CN110184303A - A method of monascorubin is prepared by raw material of yellow serofluid - Google Patents
A method of monascorubin is prepared by raw material of yellow serofluid Download PDFInfo
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- CN110184303A CN110184303A CN201910536605.5A CN201910536605A CN110184303A CN 110184303 A CN110184303 A CN 110184303A CN 201910536605 A CN201910536605 A CN 201910536605A CN 110184303 A CN110184303 A CN 110184303A
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
The present invention provides a kind of methods for preparing monascorubin as raw material using yellow serofluid, it is raw material using yellow serofluid, addition different carbon source substance is made fermentation liquid and prepares monascorubin respectively, when adding glucose is carbon source, it is 862.8 ± 13.6U/g that fermentation, which measures glucose and is converted into the maximum yield of pigment,；When adding soluble starch is carbon source, it is 739.4 ± 7.48U/g that fermentation, which measures soluble starch and is converted into the maximum yield of pigment,；When adding old rice powder is carbon source, it is 1066 ± 18.75U/g that fermentation, which measures old rice powder and is converted into the maximum yield of pigment,.The present invention adds different carbon source as raw material using yellow serofluid and produces monascorubin, preferably solves using yellow serofluid as the wasting of resources and problem of environmental pollution caused by waste discharge, extends industrial chain, realizing turns waste into wealth.Preparation process of the present invention is simple, low in cost, and waste utilization is economic and environment-friendly, high production efficiency, has significant economic benefit.
Description
Technical field
The present invention relates to monascorubin production technical fields, particularly, are related to one kind by raw material of yellow serofluid and prepare red yeast rice
The method of pigment.
Background technique
Monascorubin is the polyketone class cometabolism generated by monascus ruber by solid state fermentation or liquid submerged fermentation
Product.It is being widely used in food color always over past more than 2,000 years, is also playing in the food industry now huge
Big effect, such as monascorubin suffer from extensively inside sausage, fermented bean curd, traditional fermentation yellow rice wine, marine product, meat products
Ground application, meanwhile, monascorubin is recognized as in Europe can substitute NO3-N and NO2-N partially for meat
The hyperchromic and storage of product, for example it is used for the hyperchromic and preservation of sausage and ham.Other than being widely used as food color, red yeast rice
Pigment has many promising applications also in cosmetics, textile, dyeing.For example, due to its chromatic colour and
The ability of the harmful black light of fabulous absorption and can be used as the additive in cosmetics.They can also be used for production printer
With ink, cotton yarn, leather and wool are contaminated, or even improves the efficiency of solar panel, as the novel increasing in solar battery
Infection material.
Yellow serofluid is one of the byproduct of bean product production industry, wherein mainly containing protein, amino acid, fat, nothing
The ingredients such as machine salt, vitamin are suitble to the growth and breeding of microorganism.But current many bean product enterprises are limited due to equipment and technology etc.
System, often using yellow serofluid as waste direct emission, this not only wastes a large amount of food resource, also causes to water body environment
Certain pollution.Therefore, correct, rationally, effectively utilize such byproduct, be of great practical significance.It utilizes
Yellow serofluid is that fermenting raw materials produce monascorubin, extends industrial chain, realizing turns waste into wealth, and reduces environmental pollution, is referring to
Lead enterprise scien` production, increase added value of product etc. and have a very important significance, be yellow serofluid further exploitation with
Using having established theoretical basis.
Article (middle figure classification number: TS202.3；Document code: A；Article number: 100-9973 (2017) 01-0044-
03) a kind of method using bean curd yellow pulp water fermentation monascorubin is disclosed, this method is to focus on to mention by optimization of fermentation conditions
High monascorubin yield, optimal conditions include carbon source, nitrogen source, inorganic salts, pH, fermentation time, many indexs such as revolving speed；This hair
It is bright only to add single carbon source, and the recycling of aged rice is combined, it can turn up to refuse reclamation and cheap substrates high level
Metaplasia produces monascorubin (compound pigment), and simple process is easy to operate, and economic cost is cheap, is the waste benefit of yellow serofluid
A kind of feasible scheme is provided with the high-valued production monascorubin with old rice.
Summary of the invention
It is an object of that present invention to provide a kind of methods for preparing monascorubin as raw material using yellow serofluid, to solve yellow serofluid battalion
Feeding components utilising is insufficient, yellow serofluid discharge causes the technical problems such as environmental pollution.
To achieve the above object, the present invention provides a kind of method for preparing monascorubin as raw material using yellow serofluid, be with
Yellow serofluid is raw material, and as carbon source fermentation medium is made, then by this reality in addition glucose, soluble starch or old rice powder
Monascus parpureus Went (Monascus.purpureus) the M183 seed liquor for testing the chromogenic element of a plant height of room preservation accesses the fermentation
In culture medium, ferment to get monascorubin；Monascus parpureus Went (Monascus.purpureus) M183, on April 1st, 2018
It is preserved in the China typical culture collection center (CCTCC) positioned at Wuhan University, deposit number is CCTCC M
2018224。
Preferably, the additive amount of carbon source is 80g/L, i.e. adds 80g carbon source in every liter of yellow serofluid.
Preferably, fermentation medium the preparation method comprises the following steps: using yellow serofluid as raw material, addition glucose, soluble starch or
Old rice powder sterilizes 20 minutes under the conditions of 121 DEG C after mixing as carbon source, and cooling is spare.
Preferably, monascin seed liquor the preparation method is as follows: after actication of culture in seed culture medium, 30 DEG C,
The culture of 150rpm isothermal vibration 3 days to get the monascin seed liquor.
It is further preferred that seed culture medium the preparation method is as follows: soluble starch 30g/L, glucose 60g/L, egg
White peptone 20g/L sterilizes 20 minutes after configuring by concentration in 121 DEG C, and cooling is spare.
It is further preferred that the method for actication of culture are as follows: the monascus parpureus Went (Monascus. that two environmental protection of picking is deposited
Purpureus) M183 is uniformly drawn on PDA culture medium surface, 30 DEG C dark culturing 7 days.
Still more preferably, PDA culture medium the preparation method is as follows: potato 200g is removed the peel, it is cooked to take juice, glucose
20g, agar 20g, adds pure water to be settled to 1000mL, sterilizes 20 minutes, is poured into sterilized culture dish while hot in 121 DEG C, cold
But it solidifies spare.
Preferably, the inoculum concentration of monascin seed liquor in the fermentation medium is 10% (V/V).
Preferably, fermentation condition are as follows: 30 DEG C, dark culturing 7 days under the conditions of 150rpm.
The invention has the following advantages:
The present invention is raw material using yellow serofluid, and addition different carbon source substance is made fermentation liquid and prepares monascorubin respectively, when
When addition glucose is carbon source, it is 862.8 ± 13.6U/g that fermentation, which measures glucose and is converted into the highest yield rate of pigment,；When adding
Add soluble starch be carbon source when, fermentation measure soluble starch be converted into pigment maximum yield be 739.4 ± 7.48U/g；
When adding old rice powder is carbon source, it is 1066 ± 18.75U/g that fermentation, which measures old rice powder and is converted into the maximum yield of pigment,.This hair
It is bright to add different carbon source production monascorubin by raw material of yellow serofluid, it preferably solves using yellow serofluid as waste discharge institute
Caused by the wasting of resources and problem of environmental pollution, extend industrial chain, realizing turns waste into wealth.Preparation process of the present invention is simple,
Low in cost, waste utilization is economic and environment-friendly, high production efficiency, has significant economic benefit.
Other than objects, features and advantages described above, there are also other objects, features and advantages by the present invention.
The present invention is further detailed explanation below.
Detailed description of the invention
Fig. 1 is all band scanning figure of fermentation liquid, illustrates the maximum absorption band feelings of monascorubin fermentation liquid in the present invention
Condition.
Specific embodiment
The embodiment of the present invention is described in detail below in conjunction with attached drawing, but the present invention can be limited according to claim
Fixed and covering multitude of different ways is implemented.
Monascorubin detection method:
Fermentation liquid is centrifuged 5 minutes in 10000rpm, supernatant dilutes suitable multiple, its extinction is measured under specific wavelength
Value, measures its extracellular color value；Precipitating plus volumetric concentration are that 70% alcohol is settled to original fermentation liquor volume, in 60 DEG C of water-bath extractions 1
Hour, 10000rpm is centrifuged 5 minutes, and supernatant dilutes suitable multiple, its light absorption value is measured under specific wavelength, it is intracellular to measure its
Color value.
Color value=(OD420+OD500) × extension rate；
Total color value=color value intracellular+extracellular color value.
Above-mentioned specific wavelength according to fermentation liquid carry out all band scanning determine, concrete operations be by fermentation liquid in
12000rpm is centrifuged 5min, takes supernatant, pure water is added suitably to be diluted.Suitable dilution gradient is selected, draws 100 μ L in enzyme
In target, using pure water as blank control, all band scanning is carried out in wave-length coverage 300-800nm, finally according to scanning result
Maximum absorption band determines the measurement wavelength of fermentation liquid color value.As shown in Figure 1, determining that fermentation liquid has maximum at 420nm and 500nm
Absorption peak.
Embodiment 1:
This example demonstrates that only adding the side that glucose prepares monascorubin as carbon source through fermentation using yellow serofluid as raw material
Method, the method is as follows:
It adds 80g/L, 100g/L, 120g/L glucose respectively in yellow serofluid and fermentation medium is made, sterilize in 121 DEG C
20 minutes, then aseptically cooled to room temperature accesses monascus ruber seed liquid by the inoculum concentration of 10% (V/V),
In dark culturing 7 days on 30 DEG C, the constant-temperature table of 150rpm condition.Then pigment fermentation liquid is centrifuged 5 minutes in 10000rpm,
Supernatant dilutes suitable multiple, finally makes light absorption value between 0.2-0.6, its extinction is measured under 420nm and 500nm wavelength
Value, measures its extracellular color value；Precipitating plus 70% alcohol are settled to original fermentation liquor volume, extract 1 hour in 60 DEG C of water-baths,
10000rpm is centrifuged 5 minutes, and supernatant dilutes suitable multiple, finally makes light absorption value between 0.2-0.6, in 420nm and 500nm
Its light absorption value is measured under wavelength, measures its color value intracellular；Fermentation liquid color value is calculated by light absorption value multiplied by extension rate (intracellular (outer)
Color value=OD420 × extension rate+OD500 × extension rate), color value intracellular is total color value (total color value plus extracellular color value
=color value+extracellular color value intracellular), fermentation liquid color value is as shown in the table:
It is that carbon source prepares monascorubin situation that table 1, which adds glucose by raw material of yellow serofluid,
From the experimental results, when adding 80g/L glucose, glucose be converted into pigment yield be 862.8 ±
13.6U/g, when adding 100g/L glucose, the yield that glucose is converted into pigment is 701 ± 7U/g, adds 120g/L glucose
When, the yield that glucose is converted into pigment is 501 ± 6.65U/g, thus when adding 80g/L glucose, transformation efficiency highest, warp
Benefit of helping is most obvious.
Embodiment 2:
This example demonstrates that only adding soluble starch using yellow serofluid as raw material as carbon source through fermentation and preparing monascorubin
Method, the method is as follows:
It adds 80g/L, 100g/L, 120g/L soluble starch respectively in yellow serofluid and fermentation medium is made, in 121 DEG C
Then aseptically sterilizing 20 minutes, cooled to room temperature access monascus specie by the inoculum concentration of 10% (V/V)
Sub- liquid, in dark culturing 7 days on 30 DEG C, the constant-temperature table of 150rpm condition.Then pigment fermentation liquid is centrifuged 5 in 10000rpm
Minute, supernatant dilutes suitable multiple, finally makes light absorption value between 0.2-0.6, measures it under 420nm and 500nm wavelength
Light absorption value measures its extracellular color value；Precipitating plus 70% alcohol are settled to original fermentation liquor volume, extract 1 hour in 60 DEG C of water-baths,
10000rpm is centrifuged 5 minutes, and supernatant dilutes suitable multiple, finally makes light absorption value between 0.2-0.6, in 420nm and 500nm
Its light absorption value is measured under wavelength, measures its color value intracellular；Fermentation liquid color value is calculated by light absorption value multiplied by extension rate (intracellular (outer)
Color value=OD420 × extension rate+OD500 × extension rate), color value intracellular is total color value (total color value plus extracellular color value
=color value+extracellular color value intracellular), fermentation liquid color value is as shown in the table:
It is that carbon source prepares monascorubin situation that table 2, which adds soluble starch by raw material of yellow serofluid,
From the experimental results, when adding 80g/L soluble starch, the yield that soluble starch is converted into pigment is
739.4 ± 7.48U/g, when adding 100g/L soluble starch, the yield that soluble starch is converted into pigment is 618.8 ± 12U/
G, when adding 120g/L soluble starch, the yield that soluble starch is converted into pigment is 431.2 ± 7.47U/g, therefore when addition
When 80g/L soluble starch, transformation efficiency highest, economic benefit is most obvious.
Embodiment 3:
This example demonstrates that only adding the side that old rice powder prepares monascorubin as carbon source through fermentation using yellow serofluid as raw material
Method, the method is as follows:
It adds 80g/L, 100g/L, 120g/L old rice powder respectively in yellow serofluid and fermentation medium, old rice powder source is made
Jinjian rice industry long-grained nonglutinous rice and natural storage 5 years or more are bought in market, crushes and simultaneously crosses 60 mesh standard sieves gained, culture is based on 121 DEG C
Then aseptically sterilizing 20 minutes, cooled to room temperature access monascus specie by the inoculum concentration of 10% (V/V)
Sub- liquid, in dark culturing 7 days on 30 DEG C, the constant-temperature table of 150rpm condition.Then pigment fermentation liquid is centrifuged 5 in 10000rpm
Minute, supernatant dilutes suitable multiple, finally makes light absorption value between 0.2-0.6, measures it under 420nm and 500nm wavelength
Light absorption value measures its extracellular color value；Precipitating plus 70% alcohol are settled to original fermentation liquor volume, extract 1 hour in 60 DEG C of water-baths,
10000rpm is centrifuged 5 minutes, and supernatant dilutes suitable multiple, finally makes light absorption value between 0.2-0.6, in 420nm and 500nm
Its light absorption value is measured under wavelength, measures its color value intracellular；Fermentation liquid color value is calculated by light absorption value multiplied by extension rate (intracellular (outer)
Color value=OD420 × extension rate+OD500 × extension rate), color value intracellular is total color value (total color value plus extracellular color value
=color value+extracellular color value intracellular), fermentation liquid color value is as shown in the table:
It is that carbon source prepares monascorubin situation that table 3, which adds old rice powder by raw material of yellow serofluid,
From the experimental results, when adding 80g/L old rice powder, old rice powder be converted into pigment yield be 1066.0 ±
18.75U/g；When old rice powder additive amount is greater than 80g/L, the post-fermentation liquid that sterilizes is unfavorable for fermenting at paste, so fermentation color value
It is low, it is unfavorable for old rice powder and is converted to pigment；Add 100g/L old rice powder when, old rice powder be converted into pigment yield be 112.5 ±
5U/g, when adding 120g/L old rice powder, the yield that old rice powder is converted into pigment is 109.6 ± 5.8U/g, therefore when addition 80g/L is old
When rice flour, transformation efficiency highest, economic benefit is most obvious.
Comparative example
This example demonstrates that not adding other any substances only using yellow serofluid as raw material as fermentation liquid and preparing monascorubin
Method, the method is as follows:
Yellow serofluid is dispensed, is sterilized 20 minutes in 121 DEG C, cooled to room temperature, then aseptically, by 10%
(V/V) inoculum concentration accesses monascus ruber seed liquid, in dark culturing 7 days on 30 DEG C, the constant-temperature table of 150rpm condition.Then
Pigment fermentation liquid is centrifuged 5 minutes in 10000rpm, supernatant dilutes suitable multiple, and finally make light absorption value between 0.2-0.6,
Its light absorption value is measured under 420nm and 500nm wavelength, measures its extracellular color value；Precipitating plus 70% alcohol are settled to original fermentation liquor
Volume extracts 1 hour in 60 DEG C of water-baths, and 10000rpm is centrifuged 5 minutes, and supernatant dilutes suitable multiple, finally light absorption value is made to exist
Between 0.2-0.6, its light absorption value is measured under 420nm and 500nm wavelength, measures its color value intracellular；Fermentation liquid color value presses extinction
Value calculates ((outer) color value=OD420 × extension rate+OD500 × extension rate intracellular) multiplied by extension rate, and color value intracellular adds
The outer color value of upper eye lid is total color value (total color value=color value intracellular+extracellular color value).
It is only 5.3 ± 0.4U/mL, 2.1 ± 0.3g/L of biomass that fermentation liquid color value is measured after fermentation, the results showed that, Huang slurry
The protein contained in water, amino acid, fat, inorganic salts, the ingredients such as vitamin, the growth and breeding of suitable Monascus, but due to
It lacks carbon source, causes Monascus that cannot sufficiently grow, and pigment synthesis lacks enough carbon source materials, leads to pigment production pole
It is low, so proper amount of carbon source can be added wherein, guarantee Monascus can normal growth wherein, and efficiently synthesize pigment.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.All within the spirits and principles of the present invention, made any to repair
Change, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (8)
1. a kind of method for preparing monascorubin as raw material using yellow serofluid, which is characterized in that be to add Portugal using yellow serofluid as raw material
As carbon source fermentation medium is made, then by deposit number CCTCC M's 2018224 in grape sugar, soluble starch or old rice powder
The monascin seed liquor that monascus parpureus Went (Monascus.purpureus) M183 is obtained accesses in the fermentation medium, hair
Ferment is to get monascorubin.
2. the method according to claim 1, wherein fermentation medium the preparation method comprises the following steps: with yellow serofluid be original
Material, addition glucose, soluble starch or old rice powder sterilize 20 minutes under the conditions of 121 DEG C after mixing as carbon source, and cooling is standby
With.
3. the method according to claim 1, wherein monascin seed liquor the preparation method is as follows: strain is living
After change in seed culture medium, 30 DEG C, the culture of 150rpm isothermal vibration 3 days to get the monascin seed liquor.
4. according to the method described in claim 3, it is characterized in that, seed culture medium the preparation method is as follows: soluble starch
30g/L, glucose 60g/L, peptone 20g/L sterilize 20 minutes after configuring by concentration in 121 DEG C, and cooling is spare.
5. according to the method described in claim 3, it is characterized in that, the method for actication of culture are as follows: the purple that two environmental protection of picking is deposited
Monascus (Monascus.purpureus) M183 is uniformly drawn on PDA culture medium surface, 30 DEG C dark culturing 7 days.
6. according to the method described in claim 5, it is characterized in that, PDA culture medium the preparation method is as follows: potato 200g is gone
Skin, cooked to take juice, glucose 20g, agar 20g adds pure water to be settled to 1000mL, sterilizes 20 minutes, is poured into while hot in 121 DEG C
In the culture dish of sterilizing, cooled and solidified is spare.
7. the method according to claim 1, wherein the inoculum concentration of monascin seed liquor in the fermentation medium
It is 10%.
8. the method according to claim 1, wherein fermentation condition are as follows: 30 DEG C, dark culturing under the conditions of 150rpm
7 days.
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| CN109234318A (en) * | 2018-09-25 | 2019-01-18 | 中南林业科技大学 | A method of improving monascus ruber exo-cell pigment |
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2019
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|---|---|---|---|---|
| US20020061547A1 (en) * | 2000-09-29 | 2002-05-23 | Yen-Lin Chen | Monascus prupureus mutant and its use in preparing yellow pigment |
| CN104498528A (en) * | 2014-12-25 | 2015-04-08 | 天津科技大学 | Monascus fermentation method for low-yield citrinin and high-yield monascorubin |
| CN108719991A (en) * | 2018-06-07 | 2018-11-02 | 南京工业大学 | A method of preparing the red yeast rice containing Bacillus natto using bean dregs and bean curd yellow pulp water |
| CN108841731A (en) * | 2018-07-23 | 2018-11-20 | 中南林业科技大学 | The Monascus Strains of high yield pigment and its application |
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