CN101168747A - Method for preparing ethanol and yeast superoxide dismutase from sweet sorghum stem liquor - Google Patents
Method for preparing ethanol and yeast superoxide dismutase from sweet sorghum stem liquor Download PDFInfo
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- CN101168747A CN101168747A CNA2007100476855A CN200710047685A CN101168747A CN 101168747 A CN101168747 A CN 101168747A CN A2007100476855 A CNA2007100476855 A CN A2007100476855A CN 200710047685 A CN200710047685 A CN 200710047685A CN 101168747 A CN101168747 A CN 101168747A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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Abstract
The invention relates to a method for producing ethyl alcohol and yeast superoxide dismutase by using juice of stems of sweet sorghum, and belongs to the technical field of the biomaterial energy engineering. The invention has the following steps: firstly, the raw material of the juice of the stems of the sweet sorghum and fermentation auxiliary materials are taken to be sterilized and cooled for standup; secondly, saccharomyces cervisiae is taken to be augmented stage by stage and cultured until the cell number per milliliter achieves above 1.0*10<8>; thirdly, the juice of the stems of the sweet sorghum and the fermentation auxiliary materials are mixed, and filled with the saccharomyces cerivisiae to perform ethyl alcohol fermentation; fourthly, after the ethyl alcohol fermentation is finished, refrigerated centrifugation to the fermentation liquid is performed, the supernatant fluid is taken to be used for distilling and extracting the ethyl alcohol, and the thallus is used for extracting the SOD; fifthly, after the collected thallus is activated, the cell wall of microzyme is damaged, and is heat insulated and extracted by phosphate buffer solution, then solid liquid separation is performed, and the obtained clear liquid is yeast SOD extracting solution. The yield rate of the ethyl alcohol can achieve 90.3 to 97.03 percent, and the output of the superoxide dismutase in each liter of fermentation liquid is 3.64*10<4>U to 6.95*10<4>U.
Description
Technical field
What the present invention relates to is a kind of method of technical field of chemical engineering, and specifically, what relate to is the method for a kind of juice of sugar grass stalks ethanol production and yeast superoxide dismutase.
Background technology
Sweet sorghum is a kind of C4 crop varieties with higher biological yield, every hm
2Sweet sorghum general energy output 2250～7500kg grain and 55～75t are rich in the stem stalk of sugar (15 °～20 ° Brix).15 °～20 ° Brix of its stem stalk sugar content, crushing juice rate are more than 55%, and being suitable for adopting the liquid state fermentation technical transform is that ethanol is used for liquid fuel, have broad prospects aspect the alcohol fuel producing, and are described as " green energy resource " of 21 century.Relevant investigator was a major objective with the fermenting alcohol productive rate once, ethanol fermentation conditions such as nitrogenous source, nutritive salt and the initial sugared concentration of fermented liquid, inoculum size, temperature, shaking table (stirring) revolution that juice of sugar grass stalks is added have been carried out more deep research, have obtained comparatively ideal alcohol yied.But because the product of sweet sorghum stalk preparing ethanol by fermentation is single, higher with the cost of preparation of ethanol by sweet Chinese sorghum stalk, its advantage does not fully manifest, and industrialization process is slower.
Superoxide-dismutase (being called for short SOD) is a kind of main enzyme of organism defence oxidative damage, is present in animals and plants and the aerobic microbiological body.SOD acts on substrate superoxide anion (O
- 2), be broken down into O
2And H
2O
2, H
2O
2Become H through superoxide enzyme and catalatic catalysis again
2O, thus 3O removed
- 2(superoxide anion) plays a protective role to the damage of biological cell.SOD product through extraction, purifying has been widely used in fields such as medicine, makeup, food.In the past, SOD mainly extracted from tissues such as animal blood, liver, since raw materials used limited, SOD quality instability.After the eighties in 20th century, the U.S. and Japan have successively developed the technology that yeast fermentation method is produced SOD, have reduced the production cost of SOD widely.At present, the research of yeast SOD is mainly concentrated on extracting and purifying method, superior strain seed selection, and the aspects such as Physiologic Studies that form of SOD.The research of producing yeast SOD in the time of with the juice of sugar grass stalks preparing ethanol by fermentation yet there are no report.
Find that through literature search Shenyang Pharmaceutical University's master thesis in 2002 discloses name and has been called: " seed selection of yeast SOD superior strain and the research of zymologic property thereof ", this technology filters out a strain Y from 9 saccharomycetes to prior art
12Yeast, through culture medium prescription and main Optimizing Conditions of Fermentation, the high-content that obtains wet thallus SOD is 1263Ug
-1, every liter of fermented liquid can obtain the 30g wet thallus, and then the production peak of every liter of fermented liquid yeast SOD is 4.789 * 10
4U.This technology only can obtain SOD product, and output is lower.
Summary of the invention
The objective of the invention is to overcome deficiency of the prior art, the method of a kind of juice of sugar grass stalks while ethanol production and yeast superoxide dismutase (SOD) is provided, have one time fermentation and can obtain ethanol and two kinds of product characteristics of SOD, preparation equipment and technology are simple, are easy to realize and the docking of existing juice of sugar grass stalks attitude preparing ethanol by fermentation technology.Can make alcohol yied reach 90.3%～97.03%, yeast superoxide dismutase (SOD) output of every liter of fermented liquid reaches 3.64 * 10
4U～6.95 * 10
4U.Reduce the production cost of sweet sorghum stalk preparing ethanol by fermentation greatly.
The present invention is achieved by the following technical solutions, may further comprise the steps:
1. former, auxiliary material is prepared: sterilize, cool off juice of sugar grass stalks and fermentation auxiliary material stoste standby.
Described juice of sugar grass stalks total sugar content is 12%～18%.
Described fermentation auxiliary material is respectively (NH)
2SO
4, K
2HPO
4, MgSO
47H
2O, Zn (CH
3COO)
22H
2O, CuSO
45H
2The aqueous solution of O.
Described sterilization, cooling are meant: in 121 ℃, 0.1MPa autoclaving 15min is cooled to below 40 ℃ again.
2. fermented bacterium is prepared: get yeast saccharomyces cerevisiae (Saccharomyces cerevisiae CICC 1308) bacterial classification step by step amplification cultivation to every ml cells number reach 1.0 * 10
8More than individual.
Yeast saccharomyces cerevisiae (Saccharomyces cerevisiae CICC 1308) can directly be bought acquisition, the purchasing channel: Chinese industrial microbial strains preservation administrative center, numbering: CICC 1308, trade name: yeast saccharomyces cerevisiae, generic name: Saccharomyces plants name: cerevisiae.
3. ethanol fermentation: under aseptic condition, sweet sorghum stalk juice and fermentation auxiliary material are mixed, insert as 2. described fermented bacterium, carry out ethanol fermentation.
The using dosage of described fermentation auxiliary material in fermented liquid is respectively:
(NH)
2SO
4 20-25g·L
-1；
K
2HPO
4 2.0-2.5g·L
-1；
MgSO
4·7H
2O 0.5-0.65g·L
-1；
Zn(CH
3COO)
2·2H
2O 5.0-6.5mg·L
-1；
CuSO
4·5H
2O 12-15mg·L
-1。
Described fermented bacterium inoculum size is 10%～15% (v/v) of fermentating liquid volume.
After described sweet sorghum stalk juice and fermentation auxiliary material mix, use 6molL
-1The HCl sterile solution is adjusted pH value to 4.0～5.0.
Described ethanol fermentation temperature is 30～35 ℃, and with stirring, stir speed (S.S.) is 110～150rmin
-1
4. ethanol distillation and microorganism collection: ethanol fermentation with the fermented liquid frozen centrifugation, is got supernatant liquor and is carried out ethanol distillation after finishing, the collecting precipitation thalline, and washing and centrifugal collection thalline are standby again.
Described fermented liquid frozen centrifugation is meant: with fermented liquid in 5～7 ℃, 3000～5000rmin
-1Centrifugal 15min.
Described washing thalline, centrifugal collection thalline are meant: with aseptic washing thalline 1～2 time, and 3000rmin
-1The centrifugal 10min of normal temperature collects thalline.
5. thalline activation and superoxide-dismutase extract: add the thalline activation solution, after vibration activates in water-bath, destroy the yeast cell wall, wet yeast adds phosphate buffered saline buffer, place the water-bath vibration to extract, collect supernatant liquor with centrifugal or suction method, be the yeast superoxide dismutase extracting solution.
Described vibration activation in water-bath is meant: 120rmin in 30 ℃ of water-baths
-1Vibration activation 30～60min;
Described phosphate buffered saline buffer, its addition is: the wet yeast of every 1g adds 15mL;
Describedly place water-bath vibration to extract, be meant: place 30 ℃ of water-bath 120rmin
-15h is extracted in vibration under the condition.
Principle of the present invention is, yeast utilizes the sugar in the sweet sorghum stalk to carry out self-reproduction, produces a large amount of somatic cells, and under anoxia condition, the somatic cells metabolism produces ethanol and CO
2, yeast cell can stimulate or adverse circumstance environment generation stress reaction, to improve the adaptive faculty of himself by the synthetic cytoprotective material that comprises superoxide-dismutase (SOD) to external world for keeping normal Metabolic activity.Within the specific limits, coercing that yeast cell is subjected to stimulates increase, and the SOD amount of generation also increases thereupon.The present invention is target according to this principle to improve ethanol and yeast SOD productive rate, selects suitable bacterial classification, adjusts nutrient environment and the technological condition for fermentation of optimizing fermented liquid, makes alcohol yied and yeast SOD productive rate all reach higher level.
The present invention is on the prior art basis, and the method by ethanol fermentation bacterial screening, nutritive salt kind and additive capacity selection and optimization for fermentation technology parameter when guaranteeing the higher ethanol productive rate, makes the content of SOD in the yeast thalline also reach higher level.Yeast SOD can significantly reduce the cost of juice of sugar grass stalks ethanol production as the ethanol fermentation byproduct with high added value.
The present invention is applicable to the juice of sugar grass stalks to be raw material, with commercially available yeast saccharomyces cerevisiae (Saccharomycescerevisiae CICC 1308) is that fermented bacterium carries out ethanol fermentation, while ethanol production and two kinds of products of yeast SOD, alcohol yied can reach 90.3%～97.03% during fermentation ends, and every liter of fermented liquid yeast SOD output can reach 3.64 * 10
4U～6.95 * 10
4U.
Embodiment
Below embodiments of the invention are elaborated.Present embodiment is being to implement under the prerequisite with the technical solution of the present invention, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
Various cultivations of adopting in following examples of the present invention and substratum all adopt operation commonly used:
Described yeast saccharomyces cerevisiae is amplification cultivation step by step, be meant with the routine operation method by the slant tube bacterial classification step by step enlarged culturing become ferment-seeded.Concrete grammar is: under aseptic technique, gets a ring slant tube bacterial classification inoculation on fresh solid slant culture base, behind 30 ℃ of cultivation 12h, gets 1～2 ring and be inoculated in the 100mL liquid seed culture medium, and 28 ℃, 120rmin
-1Shaking table is cultivated 12-18h, gets 5mL and transfers cultivate 12～18h in the 500mL liquid seed culture medium, when cell count reaches 1.0 * 10
8Can be used for ethanol fermentation when above.
Described liquid seed culture medium and solid slant culture base are that yeast is cultivated common prescription, and concrete component sees Table 1.
Table 1 liquid seed culture medium and solid slant culture base component
| Medium component | The liquid slant medium | The solid seed culture medium |
| Glucose or sucrose (g) yeast extract paste (g) peptone (g) K 2HPO 4(g) MgSO 4·7H 2O(g) | 5.0 0.5 0.5 0.1 0.1 | 5.0 0.5 0.5 0.1 0.1 |
| Gelatin sterilized water (mL) | 0 100 | 2.0 100 |
Described thalline activation solution component is identical with the liquid seed culture medium component, and addition is 10% (w/w) of wet thallus weight.
Described phosphate buffered saline buffer, the pH value that is meant according to a conventional method preparation are 7.8 and contain 0.1mmolL
-1The aqueous solution of ethylenediamine tetraacetic ethylhexyldithiophosphoric acid disodium (EDTA-2Na), its addition is: the wet yeast of every 1g adds 15mL.
Embodiment 1
Measure the sweet sorghum stalk juice (total sugar content is 12%) after the sterilization, under aseptic condition, add (NH)
2SO
420gL
-1, K
2HPO
42.0gL
-1, MgSO
47H
2O 0.5gL
-1, Zn (CH
3COO)
22H
2O5.0mgL
-1, CuSO
45H
2O 12mgL
-1Use 6molL
-1It is 4.0 that the HCl sterile solution is adjusted the pH value; 10% (v/v) that presses the fermented liquid cumulative volume inserts barms---yeast saccharomyces cerevisiae.At 30 ℃, 110rmin
-1The stir speed (S.S.) bottom fermentation.After ethanol fermentation finishes, with fermented liquid in 5 ℃, 5000rmin
-1Centrifugal 15min gets supernatant liquor and is used for distillation extraction ethanol.The precipitation thalline is with aseptic washing 1 time, 3000rmin
-1The centrifugal collection thalline of normal temperature.10% (w/w) that presses wet thallus weight adds thalline activation solution, 120rmin in 30 ℃ of water-baths
-1Behind the vibration activation 60min, destroy the yeast cell wall with ordinary method, the phosphate buffered saline buffer that adds 15mLpH7.8 by the wet yeast of every 1g (contains 0.1mmolL
-1EDTA-2Na), put vibration (120rmin in 30 ℃ of water-baths
-1) extract 5h, collect supernatant liquor with centrifuging, be yeast SOD extracting solution.Alcohol yied is 97.03%, and yeast SOD output is 6.417 * 10
4UL
-1Fermented liquid.
Embodiment 2
Measure the sweet sorghum stalk juice (total sugar content is 12%) after the sterilization, under aseptic condition, add (NH)
2SO
425gL
-1, K
2HPO
42.5gL
-1, MgSO
47H
2O 0.65gL
-1, Zn (CH
3COO)
22H
2O5.7mgL
-1, CuSO
45H
2O 12mgL
-1Use 6molL
-1It is 4.0 that the HCl sterile solution is adjusted the pH value; 10% (v/v) that presses the fermented liquid cumulative volume inserts barms---yeast saccharomyces cerevisiae.At 30 ℃, 130rmin
-1The stir speed (S.S.) bottom fermentation.After the fermentation ends, with fermented liquid in 7 ℃, 3000rmin
-1Centrifugal 15min gets supernatant liquor and carries out alcohol distillation.With aseptic washing thalline 2 times, 3000rmin
-1The centrifugal collection thalline of normal temperature.10% (w/w) that presses wet thallus weight adds thalline activation solution, 120rmin in 30 ℃ of water-baths
-1Behind the vibration activation 30min, destroy the yeast cell wall with ordinary method, the phosphate buffered saline buffer that adds 15mLpH7.8 by the wet yeast of every 1g (contains 0.1mmolL
-1EDTA-2Na), put vibration (120rmin in 30 ℃ of water-baths
-1) extract 5h, collect supernatant liquor with centrifuging, be yeast SOD extracting solution.Alcohol yied is 91.59%, and yeast SOD output is 6.95 * 10
4UL
-1Fermented liquid.
Embodiment 3
Measure the sweet sorghum stalk juice (total sugar content is 15%) after the sterilization, under aseptic condition, add (NH)
2SO
422.5gL
-1, K
2HPO
42.0gL
-1, MgSO
47H
2O 0.6gL
-1, Zn (CH
3COO)
22H
2O5.0mgL
-1, CuSO
45H
2O 15mgL
-1Use 6molL
-1It is 5.0 that the HCl sterile solution is adjusted the pH value; 15% (v/v) that presses the fermented liquid cumulative volume inserts barms---yeast saccharomyces cerevisiae.At 35 ℃, 150rmin
-1The stir speed (S.S.) bottom fermentation.After the fermentation ends, with fermented liquid in 5 ℃, 5000rmin
-1Centrifugal 15min gets supernatant liquor and carries out alcohol distillation.With aseptic washing thalline 2 times, 3000rmin
-1The centrifugal collection thalline of normal temperature.10% (w/w) that presses wet thallus weight adds thalline activation solution, 120rmin in 30 ℃ of water-baths
-1Behind the vibration activation 60min, destroy the yeast cell wall with ordinary method, the phosphate buffered saline buffer that adds 15mLpH7.8 by the wet yeast of every 1g (contains 0.1mmolL
-1EDTA-2Na), put vibration (120rmin in 30 ℃ of water-baths
-1) extract 5h, collect filtrate with suction method, be yeast SOD extracting solution.Alcohol yied is 94.33%, and yeast SOD output is 5.92 * 10
4UL
-1Fermented liquid.
Embodiment 4
Measure the sweet sorghum stalk juice (total sugar content is 18%) after the sterilization, under aseptic condition, add (NH)
2SO
425gL
-1, K
2HPO
42.5gL
-1, MgSO
47H
2O 0.65gL
-1, Zn (CH
3COO)
22H
2O6.5mgL
-1, CuSO
45H
2O 15mgL
-1Use 6molL
-1It is 4.5 that the HCl sterile solution is adjusted the pH value; 15% (v/v) that presses the fermented liquid cumulative volume inserts barms---yeast saccharomyces cerevisiae.At 32.5 ℃, 150rmin
-1The stir speed (S.S.) bottom fermentation.After the fermentation ends, with fermented liquid in 5 ℃, 5000rmin
-1Centrifugal 15min gets supernatant liquor and carries out alcohol distillation.With aseptic washing thalline 2 times, 3000rmin
-1The centrifugal collection thalline of normal temperature.10% (w/w) that presses wet thallus weight adds thalline activation solution, 120rmin in 30 ℃ of water-baths
-1Behind the vibration activation 45min, destroy the yeast cell wall with ordinary method, the phosphate buffered saline buffer that adds 15mLpH7.8 by the wet yeast of every 1g (contains 0.1mmolL
-1EDTA-2Na), put vibration (120rmin in 30 ℃ of water-baths
-1) extract 5h, collect yeast SOD extracting solution with centrifuging.Alcohol yied is 90.3%, and yeast SOD output is 6.92 * 10
4UL
-1Fermented liquid.
Embodiment 5
Measure the sweet sorghum stalk juice (total sugar content is 15%) after the sterilization, under aseptic condition, add (NH)
2SO
422.5gL
-1, K
2HPO
42.25gL
-1, MgSO
47H
2O 0.575gL
-1, Zn (CH
3COO)
22H
2O5.75mgL
-1, CuSO
45H
2O 13.5mgL
-1Use 6molL
-1It is 4.5 that the HCl sterile solution is adjusted the pH value; 12.5% (v/v) that presses the fermented liquid cumulative volume inserts barms---yeast saccharomyces cerevisiae.At 35 ℃, 150rmin
-1The stir speed (S.S.) bottom fermentation.After the fermentation ends, with fermented liquid in 5 ℃, 4000rmin
-1Centrifugal 15min gets supernatant liquor and carries out alcohol distillation.With aseptic washing thalline 1 time, 3000rmin
-1The centrifugal collection thalline of normal temperature.10% (w/w) that presses wet thallus weight adds thalline activation solution, 120rmin in 30 ℃ of water-baths
-1Behind the vibration activation 60min, destroy the yeast cell wall with ordinary method, the phosphate buffered saline buffer that adds 15mLpH7.8 by the wet yeast of every 1g (contains 0.1mmolL
-1EDTA-2Na), put vibration (120rmin in 30 ℃ of water-baths
-1) extract 5h, collect yeast SOD extracting solution with centrifuging.Alcohol yied is 93.40%, and yeast SOD output is 3.92 * 10
4UL
-1Fermented liquid.
Embodiment 6
Measure the sweet sorghum stalk juice (total sugar content is 18%) after the sterilization, under aseptic condition, add (NH)
2SO
420gL
-1, K
2HPO
42.5gL
-1, MgSO
47H
2O 0.5gL
-1, Zn (CH
3COO)
22H
2O5.75mgL
-1, CuSO
45H
2O 12mgL
-1Use 6molL
-1It is 4.5 that the HCl sterile solution is adjusted the pH value; 15% (v/v) that presses the fermented liquid cumulative volume inserts barms---yeast saccharomyces cerevisiae.At 30 ℃, 130rmin
-1The stir speed (S.S.) bottom fermentation.After the fermentation ends, with fermented liquid in 6 ℃, 4000rmin
-1Centrifugal 15min gets supernatant liquor and carries out alcohol distillation.With aseptic washing thalline 2 times, 3000rmin
-1The centrifugal collection thalline of normal temperature.10% (w/w) that presses wet thallus weight adds thalline activation solution, 120rmin in 30 ℃ of water-baths
-1Behind the vibration activation 50min, destroy the yeast cell wall with ordinary method, the phosphate buffered saline buffer that adds 15mLpH7.8 by the wet yeast of every 1g (contains 0.1mmolL
-1EDTA-2Na), put vibration (120rmin in 30 ℃ of water-baths
-1) extract 5h, collect yeast SOD extracting solution with centrifuging.Alcohol yied is 91.12%, and yeast SOD output is 3.64 * 10
4UL
-1Fermented liquid.
Embodiment 7
Measure the sweet sorghum stalk juice (total sugar content is 12.2%) after the sterilization, under aseptic condition, add (NH)
2SO
424.8gL
-1, K
2HPO
42.25gL
-1, MgSO
47H
2O 0.56gL
-1, Zn (CH
3COO)
22H
2O 5.8mgL
-1, CuSO
45H
2O 13.5mgL
-1Use 6molL
-1It is 4.5 that the HCl sterile solution is adjusted the pH value; 13.1% (v/v) that presses the fermented liquid cumulative volume inserts barms---yeast saccharomyces cerevisiae.At 32.5 ℃, 130rmin
-1The stir speed (S.S.) bottom fermentation.After the fermentation ends, with fermented liquid in 6 ℃, 5000rmin
-1Centrifugal 15min gets supernatant liquor and carries out alcohol distillation.With aseptic washing thalline 2 times, 3000rmin
-1The centrifugal collection thalline of normal temperature.10% (w/w) that presses wet thallus weight adds thalline activation solution, 120rmin in 30 ℃ of water-baths
-1Behind the vibration activation 60min, destroy the yeast cell wall with ordinary method, the phosphate buffered saline buffer that adds 15mLpH7.8 by the wet yeast of every 1g (contains 0.1mmolL
-1EDTA-2Na), put vibration (120rmin in 30 ℃ of water-baths
-1) extract 5h, collect yeast SOD extracting solution with centrifuging.Alcohol yied is 90.6%, and yeast SOD output is 6.91 * 10
4UL
-1Fermented liquid.
Claims (10)
1. the method for juice of sugar grass stalks ethanol production and yeast superoxide dismutase is characterized in that, may further comprise the steps:
1. with juice of sugar grass stalks and fermentation auxiliary material stoste, sterilize, cool off standby;
2. get yeast saccharomyces cerevisiae step by step amplification cultivation reach 1.0 * 10 to every ml cells number
8More than individual;
3. under aseptic condition, sweet sorghum stalk juice and fermentation auxiliary material are mixed, insert as 2. described yeast saccharomyces cerevisiae, carry out ethanol fermentation;
4. after ethanol fermentation finishes,, get supernatant liquor and carry out ethanol distillation the fermented liquid frozen centrifugation, the collecting precipitation thalline, washing and centrifugal collection thalline are standby again;
5. add the thalline activation solution, after the vibration activation, destroy the yeast cell wall in water-bath, wet yeast adds phosphate buffered saline buffer, places the water-bath vibration to extract, and collects supernatant liquor with centrifugal or suction method, is the yeast superoxide dismutase extracting solution.
2. the method for juice of sugar grass stalks ethanol production according to claim 1 and yeast superoxide dismutase is characterized in that, described juice of sugar grass stalks total sugar content is 12%～18%.
3. the method for juice of sugar grass stalks ethanol production according to claim 1 and yeast superoxide dismutase is characterized in that, described fermentation auxiliary material is respectively (NH)
2SO
4, K
2HPO
4, MgSO
47H
2O, Zn (CH
3COO)
22H
2O, CuSO
45H
2The O aqueous solution.
4. the method for juice of sugar grass stalks ethanol production according to claim 3 and yeast superoxide dismutase is characterized in that, the using dosage of described fermentation auxiliary material in fermented liquid is respectively:
(NH)
2SO
4 20-25g·L
-1；
K
2HPO
4 2.0-2.5g·L
-1；
MgSO
4·7H
2O 0.5-0.65g·L
-1；
Zn(CH
3COO)
2·2H
2O 5.0-6.5mg·L
-1；
CuSO
4·5H
2O 12-15mg·L
-1。
5. the method for juice of sugar grass stalks ethanol production according to claim 1 and yeast superoxide dismutase is characterized in that, described yeast saccharomyces cerevisiae inoculum size is 10%～15% of a fermentating liquid volume.
6. the method for juice of sugar grass stalks ethanol production according to claim 1 and yeast superoxide dismutase is characterized in that, after described sweet sorghum stalk juice and fermentation auxiliary material mix, uses 6molL
-1It is 4.0～5.0 that the HCl sterile solution is adjusted the pH value.
7. the method for juice of sugar grass stalks ethanol production according to claim 1 and yeast superoxide dismutase is characterized in that, the described ethanol fermentation that carries out, and its leavening temperature is 30～35 ℃, and with stirring, stir speed (S.S.) is 110～150rmin
-1
8. the method for juice of sugar grass stalks ethanol production according to claim 1 and yeast superoxide dismutase is characterized in that, described fermented liquid frozen centrifugation is meant: with fermented liquid in 5～7 ℃, 3000～5000rmin
-1Centrifugal 15min;
Described washing thalline, centrifugal collection thalline are meant: with aseptic washing thalline 1～2 time, and 3000rmin
-1The centrifugal 10min of normal temperature collects thalline.
9. the method for juice of sugar grass stalks ethanol production according to claim 1 and yeast superoxide dismutase is characterized in that, described thalline activation solution consists of, glucose or sucrose 5.0g, yeast extract paste 0.5g, peptone 0.5g, K
2HPO
40.1g, MgSO
47H
2O 0.1g, sterilized water 100mL;
Described vibration activation in water-bath is meant: 120rmin in 30 ℃ of water-baths
-1Vibration activation 30～60min.
10. the method for juice of sugar grass stalks ethanol production according to claim 1 and yeast superoxide dismutase is characterized in that, described phosphate buffered saline buffer, and its addition is: the wet yeast of every 1g adds 15mL;
Describedly place water-bath vibration to extract, be meant: place 30 ℃ of water-bath 120rmin
-15h is extracted in vibration under the condition.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102391998A (en) * | 2011-10-31 | 2012-03-28 | 中兴能源（天津）有限公司 | Method for increasing content of adenosylmethionine and superoxide dismutase in ethanol waste saccharomyces cerevisiae as sorgo fuel |
| CN102504000A (en) * | 2011-10-31 | 2012-06-20 | 中兴能源（天津）有限公司 | Method for comprehensively extracting S-adenosine-L-methionine and superoxide dismutase from saccharomyces cerevisiae |
| CN103131750A (en) * | 2013-02-05 | 2013-06-05 | 大连工业大学 | Method for detecting aging of yeast |
| CN104593337A (en) * | 2014-12-31 | 2015-05-06 | 唯美度科技（北京）有限公司 | Orgotein extracted from beer yeast |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1699583B (en) * | 2005-06-20 | 2011-07-27 | 郎咏梅 | Process for preparing ethanol by solid fermentation and distillation of sweet sorghum stalks |
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2007
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102391998A (en) * | 2011-10-31 | 2012-03-28 | 中兴能源（天津）有限公司 | Method for increasing content of adenosylmethionine and superoxide dismutase in ethanol waste saccharomyces cerevisiae as sorgo fuel |
| CN102504000A (en) * | 2011-10-31 | 2012-06-20 | 中兴能源（天津）有限公司 | Method for comprehensively extracting S-adenosine-L-methionine and superoxide dismutase from saccharomyces cerevisiae |
| CN102504000B (en) * | 2011-10-31 | 2014-05-21 | 中兴能源（天津）有限公司 | Method for comprehensively extracting S-adenosine-L-methionine and superoxide dismutase from saccharomyces cerevisiae |
| CN102391998B (en) * | 2011-10-31 | 2015-07-22 | 中兴能源（天津）有限公司 | Method for increasing content of adenosylmethionine and superoxide dismutase in ethanol waste saccharomyces cerevisiae as sorgo fuel |
| CN103131750A (en) * | 2013-02-05 | 2013-06-05 | 大连工业大学 | Method for detecting aging of yeast |
| CN104593337A (en) * | 2014-12-31 | 2015-05-06 | 唯美度科技（北京）有限公司 | Orgotein extracted from beer yeast |
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| Publication number | Publication date |
|---|---|
| CN101168747B (en) | 2010-08-18 |
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