CN102154388B - Method for producing succinic acid by fermentation of crop straw - Google Patents
Method for producing succinic acid by fermentation of crop straw Download PDFInfo
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- CN102154388B CN102154388B CN 201010610881 CN201010610881A CN102154388B CN 102154388 B CN102154388 B CN 102154388B CN 201010610881 CN201010610881 CN 201010610881 CN 201010610881 A CN201010610881 A CN 201010610881A CN 102154388 B CN102154388 B CN 102154388B
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Abstract
The invention provides a method for producing succinic acid by fermentation of crop straw, which is to produce succinic acid by fermentation with mixed sugar solution of straw enzymatic hydrolysis sugar solution and xylose mother liquor as a carbon source, wherein in a fermentation process, sugar is supplied by batch feeding, the sugar concentration of the fermentation liquid is controlled to be 3 to 5 g/L, the pH value of the fermentation liquid is controlled to be 6.5 to 7.0, the temperature is 35 to 40 DEG, and the fermentation is performed for 30 to 60 hours. The method provided by the invention has the advantages of wide raw material source and low production cost, and meanwhile, a new method is provided for processing the xylose mother liquor which is difficult to process.
Description
Technical field
The present invention relates to a kind of method of comprehensive utilization of agricultural crop straw, specifically, relate to a kind of method of utilizing stalk stalk enzymolysis sugar and xylose mother liquid to prepare succinic acid.
Background technology
Succinic acid is called again succsinic acid, is a kind of binary organic acid, and it is a kind of important C
4The hardware and software platform compound can be used as the intermediate that synthesizes complex organic compound, is widely used in pharmacy, food-processing, synthetic plastics, rubber, protective coating, dyestuff and other industry.
The method of producing succinic acid mainly contains two kinds: chemical synthesis and microbe fermentation method.Traditional chemical synthesis is because using the reasons such as fossil feedstock, environmental pollution to make the industrial scale of succinic acid and application be subject to larger restriction.
The many merits such as the Production by Microorganism Fermentation succinic acid is high due to efficient, pollution is little are studied extensively and profoundly.But in food crop, starch is raw material because present Production by Microorganism Fermentation succinic acid all utilizes, and a large amount of grain of need of production consumption of succinic acid forms the situation of striving grain with the people, to the mankind's the certain threat of existence formation.
China is a large agricultural country, and agricultural crop straw is the renewable resource of great potential, especially paid attention to widely as the raw material of the renewable energy resources and Chemicals, but at present except small portion as fuel combustion, major part is not utilized.
Therefore, need to seek a kind of succinic acid production method of not using petrochemicals, can be mass-produced, do not strive grain, starting material wide material sources with the people.
Summary of the invention
The object of the present invention is to provide a kind of method of utilizing fermenting crops straws to produce succinic acid.
The method of utilizing stalk fermentation to produce succinic acid provided by the invention, take the mixed sugar liquid of stalk enzymolysis liquid glucose and xylose mother liquid as the carbon source through fermentation succinic acid, stream adds benefit sugar during the fermentation, the sugared concentration of controlled fermentation liquid is 3～5g/L, controlling pH is 6.5～7.0,35～40 ℃, fermentation 30～60h.
Wherein, in the liquid glucose that is mixed of stalk enzymolysis liquid glucose and xylose mother liquid, the mass ratio of glucose and xylose is 1～2: 1, is preferably 4/3～3/2: 1.
According to the present invention, the concentration of fermentation end sugar is 10～30g/L, is preferably 20g/L, and the concentration of fermentation stream sugaring is 300～400g/L.
According to the present invention, in the succinic acid fermenting process, begin stream the end of when when sugar concentration is lower than 3g/L and add benefit sugar, in controlled fermentation liquid, sugared concentration is in 3～5g/L scope.
According to the present invention, stalk enzymolysis liquid glucose is the enzymolysis solution that agricultural crop straw obtains through soda boiling, solid-liquid separation, enzymolysis, solid-liquid separation.
Wherein, described stalk is agricultural crop straw, comprises Wheat Straw, rice straw or other agricultural crop straws.
Described agricultural crop straw can carry out pre-treatment before soda boiling, general through pulverizing and washing standby.
According to the present invention, xylose mother liquid is the mother liquor that corn cob obtains after washing, acidolysis, solid-liquid separation, crystallization.
According to the present invention, method of the present invention is applicable to the bacterial strain of suitable succinic acid-producing well known in the art, is preferably Actinobacillus succinogenes CGMCCNo.1716.
The production method of succinic acid of the present invention, its advantage is:
1. starting material wide material sources: China's agricultural straw resource is very abundant.
2. utilization rate of raw materials is high: succinic acid is produced bacterial strain can utilize pentose and hexose simultaneously, and in the agricultural crop straw raw material, main component is pentose and hexose.
3. production cost is low: agricultural crop straw raw material and xylose mother liquid raw material are cheap than starchy material, and the succinic acid production cost is low.
At present, utilize the fermenting crops straws succinic acid, fermentation period is long, residual sugar is high, is unfavorable for the scale operation of succinic acid.The present invention's trim point of glucose and xylose concentration from seek the stalk hydrolysis sugar is set about, and keeps low sugar concn in fermenting process, thereby has solved this drawback.
Embodiment
Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.
Embodiment 1 preparation stalk enzymolysis liquid glucose
1) maize straw is pulverized and washed;
2) maize straw soda boiling: the agricultural crop straw after will pulverizing and wash is in 16g/L NaOH solution, 100 ℃ of lower boilings 2 hours;
3) solid-liquid separation: the soda boiling solid and liquid is separated, and obtains the soda boiling slag;
4) soda boiling slag enzymolysis: with every gram soda boiling slag 20IU cellulase, 10IU cellobiase, 15IU zytase; In enzymolysis process, pH is 6.0, and temperature is 40 ℃, and enzymolysis time is 40 hours, gets enzymolysis solution;
5) solid-liquid separation: the enzymolysis solution solid-liquid separation obtains being rich in glucose, wood sugar and stalk enzymolysis liquid glucoses of other assorted sugar and the Production With Residue of Enzymatic Hydrolysis that are rich in xylogen on a small quantity.
Embodiment 2 preparation stalk enzymolysis liquid glucoses
1) maize straw is pulverized and washed;
2) maize straw soda boiling: the agricultural crop straw after will pulverizing and wash is in the 20g/L sodium hydroxide solution, 120 ℃ of lower boilings 1 hour;
3) solid-liquid separation: the soda boiling solid and liquid is separated, and obtains the soda boiling slag;
4) soda boiling slag enzymolysis: with every gram soda boiling slag 30IU cellulase, 20IU zytase; In enzymolysis process, pH is 6.5, and temperature is 45 ℃, and enzymolysis time is 45 hours, gets enzymolysis solution;
5) solid-liquid separation: the enzymolysis solution solid-liquid separation obtains being rich in glucose, wood sugar and stalk enzymolysis liquid glucoses of other assorted sugar and the Production With Residue of Enzymatic Hydrolysis that are rich in xylogen on a small quantity.
Embodiment 3 preparation xylose mother liquids
Corn cob is washed;
Corncob acid hydrolysis: after washing corn cob in 12g/L sulfuric acid, 125 ℃ of acid treatment 2 hours;
Solid-liquid separation: the corn cob pickling agent carries out solid-liquid separation, gets acid hydrolysis solution and acidolysis slag;
Crystallization: with acid hydrolysis solution purifying, concentrated, crystallization, get wood sugar and xylose mother liquid.
Embodiment 4 preparation Actinobacillus succinogenes seed liquor
Adopt the dull and stereotyped Actinobacillus succinogenes bacterial classification of cultivating (buy Nanjing University of Technology and preserve the Actinobacillus succinogenes that is numbered CGMCCNo.1716) to be inoculated into glucose 20g/L, yeast extract paste 4g/L, peptone 3g/L, corn steep liquor 4g/L, Na
2CO
312g/L, NaH
2PO
49g/L, K
2HPO
415g/L, Na
2HPO
43g/L, NaCl2g/L, the pH nature is cultivated in the shake-flask seed substratum of tap water preparation, cultivates 18 hours for 35 ℃, and pH is controlled at 6.5～7.0, shaking speed 180rpm.Passing into carbonic acid gas in the seeding tank that cultured shake-flask seed access is identical with the shake-flask seed substratum again cultivates, cultivated 15 hours for 35 ℃, pH is controlled at 6.7, mixing speed 175rpm, ventilating ratio (ratio of volume of material and air capacity, lower same) 1: 0.4.
Embodiment 5
The fermention medium concentration proportioning is: stalk enzymolysis liquid glucose (according to embodiment 1 preparation) is 10g/L with xylose mother liquid (according to the embodiment 3 preparations) total sugar concentration that is mixed, wherein, being mixed, the mass ratio of glucose and xylose is 2: 1 in total reducing sugar, yeast extract paste 12g/L, peptone 4g/L, corn steep liquor 15g/L, NaAc 1.2g/L, NaH
2PO
40.25g/L, K
2HPO
43g/L, Na
2HPO
40.3g/L, NaCl 0.3g/L, CaCl
20.3g/L, MgCl
20.3g/L, pH nature, tap water preparation.
The supplemented medium concentration proportioning is: stalk enzymolysis liquid glucose (according to embodiment 1 preparation) and xylose mother liquid (according to the embodiment 3 preparations) total sugar concentration that is mixed 300g/L, wherein, being mixed, the mass ratio of glucose and xylose is 2: 1 in total reducing sugar, yeast extract paste 12g/L, peptone 4g/L, corn steep liquor 15g/L, NaAc 1.2g/L, NaH
2PO
40.25g/L, K
2HPO
43g/L, Na
2HPO
40.3g/L, NaCl 0.3g/L, CaCl
20.3g/L, MgCl
20.3g/L, pH nature, tap water preparation.
Select the 50L fermentor tank, sterilized 15 minutes for 121 ℃, access the Actinobacillus succinogenes seed of cultured embodiment 4 preparations when being cooled to 37 ℃.In fermenting process, flow feeding is controlled substratum sugar concentration at 3～5g/L, stream adds aseptic sodium hydroxide the pH value is controlled at 6.7 simultaneously, carbon dioxide flows in the fermention medium of fermentor tank take the rate of venting of ventilating ratio as 1: 0.2, mixing speed is 300rpm, temperature is controlled at 35 ℃, anaerobism was cultivated 40 hours, and the succinic acid productive rate reaches 47.3g/L.
Embodiment 6
The fermention medium concentration proportioning is: stalk enzymolysis liquid glucose (according to embodiment 2 preparations) is 10g/L with xylose mother liquid (according to the embodiment 3 preparations) total sugar concentration that is mixed, wherein, being mixed, the mass ratio of glucose and xylose is 1: 1 in total reducing sugar, yeast extract paste 12g/L, peptone 4g/L, corn steep liquor 15g/L, NaAc 1.2g/L, NaH
2PO
40.25g/L, K
2HPO
43g/L, Na
2HPO
40.3g/L, NaCl 0.3g/L, CaCl
20.3g/L, MgCl
20.3g/L, pH nature, tap water preparation.
The supplemented medium concentration proportioning is: stalk enzymolysis liquid glucose (according to embodiment 2 preparations) is 300g/L with xylose mother liquid (according to the embodiment 3 preparations) total sugar concentration that is mixed, wherein, being mixed, the mass ratio of glucose and xylose is 1: 1 in total reducing sugar, yeast extract paste 12g/L, peptone 4g/L, corn steep liquor 15g/L, NaAc 1.2g/L, NaH
2PO
40.25g/L, K
2HPO
43g/L, Na
2HPO
40.3g/L, NaCl 0.3g/L, CaCl
20.3g/L, MgCl
20.3g/L, pH nature, tap water preparation.
Select the 50L fermentor tank, sterilized 15 minutes for 121 ℃, access the Actinobacillus succinogenes seed of cultured embodiment 4 preparations when being cooled to 37 ℃.In fermenting process, flow feeding is controlled substratum sugar concentration at 3～5g/L, stream adds aseptic magnesium hydroxide the pH value is controlled at 7.0 simultaneously, carbon dioxide flows in the fermention medium of fermentor tank take the rate of venting of ventilating ratio as 1: 0.2, mixing speed is 300rpm, temperature is controlled at 35 ℃, anaerobism was cultivated 50 hours, and the succinic acid productive rate reaches 44.9g/L.
Embodiment 7
The fermention medium concentration proportioning is: stalk enzymolysis liquid glucose (according to embodiment 2 preparations) is 30g/L with xylose mother liquid (according to the embodiment 3 preparations) total sugar concentration that is mixed, wherein, being mixed, the mass ratio of glucose and xylose is 3: 2 in total reducing sugar, yeast extract paste 12g/L, peptone 4g/L, corn steep liquor 15g/L, NaAc 1.2g/L, NaH
2PO
40.25g/L, K
2HPO
43g/L, Na
2HPO
40.3g/L, NaCl 0.3g/L, CaCl
20.3g/L, MgCl
20.3g/L, pH nature, tap water preparation.
The supplemented medium concentration proportioning is: stalk enzymolysis liquid glucose (according to embodiment 2 preparations) is 400g/L with xylose mother liquid (according to the embodiment 3 preparations) total sugar concentration that is mixed, wherein, being mixed, the mass ratio of glucose and xylose is 3: 2 in total reducing sugar, yeast extract paste 12g/L, peptone 4g/L, corn steep liquor 15g/L, NaAc 1.2g/L, NaH
2PO
40.25g/L, K
2HPO
43g/L, Na
2HPO
40.3g/L, NaCl 0.3g/L, CaCl
20.3g/L, MgCl
20.3g/L, pH nature, tap water preparation.
Select the 50L fermentor tank, sterilized 15 minutes for 121 ℃, access the Actinobacillus succinogenes seed of cultured embodiment 4 preparations when being cooled to 37 ℃.In fermenting process, flow feeding is controlled substratum sugar concentration at 3～5g/L, stream adds aseptic magnesiumcarbonate the pH value is controlled at 6.5 simultaneously, carbon dioxide flows in the fermention medium of fermentor tank take the rate of venting of ventilating ratio as 1: 0.2, mixing speed is 300rpm, temperature is controlled at 35 ℃, anaerobism was cultivated 60 hours, and the succinic acid productive rate reaches 59.7g/L.
Embodiment 8
Stalk enzymolysis liquid glucose (according to embodiment 1 preparation) is 20g/L with xylose mother liquid (according to the embodiment 3 preparations) total sugar concentration that is mixed, and wherein, in the total reducing sugar that is mixed, the mass ratio of glucose and xylose is 4: 3, yeast extract paste 12g/L, peptone 4g/L, corn steep liquor 15g/L, NaAc 1.2g/L, NaH
2PO
40.25g/L, K
2HPO
43gL, Na
2HPO
40.3g/L, NaCl 0.3g/L, CaCl
20.3g/L, MgCl
20.3g/L, pH nature, tap water preparation.
The supplemented medium concentration proportioning is: stalk enzymolysis liquid glucose (according to embodiment 1 preparation) is 400g/L with xylose mother liquid (according to the embodiment 3 preparations) total sugar concentration that is mixed, wherein, being mixed, the mass ratio of glucose and xylose is 4: 3 in total reducing sugar, yeast extract paste 12g/L, peptone 4g/L, corn steep liquor 15g/L, NaAc 1.2g/L, NaH
2PO
40.25g/L, K
2HPO
43g/L, Na
2HPO
40.3g/L, NaCl 0.3g/L, CaCl
20.3g/L, MgCl
20.3g/L, pH nature, tap water preparation.
Select the 50L fermentor tank, sterilized 15 minutes for 121 ℃, access the Actinobacillus succinogenes seed of cultured embodiment 4 preparations when being cooled to 37 ℃.In fermenting process, flow feeding is controlled substratum sugar concentration at 3～5g/L, stream adds aseptic magnesium hydroxide the pH value is controlled at 6.7 simultaneously, carbon dioxide flows in the fermention medium of fermentor tank take the rate of venting of ventilating ratio as 1: 0.2, mixing speed is 300rpm, temperature is controlled at 35 ℃, anaerobism was cultivated 60 hours, and the succinic acid productive rate reaches 60.4g/L.
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements, all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (4)
1. method of utilizing the crop straws for producing succinic acid, it is characterized in that, take stalk enzymolysis liquid glucose and xylose mother liquid mixed sugar liquid as the carbon source through fermentation succinic acid, stream adds benefit sugar during the fermentation, the sugared concentration of controlled fermentation liquid is 3～5g/L, controlling pH is 6.5～7.0,35～40 ℃ of temperature, fermentation 30～60h;
Wherein, in described stalk enzymolysis liquid glucose and xylose mother liquid mixed sugar liquid, the mass ratio of glucose and xylose is 1～2:1;
Wherein, described stalk enzymolysis liquid glucose is the enzymolysis solution that agricultural crop straw obtains through soda boiling, solid-liquid separation, enzymolysis, solid-liquid separation; Described stalk is agricultural crop straw, comprises Wheat Straw, rice straw or other agricultural crop straws; Described agricultural crop straw carried out pre-treatment before soda boiling, through pulverizing and washing standby;
Wherein, described xylose mother liquid is the mother liquor that corn cob obtains after washing, acidolysis, solid-liquid separation, crystallization;
Wherein, the bacterial strain of described fermentation succinic acid be Actinobacillus succinogenes CGMCCN o.1716;
The proportioning of fermention medium concentration is: in described stalk enzymolysis liquid glucose and xylose mother liquid mixed sugar liquid, the mass ratio of glucose and xylose is 1～2:1, and fermentation end sugar concentration is 10～30g/L, and it is 300～400g/L that stream adds the concentration of mending sugar;
The component that fermention medium is mixed outside total reducing sugar is yeast extract paste 12g/L, peptone 4g/L, corn steep liquor 15g/L, NaAc1.2g/L, NaH
2PO
40.25g/L, K
2HPO
43g/L, Na
2HPO
40.3g/L, NaCl0.3g/L, CaCl
20.3g/L, MgCl
20.3g/L, pH nature, tap water preparation;
Component outside the total reducing sugar that is mixed of benefit sugar culture-medium is: yeast extract paste 12g/L, peptone 4g/L, corn steep liquor 15g/L, NaAc1.2g/L, NaH
2PO
40.25g/L, K
2HPO
43g/L, Na
2HPO
40.3g/L, NaCl0.3g/L, CaCl
20.3g/L, MgCl
20.3g/L, pH nature, tap water preparation.
2. the method for claim 1, is characterized in that, begins stream and add benefit sugar when in fermented liquid, sugared concentration is lower than 3g/L, and in controlled fermentation liquid, sugared concentration is in 3～5g/L scope.
3. the method for claim 1, is characterized in that, adds aseptic neutralizing agent by stream the pH value of fermented liquid is controlled at 6.5～7.0.
4. method as claimed in claim 3, is characterized in that, described aseptic neutralizing agent is magnesiumcarbonate, magnesium hydroxide or sodium hydroxide.
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| CN112028224B (en) * | 2020-09-16 | 2021-10-12 | 广东恩维乐环境科技有限公司 | Processing technology for preparing carbon source for high-quality sewage treatment from waste sugar |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101603058A (en) * | 2008-06-11 | 2009-12-16 | 浙江杭州鑫富药业股份有限公司 | Utilize fermenting sugar grass raw material to produce the method for Succinic Acid |
| CN101608192A (en) * | 2009-07-22 | 2009-12-23 | 安徽丰原发酵技术工程研究有限公司 | A kind of method of utilizing corn cob to produce Succinic Acid |
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| US20110091941A1 (en) * | 2008-07-03 | 2011-04-21 | Novozymes A/S | Process For Producing Fermentation Products |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101603058A (en) * | 2008-06-11 | 2009-12-16 | 浙江杭州鑫富药业股份有限公司 | Utilize fermenting sugar grass raw material to produce the method for Succinic Acid |
| CN101608192A (en) * | 2009-07-22 | 2009-12-23 | 安徽丰原发酵技术工程研究有限公司 | A kind of method of utilizing corn cob to produce Succinic Acid |
Non-Patent Citations (4)
| Title |
|---|
| Efficient conversion of crop stalk wastes into succinic acid production by Actinobacillus succinogenes;Li Qiang et al;《Bioresource Technology》;20100112;第101卷;3292-3294 * |
| Li Qiang et al.Efficient conversion of crop stalk wastes into succinic acid production by Actinobacillus succinogenes.《Bioresource Technology》.2010,第101卷3292-3294. |
| 李兴江等.秸秆两步发酵法制备丁二酸研究.《食品科学》.2009,第30卷(第8期),72-75. |
| 秸秆两步发酵法制备丁二酸研究;李兴江等;《食品科学》;20091231;第30卷(第8期);72-75 * |
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