CN103820511A - Xylitol fermentation output improvement method - Google Patents
Xylitol fermentation output improvement method Download PDFInfo
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- CN103820511A CN103820511A CN201410060877.XA CN201410060877A CN103820511A CN 103820511 A CN103820511 A CN 103820511A CN 201410060877 A CN201410060877 A CN 201410060877A CN 103820511 A CN103820511 A CN 103820511A
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Abstract
The invention provides a xylitol fermentation output improvement method. The xylitol fermentation output is improved by adding sodium gluconate with Candida tropicalis as a fermenting strain. The addition of 5-10g/L of sodium gluconate in an initial medium improves the flux of a phosphopentose pathway in the fermentation process, increases the supply of NADP and enables the conversion rate of xyloseas fermented as a substrate to produce xylitol to reach 96.5% and the xylitol output to reach 96.5g/L. The method has the advantages of simplicity, easy implementation, low cost and less energy consumption, and can be used for the industrialized large-scale production of xylitol.
Description
Technical field
The invention belongs to technical field of bioengineering, be specifically related to a kind of method that improves xylitol fermentation output.
Background technology
Xylitol is a kind of important five-carbon sugar alcohol, is the sweeting agent that extracts and pass through chemical process from other natural plants such as corn cob.Estimation according to the insiders, current global Xylitol turnout has reached ten thousand tons of 5-6, and wherein China has accounted for more than 60%, ten thousand tons of about 3-4.And global Xylitol market demand may reach more than 100,000 tons in next three years, potential market demand will be up to 40%.But the competition of Xylitol industry is also more and more fierce, controlling cost and ensuring the quality of products is the basic assurance that obtains competitive edge.
Xylose mother liquid is the saturate thick liquid staying after xylose crystalline, the assorted sugar such as wood sugar, L-arabinose, glucose, semi-lactosi, seminose are contained in the inside, wherein the ratio of wood sugar, pectinose, glucose is generally 2:1:1, and other assorted sugar (semi-lactosi seminose) content is below 8%.The different manufacturers of xylose mother liquid and different lot numbers containing total reducing sugar amount difference, general total reducing sugar is between 60-75%.Wood sugar accounts between the 50-70% of total reducing sugar.Xylose mother liquid, because the assorted sugared content of the inside is high, causes the solubleness of wood sugar wherein cannot use conventional methods and crystallize out greatly.If adopt the method for chromatographic separation, sugar different the inside can be separated.Adopt chromatographic separation technology to separate various monose, comprise Xylitol, this explained hereafter cost is high.
Produce at present Xylitol and have two kinds of chemical hydrogenation method and biological fermentation process.Because xylose mother liquid is containing multiple assorted sugar, cannot utilize chemical hydrogenation method to produce Xylitol.Production by Microorganism Fermentation Xylitol can be avoided violent chemical reaction, and fermentation condition gentleness does not need special corrosion-resistant anti-high-tension apparatus; The anti-adversity ability of cell can adapt to more extensive raw material; The Substratspezifitaet of biological catalyst guarantees to only have Xylitol in product, and purification procedures is simple, can significantly reduce production costs, and is considered to have most the Xylitol production method of development potentiality.Occurring in nature can utilize the microorganism of wood sugar generation Xylitol a lot, and from conversion capability, the ability of yeast production Xylitol is the most superior, candida tropicalis (C.tropicalis) especially wherein.
Summary of the invention
The object of this invention is to provide a kind of method of adding Sunmorl N 60S and improve Xylitol output in fermention medium.The present invention, take wood sugar as fermentation substrate, uses candida tropicalis strain, adds Sunmorl N 60S in fermentation culture, carries out the production of Xylitol by batch fermentation.
The party's ratio juris is that gluconic acid can be induced the synthetic of gluconokinase, make gluconic acid synthetic 6-p-gluconic acid under the effect of gluconokinase, 6-p-gluconic acid generates ribose 5-phosphate again under the effect of 6-p-gluconate dehydrogenase, has increased the flux of phosphopentose pathway (HMP approach).And key enzyme Xylose reductase in xylitol fermentation is the single-minded dependent form of DPNH I (NADPH), and NADPH is generated in a large number by HMP approach.The enhancing of HMP approach has increased the supply of NADPH greatly, thereby has promoted the fermentation of Xylitol.
In the present invention, the formula of fermentation culture is: corn steep liquor 3-6g/L, yeast powder 3-6g/L, potassium primary phosphate 6-9g/L, MgSO
47H
2o0.1-0.4g/L, urea 3-6g/L, wood sugar 90-120g/L, Sunmorl N 60S 5-10g/L, prepares with water.
Further, the formula of fermentation culture is: corn steep liquor 3-6g/L, yeast powder 3-6g/L, potassium primary phosphate 6-9g/L, MgSO
47H
2o0.1-0.4g/L, urea 3-6g/L, wood sugar 90-120g/L, Sunmorl N 60S 5-8g/L, prepares with water.
Preferably, the formula of fermentation culture is: corn steep liquor 3.5g/L, yeast powder 3.5g/L, potassium primary phosphate 6g/L, MgSO
47H
2o0.2g/L, urea 3g/L, wood sugar 110g/L, Sunmorl N 60S 6g/L, prepares with water.
Yeast described in the inventive method is the yeast in mycocandida.
Preferably, described yeast is candida tropicalis (C.tropicalis).
In the inventive method, leavening temperature is 30-35 ℃, initial pH4-5, OD
600nmit is 20%-30% that value is controlled dissolved oxygen before reaching 20, OD
600nmvalue reaches 20 rear reduction dissolved oxygens to 0%-5%, and fermentation period is 60-70 hour.
Preferably, leavening temperature is 33 ℃, initial pH4-5, OD
600nmit is 20%-30% that value is controlled dissolved oxygen before reaching 20, OD
600nmvalue reaches 20 rear reduction dissolved oxygens to 0%-5%, and fermentation period is 60-65 hour.
The invention provides Sunmorl N 60S in the application improving in xylitol fermentation output.
Above-mentioned application refers to that in fermentation culture, gluconic acid na concn is 5-10g/L.
Particularly, can adopt the impact of Sunmorl N 60S on Xylitol output in following methods checking real attenuation:
Utilize 50L fermentor tank, carry out fermenting experiment using candida tropicalis as fermentation strain, in initial medium, add Sunmorl N 60S, make that its final concentration reaches respectively 0,3,6,9g/L, other nutritive ingredients are identical, are 4-5 at the fermentation initial stage with NaOH adjust pH, temperature control is 30-35 ℃, OD
600nmbefore reaching 20, be 20-30% by adjusting stirring velocity and air flow control dissolved oxygen, work as OD
600nmreach after 20, be reduced to the hypoxia of 0-5%, and be maintained to fermentation ends by reducing stirring velocity and air flow control dissolved oxygen, fermentation period is 60-70 hour.
Experimental result shows, does not add Sunmorl N 60S, and in the time that initial Xylose Content is 100g/L, last 64h Determination of Xylitol is only 82.1g/L, and sugar alcohol transformation efficiency is 82.1%.In initial incubation liquid, the Sunmorl N 60S of 6g/L has best effect to xylitol fermentation, makes fermentation period foreshorten to 61 hours, and finally, in the time that initial Xylose Content is 100g/L, Xylitol output reaches 96.5g/L, and sugar alcohol transformation efficiency has reached 96.5%.In addition, as shown in Figure 1, the addition (9g/L) that further increases Sunmorl N 60S is not significantly improved to Xylitol output and transformation efficiency, has but additionally increased production cost.In sum, add 6g/L Sunmorl N 60S the fermentation of Xylitol is had to best effect.
The method that improves Xylitol output by add Sunmorl N 60S in fermention medium provided by the invention, the method is simply effective, because industrial glucose acid sodium consumption is less, scale operation cost, without obvious increase, is therefore applicable to the heavy industrialization fermentative production of Xylitol.
Accompanying drawing explanation
Fig. 1 adds the affect figure of different glucose acid sodium on xylitol fermentation output.
Embodiment
Following examples further illustrate content of the present invention, but should not be construed as limitation of the present invention.Without departing from the spirit and substance of the case in the present invention, the modification that the inventive method, step or condition are done or replacement, all belong to scope of the present invention.
If do not specialize, the conventional means that in embodiment, technique means used is well known to those skilled in the art, in embodiment 1-4, adds each raw material except special instruction, is commercially available.Candida tropicalis (C.tropicalis) deposit number using in the present invention is CICC1316, purchased from Chinese industrial microbial strains preservation center.
Embodiment 1 Sunmorl N 60S addition is 0g/L
In 50L fermentor tank, initial fermentation culture is settled to 27L, does not add Sunmorl N 60S in nutrient solution.Nutrient solution formula is: corn steep liquor 5g/L, yeast powder 4g/L, potassium primary phosphate 8g/L, MgSO
47H
2o0.2g/L, urea 5g/L, wood sugar 100g/L, prepare with water.
Candida tropicalis kind liquid, according to 10% inoculum size (about 3L), is accessed in sterilized initial incubation liquid and fermented.Initial Xylose Content is pressed 100g/L preparation, and before kind, first regulating initial pH value with NaOH is 4.5, and during the fermentation, leavening temperature maintains 34 ℃, the not automatic control of fermented liquid pH value.Initial sterile air ventilating ratio is 1:0.6, initial mixing speed is 400rpm, and initial tank pressure is 0.04Mpa, and initial dissolved oxygen is set to 100%, along with the carrying out of fermentation, dissolved oxygen starts to decline, and cell concentration starts continuous increase, when the dense OD of bacterium (600nm) reached before 20, control dissolved oxygen in 27% left and right, in the time that the dense OD of bacterium (600nm) reaches 20, reduce and be stirred to 150rpm, make dissolved oxygen be reduced to 3% left and right.Through fermentation in 64 hours, Determination of Xylitol reached 82.1g/L, and wood sugar reaches 82.1% to the quality transformation efficiency of Xylitol.
Embodiment 2 Sunmorl N 60S additions are 6g/L
In 50L fermentor tank, initial fermention medium is settled to 27L, wherein adds Sunmorl N 60S and makes concentration reach 6g/L.Nutrient solution formula is: corn steep liquor 5g/L, yeast powder 4g/L, potassium primary phosphate 8g/L, MgSO
47H
2o0.3g/L, urea 3-6g/L, wood sugar 100g/L, Sunmorl N 60S 6g/L, prepares with water.
Candida tropicalis kind liquid, according to 10% inoculum size (about 3L), is accessed in sterilized initial medium and fermented.Initial Xylose Content is pressed 100g/L preparation, and before kind, first regulating initial pH value with NaOH is 4.5, and during the fermentation, leavening temperature maintains 32 ℃, the not automatic control of fermented liquid pH value.Initial sterile air ventilating ratio is 1:0.6, initial mixing speed is 400rpm, and initial tank pressure is 0.04Mpa, and initial dissolved oxygen is set to 100%, along with the carrying out of fermentation, dissolved oxygen starts to decline, and cell concentration starts continuous increase, when the dense OD of bacterium (600nm) reached before 20, control dissolved oxygen in 26% left and right, in the time that the dense OD of bacterium (600nm) reaches 20, reduce and be stirred to 150rpm, make dissolved oxygen be reduced to 2%.Through fermentation in 60 hours, Determination of Xylitol reached 95.1g/L, reaches 95.1% by wood sugar to the quality transformation efficiency of Xylitol.
Embodiment 3 Sunmorl N 60S additions are 5g/L
In 500L fermentor tank, initial fermention medium is settled to 270L, wherein adds Sunmorl N 60S and makes concentration reach 5g/L.The formula of fermentation culture is: corn steep liquor 3g/L, yeast powder 3g/L, potassium primary phosphate 6g/L, MgSO
47H
2o0.1g/L, urea 3g/L, wood sugar 100g/L, Sunmorl N 60S 5g/L, prepares with water.
Candida tropicalis kind liquid, according to 10% inoculum size (about 30L), is accessed in sterilized initial medium and fermented.Initial Xylose Content is pressed 100g/L preparation, and before kind, first regulating initial pH value with NaOH is 4.5, and during the fermentation, leavening temperature maintains 35 ℃, the not automatic control of fermented liquid pH value.Initial sterile air ventilating ratio is 1:0.4, initial mixing speed is 300rpm, and initial tank pressure is 0.04Mpa, and initial dissolved oxygen is set to 100%, along with the carrying out of fermentation, dissolved oxygen starts to decline, and cell concentration starts continuous increase, when the dense OD of bacterium (600nm) reached before 20, control dissolved oxygen in 22% left and right, after the dense OD of bacterium (600nm) reaches 20, reduce and be stirred to 100rpm, make dissolved oxygen be reduced to 5%.Through fermentation in 62 hours, Determination of Xylitol reached 94.5g/L, and wood sugar reaches 94.5% to the quality transformation efficiency of Xylitol.
Embodiment 4 Sunmorl N 60S additions are 9g/L
At 60m
3in fermentor tank, initial fermention medium is settled to 27m
3, wherein add Sunmorl N 60S and make concentration reach 9g/L.The formula of fermentation culture is: corn steep liquor 6g/L, yeast powder 6g/L, potassium primary phosphate 9g/L, MgSO
47H
2o0.4g/L, urea 6g/L, wood sugar 100g/L, Sunmorl N 60S 9g/L, prepares with water.
Candida tropicalis kind liquid, according to 10% inoculum size (about 30L), is accessed in sterilized initial medium and fermented.Initial Xylose Content is pressed 100g/L preparation, and before kind, first regulating initial pH value with NaOH is 4.5, and during the fermentation, leavening temperature maintains 33 ℃, the not automatic control of fermented liquid pH value.Initial sterile air ventilating ratio is 1:0.2, initial mixing speed is 250rpm, and initial tank pressure is 0.02Mpa, and initial dissolved oxygen is set to 100%, along with the carrying out of fermentation, dissolved oxygen starts to decline, and cell concentration starts continuous increase, when the dense OD of bacterium (600nm) reached before 20, control dissolved oxygen in 20% left and right, in the time that the dense OD of bacterium (600nm) reaches 20, reduce and be stirred to 100rpm, make dissolved oxygen be reduced to 3% left and right.Through fermentation in 61.5 hours, Determination of Xylitol reached 95.5g/L, and wood sugar reaches 95.5% to the quality transformation efficiency of Xylitol.
Embodiment 5 Sunmorl N 60S additions are 3g/L
In 500L fermentor tank, initial fermention medium is settled to 270L, wherein adds Sunmorl N 60S and makes concentration reach 3g/L.The formula of fermentation culture is: corn steep liquor 5g/L, yeast powder 4g/L, potassium primary phosphate 5g/L, MgSO
47H
2o0.2g/L, urea 4g/L, wood sugar 100g/L, Sunmorl N 60S 3g/L, prepares with water.
Candida tropicalis kind liquid, according to 10% inoculum size (about 30L), is accessed in sterilized initial medium and fermented.Initial Xylose Content is pressed 100g/L preparation, and before kind, first regulating initial pH value with NaOH is 4.5, and during the fermentation, leavening temperature maintains 30 ℃, the not automatic control of fermented liquid pH value.Initial sterile air ventilating ratio is 1:0.4, initial mixing speed is 300rpm, and initial tank pressure is 0.04Mpa, and initial dissolved oxygen is set to 100%, along with the carrying out of fermentation, dissolved oxygen starts to decline, and cell concentration starts continuous increase, when the dense OD of bacterium (600nm) reached before 20, control dissolved oxygen in 29% left and right, in the time that the dense OD of bacterium (600nm) reaches 20, reduce and be stirred to 100rpm, make dissolved oxygen be reduced to 0.Through fermentation in 66 hours, Determination of Xylitol reached 86.5g/L, and wood sugar reaches 86.5% to the quality transformation efficiency of Xylitol.
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 without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (10)
1. a method that improves xylitol fermentation output, is characterized in that, take wood sugar as fermentation substrate, adds Sunmorl N 60S in fermentation culture, produces Xylitol by yeast fermentation.
2. the method for claim 1, is characterized in that, the formula of fermentation culture is: corn steep liquor 3-6g/L, yeast powder 3-6g/L, potassium primary phosphate 6-9g/L, MgSO
47H
2o0.1-0.4g/L, urea 3-6g/L, wood sugar 90-120g/L, Sunmorl N 60S 5-10g/L, prepares with water.
3. method as claimed in claim 2, is characterized in that, the formula of fermentation culture is: corn steep liquor 3-6g/L, yeast powder 3-6g/L, potassium primary phosphate 6-9g/L, MgSO
47H
2o0.1-0.4g/L, urea 3-6g/L, wood sugar 90-120g/L, Sunmorl N 60S 5-8g/L, prepares with water.
4. method as claimed in claim 2, is characterized in that, the formula of fermentation culture is: corn steep liquor 3.5g/L, yeast powder 3.5g/L, potassium primary phosphate 6g/L, MgSO
47H
2o0.2g/L, urea 3g/L, wood sugar 110g/L, Sunmorl N 60S 6g/L, prepares with water.
5. the method as described in as arbitrary in claim 1-4, is characterized in that, described yeast is the yeast in mycocandida.
6. method as claimed in claim 5, is characterized in that, described yeast is candida tropicalis (C.tropicalis).
7. the method as described in as arbitrary in claim 1-4, is characterized in that, leavening temperature is 30-35 ℃, initial pH4-5, OD
600nmbefore reaching 20, controlling dissolved oxygen is 20%-30%, OD
600nmvalue reaches 20 rear reduction dissolved oxygens to 0%-5%, and fermentation period is 60-70 hour.
8. method as claimed in claim 7, is characterized in that, leavening temperature is 33 ℃, initial pH4-5, OD
600nmbefore reaching 20, controlling dissolved oxygen is 20%-30%, OD
600nmvalue reaches 20 rear reduction dissolved oxygens to 0%-5%, and fermentation period is 60-65 hour.
9. Sunmorl N 60S is in the application improving in xylitol fermentation output.
10. application as claimed in claim 9, is characterized in that, in fermentation culture, gluconic acid na concn is 5-10g/L.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104357339A (en) * | 2014-11-05 | 2015-02-18 | 江南大学 | Strain for producing xylitol and method for producing xylitol |
| CN106148428A (en) * | 2015-03-23 | 2016-11-23 | 河南工业大学 | A kind of immobilized cell multibatches produces the new technique of xylitol |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101643753A (en) * | 2009-09-11 | 2010-02-10 | 安徽丰原发酵技术工程研究有限公司 | Preparation method for klinint |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101643753A (en) * | 2009-09-11 | 2010-02-10 | 安徽丰原发酵技术工程研究有限公司 | Preparation method for klinint |
Non-Patent Citations (2)
| Title |
|---|
| S RANGASWAMY ET AL: "Screening of facultative anaerobic bacteria utilizing D-xylose for xylitol production", 《APPL MICROBIOL BIOTECHNOL》, vol. 60, 8 August 2002 (2002-08-08), pages 88 - 93 * |
| 林燕清: "葡萄糖酸氧化杆菌产木糖醇发酵条件优化及关键酶基因的克隆与表达", 《中国优秀硕士学位论文全文数据库工程科技I辑》, 15 September 2012 (2012-09-15) * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104357339A (en) * | 2014-11-05 | 2015-02-18 | 江南大学 | Strain for producing xylitol and method for producing xylitol |
| CN106148428A (en) * | 2015-03-23 | 2016-11-23 | 河南工业大学 | A kind of immobilized cell multibatches produces the new technique of xylitol |
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