CN106191130A - A kind of production technology of Natural Pigments Produced by Microorganisms - Google Patents
A kind of production technology of Natural Pigments Produced by Microorganisms Download PDFInfo
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- CN106191130A CN106191130A CN201610575195.1A CN201610575195A CN106191130A CN 106191130 A CN106191130 A CN 106191130A CN 201610575195 A CN201610575195 A CN 201610575195A CN 106191130 A CN106191130 A CN 106191130A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B61/00—Dyes of natural origin prepared from natural sources, e.g. vegetable sources
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B67/00—Influencing the physical, e.g. the dyeing or printing properties of dyestuffs without chemical reactions, e.g. by treating with solvents grinding or grinding assistants, coating of pigments or dyes; Process features in the making of dyestuff preparations; Dyestuff preparations of a special physical nature, e.g. tablets, films
- C09B67/0096—Purification; Precipitation; Filtration
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
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- General Engineering & Computer Science (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses the production technology of a kind of Natural Pigments Produced by Microorganisms, pigment, purification refine pigment and nine steps of concentration are slightly put forward including strain culturing, temperature setting, time setting, mycelia inactivation, dissolving pigment, pigment test, dissolving, compared with prior art, present invention process is applied widely；Technological operation is simple, good product quality, production efficiency are high；The profit that technique production cost is low, produce is high；Technique production procedure economizes on resources, pollution-free, Environmental compatibility good；Technique can meet laboratory research needs, it is also possible to meets the needs of enterprise's factorial praluction.To a certain extent solve utilize microbial fungi, actinomycetes, antibacterial produce natural pigment typically face the problem how improving yield, how the problem of purification refine pigment, the problem how reducing cost, the stability of pigment produced, safety etc. require basic problem.
Description
Technical field
The present invention relates to microorganism field, particularly relate to the production technology of a kind of Natural Pigments Produced by Microorganisms.
Background technology
At present, the domestic and international commonly used method directly extracting natural pigment from plant tissue produces pigment, uses micro-
Although the research that biological culture produces natural pigment early has started to, and it is fruitful and has applied to human being's production life.But
Because microbe species is various, biological characteristics is ever-changing, and natural pigment complicated component, character is various, whether real
Test in room research or production application, for separate sources, natural pigment that character is different, general use extraction, produce work
Skill is different.So, up to the present, the most ununified, that be applicable to most chromogenic element microbial strains, for
Microorganism culturing produces the technological standards of natural pigment.
Summary of the invention
The purpose of the present invention is that the production providing a kind of Natural Pigments Produced by Microorganisms in order to solve the problems referred to above
Technique.
The present invention is achieved through the following technical solutions above-mentioned purpose:
The present invention comprises the following steps:
(1) use solid microbe culture medium that the secretion fungus of natural pigment or actinomycetes or bacterial isolates are cultivated, Gu
The thickness of body culture medium should be set according to the ability of required incubation time and bacterial strain production pigment, culture medium thickness
It is preferred for containing the 1/2 of the vessel volume of culture medium；
(2) spy of the pigment that the condition such as cultivation temperature, humidity should be produced according to the growth characteristic of the bacterial strain cultivated and bacterial strain
Property set, fungus, actinomycetes cultivation temperature are 28 DEG C, and antibacterial is 37 DEG C；
(3) according to the different abilities of the chromogenic element of each bacterial strain, the optimal incubation time cultivated needed for bacterial strain production pigment is measured, so
Each bacterial strain is cultivated by the optimal incubation time of rear employing, and incubation time is more than 5 days；
(4) after cultivation terminates, separating mycelia and the solid medium of full pigment, mycelia inactivates or saves backup；It is full of color
The solid medium of element is continued to employ, and is stored in the environment identical with strain culturing temperature；
(5) method of the solid medium machinery being full of pigment is fully pulverized, and makes that pigment is quick, be completely dissolved in pigmentolysis
Agent；
(6) take the culture medium crushed on a small quantity, test the dissolution properties of pigment therein, mainly determine that pigment water solublity, alcohol are molten
Property, ester dissolubility etc., in order to dissolve and slightly carry pigment；
(7) dissolving slightly carries pigment: the solid medium containing pigment crushed and pigmentolysis agent are mixed by 1:50 volume ratio
Even and shake more than 2 hours, allow pigment be completely dissolved in extractant, slightly carry pigment, separate pigment crude extract and solid training
Support base, repeat to extract repeatedly, mixed extract；
(8) purification refine pigment, purification process has the extractant extraction pigment that two kinds: A employing is immiscible with pigmentolysis agent
Crude extract, B uses the technological means purification pigments such as chromatograph, obtains the relatively good pigment of purification effect；
(9) rotatory vacuum evaporation, lyophilisation, constant temperature baking technology is used to be concentrated by pigment solution good for purification or system
Become dry product.
In described step (1), solid microbe culture medium is solid PDA medium, solid Czapek's medium, solid LB training
Support base, the one of solid beef-protein medium.
The solid medium being full of pigment in described step (5) contains moisture in the case of not drying, and is to be ground into
Powder, if the most broken the most permissible；Solid medium containing moisture and pigment is being cultivated temperature with microbial strains
Spend constant temperature drying under identical temperature conditions or lower temperature, powder can be ground into.
In described step (7), pigmentolysis agent is distilled water, dehydrated alcohol or ethyl acetate.
The beneficial effects of the present invention is:
The present invention is the production technology of a kind of Natural Pigments Produced by Microorganisms, and compared with prior art, present invention process is suitable for model
Enclose extensively；Technological operation is simple, good product quality, production efficiency are high；The profit that technique production cost is low, produce is high；Technique is raw
Runoff yield journey economizes on resources, pollution-free, Environmental compatibility good；Technique can meet laboratory research needs, it is also possible to meets enterprise
The needs of factorial praluction.Solve to a certain extent to utilize microbial fungi, actinomycetes, antibacterial to produce the general face of natural pigment
The problem how improving yield faced, the how problem of purification refine pigment, the problem how reducing cost, the pigment that produced
Stability, safety etc. require basic problem.
Detailed description of the invention
The invention will be further described below:
The present invention comprises the following steps:
(1) use solid microbe culture medium that the secretion fungus of natural pigment or actinomycetes or bacterial isolates are cultivated, Gu
The thickness of body culture medium should be set according to the ability of required incubation time and bacterial strain production pigment, culture medium thickness
It is preferred for containing the 1/2 of the vessel volume of culture medium；
(2) spy of the pigment that the condition such as cultivation temperature, humidity should be produced according to the growth characteristic of the bacterial strain cultivated and bacterial strain
Property set, fungus, actinomycetes cultivation temperature are 28 DEG C, and antibacterial is 37 DEG C；
(3) according to the different abilities of the chromogenic element of each bacterial strain, the optimal incubation time cultivated needed for bacterial strain production pigment is measured, so
Each bacterial strain is cultivated by the optimal incubation time of rear employing, and incubation time is more than 5 days；
(4) after cultivation terminates, separating mycelia and the solid medium of full pigment, mycelia inactivates or saves backup；It is full of color
The solid medium of element is continued to employ, and is stored in the environment identical with strain culturing temperature；
(5) method of the solid medium machinery being full of pigment is fully pulverized, and makes that pigment is quick, be completely dissolved in pigmentolysis
Agent；
(6) take the culture medium crushed on a small quantity, test the dissolution properties of pigment therein, mainly determine that pigment water solublity, alcohol are molten
Property, ester dissolubility etc., in order to dissolve and slightly carry pigment；
(7) dissolving slightly carries pigment: the solid medium containing pigment crushed and pigmentolysis agent are mixed by 1:50 volume ratio
Even and shake more than 2 hours, allow pigment be completely dissolved in extractant, slightly carry pigment, different pigmentolysis character are different, mixed
Close liquid and reach the time difference of degree of dissolution saturation, so the concussion time should reach according to pigment to be produced in lytic agent
The situation of degree of dissolution saturation sets.Separate pigment crude extract and solid medium, repeat to extract repeatedly, mixed extract；
(8) purification refine pigment, purification process has the extractant extraction pigment that two kinds: A employing is immiscible with pigmentolysis agent
Crude extract, the method purification effect is relatively poor；B uses the technological means purification pigments such as chromatograph, obtains purification effect relatively
Good pigment；Because natural pigment complicated component, even if using the state-of-the-art technology means such as chromatograph the most simply to make pigment reach one
Of a relatively high purity, is not to reach absolutely purity；
(9) rotatory vacuum evaporation, lyophilisation, constant temperature baking technology is used to be concentrated by pigment solution good for purification or system
Become dry product.Depending on this is according to experiment or Production requirement or requirement, the temperature concentrate, being dried not can exceed that microbial strains cultivates temperature
The optimum temperature that degree and pigment stability require.Arriving this, the employing solid medium cultivating microorganism of present invention design produces sky
So all technological processes of pigment just finish.
In described step (1), solid microbe culture medium is solid PDA medium, solid Czapek's medium, solid LB training
Support base, the one of solid beef-protein medium.
The solid medium being full of pigment in described step (5) contains moisture in the case of not drying, and is to be ground into
Powder, if the most broken the most permissible；Solid medium containing moisture and pigment is being cultivated temperature with microbial strains
Spend constant temperature drying under identical temperature conditions or lower temperature, powder can be ground into.
In described step (7), pigmentolysis agent is distilled water, dehydrated alcohol or ethyl acetate.
The present invention design a set of applied widely, efficiency is high, low cost, simple to operate utilize microbial fungi,
Microorganism actinomycetes, microbial bacterial solid culture produce natural pigment production technology:
1. fungus, actinomycetes, antibacterial are the major microorganisms producing natural pigment.Fungus, actinomycetes, antibacterial can be from nature
Biology, abiotic component separate obtain, purification is simple, and culture environment is required relatively low by it, and growth cycle is short,
Secretion pigment ability is strong, can be suitable for being applied to factory of enterprise metaplasia with manual control fungus, actinomycetes, bacterial growth and metabolism
Produce natural pigment.
2. fungus, actinomycetes, bacterial micro-organism can secrete the pigment of different colours.Wherein most pigments is steady
Fixed, use safety.The pigment of these three Institute of Micro-biology secretion comprises the compositions such as saccharide, ketone, terpenoid, quinones, many one-tenth more
Divide and there is nutrition and pharmacological action, when pigment uses as coloring agent, have both the most simultaneously and the function of nutrition and health care is provided
Or the effect of disease preventing and treating.
3. produce natural pigment typically face 4 big class problems currently with microbial fungi, actinomycetes, antibacterial: (1) micro-life
Thing fungus, actinomycetes, antibacterial culturing produce the problem how improving yield in pigment technique；(2) microbial fungi, actinomycetes,
Antibacterial culturing produces in pigment technique how problem (3) microbial fungi of purification refine pigment, actinomycetes, antibacterial culturing and produces
How pigment technique reduces problem (4) microbial fungi of cost, actinomycetes, the stability of pigment of antibacterial production, peace
Quan Xing etc. require problem.Front 3 problems all fully belong to production technology problem, and the 4th problem relates to the stability problem of product.
The ingenious part of the production technology of present invention design is that and can solve this 4 problems to a certain extent: first, the present invention
Use solid microbe culture medium culturing micro-organisms pigment: (a) uses solid microbe culture medium culturing micro-organisms
Pigment, solid culture base density is big, when being filled with pigment, cultivates compared with the microbial liquid of the same volume of full pigment
The amount of the pigment that base contains is many, so productivity is high；B the solid during () microbial mycelial inherently pigment production is miscellaneous
Matter, during producing pigment by solid medium cultivating microorganism, microorganism mycelia is constantly in the upper table of solid medium
Face, separates mycelium and solid medium is simple and abundant, just just with the solid of full pigment in pigment production technique
Culture medium part, reduces impurity；(c) many microbial mycelial inherently have color (particularly fungus and actinomycetes) and
How different mycelium color and the color of pigment produced by it be, if producing pigment by fluid medium cultivating microorganism,
The pigment of microbial mycelial itself also can dissolve entrance culture fluid, affects pigment quality, uses solid medium to cultivate micro-life
Thing, microbial mycelial and its pigment relative separation produced, so its pigment production the most well avoids microorganism mycelia
The pigmentolysis of body itself enters this defect of culture fluid.D () culture medium is during cultivating microorganism, its composition is not
Being absorbed by the micro-organisms completely, if producing pigment by liquid medium cultivating microorganism, the medium component amount of being is maximum " miscellaneous
Matter ".The present invention uses solid medium cultivating microorganism to produce pigment, and in solid medium, the agarose of condensation dissolves hardly
In common lytic agent, and some compositions in the solid medium condensed are fixed in agarose, seldom dissolve entrance
Pigmentolysis agent, so pigment purity is high.Secondly, the present invention uses at microorganism culturing temperature or lower temperature purification, dense
Contracting, dry pigment, it is to avoid the higher temperature impact on pigment.Finally, the present invention use purification, concentrate, be dried pigment skill
Art is the Modern News such as liquid-liquid extraction, chromatograph, rotatory vacuum evaporation, lyophilisation, ripe technology, effective, efficiency
High.
The ultimate principle of the present invention and principal character and advantages of the present invention have more than been shown and described.The technology of the industry
Personnel, it should be appreciated that the present invention is not restricted to the described embodiments, simply illustrating this described in above-described embodiment and description
The principle of invention, without departing from the spirit and scope of the present invention, the present invention also has various changes and modifications, and these become
Change and improvement both falls within scope of the claimed invention.Claimed scope by appending claims and
Equivalent defines.
Claims (4)
1. the production technology of a Natural Pigments Produced by Microorganisms, it is characterised in that comprise the following steps:
(1) use solid microbe culture medium that the secretion fungus of natural pigment or actinomycetes or bacterial isolates are cultivated, Gu
The thickness of body culture medium should be set according to the ability of required incubation time and bacterial strain production pigment, culture medium thickness
To contain 1/2 being preferred of the vessel volume of culture medium；
(2) spy of the pigment that the condition such as cultivation temperature, humidity should be produced according to the growth characteristic of the bacterial strain cultivated and bacterial strain
Property set, fungus, actinomycetes cultivation temperature are 28 DEG C, and antibacterial is 37 DEG C；
(3) according to the different abilities of the chromogenic element of each bacterial strain, the optimal incubation time cultivated needed for bacterial strain production pigment is measured, so
Each bacterial strain is cultivated by the optimal incubation time of rear employing, and fungus, actinomycetic incubation time were more than 5 days；
(4) after cultivation terminates, separating mycelia and the solid medium of full pigment, mycelia inactivates or saves backup；It is full of color
The solid medium of element is continued to employ, and is stored in the environment identical with strain culturing temperature；
(5) method of the solid medium machinery being full of pigment is fully pulverized, and makes that pigment is quick, be completely dissolved in pigmentolysis
Agent；
(6) take the culture medium crushed on a small quantity, test the dissolution properties of pigment therein, mainly determine that pigment water solublity, alcohol are molten
Property, ester dissolubility etc., in order to dissolve and slightly carry pigment；
(7) dissolving slightly carries pigment: the solid medium containing pigment crushed and pigmentolysis agent are mixed by 1:50 volume ratio
Even and shake more than 2 hours, allow pigment be completely dissolved in extractant, slightly carry pigment, separate pigment crude extract and solid training
Support base, repeat to extract repeatedly, mixed extract；
(8) purification refine pigment, purification process has the extractant extraction pigment that two kinds: A employing is immiscible with pigmentolysis agent
Crude extract, B uses the technological means purification pigments such as chromatograph, obtains the relatively good pigment of purification effect；
(9) rotatory vacuum evaporation, lyophilisation, constant temperature baking technology is used to be concentrated by pigment solution good for purification or system
Become dry product.
The production technology of Natural Pigments Produced by Microorganisms the most according to claim 1, it is characterised in that: described step (1)
Middle solid microbe culture medium is solid PDA medium, solid Czapek's medium, solid LB media, solid Carnis Bovis seu Bubali cream albumen
The one of peptone culture medium.
The production technology of Natural Pigments Produced by Microorganisms the most according to claim 1, it is characterised in that: described step (5)
In be full of the solid medium of pigment in the case of not drying containing moisture, be cannot pulverize Powdered, if the most broken
Broken the most permissible；By containing moisture and the solid medium of pigment in the temperature conditions identical with microbial strains cultivation temperature or more
Constant temperature drying under low temperature, can be ground into powder.
The production technology of Natural Pigments Produced by Microorganisms the most according to claim 1, it is characterised in that: described step (7)
Middle pigmentolysis agent is distilled water, dehydrated alcohol or ethyl acetate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610575195.1A CN106191130A (en) | 2016-07-20 | 2016-07-20 | A kind of production technology of Natural Pigments Produced by Microorganisms |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610575195.1A CN106191130A (en) | 2016-07-20 | 2016-07-20 | A kind of production technology of Natural Pigments Produced by Microorganisms |
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| CN106191130A true CN106191130A (en) | 2016-12-07 |
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| CN201610575195.1A Pending CN106191130A (en) | 2016-07-20 | 2016-07-20 | A kind of production technology of Natural Pigments Produced by Microorganisms |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110983816A (en) * | 2019-11-07 | 2020-04-10 | 浙江理工大学 | Application of actinomycin |
| CN112760240A (en) * | 2021-03-23 | 2021-05-07 | 安徽农业大学 | Method for extracting natural green pigment from penicillium fungi |
| CN112778792A (en) * | 2021-03-08 | 2021-05-11 | 安徽农业大学 | Method for extracting natural green pigment from trichoderma fungus |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101235353A (en) * | 2007-12-14 | 2008-08-06 | 华南理工大学 | Monascus mutant and method for preparing flavochrome by fermenting the same |
| CN101942397A (en) * | 2010-09-13 | 2011-01-12 | 东莞市天益生物工程有限公司 | Monascus mutant strain and method for preparing monascus yellow pigment by submerged fermentation thereof and article thereof |
-
2016
- 2016-07-20 CN CN201610575195.1A patent/CN106191130A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101235353A (en) * | 2007-12-14 | 2008-08-06 | 华南理工大学 | Monascus mutant and method for preparing flavochrome by fermenting the same |
| CN101942397A (en) * | 2010-09-13 | 2011-01-12 | 东莞市天益生物工程有限公司 | Monascus mutant strain and method for preparing monascus yellow pigment by submerged fermentation thereof and article thereof |
Non-Patent Citations (1)
| Title |
|---|
| 牛国强: "红曲菌橙、黄色素的发酵及提取技术研究", 《中国优秀硕士学位论文全文数据库（工程科技Ⅰ辑）》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110983816A (en) * | 2019-11-07 | 2020-04-10 | 浙江理工大学 | Application of actinomycin |
| CN110983816B (en) * | 2019-11-07 | 2021-05-18 | 浙江理工大学 | Application of actinomycin |
| CN112778792A (en) * | 2021-03-08 | 2021-05-11 | 安徽农业大学 | Method for extracting natural green pigment from trichoderma fungus |
| CN112778792B (en) * | 2021-03-08 | 2022-11-11 | 安徽农业大学 | Method for extracting natural green pigment from trichoderma fungus |
| CN112760240A (en) * | 2021-03-23 | 2021-05-07 | 安徽农业大学 | Method for extracting natural green pigment from penicillium fungi |
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Application publication date: 20161207 |