CN101824439A - Method for fermentation preparation of L-lactic acid after microwave-alkali coupling pretreatment of distilled grain - Google Patents
Method for fermentation preparation of L-lactic acid after microwave-alkali coupling pretreatment of distilled grain Download PDFInfo
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- CN101824439A CN101824439A CN 201010158290 CN201010158290A CN101824439A CN 101824439 A CN101824439 A CN 101824439A CN 201010158290 CN201010158290 CN 201010158290 CN 201010158290 A CN201010158290 A CN 201010158290A CN 101824439 A CN101824439 A CN 101824439A
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Abstract
The invention relates to a method for fermentation preparation of L-lactic acid after microwave-alkali coupling pretreatment of distilled grains. The method has the steps that fresh distilled grains are pulverized and water content thereof is moderated to 70% to 90% (w/w); alkali is added as 1% to 10 % (w/w) of the dry weight of the distilled grains; and the mixture is treated for 4 to 10 minutes in a microwave reactor with the power of 200 to 500 W. The pretreated distilled grains are taken out and solid-to-liquid ratio thereof is moderated; 3000 to 5000U of cellulose and 1 to 3g of L-lactic acid bacteria are added into every liter of the fermented liquor; the mixture is synchronously saccharified and fermented for 48 to 72 hours at the temperature of 35 to 40 DEG C; and then lactic acid produced by fermentation is separated. The microwave-alkali coupling pretreatment breaks crystallizing zones of cellulose in the distilled grains, which is favorable for enzymolysis reaction, greatly shortens production cycle and improves the production rate of L- lactic acid.
Description
Technical field
What the present invention relates to a kind of solid vinasse utilizes method, the method for particularly a kind of microwave-alkali coupling pretreatment of distilled grain secondary fermentation preparation of L-lactic acid again.
Background technology
The solid vinasse are by products of wine brewing, and China various places are annual to produce a large amount of solid vinasse, and vinasse also have crushed materials such as rice husk or stalk except that containing grain residue such as undecomposed starch.Contain amino acid, reducing sugar and organic acid nutritive substance in the vinasse, especially contain a large amount of robust fibre class materials, account for 24.8～33% of dry weight greatly, have the value of recycling.
Present industrial vinasse are mainly used in produces environmental fertilizer, single-cell protein feed etc., and added value is lower.And plant fiber material is transformed, studying morely both at home and abroad to be with ethanol or methane is target product, but because cellulosic trans-utilization rate is low, causes yielding poorly of target product, thereby limited process of industrialization.
The cellulose substances alkali pre-treatment of development in recent years and Microwave Pretreatment technology, reduce cellulosic degree of crystallinity at the destruction cellulosic structure, slough xylogen, the sponginess that increases raw material with increase cellulase system with cellulosic effective contact etc. that the aspect obtains must effect, thereby the output of raising enzymolysis efficiency and succeeding target product.
Publication number is the regulate and control method that the Chinese invention patent application of CN101671701 discloses a kind of producing L-lactic acid via opened fermented food waste, wherein utilizes Chinese liquor distiller grains inoculation lactobacillus johnsonii to make L-lactic acid.But vinasse are a kind of as cellulose class refuse, and the quality of its pretreating effect is directly connected to the size of productive rate.Therefore, research high-cellulose transformation efficiency, for follow-up lactic fermentation provides the preconditioning technique that enriches sugared source, most important.
Summary of the invention
The method that the purpose of this invention is to provide a kind of microwave-alkali coupling pretreatment of distilled grain secondary fermentation preparation of L-lactic acid, adopt the pretreated vinasse of this method can directly carry out simultaneous saccharification and fermentation and produce L-lactic acid, solved single treatment technology length consuming time, treatment effect is not obvious and the cellulose utilization rate is low, problems such as saccharification result is poor, and lactic acid yield is low.
Technical scheme of the present invention is as follows: the method for a kind of microwave-alkali coupling pretreatment of distilled grain secondary fermentation preparation of L-lactic acid, and its step is as follows:
(1) pre-treatment: after fresh vinasse pulverizing, regulating the vinasse water ratio is 70～90%wt, add alkali, mix with vinasse, vinasse are in be fully immersed in the state in the lysed alkali lye, the quality of alkali accounts for 1～10% (w/w) of fresh vinasse dry weight, and alkali lye is fully contacted with vinasse, and is to handle 4～10min under the microwave state of 200～500W to obtain fermented liquid at power.Alkali can be sodium hydroxide or calcium hydroxide.The ammonia soln that perhaps can directly add 8%wt makes that the quality of ammonia accounts for 1～10% of distiller's dried grain quality in the ammoniacal liquor, also can satisfy production requirement even so regulate the water ratio of vinasse earlier, and vinasse are fully immersed in the ammoniacal liquor.
(2) simultaneous saccharification and fermentation produces L-lactic acid: the pretreated fermented liquid of step (1) is regulated solid-to-liquid ratio is: according to the fresh vinasse of raw material that add in the 1g step (1): the amount of 4～8g water is regulated, the L-milk-acid bacteria that in every L fermented liquid, adds cellulase and the 1～3g of 3000～5000U, under 35～40 ℃ of conditions, carry out anaerobically fermenting 48～72h, the L-lactic acid that separate fermentation produces.
Shortcomings such as the present invention is with the cellulosic materials raw material of vinasse as fermenting lactic acid, and, lactic acid production low at cellulosic materials fermenting raw materials lactic acid producing saccharification efficient is few adopt microwave-alkali coupled method that vinasse are carried out pre-treatment, have following characteristics and advantage:
(1) the present invention adopts microwave to carry out pre-treatment in the presence of alkali lye, makes to need only 4～10min by preprocessing process weak point consuming time, and cellulose conversion is the efficient height of sugar, has shortened the fermentative production cycle.
(2) the present invention has brought into play the chemical action of alkali and the physical action of microwave, destroy cellulosic structure and reduce cellulosic degree of crystallinity, slough xylogen, can increase sponginess and the cellulase system and cellulosic effective contact of raw material, thereby improve enzymolysis efficiency and follow-up L-lactic acid yield.
(3) the present invention can directly carry out simultaneous saccharification and fermentation after pretreated vinasse add cellulase and L-milk-acid bacteria, need not sterilize, do not need to add in addition nutritive substance, when efficiently fermenting, reduced fermentation costs, help the high value added utilization of vinasse.
Embodiment
Below by embodiment technical scheme of the present invention is described further.
Embodiment 1 (microwave-alkali coupling pre-treatment): get distiller's dried grain 20g or fresh vinasse 57.1g, be ground into fine particle, the adjusting water ratio is 90%wt, add sodium hydroxide and mix, dissolution of sodium hydroxide is fully immersed in the sodium hydroxide solution vinasse in the water that vinasse contain, wherein the quality of sodium hydroxide accounts for the quality 6% of distiller's dried grain, uniform mixing obtains fermented liquid, puts into microwave, microwave 8min under 328W.Take out pretreated material, the adjusting solid-to-liquid ratio is: according to 1g raw material fresh vinasse: the ratio of 4g water is regulated, in every liter fermented liquid, add the cellulase of 4000U and the L-milk-acid bacteria of 1.4g, under 35 ℃ of conditions, carry out anaerobically fermenting 48h, survey L-lactic acid yield.Obtain L-lactic acid/g distiller's dried grain of 0.2341g, L-lactic acid/100g fermented product of 1.64g.
Embodiment 2 (single alkali pre-treatment): get distiller's dried grain 20g or fresh vinasse 57.1g, be ground into fine particle, the adjusting water ratio is 90%wt, adding sodium hydroxide is fully immersed in the solution vinasse, wherein the quality of sodium hydroxide account for distiller's dried grain heavy 6%, uniform mixing obtains fermented liquid, the solid-to-liquid ratio of regulating fermented liquid is: according to 1g raw material fresh vinasse: the ratio of 4g water is regulated, in every liter fermented liquid, add cellulase 4000U and 1.4g milk-acid bacteria, under 35 ℃ of conditions, carry out anaerobically fermenting 48h, survey L-lactic acid yield.Obtain L-lactic acid/g distiller's dried grain of 0.0542g, L-lactic acid/100g fermented product of 0.38g.
Embodiment 3 (single Microwave Pretreatment): get distiller's dried grain 20g or fresh vinasse 57.1g, be ground into fine particle, the adjusting water ratio is 90%wt, put into microwave, microwave 8min under 328W, take out fermented liquid and regulate solid-to-liquid ratio and be: according to 1g raw material fresh vinasse: the ratio of 4g water is regulated, and adds the cellulase of 4000U and the L-milk-acid bacteria of 1.4g in every liter of fermented liquid, under 35 ℃ of conditions, carry out anaerobically fermenting 48h, survey L-lactic acid yield.Obtain L-lactic acid/g distiller's dried grain of 0.077g, L-lactic acid/100g fermented product of 0.54g.
Embodiment 4 is identical with embodiment 1, and just the lactic fermentation time lengthening the results are shown in following table to 72h.
Embodiment 5 is identical with embodiment 2, and just the lactic fermentation time lengthening the results are shown in following table to 72h.
Embodiment 6 is identical with embodiment 3, and just the lactic fermentation time lengthening the results are shown in following table to 72h.
| Embodiment | Every gram distiller's dried grain produces L-lactic acid production (g) | Every 100g fermentation produce L-lactic acid productions (g) |
| Embodiment 4 (microwave-alkali coupling pre-treatment) | ??0.2627 | ??1.84 |
| Embodiment 5 (single alkali pre-treatment) | ??0.0728 | ??0.51 |
| Embodiment 6 (single Microwave Pretreatment) | ??0.0799 | ??0.59 |
The L-lactic acid yield that can obviously be found out the preparation of L-lactic acid that ferments again behind employing microwave-alkali coupling pretreatment of distilled grain by above embodiment is the highest, and effect is best.
Embodiment 7 (microwave-alkali coupling pre-treatment): get distiller's dried grain 20g, be ground into fine particle, the adjusting water ratio is 70%wt, adding calcium hydroxide is fully immersed in the alkali lye vinasse, wherein the quality of calcium hydroxide accounts for distiller's dried grain quality 1%, uniform mixing obtains fermented liquid, puts into microwave, microwave 4min under 500W.Take out pretreated material, the adjusting solid-to-liquid ratio is: according to 1g raw material fresh vinasse: the ratio of 6g water is regulated, in every liter fermented liquid, add the cellulase of 5000U and the L-milk-acid bacteria of 1g, under 40 ℃ of conditions, carry out anaerobically fermenting 60h, the L-lactic acid that separate fermentation produces.
Embodiment 1 (microwave-alkali coupling pre-treatment): get dried fresh vinasse 57.1g, be ground into fine particle, ammonia soln 10.9～the 109ml that can directly add 8%wt, just wherein in the ammoniacal liquor quality of ammonia account for distiller's dried grain quality 1～10%, even the water ratio that uncomfortable like this sobriety is poor also can meet the demands, vinasse are fully immersed in the ammoniacal liquor, uniform mixing obtains fermented liquid, put into microwave, microwave 10min under 200W.Take out pretreated material, the adjusting solid-to-liquid ratio is: according to the 1g fresh vinasse: the ratio of 8g water is regulated, in every liter fermented liquid, add the cellulase of 3000U and the L-milk-acid bacteria of 3g, under 38 ℃ of conditions, carry out anaerobically fermenting 58h, the L-lactic acid that separate fermentation produces.
Claims (7)
1. the method for microwave-alkali coupling pretreatment of distilled grain secondary fermentation preparation of L-lactic acid is characterized in that step is as follows:
The first step, pre-treatment: after the vinasse pulverizing, regulating the vinasse water ratio is 70～90%wt, adds alkali, mixes, and vinasse are fully immersed in the alkali lye, and microwave treatment 4～10min obtains fermented liquid then;
Second step, simultaneous saccharification and fermentation produces L-lactic acid: it is the 1g vinasse that the pretreated fermented liquid of the first step is regulated solid-to-liquid ratio: 4～8g water, add cellulase and milk-acid bacteria, under 35～40 ℃ of conditions, carried out anaerobically fermenting 48～72 hours, the L-lactic acid that separate fermentation produces.
2. the method for a kind of microwave according to claim 1-alkali coupling pretreatment of distilled grain secondary fermentation preparation of L-lactic acid is characterized in that the quality of alkali is 1～10% of a vinasse dry weight in the first step pre-treatment.
3. the method for a kind of microwave according to claim 1 and 2-alkali coupling pretreatment of distilled grain secondary fermentation preparation of L-lactic acid is characterized in that alkali is sodium hydroxide or calcium hydroxide in the first step pre-treatment.
4. the method for a kind of microwave according to claim 1 and 2-alkali coupling pretreatment of distilled grain secondary fermentation preparation of L-lactic acid is characterized in that alkali is the ammoniacal liquor of 8%wt in the first step pre-treatment, makes that the quality of ammonia accounts for 1～10% of vinasse dry weight in the ammoniacal liquor.
5. the method for a kind of microwave according to claim 1-alkali coupling pretreatment of distilled grain secondary fermentation preparation of L-lactic acid is characterized in that the power of microwave treatment is 200～500W in the first step.
6. the method for a kind of microwave according to claim 1-alkali coupling pretreatment of distilled grain secondary fermentation preparation of L-lactic acid is characterized in that, in second step, adds the cellulase of 3000～5000U in every liter fermented liquid.
7. the method for a kind of microwave according to claim 1-alkali coupling pretreatment of distilled grain secondary fermentation preparation of L-lactic acid is characterized in that, in second step, adds the L-milk-acid bacteria of 1～3g in every liter fermented liquid.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104431295A (en) * | 2014-12-19 | 2015-03-25 | 四川省食品发酵工业研究设计院 | Method for preparing livestock feed from vinasse |
| CN113621674A (en) * | 2021-08-27 | 2021-11-09 | 泸州老窖股份有限公司 | Method for producing L-lactic acid by using liquor distiller grains |
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| CN101155925A (en) * | 2005-04-12 | 2008-04-02 | 纳幕尔杜邦公司 | System and process for biomass treatment |
| CN101230547A (en) * | 2008-02-26 | 2008-07-30 | 广西壮族自治区中医药研究院 | Method for preparing cellulose and carboxymethyl cethylose by wood fibre material |
| JP2009001733A (en) * | 2007-06-25 | 2009-01-08 | Purex:Kk | Method for decomposing/liquefying solid cellulose, and resultant decomposed product |
| CN101671701A (en) * | 2009-10-14 | 2010-03-17 | 北京科技大学 | Adjusting control method for producing L-lactic acid via opened fermented food waste |
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2010
- 2010-04-28 CN CN 201010158290 patent/CN101824439A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1431231A (en) * | 2003-01-29 | 2003-07-23 | 广西壮族自治区中国科学院广西植物研究所 | Method for extracting xylan from woody fiber material |
| CN101155925A (en) * | 2005-04-12 | 2008-04-02 | 纳幕尔杜邦公司 | System and process for biomass treatment |
| JP2009001733A (en) * | 2007-06-25 | 2009-01-08 | Purex:Kk | Method for decomposing/liquefying solid cellulose, and resultant decomposed product |
| CN101230547A (en) * | 2008-02-26 | 2008-07-30 | 广西壮族自治区中医药研究院 | Method for preparing cellulose and carboxymethyl cethylose by wood fibre material |
| CN101671701A (en) * | 2009-10-14 | 2010-03-17 | 北京科技大学 | Adjusting control method for producing L-lactic acid via opened fermented food waste |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104431295A (en) * | 2014-12-19 | 2015-03-25 | 四川省食品发酵工业研究设计院 | Method for preparing livestock feed from vinasse |
| CN104431295B (en) * | 2014-12-19 | 2017-05-03 | 四川省食品发酵工业研究设计院 | Method for preparing livestock feed from vinasse |
| CN113621674A (en) * | 2021-08-27 | 2021-11-09 | 泸州老窖股份有限公司 | Method for producing L-lactic acid by using liquor distiller grains |
| CN113621674B (en) * | 2021-08-27 | 2022-06-21 | 泸州老窖股份有限公司 | Method for producing L-lactic acid by using liquor distiller grains |
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