CN1230549C - Method for increasing hyperosmotic-resistant yeast-fermented glycerol concentration by adding amphotericin fermented broth - Google Patents
Method for increasing hyperosmotic-resistant yeast-fermented glycerol concentration by adding amphotericin fermented broth Download PDFInfo
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- CN1230549C CN1230549C CN 200310100271 CN200310100271A CN1230549C CN 1230549 C CN1230549 C CN 1230549C CN 200310100271 CN200310100271 CN 200310100271 CN 200310100271 A CN200310100271 A CN 200310100271A CN 1230549 C CN1230549 C CN 1230549C
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- amphotericin
- glycerol
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- glucose
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Abstract
The present invention discloses a method for increasing the fermented glycerol concentration by adding amphotericin fermented broth, which belongs to the technical field of glycerol preparation. Hyperosmotic-resistant yeast strains are inoculated to a glycerol fermentation culture medium, and simultaneously, in the process of fermenting glycerol, amphotericin fermented broth is added to continue glycerol fermentation. By adding the amphotericin fermented broth, the membrane permeability of hyperosmotic-resistant yeast cells is improved, and the osmotic pressure is regulated, glycerol in the cells is secreted outside the cells, so that the concentration of the fermented glycerol is improved, and the concentration of the fermented glycerol reaches 10% to 90%. The technological process has the advantages of simplicity, easy control and lower cost, and is easy to realize industrialization application.
Description
Technical field
The invention belongs to the glycerine preparing technical field.Be particularly related to a kind of method of adding amphotericin fermented liquid raising ferment glycerin concentration that adopts in the osmophilic yeast fermentative production glycerine process.
Background technology
Fermentative Production glycerine comes from during the World War I, and Germany needed a large amount of glycerine to make explosive at that time, has at first invented and add the technology that sulphite is produced glycerine in the yeast fermentation mash, and founded the factory and produce in enormous quantities.Fail to proceed to produce because this technology is unreasonable, environmental pollution is serious, production level is low, production cost is high, just stopped after the war.Afterwards, many countries particularly developed country attach great importance to the research of ferment glycerin, have carried out a large amount of experimental study work, but low because of fermentation production rate, cost is high, is difficult to large-scale industrial production.Along with the increase day by day of glycerine demand, middle fifties, software engineering researchers invent has gone out to utilize the technology of osmophilic yeast at the condition bottom fermentation production high density glycerine of high glucose concentration and ventilation.This technology is similar to the production process of conventional leavened prod, compares with the zymotechnique that adds sulphite, does not cause environmental pollution substantially, and the glycerol content in the fermented liquid is enhanced, thereby is subjected to people's height favor.
Use the osmophilic yeast ferment glycerin, its cell leakage influences the transportation of substrate and product, and the structure of cytolemma has influenced cell leakage.At document 1) .Son Tran-Dinh, Martine Herve, OdetteLebourguais, Muriel Jerome and Jaime Wietzerbin, 1991, Effects ofamphotericin B on the glucose metabolism in Saccharomyces cerevisiae cells, European Journal of Biochemistry, 197 (1): 271-279) with document 2) .H.Ramos, A.Attias de Murciano, B.E.Cohen and J.Bolard, 1989, The polyene antibioticamphoteric in B acts as a Ca
2+Ionophore in sterol-containing liposomes, Biochimica et Biophysica Acta, 982:303-306. .Wilfred N.Arnoldand Brenda P.Johnson and document 3), 1982, Effects of polyenes, Detergents, and OtherPotential Membrane Perturbants on an Osmotolerent Yeast, Saccharomycesrouxii, Applied and Environmental Microbiology, 43 (2): 311-318 has reported the influence of amphotericin pair cell membrane permeability respectively.Utilize amphotericin to combine and form the membrane molecule passage, can improve the membrane permeability of cell, improve the concentration of tunning with cytolemma lipid composition.Yet because amphotericin pharmaceutical grade reagent costs an arm and a leg, using in suitability for industrialized production to increase production cost.
Summary of the invention
The purpose of this invention is to provide a kind of method that the amphotericin fermented liquid improves osmophilic yeast ferment glycerin concentration of adding.It is characterized in that: in the process of osmophilic yeast ferment glycerin, add the membrane permeability that the amphotericin fermented liquid improves the osmophilic yeast cell, regulate osmotic pressure, intracellular glycerol is secreted into outside the born of the same parents, and then improve the concentration of ferment glycerin.Specific implementation process is to add the glycerol fermentation substratum of 50-100ml in triangular flask, insert the osmophilic yeast bacterial classification of volume percent 5-10%, in temperature be under 30 ℃, 32 ℃, 34 ℃ or 35 ℃, shaking speed 140-160r/min bottom fermentation produces glycerine, simultaneously, in the glycerol fermentation process, the amphotericin fermented liquid that added volume percent 0.5-2% between 36-96 hour is proceeded glycerol fermentation.
The substratum composition of described fermentative production glycerine comprises: slant medium consists of 20% glucose, 0.3% corn steep liquor, 0.3% urea, 2% agar, pH are that 4.0-4.5, seed culture medium consist of 10% glucose, 0.4% corn steep liquor, 0.3% urea, pH are that 4.0-4.5 and fermention medium consist of 30% glucose, 0.25% corn steep liquor, 0.25% urea, pH are 4.0-4.5; More than each substratum all in 110 ℃, sterilized 20 minutes.
The medium component of described interpolation amphotericin fermented liquid fermentative production comprises: slant medium consists of yeast extract: 5g, glucose: 5g, CaCO
3: 0.05g, K
2HPO
4: 0.25g, agar: 7.8g, pH are that 7.2-7.4, seed culture medium consist of yeast extract: 5g, glucose: 5g, CaCO
3: 0.05g, pH are that 7.2-7.4 and fermention medium consist of glucose: 25g, peptone: 5g, CaCO
3: 0.5g, MnCl
24H
2O:0.005g, FeSO
47H
2O:0.005g, pH are 7.2-7.4.More than each substratum all in 110 ℃, sterilized 20 minutes.
The bacterial classification of described interpolation amphotericin fermented liquid fermentative production is a streptomyces nodocus.
The temperature of described interpolation amphotericin fermented liquid fermentative production is 28-30 ℃, and shaking speed is 220-250r/min.
The invention has the beneficial effects as follows and add the concentration that can increase ferment glycerin after the amphotericin fermented liquid.This technological process is simple, be easy to control, cost is lower.Be easy to realize industrial applications.
Embodiment
The present invention is for adding the method for amphotericin fermented liquid raising osmophilic yeast ferment glycerin concentration.In the process of osmophilic yeast ferment glycerin, add the membrane permeability that the amphotericin fermented liquid improves the osmophilic yeast cell, regulate osmotic pressure, intracellular glycerol is secreted into outside the born of the same parents, and then improves the concentration of ferment glycerin.Specific implementation process is to add the glycerol fermentation substratum of 50-100ml in triangular flask, insert the osmophilic yeast bacterial classification of volume percent 5-10%, at temperature 30-35 ℃, shaking speed 140-160r/min bottom fermentation production glycerine, simultaneously, in the glycerol fermentation process, the amphotericin fermented liquid that added volume percent 0.5-2% between 36-96 hour is proceeded glycerol fermentation.The bacterial classification of fermentative production glycerine is an osmophilic yeast.The substratum of fermentative production glycerine comprises: slant medium is: 20% glucose, 0.3% corn steep liquor, 0.3% urea, 2% agar, pH are that 4.0-4.5, seed culture medium are: 10% glucose, 0.4% corn steep liquor, 0.3% urea, pH are that 4.0-4.5 and fermention medium are: 30% glucose, 0.25% corn steep liquor, 0.25% urea, pH are 4.0-4.5; More than each substratum all in 110 ℃, sterilized 20 minutes.
The medium component of the fermentative production of above-mentioned interpolation amphotericin fermented liquid comprises: slant medium is formed and is comprised yeast extract: 5g, glucose: 5g, CaCO
3: 0.05g, K
2HPO
4: 0.25g, agar: 7.8g, pH are that 7.2-7.4, seed culture medium consist of yeast extract: 5g, glucose: 5g, CaCO
3: 0.05g, pH are that 7.2-7.4 and fermention medium consist of glucose: 25g, peptone: 5g, CaCO
3: 0.5g, MnCl
24H
2O:0.005g, FeSO
47H
2O:0.005g, pH are 7.2-7.4.More than each substratum all in 110 ℃, sterilized 20 minutes.Fermentative production inserts the streptomyces nodocus bacterial classification, is 28-30 ℃ in temperature again, and shaking speed is the production of 220-250r/min bottom fermentation.
For embodiment the present invention is further specified again below.
Embodiment 1
50ml glycerol fermentation substratum is placed triangular flask, connect the osmophilic yeast bacterial classification of 5ml, 35 ℃ of temperature, shaking speed 160r/min bottom fermentation is cultured to 24 hours, the amphotericin fermented liquid that adds 0.25ml continues fermentation culture to 36 hour, compare with the ferment glycerin concentration of not adding the amphotericin fermented liquid, the ferment glycerin concentration of adding the amphotericin fermented liquid has increased by 41%.
Embodiment 2
50ml glycerol fermentation substratum is placed triangular flask, connect the osmophilic yeast bacterial classification of 5ml, 34 ℃ of temperature, shaking speed 160r/min bottom fermentation is cultured to 24 hours, the amphotericin fermented liquid that adds 0.5ml continues fermentation culture to 72 hour, compare with the ferment glycerin concentration of not adding the amphotericin fermented liquid, the ferment glycerin concentration of adding the amphotericin fermented liquid has increased by 43%.
Embodiment 3
50ml glycerol fermentation substratum is placed triangular flask, connect the osmophilic yeast bacterial classification of 5ml, 33 ℃ of temperature, shaking speed 160r/min bottom fermentation is cultured to 24 hours, the amphotericin fermented liquid that adds 1ml continues fermentation culture to 36 hour, compare with the ferment glycerin concentration of not adding the amphotericin fermented liquid, the ferment glycerin concentration of adding the amphotericin fermented liquid has increased by 34%.
Embodiment 4
50ml glycerol fermentation substratum is placed triangular flask, connect the osmophilic yeast bacterial classification of 5ml, 32 ℃ of temperature, shaking speed 160r/min bottom fermentation is cultured to 24 hours, the amphotericin fermented liquid that adds 0.5ml continues fermentation culture to 60 hour, compare with the ferment glycerin concentration of not adding the amphotericin fermented liquid, the ferment glycerin concentration of adding the amphotericin fermented liquid has increased by 55%.
Embodiment 5
50ml glycerol fermentation substratum is placed triangular flask, connect the osmophilic yeast bacterial classification of 5ml, 30 ℃ of temperature, shaking speed 160r/min bottom fermentation is cultured to 60 hours, the amphotericin fermented liquid that adds 0.5ml continues fermentation culture to 96 hour, compare with the ferment glycerin concentration of not adding the amphotericin fermented liquid, the ferment glycerin concentration of adding the amphotericin fermented liquid has increased by 13%.
Claims (3)
1. one kind is added the method that the amphotericin fermented liquid improves osmophilic yeast ferment glycerin concentration, it is characterized in that: in the process of osmophilic yeast ferment glycerin, interpolation amphotericin fermented liquid improves the membrane permeability of osmophilic yeast cell, regulate osmotic pressure, intracellular glycerol is secreted into outside the born of the same parents, and then improves the concentration of ferment glycerin; Specific implementation process is to add the glycerol fermentation substratum of 50-100ml in triangular flask, insert the osmophilic yeast bacterial classification of volume percent 5-10%, in temperature is under 30 ℃, 32 ℃, 34 ℃ or 35 ℃, shaking speed 140-160r/min bottom fermentation is produced glycerine, simultaneously, in the glycerol fermentation process, the amphotericin fermented liquid that added volume percent 0.5-2% between 24-96 hour is proceeded glycerol fermentation.
2. improve the method for osmophilic yeast ferment glycerin concentration according to the described interpolation amphotericin of claim 1 fermented liquid, it is characterized in that: the substratum of described fermentative production glycerine comprises: slant medium consists of 20% glucose, 0.3% corn steep liquor, 0.3% urea, 2% agar, pH is that 4.0-4.5, seed culture medium consist of 10% glucose, 0.4% corn steep liquor, 0.3% urea, pH is that 4.0-4.5 and fermention medium consist of 30% glucose, 0.25% corn steep liquor, 0.25% urea, pH are 4.0-4.5; More than each substratum all in 110 ℃, sterilized 20 minutes.
3. improve the method for osmophilic yeast ferment glycerin concentration according to the described interpolation amphotericin of claim 1 fermented liquid, it is characterized in that: the medium component of the fermentative production of described amphotericin fermented liquid comprises: slant medium consists of yeast extract: 5g, glucose: 5g, CaCO
3: 0.05g, K
2HPO
4: 0.25g, agar: 7.8g, pH are that 7.2-7.4, seed culture medium consist of yeast extract: 5g, glucose: 5g, CaCO
3: 0.05g, pH are that 7.2-7.4 and fermention medium consist of glucose: 25g, peptone: 5g, CaCO
3: 0.5g, MnCl
24H
2O:0.005g, FeSO
47H
2O:0.005g, pH are 7.2-7.4; More than each substratum all in 110 ℃, sterilized 20 minutes.
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|---|---|---|---|
| CN 200310100271 CN1230549C (en) | 2003-10-17 | 2003-10-17 | Method for increasing hyperosmotic-resistant yeast-fermented glycerol concentration by adding amphotericin fermented broth |
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| CN 200310100271 CN1230549C (en) | 2003-10-17 | 2003-10-17 | Method for increasing hyperosmotic-resistant yeast-fermented glycerol concentration by adding amphotericin fermented broth |
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| CN1230549C true CN1230549C (en) | 2005-12-07 |
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