementation result: the present embodiment product can get through the thin layer chromatography analysis result: thin layer chromatography analysis can obviously detect the existence of Citrinin in the tensio-active agent dilute phase, the tensio-active agent concentrated phase almost detect less than.
Embodiment 6
Present embodiment relates to a kind of extractive fermentation and regulates and control the method that pH prepares monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium (100ml, peptone 2g, yeast powder 2g, glucose 2g) cultivated 32 hours in, seed liquor, described monascus ruber for Chinese industrial microorganism strains preservation center provide (Monascus anka, CICC5013);
Step 2, it is 3.5 that described seed liquor is inoculated into initial pH value, is in the fermentation culture of nitrogenous source with the Sodium Glutamate, carries out fermentation culture after 6 days, regulates the pH value of fermentation culture, centrifugal, gets the monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, detect fermented liquid pH value, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell; Wherein, described pre-treatment is specially: get the 0.5g wet cell at isopyknic fermented liquid 70%(V/V) ethanol (pH=2) solution in soaked 1 hour;
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 5,2.5 and 5 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 21,33 and 17 absorbance units.
The fermented liquid final pH is 3.7, and the inside and outside citrinin content of born of the same parents shows the content height through thin layer chromatography analysis, and obvious distribution is all arranged inside and outside born of the same parents, is more prone to be distributed in cell free fermentation liquid.Thin-layer chromatography shows that the monascorubin main component is the red colouring agent for food, also used as a Chinese medicine citraurin.
Step 3 with the cell 0.5g of fermentation culture, adds initial pH and is 3 fermention medium (water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, Sodium Glutamate 15g/100ml, Triton X-100 3g/100ml) and middle the cultivation after 3 days, the absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 60,35 and 20 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 25,10 and 8 absorbance units.The fermentation final pH is 3.5, and there is a small amount of Citrinin in the inside and outside citrinin content of born of the same parents through thin layer chromatography analysis inside and outside cell, and Citrinin is more prone to be distributed in outside the born of the same parents.Thin-layer chromatography shows that the monascorubin main component is the red colouring agent for food, also used as a Chinese medicine citraurin.Further cloud point extraction cell free fermentation liquid in 70 degree water-baths, in the tensio-active agent concentrated phase 410,470 and the absorbancy of 510nm place monascorubin be respectively 110,75 and 35 absorbance units; In the tensio-active agent dilute phase 410,470 and the absorbancy at 510nm place be respectively 5,2 and 1 absorbance unit.
Implementation result: the present embodiment product can get through the thin layer chromatography analysis result: thin layer chromatography analysis can obviously detect the existence of Citrinin in the tensio-active agent dilute phase, the tensio-active agent concentrated phase almost detect less than.
Embodiment 7
Present embodiment relates to a kind of extractive fermentation and regulates and control the method that pH prepares monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium (100ml, peptone 2g, yeast powder 2g, glucose 2g) cultivated 32 hours in, seed liquor, described monascus ruber for Chinese industrial microorganism strains preservation center provide (Monascus anka, CICC5013);
Step 2, it is 2.5 that described seed liquor is inoculated into initial pH value, is in the fermentation culture of nitrogenous source with the Sodium Glutamate, carries out fermentation culture after 6 days, regulates the pH value of fermentation culture, centrifugal, gets the monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, detect fermented liquid pH value, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell; Wherein, described pre-treatment is specially: get the 0.5g wet cell at isopyknic fermented liquid 70%(V/V) ethanol (pH=2) solution in soaked 1 hour;
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 3,1 and 1 absorbance unit; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 24,38 and 18 absorbance units.
The fermented liquid final pH is 2.8, and the inside and outside citrinin content of born of the same parents shows that through thin layer chromatography analysis content obviously with low, all has obvious distribution inside and outside born of the same parents.Thin-layer chromatography shows that the monascorubin main component is the red colouring agent for food, also used as a Chinese medicine citraurin.
Step 3 with the cell 0.5g of fermentation culture, adds initial pH and is 5 fermention medium (water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, rice meal 5g/100ml, Triton X-100 3g/100ml) and middle the cultivation after 3 days, the absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 54,40 and 51 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 16,9 and 12 absorbance units.The fermentation final pH is 6.2, and obviously there is Citrinin in the inside and outside citrinin content of born of the same parents through thin layer chromatography analysis inside and outside cell, and Citrinin is more prone to be distributed in outside the born of the same parents.Thin-layer chromatography shows that the monascorubin main component is a monascorubin.Further cloud point extraction cell free fermentation liquid in 70 degree water-baths, in the tensio-active agent concentrated phase 410,470 and the absorbancy of 510nm place monascorubin be respectively 100,80 and 108 absorbance units; In the tensio-active agent dilute phase 410,470 and the absorbancy at 510nm place be respectively 5,2 and 5 absorbance units.
Implementation result: the present embodiment product can get through the thin layer chromatography analysis result: thin layer chromatography analysis can obviously detect the existence of Citrinin in the tensio-active agent dilute phase, the tensio-active agent concentrated phase almost detect less than.
Embodiment 8
Present embodiment relates to a kind of extractive fermentation and regulates and control the method that pH prepares monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium (100ml, peptone 2g, yeast powder 2g, glucose 2g) cultivated 32 hours in, seed liquor, described monascus ruber for Chinese industrial microorganism strains preservation center provide (Monascus anka, CICC5013);
Step 2, it is 6 that described seed liquor is inoculated into initial pH value, is in the fermentation culture of nitrogenous source with the Sodium Glutamate, carries out fermentation culture after 6 days, regulates the pH value of fermentation culture, centrifugal, gets the monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, detect fermented liquid pH value, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell; Wherein, described pre-treatment is specially: get the 0.5g wet cell at isopyknic fermented liquid 70%(V/V) ethanol (pH=2) solution in soaked 1 hour;
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 5,2 and 6 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 21,30 and 24 absorbance units.
The fermented liquid final pH is 4.3, and the inside and outside citrinin content of born of the same parents shows the content height through thin layer chromatography analysis, and obvious distribution is all arranged inside and outside born of the same parents, is more prone to be distributed in cell free fermentation liquid.Thin-layer chromatography shows that the monascorubin main component is the red colouring agent for food, also used as a Chinese medicine citraurin.
Step 3 with the cell 0.5g of fermentation culture, adds initial pH and is 5 fermention medium (water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, rice meal 5g/100ml, Triton X-100 3g/100ml) and middle the cultivation after 3 days, the absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 54,40 and 51 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 16,9 and 12 absorbance units.The fermentation final pH is 6.2, and obviously there is Citrinin in the inside and outside citrinin content of born of the same parents through thin layer chromatography analysis inside and outside cell, and Citrinin is more prone to be distributed in outside the born of the same parents.Thin-layer chromatography shows that the monascorubin main component is a monascorubin.Further cloud point extraction cell free fermentation liquid in 70 degree water-baths, in the tensio-active agent concentrated phase 410,470 and the absorbancy of 510nm place monascorubin be respectively 100,80 and 108 absorbance units; In the tensio-active agent dilute phase 410,470 and the absorbancy at 510nm place be respectively 5,2 and 5 absorbance units.
Implementation result: the present embodiment product can get through the thin layer chromatography analysis result: thin layer chromatography analysis can obviously detect the existence of Citrinin in the tensio-active agent dilute phase, the tensio-active agent concentrated phase almost detect less than.
Embodiment 9
Present embodiment relates to a kind of extractive fermentation and regulates and control the method that pH prepares monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium (100ml, peptone 2g, yeast powder 2g, glucose 2g) cultivated 32 hours in, seed liquor, described monascus ruber for Chinese industrial microorganism strains preservation center provide (Monascus anka, CICC5013);
Step 2, it is 4 that described seed liquor is inoculated into initial pH value, is in the fermentation culture of nitrogenous source with the Sodium Glutamate, carries out fermentation culture after 6 days, regulates the pH value of fermentation culture, centrifugal, gets the monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, detect fermented liquid pH, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell; Wherein, described pre-treatment is specially: get the 0.5g wet cell at isopyknic fermented liquid 70%(V/V) ethanol (pH=2) solution in soaked 1 hour;
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 4,1.5 and 0.8 absorbance unit; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 45,35 and 30 absorbance units.
The fermented liquid final pH is 2.3, and the inside and outside citrinin content of born of the same parents shows through thin layer chromatography analysis and do not contain Citrinin substantially.Thin-layer chromatography shows that the monascorubin main component is the red colouring agent for food, also used as a Chinese medicine citraurin.
Step 3 with the cell 0.5g of fermentation culture, adds initial pH and is 4 fermention medium (water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, Semen Maydis powder 15g/100ml, Triton X-100 3g/100ml) and middle the cultivation after 3 days, the absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 35,15 and 10 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 15,10 and 7 absorbance units.The fermentation final pH is 2.2, and the inside and outside citrinin content of born of the same parents shows through thin layer chromatography analysis and do not contain Citrinin substantially.Thin-layer chromatography shows that the monascorubin main component is a monascus yellow pigment.Further cloud point extraction cell free fermentation liquid in 70 degree water-baths, in the tensio-active agent concentrated phase 410,470 and the absorbancy of 510nm place monascorubin be respectively 80,30 and 10 absorbance units; In the tensio-active agent dilute phase 410,470 and the absorbancy at 510nm place be respectively 15,8 and 7 absorbance units.
Implementation result: the present embodiment product can get through the thin layer chromatography analysis result: thin layer chromatography analysis can detect the existence of Citrinin in the tensio-active agent dilute phase, the tensio-active agent concentrated phase detect less than.
Embodiment 10
Present embodiment relates to a kind of extractive fermentation and regulates and control the method that pH prepares monascorubin, and described method comprises the steps:
Step 1, monascus ruber is at seed culture medium (100ml, peptone 2g, yeast powder 2g, glucose 2g) cultivated 32 hours in, seed liquor, described monascus ruber for Chinese industrial microorganism strains preservation center provide (Monascus anka, CICC5013);
Step 2, it is 4 that described seed liquor is inoculated into initial pH value, is in the fermentation culture of nitrogenous source with the Sodium Glutamate, carries out fermentation culture after 6 days, regulates the pH value of fermentation culture, centrifugal, gets the monascus ruber wet cell; Get the pre-treatment of monascus ruber wet cell, discharge intracellular product, detect fermented liquid pH value, inside and outside pigment concentration, the inside and outside citrinin content of cell of cell; Wherein, described pre-treatment is specially: get the 0.5g wet cell at isopyknic fermented liquid 70%(V/V) ethanol (pH=2) solution in soaked 1 hour;
The absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 4,1.5 and 0.8 absorbance unit; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 45,35 and 30 absorbance units.
The fermented liquid final pH is 6.3, and the inside and outside citrinin content of born of the same parents shows through thin layer chromatography analysis and do not contain Citrinin substantially.Thin-layer chromatography shows that the monascorubin main component is the red colouring agent for food, also used as a Chinese medicine citraurin.
Step 3 with the cell 0.5g of fermentation culture, adds initial pH and is 4 fermention medium (water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, analysis for soybean powder 15g/100ml, Triton X-100 3g/100ml) and middle the cultivation after 3 days, the absorbancy of analyzing cell free fermentation liquid 410,470 and 510nm place monascorubin is respectively 45,25 and 15 absorbance units; Parachrome after ethanolic soln extracts 410,470 and the absorbancy at 510nm place be respectively 18,13 and 6 absorbance units.The fermentation final pH is 2.2, and the inside and outside citrinin content of born of the same parents shows through thin layer chromatography analysis and do not contain Citrinin substantially.Thin-layer chromatography shows that the monascorubin main component is a monascus yellow pigment.Further cloud point extraction cell free fermentation liquid in 70 degree water-baths, in the tensio-active agent concentrated phase 410,470 and the absorbancy of 510nm place monascorubin be respectively 90,35 and 15 absorbance units; In the tensio-active agent dilute phase 410,470 and the absorbancy at 510nm place be respectively 15,8 and 7 absorbance units.
Implementation result: the present embodiment product can get through the thin layer chromatography analysis result: thin layer chromatography analysis can detect the existence of Citrinin in the tensio-active agent dilute phase, the tensio-active agent concentrated phase detect less than.
The present invention combines with regulation and control pH technology by the extractive fermentation technology, realized microbial fermentation produce the low and monascus pigment component of citrinin content single relatively water-soluble/serial monascorubin products such as hydrophobicity monascorubin, hydrophobicity red colouring agent for food, also used as a Chinese medicine citraurin, hydrophobicity monascus yellow pigment.Adopt the extractive fermentation technology, microorganism in nonionogenic tenside micellar solution fermentation culture can be with born of the same parents in monascorubin be discharged into born of the same parents' external surfactants micellar solution, and under the low pH condition of control fermented liquid, produce monascus yellow pigment outside the low born of the same parents of citrinin content.Elevated temperature can be realized cloud point extraction, and cloud point extraction can make Citrinin be distributed in the tensio-active agent dilute phase and monascorubin is distributed in the tensio-active agent concentrated phase.When low pH extractive fermentation, form the outer yellow pigment fermented liquid of the low born of the same parents of citrinin content, cloud point extraction can further be removed Citrinin wherein and obtain the low hydrophobicity monascus yellow pigment of citrinin content.When high pH extractive fermentation, form the high water-soluble/hydrophobicity monascorubin fermented liquid of citrinin content, cloud point extraction also can be removed Citrinin wherein and obtain the low water-soluble/hydrophobicity monascorubin of citrinin content.
It more than is the description of the preferred embodiment of the present invention, should be understood that, enforcement of the present invention is not limited to above-mentioned situation, the present invention is particularly suitable for the fermenting process of product in the cell, as the production of oil compounds in the production of organic molecule material in the production of microbial enzyme in the cell, the cell and the cell etc., can remove intracellular product effectively and suppress, improve the concentration of microbial fermentation product and the composition of adjusting microbial fermentation secondary metabolite.Thereby the volume productivity and the efficient that has improved processes such as intracellular product fermentation, product release of microbial fermentation have been improved.
More than specific embodiments of the invention are described.It will be appreciated that the present invention is not limited to above-mentioned specific implementations, those skilled in the art can make various distortion or modification within the scope of the claims, and this does not influence flesh and blood of the present invention.
Claims (9)
1. extractive fermentation and regulation and control pH prepare the method for monascorubin, it is characterized in that, described method combines with regulation and control pH technology by the extractive fermentation technology, prepares the low and single relatively monascorubin of monascus pigment component of citrinin content; Described method comprises the steps:
Step 1, monascus ruber is cultivated in seed culture medium, gets seed liquor;
Step 2 is inoculated into described seed liquor in the fermentation culture that contains nitrogenous source, carries out fermentation culture, regulates the pH value of fermentation culture, and is centrifugal, gets the monascus ruber wet cell;
Step 3 is inoculated into described monascus ruber wet cell in the fermention medium that contains nonionogenic tenside, cultivates, and regulates the pH value of fermention medium, and cloud point extraction separates, and gets final product.
2. extractive fermentation according to claim 1 and regulation and control pH prepare the method for monascorubin, it is characterized in that, in the step 2, the final pH value of described fermentation culture is 2.5 when following, microbial fermentation generation red colouring agent for food, also used as a Chinese medicine citraurin, and citrinin content is low.
3. extractive fermentation according to claim 1 and 2 and regulation and control pH prepare the method for monascorubin, it is characterized in that in the step 2, the initial pH value of described fermentation culture is 2.5～6, and the final pH value is 2.3～7.
4. extractive fermentation according to claim 1 and regulation and control pH prepare the method for monascorubin, it is characterized in that, in the step 3, the final pH value of described fermention medium is 2.5 when following, microbial fermentation generation monascus yellow pigment, and citrinin content is low.
5. prepare the method for monascorubin according to claim 1 or 4 described extractive fermentations and regulation and control pH, it is characterized in that in the step 3, the initial pH value of described fermention medium is 3～5, the final pH value is 2.2～6.2.
6. prepare the method for monascorubin according to claim 1 or 4 described extractive fermentations and regulation and control pH, it is characterized in that in the step 3, described cultivation is meant that in temperature be 30 ℃, carries out in the shaking table of 200rpm, incubation time is 3～4 days; Described separation is meant left standstill in 70 ℃ water-bath 2 hours.
7. extractive fermentation according to claim 1 and regulation and control pH prepare the method for monascorubin, it is characterized in that, in the step 3, are made up of each component of following content in the described fermention medium: water 100ml, glucose 5g, KH
2PO
40.4g, MgSO
40.1g, CaCl
20.005g, nonionogenic tenside 1～4g/100ml, nitrogenous source 5g/100～15g/100ml.
8. prepare the method for monascorubin according to claim 1 or 7 described extractive fermentations and regulation and control pH, it is characterized in that described nitrogenous source is rice meal, Semen Maydis powder, analysis for soybean powder, Sodium Glutamate, ammonium sulfate, ammonium chloride, SODIUMNITRATE.
9. prepare the method for monascorubin according to claim 1 or 7 described extractive fermentations and regulation and control pH, it is characterized in that described nonionogenic tenside is Triton X-100.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104195178A (en) * | 2014-07-08 | 2014-12-10 | 上海交通大学 | High tone monascus red pigment preparation method |
| CN107723320A (en) * | 2017-09-08 | 2018-02-23 | 广东科隆生物科技有限公司 | A kind of method for directly preparing crystal red yeast rice citraurin |
| CN109371053A (en) * | 2018-12-24 | 2019-02-22 | 江西科技师范大学 | A kind of High-productive Monascus Pigment Strain construction method |
| CN110592158A (en) * | 2019-09-19 | 2019-12-20 | 长江大学 | Liquid fermentation method for improving purity of monascus yellow pigment Monascin and Ankaflavin |
| CN112042858A (en) * | 2020-09-15 | 2020-12-08 | 江西理工大学 | Method for extracting biosurfactant of monascus pigment |
| CN112430635A (en) * | 2020-12-01 | 2021-03-02 | 广东肇庆星湖生物科技股份有限公司 | Fermentation method for low-yield citrinin and high-yield water-soluble monascus yellow pigment |
| CN112481327A (en) * | 2020-12-01 | 2021-03-12 | 广东肇庆星湖生物科技股份有限公司 | Fermentation regulation and control method of water-soluble monascus yellow pigment |
| CN114507698A (en) * | 2020-11-17 | 2022-05-17 | 天津科技大学 | Liquid state fermentation method for increasing monascus orange pigment yield |
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| CN102443605A (en) * | 2011-11-04 | 2012-05-09 | 上海交通大学 | Method for perstraction of fermented microbial intracellular product with non-ionic surfactant |
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Patent Citations (1)
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| CN102443605A (en) * | 2011-11-04 | 2012-05-09 | 上海交通大学 | Method for perstraction of fermented microbial intracellular product with non-ionic surfactant |
Non-Patent Citations (2)
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| BIYU K ET AL: "Effect of pH and nonionic surfactant on profile of intracellular and extracellular monascus pigments", 《PROCESS BIOCHEMISTRY》 * |
| SANDRA FERNANDA BILBAO OROZCO等: "Effect of pH on citrinin and red pigments production by Monascus purpureus CCT3802", 《WORLD J MICROBIOL BIOTECHNOL》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104195178A (en) * | 2014-07-08 | 2014-12-10 | 上海交通大学 | High tone monascus red pigment preparation method |
| CN107723320A (en) * | 2017-09-08 | 2018-02-23 | 广东科隆生物科技有限公司 | A kind of method for directly preparing crystal red yeast rice citraurin |
| CN107723320B (en) * | 2017-09-08 | 2020-09-18 | 广东科隆生物科技有限公司 | Method for directly preparing crystal monascus orange pigment |
| CN109371053A (en) * | 2018-12-24 | 2019-02-22 | 江西科技师范大学 | A kind of High-productive Monascus Pigment Strain construction method |
| CN110592158A (en) * | 2019-09-19 | 2019-12-20 | 长江大学 | Liquid fermentation method for improving purity of monascus yellow pigment Monascin and Ankaflavin |
| CN112042858A (en) * | 2020-09-15 | 2020-12-08 | 江西理工大学 | Method for extracting biosurfactant of monascus pigment |
| CN112042858B (en) * | 2020-09-15 | 2022-11-29 | 江西理工大学 | Method for extracting biosurfactant of monascus pigment |
| CN114507698A (en) * | 2020-11-17 | 2022-05-17 | 天津科技大学 | Liquid state fermentation method for increasing monascus orange pigment yield |
| CN112430635A (en) * | 2020-12-01 | 2021-03-02 | 广东肇庆星湖生物科技股份有限公司 | Fermentation method for low-yield citrinin and high-yield water-soluble monascus yellow pigment |
| CN112481327A (en) * | 2020-12-01 | 2021-03-12 | 广东肇庆星湖生物科技股份有限公司 | Fermentation regulation and control method of water-soluble monascus yellow pigment |
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| CN103224958B (en) | 2016-02-10 |
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