CN104946699A - Double-liquid-phase fermentation method of monascus yellow pigment by coupled in-situ fermentation-extraction - Google Patents
Double-liquid-phase fermentation method of monascus yellow pigment by coupled in-situ fermentation-extraction Download PDFInfo
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Abstract
The invention discloses a double-liquid-phase fermentation method of monascus yellow pigment by coupled in-situ fermentation-extraction, belonging to the field of fermentation engineering. The varieties and contents of carbon and nitrogen sources in the fermentation culture medium are controlled to effectively control the monascus pigment biosynthesis pathway, so that citraurin generated by the monascus is no longer converted into the monascus red pigment; and the double-liquid-phase extraction-fermentation coupled system can ensure the normal growth and fermentation of the monascus, is beneficial to enhancing the dissolved oxygen level, and is capable of promoting the secretion of the monascus pigment to the outside of cells and lowering the negative feedback inhibiting action of the monascus fermentation product, thereby greatly enhancing the fermentation level of the monascus yellow pigment and shortening the fermentation time. After the fermentation finishes, the product can be easily separated. The method can greatly enhance the yield and production efficiency of the monascus yellow pigment and lower the production cost.
Description
Technical field
The present invention relates to the Two Liquid Phases fermentation process of a kind of coupling situ extracting fermentation monascus yellow pigment, belong to field of fermentation engineering.
Background technology
Natural yellow pigment is divided into plant milk extract and microbial fermentation product.Natural yellow pigment mainly carotenoid and the monascus yellow pigment that the fermentation of current energy Large-scale microbial produces.Monascus yellow pigment is divided into two kinds, and one is Monascin (monascine), and another kind is Monascus anka flavine (ankaflavine).The wavelength of the maximum absorption band of monascus yellow pigment is generally at 400-440nm.In addition market also have a kind of also from monascorubin but through the yellow pigment of chemically modified gained, namely with natural monascorubin for raw material, itself and V-Brite B are reacted in basic solution, make Ester hydrolysis in monascorubin, in pigmentary structures, introduce sulfonic group simultaneously, obtain " water-soluble monascus yellow pigment ", the wavelength that the maximum absorption band of this pigment is corresponding is 476nm.From the structure of production method and pigment, this so-called monascus yellow pigment is a kind of semi-natural monascus yellow pigment.
The natural monascus yellow pigment of Monascus anka Nakazawa et sato Production by Microorganism Fermentation is utilized to be the research and development emphasis of current monascorubin industry.For improving monascus yellow pigment production level, the emphasis that the paper of domestic and foreign literature report or patent are paid close attention to mainly concentrates on the mutagenesis screening of high yield monascus bacterial classification and these two aspects of improvement of regular fermentation process condition, but it is little to produce effects.The Research Team of such as China Ma Meirong, Thailand Yongsmith from since the nineties in 20th century by the Monascus anka Nakazawa et sato of mutagenic and breeding to per unit area yield yellow pigment, be applied to liquid state fermentation and solid state fermentation produces monascus yellow pigment.The wavelength that the maximum absorption band of the monascus yellow pigment that this bacterial strain is produced is corresponding is 370nm.But liquid fermentation method yellow pigment production level is very low, not yet realizes suitability for industrialized production so far.Red colouring agent for food, also used as a Chinese medicine Huang (rice) the look valency that this bacterium solid state fermentation is produced for 12 days reaches 2224.63U/gdw, although pigment production level is higher, fermentation time is long.The people such as domestic cycle have gone out monascus yellow pigment producing strains by mutagenic and breeding, and the wavelength that the maximum absorption band of the monascus yellow pigment that this bacterial strain is produced is corresponding is 410nm, and the yellow plain color of the liquid state fermentation fermented liquid of 7 days is worth and only reaches 110U/mL.Above a few example due to fermentation pigment concentration on the low side, fermentation time is partially long, does not all meet the requirement of suitability for industrialized production.
Summary of the invention
The problem to be solved in the present invention is to provide the Two Liquid Phases fermentation process of a kind of coupling situ extracting fermentation monascus yellow pigment, is that monascus yellow pigment is produced in fermentation by Monascus anka Nakazawa et sato liquid seeds access fermention medium; Described fermention medium is the double liquid phase extraction fermentation system of aqueous phase and extraction phase composition, and described aqueous phase is the minimum medium containing Monascus anka Nakazawa et sato growth desired nutritional composition, and described extraction phase contains glycerides.
Described red colouring agent for food, also used as a Chinese medicine bacterial classification is the red colouring agent for food, also used as a Chinese medicine bacterial classification that can produce monascus yellow pigment.
Described minimum medium contains W-Gum 20-80g/L, analysis for soybean powder 0.05-10g/L, ammonium sulfate 3-10g/L, NaNO
32-6g/L, yeast extract paste 0.05-5g/L, corn starch 0.05-10g/L, zinc sulfate 0.1-0.5g/L, magnesium sulfate 0.5-1.5g/L, potassium primary phosphate 0.5-1.5g/L.
Glycerides in described extraction phase is improved the effect of dissolved oxygen as extraction agent, can by be attached in fermented liquid and the pigment extraction on Monascus anka Nakazawa et sato ball surface be enriched to extraction agent mutually in, increase substantially concentration and the productive rate of red colouring agent for food, also used as a Chinese medicine liquid state fermentation synthetic yellow pigment product.
Described glycerides is triglyceride material, and consumption volume accounts for the 10-40% of minimum medium volume.
Described triglyceride material is caprylin, three isocaprylic acid glyceryl ester, vegetables oil or its mixture.Except plant oil, all the other triglyceride materials add the time all after the rear 24-72h of red colouring agent for food, also used as a Chinese medicine seed liquor inoculation.
Described glycerides, except triglyceride material, also comprises di-glycerides material, monoglyceride class material or its mixture simultaneously.The interpolation time of described di-glycerides material, monoglyceride class material is after red colouring agent for food, also used as a Chinese medicine seed liquor inoculates 24-72h.When independent interpolation di-glycerides material or monoglyceride class material, the volume of interpolation accounts for the 0.1-5% of minimum medium volume; When mixing is added, cumulative volume accounts for the 0.1-5% of minimum medium volume.
Described di-glycerides material is 1,3-DAG, 1,2-triglyceride, two caprylins or its mixture.
Described monoglyceride class material is Capmul MCM C8.
The Monascus anka Nakazawa et sato liquid seeds being in logarithmic phase is equipped with in the 500mL triangular flask of 100mL fermention medium with the access of the inoculum size of volume ratio 5-10% by the condition optimization that monascus yellow pigment is produced in described fermentation, be 80 ~ 180r/min at rotating speed, under temperature 26 ~ 32 DEG C of conditions, shake flask fermentation cultivates 4-6d.The interpolation time of partial glycerol three Ester of thalli growth, triglyceride, monoglyceride can be suppressed to be 24 ~ 72h after the inoculation of red colouring agent for food, also used as a Chinese medicine seed liquor.
First synthesize citraurin during Monascus anka Nakazawa et sato fermentation, the metabolism of red colouring agent for food, also used as a Chinese medicine citraurin is divided into two branch's approach, and an approach citraurin is reduced to monascus yellow pigment, and another approach is then the better monascorubin of water soluble characteristic with amino acid converting in aqueous phase.If block red colouring agent for food, also used as a Chinese medicine citraurin to be converted into monascorubin, the citraurin generated just major part even all may be converted into monascus yellow pigment.
The present invention is by carbon nitrogen source component reasonable in design, the content controlling substratum Free Amino Acids and Two Liquid Phases fermentation system, block the conversion of red colouring agent for food, also used as a Chinese medicine citraurin to monascorubin, and the monascorubin of synthesis is enriched to rapidly in extraction phase, thus improving the production efficiency of monascus yellow pigment, gained monascus yellow pigment look valency reaches as high as 250U/mL.Adopt common grease as the extraction agent of Two Liquid Phases fermentation system, on the one hand because grease has the effect increasing dissolved oxygen, this is for guarantee oxygen supply, and the output improving pigment is useful; Adopt on the other hand vegetables oil as extraction agent, be conducive to pigment as the special tinting material of vegetables oil.In addition, glycerolipid substance improves somatic cells membrane permeability, and the monascorubin of synthesis is secreted into outside mycelium in time, reduces the negative feedback inhibition effect of monascorubin product.The purity of monascus yellow pigment of the present invention is higher, and fermentation time is short, realizes efficient, high yield monascus yellow pigment, increases economic efficiency.
Accompanying drawing explanation
The monascus yellow pigment full wavelength scanner figure extracted in Fig. 1 fermented liquid
Embodiment
Following examples bacterial strain uses therefor and fermentation mode, according to described in such scheme, adopt coupling situ extracting fermentative production monascus yellow pigment product, further illustrate embodiments of the invention below.
The measuring method of yellow plain color valency:
(1) process of fermented liquid
The mensuration of fermented liquid weight and volume and the determination of scale-up factor: fermentation triangular flask (jumping a queue) itself and the triangular flask after adding substratum (comprising oil phase) are weighed and (is recorded as W respectively
0, W
2), show that substratum weight (is recorded as W
1); Measure culture volume (V
1); Calculate the scale-up factor (mL/g) of volume and substratum weight.
After fermentation ends, fermentation flask and fermented liquid are weighed, complement to the weight before fermentation with water.Monascus anka Nakazawa et sato fermented liquid (oil-containing) is all transferred in paste roller mill, wears into homogenate.Get a beaker and a merging of metal spoon is weighed, be equivalent to the fermented liquid of about 10mL with metal bale-out, then by beaker+metal spoon and samples weighing, draw the actual weight of sample, and be converted into the volume of fermented liquid.Get raw spirit and be about 20mL, join in beaker, after fermentation broth agitation is even, transfer in 50mL colorimetric cylinder, then secondary respectively gets 10mL absolute ethanol washing beaker respectively, the remaining fermented liquid in beaker is all transferred in colorimetric cylinder, and constant volume is to 50mL, 55 DEG C of water-baths extraction 1h, middle interval shakes 2 times, is cooled to the ethanol dilutions that room temperature gets the above-mentioned fermented liquid of 10mL as required and once dilutes according to the method described above again after water-bath terminates.
(2) yellow valency measures and calculates
With dehydrated alcohol to above-mentioned diluent redilution suitable multiple (OD value is controlled between 0.2-0.8) as requested, and be that blank carries out full wavelength scanner within the scope of 200-600nm with dehydrated alcohol, measure the OD value of peak value between 400-440nm.
Yellow valency=OD
420nm× total extension rate (U/mL)
Seed culture condition:
Slant medium: potato 200g/L, glucose 20g/L, magnesium sulfate 0.1g/L, potassium primary phosphate 0.5g/L, peptone 5g/L, agar 20g/L.
Liquid seed culture medium: W-Gum 20g/L, analysis for soybean powder 2.5g/L, ammonium sulfate 3g/L, NaNO
36g/L, yeast extract paste 2g/L, corn starch 2g/L, magnesium sulfate 0.8g/L, potassium primary phosphate 0.6g/L.
Red colouring agent for food, also used as a Chinese medicine bacterial classification is transferred in slant medium and cultivates, culture temperature 30 DEG C, cultivates 1 week, obtains fresh Monascus anka Nakazawa et sato, aseptic technique, wash lower spore with a certain amount of sterilized water from inclined-plane, prepare spore suspension, be equipped with in the 500mL triangular flask of 100mL liquid seed culture medium with the access of the inoculum size of volume ratio 10%, be placed in 30 DEG C of constant-temperature tables, rotating speed is 120r/min, cultivates 2-3d, obtains the red colouring agent for food, also used as a Chinese medicine seed liquor being in growth logarithmic phase.
Embodiment 1
Monascus anka Nakazawa et sato JJLY-4A is selected to be shaking flask bacterial classification.Fermention medium is made up of minimum medium and extraction phase.Minimum medium (g/L): W-Gum 60, analysis for soybean powder 2, ammonium sulfate 8, NaNO36, yeast extract paste 0.2, corn starch 0.2, zinc sulfate 0.2, magnesium sulfate 0.8, potassium primary phosphate 1.2; Extraction phase (with the volume ratio of substratum): soybean oil 10%, Semen Maydis oil 8%, caprylin 1%, two caprylin 0.3%, caprylin 0.2%.In extraction phase except soybean oil and Semen Maydis oil, remaining glycerides all adds substratum after fermentation starts 48h.
The Monascus anka Nakazawa et sato liquid seeds being in logarithmic phase is equipped with in the 500mL triangular flask of 100mL fermention medium with the access of the inoculum size of volume ratio 10%, is 180r/min at rotating speed, and under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 220-230U/mL.Pigment detection wavelength is between 400-440nm nm.
Embodiment 2
Identical with above-described embodiment 1 experimental procedure and fermention medium, difference is: experimental strain is Monascus anka Nakazawa et sato ZH6.Being equipped with in the 500mL triangular flask of 100mL fermention medium by the Monascus anka Nakazawa et sato liquid seeds being in logarithmic phase with the access of the inoculum size of volume ratio 10%, is 180r/min at rotating speed, and under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 160-170U/mL.
Embodiment 3
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: in fermention medium, W-Gum is 40g/L.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 150-160U/mL.
Embodiment 4
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: in fermention medium, W-Gum is 70g/L.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 240-250U/mL.
Embodiment 5
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: nitrogen source ammonium sulfate 3g/L, NaNO33g/L in fermention medium, analysis for soybean powder 6, yeast extract paste 6g/L.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 160-170U/mL.
Embodiment 6
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: nitrogen source ammonium sulfate 8g/L, NaNO36g/L in fermention medium, analysis for soybean powder 2, yeast extract paste 2g/L.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 200-210U/mL.
Embodiment 7
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: extraction phase (with the volume ratio of substratum) proportioning is: soybean oil 6%, Semen Maydis oil 6%, caprylin 1%, two caprylin 0.2%, caprylin 0.1%.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 190-200U/mL.
Embodiment 8
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: extraction phase (with the volume ratio of substratum) proportioning is: soybean oil 12%, Semen Maydis oil 12%, caprylin 2%, two caprylin 0.5%, caprylin 0.3%.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 230-240U/mL.
Embodiment 9
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: extraction phase (with the volume ratio of substratum) proportioning is: rapeseed oil 10%, peanut oil 8%, caprylin 0.5%, two caprylin 0.3%, caprylin 0.2%.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 220-230U/mL.
Embodiment 10
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: extraction phase (with the volume ratio of substratum) proportioning is: Oleum Gossypii semen 10%, soybean oil 8.5%, two caprylin 0.3%, caprylin 0.2%.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 220-230U/mL.
Embodiment 11
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: extraction phase (with the volume ratio of substratum) proportioning is: caprylin 9.5%, three isocaprylic acid glyceryl ester 9%, two caprylin 0.3%, caprylin 0.2%.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 120-130U/mL.
Embodiment 12
With Monascus anka Nakazawa et sato JJLY-4 for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: extraction phase (with the volume ratio of substratum): soybean oil 10%, Semen Maydis oil 8%, caprylin 0.5%, 1,3-DAG 0.3%, caprylin 0.2%.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 220-230U/mL.
Embodiment 13
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: extraction phase (with the volume ratio of substratum): soybean oil 10%, Semen Maydis oil 8%, caprylin 0.5%, 1,2-triglyceride 0.2%, two caprylin 0.1%, caprylin 0.2%.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 220-230U/mL.
Embodiment 14
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: extraction phase (with the volume ratio of substratum): soybean oil 10%, Semen Maydis oil 8%, caprylin 0.5%, 1,3-triglyceride 0.1%, 1,2-triglyceride 0.1%, two caprylin 0.1%, caprylin 0.2%.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 220-230U/mL.
Embodiment 15
With Monascus anka Nakazawa et sato JJLY-4A for shake flask fermentation bacterial classification.With above-described embodiment 1 step and culture medium prescription substantially identical, difference is: extraction phase (with the volume ratio of substratum) proportioning is: Semen Maydis oil 20%.Be 180r/min at rotating speed, under temperature 30 DEG C of conditions, shake flask fermentation cultivates 5d.The look valency recording the monascus yellow pigment being folded to minimum medium volume after end is between 210-220U/mL.
Monascus yellow pigment is produced in the fermentation of embodiment 16 common fermentation processes
Monascus yellow pigment liquid state fermentation process: PDA inclined-plane → make spore suspension → be seeded to seed liquor → cultivation 48h, access fermention medium, inoculum size is 5% → cultivate after 5 days, fermentation liquor pretreatment, colour examining valency.
Slant medium (PDA substratum): potato 200g adds water boil 30min, glucose 20g, agar 15 ~ 20g, constant volume 1000mL, natural pH.
Seed culture medium (g/L): W-Gum 40, ammonium sulfate 4, KH
2pO
4, 2.0; K
2hPO
4, 2.0; MgSO
47H
2o, 0.5; CaCl
2, 0.1; FeSO
47H
2o, 0.01; ZnSO
47H
2o, 0.01; MnSO
4h
2o, 0.03; NaNO
3, 2.0; PH5.0.
Liquid Medium of shaking flask fermentation (g/L): W-Gum, 60.0; Ammonium sulfate, 4.0; KH
2pO
4, 2.0; K
2hPO
4, 2.0; MgSO
47H
2o, 0.5; CaCl
2, 0.1; FeSO
47H
2o, 0.01; ZnSO
47H
2o, 0.01; MnSO
4h
2o, 0.03; NaNO
3, 2.0; Initial ph value is 5.0; Fermentation time is 4.0d.
Cultural method:
The preparation of spore suspension: get slant strains one, adds appropriate amounts of sterilized water and a small amount of granulated glass sphere, fully filters with aseptic filter paper after vibration, then with blood counting chamber counting, makes spore concentration reach 10
6individual/mL.
Monascus anka Nakazawa et sato seed culture: 250mL triangular flask liquid amount 45mL, inoculum size: spore suspension 2mL.30 DEG C of reciprocal shaker, stroke 10cm, 100r/min shaking culture 48h.
Monascus anka Nakazawa et sato shaking flask liquid state is cultivated: liquid amount 100mL(500mL triangular flask), inoculum size 5%.30 DEG C, reciprocal shaker 100r/min, incubation time 5 days.Shake flask fermentation look is worth: 50.65U/mL.
From above-described embodiment, in the Two Liquid Phases fermentation process of coupling situ extracting fermentation monascus yellow pigment, under equal fermentation condition, the key factor affecting monascus yellow pigment output and productive rate is the Two Liquid Phases coupling extractive fermentation system that medium component proportioning and extraction phase and minimum medium form.In Medium Proportion, higher carbon source concentration, higher inorganic nitrogen-sourced concentration can improve the yellow plain color valency of fermented liquid, and organic nitrogen source excessive concentration reduces yellow pigment output.Yellow plain color valency can be significantly improved when adding the extraction agent of more amount in fermention medium.Obtaining preferred process by research is with Monascus anka Nakazawa et sato JJLY-4A for fermented bacterium, when minimum medium (g/L) is: W-Gum 70, analysis for soybean powder 2, ammonium sulfate 6, NaNO34, yeast extract paste 2, corn starch 2, magnesium sulfate 0.8, potassium primary phosphate 1.2; Extraction phase (with the volume ratio of substratum): when soybean oil 10%, Semen Maydis oil 8%, caprylin 1%, two caprylin 0.3%, caprylin 0.2%, ferment 5 days, yellow plain color valency can reach 240-250U/mL.In addition, confirm after deliberation, method of the present invention can be applicable to all Monascus anka Nakazawa et sato (such as purple Monascus anka Nakazawa et sato Monascus purpureus CICC5022 that can produce monascus yellow pigment, Monascus ruber Monascus rubber CICC40711), but for different bacterial classifications, method of the present invention promotes that the degree of output is different.The output of Monascus anka Nakazawa et sato JJLY-4A fermentation monascus yellow pigment is higher than Monascus anka Nakazawa et sato ZH6's.Adopt this new fermentation process gained monascus yellow pigment look valency to be significantly higher than domestic existing level (110U/mL), and fermentation time reduction is 5 days.The method is while Monascus anka Nakazawa et sato fermentation synthetic colour product, by extraction agent by product monascus yellow pigment and thalline and separation of fermentative broth, timely releasing product negative feedback inhibition, thus significantly improve productive rate and the output of monascus yellow pigment, also be convenient to the separation and Extraction of later stage pigment simultaneously, extraction agent is recyclable recycling also, thus reduces production cost.In sum, method of the present invention is a kind of efficient, economic, monascus yellow pigment liquid state fermentation method of possessing novelty, has important practical value and economic worth at monascus yellow pigment production field.
Although the present invention with preferred embodiment openly as above; but it is also not used to limit the present invention, any person skilled in the art, without departing from the spirit and scope of the present invention; all can do various changes and modification, what therefore protection scope of the present invention should define with claims is as the criterion.
Claims (11)
1. a Two Liquid Phases fermentation process for coupling situ extracting fermentation monascus yellow pigment, is characterized in that, is that monascus yellow pigment is produced in fermentation by Monascus anka Nakazawa et sato seed access fermention medium; Described fermention medium is the double liquid phase extraction fermentation system of aqueous phase and extraction phase composition, and described aqueous phase is the minimum medium containing Monascus anka Nakazawa et sato growth desired nutritional composition, and described extraction phase contains glycerides.
2. method according to claim 1, is characterized in that, described minimum medium contains W-Gum 20-80g/L, analysis for soybean powder 0.05-10g/L, ammonium sulfate 3-10g/L, NaNO
32-6g/L, yeast extract paste 0.05-5g/L, corn starch 0.05-10g/L, zinc sulfate 0.1-0.5g/L, magnesium sulfate 0.5-1.5g/L, potassium primary phosphate 0.5-1.5g/L.
3. method according to claim 1, is characterized in that, described glycerides is triglyceride material, and the volume of interpolation accounts for the 10-40% of minimum medium volume.
4. method according to claim 3, is characterized in that, described triglyceride material is caprylin, three isocaprylic acid glyceryl ester, vegetables oil or its mixture.
5. method according to claim 3, is characterized in that, described glycerides also comprises di-glycerides material, and the interpolation time is that after red colouring agent for food, also used as a Chinese medicine seed liquor inoculates 24-72h, volume accounts for the 0.1-5% of minimum medium volume.
6. method according to claim 3, is characterized in that, described glycerides also comprises monoglyceride class material, and the interpolation time is that after red colouring agent for food, also used as a Chinese medicine seed liquor inoculates 24-72h, volume accounts for the 0.1-5% of minimum medium volume.
7. method according to claim 3, it is characterized in that, described glycerides also comprises di-glycerides and monoglyceride class material, and the interpolation time is that after red colouring agent for food, also used as a Chinese medicine seed liquor inoculates 24-72h, the cumulative volume of triglyceride and monoglyceride accounts for the 0.1-5% of minimum medium volume.
8. the method according to claim 5 or 7, is characterized in that, described di-glycerides material is in 1,3-DAG, 1,2-triglyceride, two caprylins or its mixture.
9. the method according to claim 6 or 7, is characterized in that, described monoglyceride class material is Capmul MCM C8.
10. method according to claim 4, is characterized in that, described vegetables oil comprises soybean oil, Semen Maydis oil, rapeseed oil, peanut oil, cottonseed meal oil, tea-seed oil or other edible oils and its mixture thereof.
11. methods according to claim 1, it is characterized in that, the condition that monascorubin is produced in described fermentation is: be equipped with in the 500mL triangular flask of 100mL fermention medium by the Monascus anka Nakazawa et sato liquid seeds being in logarithmic phase with the access of the inoculum size of volume ratio 5-10%, be 80 ~ 180r/min at rotating speed, under temperature 26 ~ 32 DEG C of conditions, shake flask fermentation cultivates 4-6d.
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| CN108273807A (en) * | 2017-12-19 | 2018-07-13 | 贵州航天乌江机电设备有限责任公司 | The cleaning method of autoclave body and pipeline in a kind of overcritical equipment |
| CN111248361A (en) * | 2020-02-05 | 2020-06-09 | 播恩生物技术股份有限公司 | Laying hen feed capable of increasing egg yolk degree and realizing accurate batching |
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| CN106755171A (en) * | 2017-02-09 | 2017-05-31 | 江南大学 | A kind of method for promoting red yeast rice strains liquid fermentation to produce red yeast rice citraurin |
| CN106755171B (en) * | 2017-02-09 | 2021-01-29 | 江南大学 | Method for promoting monascus liquid state fermentation to produce monascus orange pigment |
| CN106967082A (en) * | 2017-04-05 | 2017-07-21 | 江苏鸣生物股份有限公司 | A kind of method for extracting monascus yellow pigment |
| CN106967082B (en) * | 2017-04-05 | 2018-12-04 | 江苏一鸣生物股份有限公司 | A method of extraction monascus yellow pigment |
| CN108273807A (en) * | 2017-12-19 | 2018-07-13 | 贵州航天乌江机电设备有限责任公司 | The cleaning method of autoclave body and pipeline in a kind of overcritical equipment |
| CN108251457A (en) * | 2017-12-23 | 2018-07-06 | 北京工商大学 | A kind of composite extractant and the method for promoting monascorubin production using composite extractant |
| CN111248361A (en) * | 2020-02-05 | 2020-06-09 | 播恩生物技术股份有限公司 | Laying hen feed capable of increasing egg yolk degree and realizing accurate batching |
| CN111248361B (en) * | 2020-02-05 | 2024-02-02 | 播恩集团股份有限公司 | Chicken feed capable of increasing yolk chromaticity and realizing accurate ingredients of fermented pigment |
| CN114350526A (en) * | 2022-01-14 | 2022-04-15 | 北京工商大学 | Method for improving monascus pigment tone |
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