CN104628879B - Method for preparing zymosan from oil-extraction Lipomycesarkeyi - Google Patents
Method for preparing zymosan from oil-extraction Lipomycesarkeyi Download PDFInfo
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- CN104628879B CN104628879B CN201510077107.0A CN201510077107A CN104628879B CN 104628879 B CN104628879 B CN 104628879B CN 201510077107 A CN201510077107 A CN 201510077107A CN 104628879 B CN104628879 B CN 104628879B
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- zymosan
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Abstract
The invention discloses a method for preparing zymosan from oil-extraction Lipomycesarkeyi. The method comprises the following steps: a. washing oil-extraction Lipomycesarkeyi respectively with a hydrophilic organic solvent and a hydrophobic organic solvent; b. treating the washed Lipomycesarkeyi in alkali liquor or water at 80-150 DEG C, and carrying out solid-liquid separation; c. adding alcohol into the liquid obtained in the step b, and carrying out solid-liquid separation to obtain the solid water-soluble or alkali-soluble polysaccharide product; and d. treating the solid obtained in the step b with acid at 80-150 DEG C for 0.5-3 hours, and carrying out solid-liquid separation to obtain the solid-insoluble polysaccharide product. The high-added-value zymosan product prepared from the oil-extraction Lipomycesarkeyi by chemical and physical means finally changes wastes into valuable substances, implements economic benefit and environmental benefit, can utilize various cheap raw materials, and saves the techniques related in the traditional Saccharomyces cerevisiae polysaccharide extraction process.
Description
Technical field：
The present invention relates to biological chemical field, and in particular to it is many that a kind of saccharomyces oleaginosuses thalline utilized after carrying oil prepares yeast
The method of sugar.
Background technology：
Saccharomyces oleaginosuses are a kind of important industrial microorganisms, its can synthesize meet industry, food, health care it is all kinds of
Microbial grease.Generally, a large amount of non-grease bacteroids can be left after saccharomyces oleaginosuses carry oil.If directly by these non-oil lipid bacterium
Body is abandoned, and can cause the huge wasting of resources, while also polluting the environment.
In general, yeast cell wall accounts for its dry cell weight 18-30%, and in cell wall, have 85% to consist of yeast
Polysaccharide material (based on glucosan and mannan).The microorganism of itself dry weight more than 50% can be accumulated after saccharomyces oleaginosuses fermentation
Oils and fatss, and remaining thalline mostly is yeast cell wall after carrying oil, therefore it is rich in zymosan.
Typically, zymosan extracts from saccharomyces cerevisiae and obtains (preparing zymosan, trehalose with beer waste yeast
And the method for yeast extract, ZL is 200910036820.5).But technique is loaded down with trivial details, the technique being related to includes cell breakage, matter wall
Separate, removing protein, grease removal etc..
The content of the invention：
It is an object of the invention to provide a kind of method for preparing zymosan using the saccharomyces oleaginosuses thalline carried after oil.
The present invention is achieved by the following technical programs：
A kind of method for preparing zymosan using the saccharomyces oleaginosuses thalline carried after oil, the method are comprised the following steps：
A, washing roguing：Carry the saccharomyces oleaginosuses thalline hydrophilic organic solvent after oil and hydrophobic organic solvent is washed respectively
Wash；
B, alkali liquor or water process：Saccharomyces oleaginosuses thalline after washing 80-150 DEG C of process 0.5-3 hour in alkali liquor or water
Solid-liquid separation obtains solid and liquid afterwards；
C, alcohol analysis：Alcohol is added in the liquid obtained toward step b, is stood, solid-liquid separation, solid is obtained for water solublity or alkali
Soluble polysaccharide product；
D, acid treatment：The solid that step b is obtained, processes solid-liquid separation after 0.5-3 hours using sour 80-150 DEG C, consolidate
Body is sour insoluble polysaccharide product.
Described saccharomyces oleaginosuses derive from this in industry and reach saccharomyces oleaginosuses (Lipomyces starkeyi), the red winter spore of circle
Yeast (Rhodosporidium toruloides), rhodotorula glutinis (Rhodotorula glutinis), light white latent ball yeast
(Cryptococcus albidus), trichosporon cutaneum (Trichosporon cutaneum), solution fat Asia sieve yeast
It is arbitrary in (Yarrowia lipolytica), Trichosporon dermatis, Trichosporn coremiiforme
Kind.
Hydrophilic organic solvent described in step a is preferably methanol or ethanol, and the hydrophobic organic solvent is preferably stone
Oily ether or normal hexane, washed with hydrophilic organic solvent and hydrophobic organic solvent respectively carry the saccharomyces oleaginosuses thalline after oil for
Remove extracellular hydrophilic and hydrophobic contaminants.
Described alkali liquor in step b selected from sodium hydrate aqueous solution, potassium hydroxide aqueous solution, calcium hydroxide aqueous solution,
Any one in ammonia aqueous solution.
In step c, alcohol is preferably methanol or ethanol, and the consumption of alcohol is 2-8 times of the liquid volume that step b is obtained, and is stood
Time is 3-8 hours, solid-liquid separating method preferably centrifugation or filtration.
Acid in step d is preferably any one in hydrochloric acid, sulphuric acid, acetic acid, formic acid.
Beneficial effects of the present invention are：
Compared with the technique of zymosan is extracted from saccharomyces cerevisiae, yeast is prepared using the saccharomyces oleaginosuses thalline after oil is carried
Polysaccharide has following advantage：
1) saccharomyces oleaginosuses can be particularly lignocellulosic material using a large amount of different types of cheap raw materials and be fermented,
Obtaining higher Biomass is used for Polyose extraction；
2) can remove the technique being related to during saccharomyces cerevisiae Polyose extraction from includes cell breakage, plasmolysis, removing protein,
Grease removal etc..
In a word, the present invention prepares the yeast of high added value by Chemical Physics means using the saccharomyces oleaginosuses thalline carried after oil
Polyose, is not only capable of achieving " turning waste into wealth ", realizes economic benefit and environmental benefit, can also be extensively sharp by oil fermentation
Lignocellulosic material is particularly with all kinds of cheap raw materials and prepares zymosan, and eliminate conventional brew zymosan and extracted
The technique being related in journey includes cell breakage, plasmolysis, removing protein, grease removal etc., so as to provide a practicable yeast
Polysaccharide preparation technology.
Specific embodiment：
The following is and the present invention is further illustrated, rather than limitation of the present invention.
Embodiment 1：
Using the method that the saccharomyces oleaginosuses thalline after oil prepares zymosan is put forward, the method is comprised the following steps：
A, with 10g oils and fatss extract after trichosporon cutaneum (Trichosporon cutaneum) thalline as raw material, use second
Alcohol, petroleum ether wash respectively after solid-liquid separation；
Saccharomyces oleaginosuses thalline after the washing that b, step a are obtained processes 1.5 in the sodium hydrate aqueous solution of 1M at 105 DEG C
Hour, it is filtrated to get solid and liquid；
8 hours are stood after the ethanol for adding its 5 times of volumes in c, the liquid obtained toward step b to filter, and solid are obtained for 2g
Alkali-soluble polysaccharide；
The solid that d, step b are obtained, is processed 1.5 hours at 105 DEG C using 1M sulphuric acid, after filtration, obtains solid for 3g
Sour insoluble polysaccharide product.
Embodiment 2：
Using the method that the saccharomyces oleaginosuses thalline after oil prepares zymosan is put forward, the method is comprised the following steps：
A, with 10g oils and fatss extract after this up to saccharomyces oleaginosuses (Lipomyces starkeyi) thalline as raw material, with methanol,
Normal hexane wash respectively after solid-liquid separation；
To process 3 at 80 DEG C little in the potassium hydroxide aqueous solution of 1M for saccharomyces oleaginosuses thalline after the washing that b, step a are obtained
When, it is filtrated to get solid and liquid；
5 hours are stood after the methanol for adding its 2 times of volumes in c, the liquid obtained toward step b to filter, obtaining solid is
0.4g alkali-soluble polysaccharides；
The solid that d, step b are obtained, is processed 3 hours at 80 DEG C using 1M hydrochloric acid, and after filtration, it is that 3.0g is sour to obtain solid
Insoluble polysaccharide.
Embodiment 3：
Using the method that the saccharomyces oleaginosuses thalline after oil prepares zymosan is put forward, the method is comprised the following steps：
A, with 10g oils and fatss extract after round rhodosporidium toruloides (Rhodosporidium toruloides) thalline as raw material,
Solid-liquid separation after being washed with ethanol, normal hexane respectively；
Saccharomyces oleaginosuses thalline after the washing that b, step a are obtained is processed at 150 DEG C in the calcium hydroxide aqueous solution of 0.1M
0.5 hour, it is filtrated to get solid and liquid；
5 hours are stood after the methanol for adding its 2 times of volumes in c, the liquid obtained toward step b to filter, obtaining solid is
0.7g alkali-soluble polysaccharides；
The solid that d, step b are obtained, is processed 1.5 hours at 95 DEG C using 2M acetic acid, after filtration, obtains solid for 2.5g
Sour insoluble polysaccharide.
Embodiment 4：
Using the method that the saccharomyces oleaginosuses thalline after oil prepares zymosan is put forward, the method is comprised the following steps：
A, with 10g oils and fatss extract after rhodotorula glutinis (Rhodotorula glutinis) thalline as raw material, with ethanol, stone
Oily ether wash respectively after solid-liquid separation；
Saccharomyces oleaginosuses thalline after the washing that b, step a are obtained is processed at 100 DEG C in the calcium hydroxide aqueous solution of 0.1M
0.8 hour, it is filtrated to get solid and liquid；
5 hours are stood after the methanol for adding its 8 times of volumes in c, the liquid obtained toward step b to filter, obtaining solid is
0.9g alkali-soluble polysaccharides；
The solid that d, step b are obtained, is processed 0.5 hour at 150 DEG C using 2M formic acid, and after filtration, obtaining solid is
2.0g acid insoluble polysaccharide.
Embodiment 5：
Using the method that the saccharomyces oleaginosuses thalline after oil prepares zymosan is put forward, the method is comprised the following steps：
A, with 10g oils and fatss extract after light white latent ball yeast (Cryptococcus albidus) thalline as raw material, use second
Alcohol, petroleum ether wash respectively after solid-liquid separation；
Saccharomyces oleaginosuses thalline after the washing that b, step a are obtained is processed 1.0 hours at 110 DEG C in the ammonia of 0.5M, mistake
Filter obtains solid and liquid；
8 hours are stood after the ethanol for adding its 8 times of volumes in c, the liquid obtained toward step b to filter, obtaining solid is
1.0g alkali-soluble polysaccharide；
The solid that d, step b are obtained, is processed 0.5 hour at 150 DEG C using 2M formic acid, after centrifugation removes supernatant, is obtained
It is 2.3g acid insoluble polysaccharide to solid.
Embodiment 6：
Using the method that the saccharomyces oleaginosuses thalline after oil prepares zymosan is put forward, the method is comprised the following steps：
A, with 10g oils and fatss extract after solution fat Asia sieve yeast (Yarrowia lipolytica) thalline as raw material, use second
Alcohol, petroleum ether wash respectively after solid-liquid separation；
Saccharomyces oleaginosuses thalline after the washing that b, step a are obtained is processed 1.0 hours at 110 DEG C in distilled water, is centrifuged
To solid and supernatant；
C, the supernatant obtained toward step b stand 3 hours after adding the methanol of its 5 times of volumes and filter, and obtaining solid is
0.8g alkali-soluble polysaccharides；
The solid that d, step b are obtained, is processed 0.5 hour at 150 DEG C using 2M formic acid, and centrifugation is obtained after removing supernatant
Solid is obtained for 2.2g acid insoluble polysaccharide.
Embodiment 7：
Using the method that the saccharomyces oleaginosuses thalline after oil prepares zymosan is put forward, the method is comprised the following steps：
A, with 10g oils and fatss extract after Trichosporon dermatis thalline as raw material, with methanol, normal hexane distinguish
Solid-liquid separation after washing；
Saccharomyces oleaginosuses thalline after the washing that b, step a are obtained processes 1.0 in the sodium hydrate aqueous solution of 1M at 110 DEG C
Hour, it is filtrated to get solid and liquid；
8 hours are stood after the ethanol for adding its 8 times of volumes in c, the liquid obtained toward step b to filter, obtaining solid is
1.0g alkali-soluble polysaccharide；
The solid that d, step b are obtained, is processed 0.5 hour at 150 DEG C using 2M formic acid, and centrifugation is obtained after removing supernatant
Solid is 2.5g acid insoluble polysaccharide.
Embodiment 8：
Using the method that the saccharomyces oleaginosuses thalline after oil prepares zymosan is put forward, the method is comprised the following steps：
A, extracted with 10g oils and fatss after Trichosporn coremiiforme thalline as raw material, with methanol, normal hexane point
Not Xi Di after solid-liquid separation；
Saccharomyces oleaginosuses thalline after the washing that b, step a are obtained is processed 2.0 hours at 110 DEG C in tap water, is filtered
To solid and liquid；
5 hours are stood after the methanol for adding its 8 times of volumes in c, the liquid obtained toward step b to filter, obtaining solid is
1.5g alkali-soluble polysaccharide；
The solid that d, step b are obtained, is processed 1.5 hours at 120 DEG C using 2M sulphuric acid, and centrifugation is obtained after removing supernatant
Solid is 2.8g acid insoluble polysaccharide.
Claims (6)
1. a kind of using putting forward the method that the saccharomyces oleaginosuses thalline after oil prepares zymosan, it is characterised in that the method include with
Lower step：
A, carry the saccharomyces oleaginosuses thalline hydrophilic organic solvent after oil and hydrophobic organic solvent is washed respectively；
In alkali liquor or water, after 80-150 DEG C of process 0.5-3 hour, solid-liquid separation obtains solid to saccharomyces oleaginosuses thalline after b, washing
And liquid；
Alcohol is added in c, the liquid obtained toward step b, is stood, solid-liquid separation, solid is obtained for water solublity or alkali-soluble polysaccharide
Product；
The solid that d, step b are obtained, processes solid-liquid separation after 0.5-3 hours using sour 80-150 DEG C, and it is that acid is insoluble to obtain solid
Property polyose.
2. the method that the saccharomyces oleaginosuses thalline utilized after carrying oil according to claim 1 prepares zymosan, its feature exist
In, the Lipomyces starkeyi of described saccharomyces oleaginosuses in industry, Rhodosporidium toruloides,
Rhodotorula glutinis、Cryptococcus albidus、Trichosporon cutaneum、Yarrowia
Any one in lipolytica, Trichosporon dermatis, Trichosporn coremiiforme.
3. the method that the saccharomyces oleaginosuses thalline utilized after carrying oil according to claim 1 and 2 prepares zymosan, its feature
Be that hydrophilic organic solvent described in step a is methanol or ethanol, the hydrophobic organic solvent be petroleum ether or just oneself
Alkane.
4. the method that the saccharomyces oleaginosuses thalline utilized after carrying oil according to claim 1 and 2 prepares zymosan, its feature
It is that the described alkali liquor in step b is selected from sodium hydrate aqueous solution, potassium hydroxide aqueous solution, calcium hydroxide aqueous solution, ammonia
In any one.
5. the method that the saccharomyces oleaginosuses thalline utilized after carrying oil according to claim 1 and 2 prepares zymosan, its feature
It is that alcohol is selected from methanol or ethanol in step c, the consumption of alcohol is 2-8 times of the liquid volume that step b is obtained, time of repose
For 3-8 hours, solid-liquid separating method is for centrifugation or filters.
6. the method that the saccharomyces oleaginosuses thalline utilized after carrying oil according to claim 1 and 2 prepares zymosan, its feature
It is that the acid in step d is selected from any one in hydrochloric acid, sulphuric acid, acetic acid, formic acid.
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| CN101481719A (en) * | 2009-01-20 | 2009-07-15 | 华南理工大学 | Method for preparing zymosan, mycose and yeast extract from beer waste yeast |
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