CN103184165A - High-temperature resistant yeast capable of producing xylitol and application thereof - Google Patents
High-temperature resistant yeast capable of producing xylitol and application thereof Download PDFInfo
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- CN103184165A CN103184165A CN2011104629095A CN201110462909A CN103184165A CN 103184165 A CN103184165 A CN 103184165A CN 2011104629095 A CN2011104629095 A CN 2011104629095A CN 201110462909 A CN201110462909 A CN 201110462909A CN 103184165 A CN103184165 A CN 103184165A
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Abstract
Xylitol is an important functional food additive and chemical raw material, and has extensive applications. According to the invention, a high-temperature adaptive evolution method is utilized to screen TIB-Y1020 of high-temperature resistant Candida maltose which is CGMCC (china general microbiological culture collection center) No. 5521 and can grow well at the temperature ranging from 38 DEG C to 42 DEG C without influencing the transformation production of the xylitol. The high-temperature resistance strain can reduce the manufacturing cost increase brought by temperature reduction, thereby having an extensive industrial application prospect.
Description
Technical field
The invention belongs to biological technical field, be specifically related to the thermotolerant yeast bacterium that Xylitol is produced in a strain, maltose candiyeast (Candida maltosa) TIB-Y1020CGMCC No5521 and the method for utilizing this bacterium bio-transformation xylose production Xylitol.
Background technology
Xylitol is natural five carbon polyols, is one of important function derived food additives.The sugariness of Xylitol is 1.05 times of sucrose, and its metabolism need not Regular Insulin, and alternative sucrose is as sweeting agent, nutrition agent and the medicament etc. of patients with diabetes mellitus.In recent years, the market requirement of Xylitol constantly enlarges, and the market of world's Xylitol is estimated at 3.6 hundred million dollars at present, and the annual value of production of domestic Xylitol is above 1,000,000,000 yuan.In addition, Xylitol is widely used in industry such as medicine, chemical industry, food, coating, leather.Xylitol also is a kind of platform chemical, and the method by biological or chemical can be converted into 1,2-propylene glycol, multiple Chemicals such as ethylene glycol.
The production of Xylitol at present mainly is the catalysis process by the wood sugar chemical hydrogenation.Chemical method need with the corn cob hydrolyzed solution concentrate, crystallization obtains behind the wood sugar dissolved dilution again, reconcentration, crystallization behind the hydrogenation, energy consumption height.Chemical method uses the metal-powder catalyzer in addition, and to product separation, particularly the Xylitol production of food grade brings higher separation costs, and product has certain risk when being used for food.Also have chemical method to need hydrogen manufacturing, hydrogenation carries out under High Temperature High Pressure, the equipment requirements height.
Produce the problem that Xylitol exists at chemical method, begin one's study from the seventies in 20th century in the world and produce Xylitol with microbial method.The microorganism ratio juris is to utilize intracellular Xylose reductase, and wood sugar is reduced to Xylitol, and hydrogen does not need hydrogen manufacturing separately from intracellular reducibility coenzyme and hydrogen ion.Microbial reaction carries out at normal temperatures and pressures, and energy consumption is low, equipment is simple, operational safety, again because the Substratspezifitaet of enzyme makes that raw material needn't be highly purified.Biological process produce Xylitol will have pollution less, the quality product advantages of higher, have development prospect.Research and development have for many years both at home and abroad obtained a series of sugar alcohol transformation efficiencys and surpassed 0.8g/g, and are indivedual even surpass the bacterial strain of 0.95g/g, make the biological industrialization of making of Xylitol become possible (Silva et al:Enzyme Microb Technol, 2012; Ahmad et al:Bioprocess Biosyst Eng, 2011; Rodrigues et al:JInd Microbiol Biotechnol, 2011; Cheng et al:Microb Cell Fact, 2011; Prakash et al:Bioresour Technol, 2011; Chen et al:Int J Biol Sci, 2010; Nair and Zhao:Metab Eng, 2010; Sasaki et al:Appl Microbiol Biotechnol, 2010; Kumar et al:J Ind Microbiol Biotechnol, 2009; Akiinerinwa et al:Curr Opin Biotechnol, 2008;
Et al:Appl Microbiol Biotechnol, 2007a;
Et al:Appl Microbiol Biotechnol, 2007b; Silva et al:Appl Biochem Biotechnol, 1998).But microbial method production and the manufacturing of Xylitol are realized in efficient, economy ground, still there is a series of gordian technique to need further to break through, wherein high-performance is produced the core that the seed selection transformation of bacterial strain is the manufacturing of realization Xylitol microorganism, and other comprise that the optimization of zymotechnique and the green separation and purification of product etc. also require further study and develop.
Summary of the invention
The purpose of this invention is to provide a strain can withstand high temperatures, produce the yeast of Xylitol and method and the application that Xylitol is produced in bio-transformation.
It is that maltose candiyeast (Candida maltosa) TIB-Y1020 has hidden on December 2nd, 2011 and protects in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) that high temperature resistant Xylitol provided by the present invention is produced yeast, and preserving number is CGMCC No 5521.
It is at present existing that to utilize yeast to carry out Xylitol production all be to carry out under the temperature about 30 ℃.Because cell will produce heat in the process of metabolism xylose production Xylitol, need to consume a large amount of water coolants and reduce temperature in Industrial processes, to keep the production process about 30 ℃.Predesigne shows the Xylitol production that utilizes 30 ℃, and 1 ton of Xylitol of every production will consume the water coolant more than 50 tons, if improve more than the production temperature to 40 ℃, then the consumption of water coolant can be reduced to below 25 tons.Can reduce cooling water amount significantly so improve the production temperature of Xylitol, reduce energy expenditure, thereby reduce the Xylitol production cost.Realize the Xylitol production of 40 ℃ of left and right sides higher temperatures, suitable high temperature resistant production bacterial strain just must be arranged.Therefore improve yeast to the tolerance of temperature, can not reduce industrial meaning very great in the production level of well-grown under the hot environment and Xylitol.
In order to improve the bacterial strain industrial Applicability As, we efficiently transform the maltose candiyeast of xylose production Xylitol from separation energy voluntarily, utilize the method for adaptive evolution (adaptive evolution) bacterial strain to be carried out the adaptability domestication of hot environment, obtaining a strain can be at 40 ℃ of energy well-grown, and does not influence the yeast maltose candiyeast TIB-Y1020 CGMCC No5521 that Xylitol is produced.The objective of the invention is to improve the temperature that Xylitol is produced in the bacterial strain bio-transformation, can enough under hot environment, adapt to the output and the productive rate that do not influence the production Xylitol when growing.
Utilize this yeast to carry out pyrolytic conversion xylose production Xylitol following advantage arranged:
1) raw material availability height.Chemical method is raw material with pure wood sugar, has the wood sugar more than 30% can't utilize in mother liquor in the corn cob hydrolyzed solution.The high temperature biotransformation method is raw material with the corn cob hydrolyzed solution directly, and has utilized waste wood sugar mother liquor, and raw material availability is improved a lot.
2) production efficiency height: this bacterial classification has repeatedly the ability of using repeatedly, so the Xylitol output of unit cell can exceed nearly 10 times of international highest level, and the Xylitol output of unit volume fermentor tank can exceed international highest level more than 20 times.
3) environmental pollution is low.Pollute because the raw materials pretreatment process can produce certain environment inevitably, also alleviated environmental pollution when therefore having improved raw material availability.In addition, biotransformation is also environmentally friendly than chemical catalysis process.
4) production unit is simple.Chemical method needs hydrogen manufacturing, and hydrogenation carries out under High Temperature High Pressure, the equipment requirements height.The fermentation method major equipment is fermentor tank, and is also complicated.Biotransformation method is only used few fermentor tank, and major equipment is simple reactive tank.
5) simple to operate, safety.Conversion process is carried out under normal temperature, normal pressure, and need not aseptic technique.Compare with chemical method or fermentation method, operation is the simplest, safest.
6) good product quality.Biological process has been avoided and the contacting of metal-powder catalyzer, and product is more suitable for as food.Bioconversion systems does not have the nutrient media components of fermentation system again, so quality product is more reliable.
Concrete enforcement
Below be listed examples, so that understand the present invention better.
The acquisition of embodiment 1 heatproof maltose candiyeast (Candida maltosa) TIB-Y1020 CGMCCNo5521
The maltose candiyeast of Fen Liing (Candida maltosa) is inoculated in the adaptive evolution substratum voluntarily, and this medium pH is 5.5, contains following component by mass percent: wood sugar 10%, extractum carnis 3%, KH
2PO
40.1%, MgSO
47H
2O0.05%.Shake in the bottle in 37 ℃, the cultivation of 220r/min at the 50ml that contains the 10mL substratum and after 12 hours, to be inoculated in the fresh same medium with 0.2 OD600 value then, with the cultivation of going down to posterity of identical method.By that analogy, after the ambient growth that makes bacterial strain adapt to 37 ℃ is stablized, improve growth temperature gradually with identical substratum and culture condition, carry out the adaptability domestication and reach 40 ℃ up to temperature, make it still can stable growth, and increment and starting strain are basic identical 30 ℃ of increments, thereby obtain maltose candiyeast (Candida maltosa) TIB-Y1020 CGMCC No5521.
TIB-Y1020CGMCC No552130 ℃ and 40 ℃ Xylitol production contrasts of embodiment 2 maltose candiyeasts (Candida maltosa)
Maltose candiyeast (Candida maltosa) TIB-Y1020CGMCC No5521 is inoculated in the seed culture medium, this medium pH is 5.5, contain following component by mass percent: wood sugar 5%, peptone 1%, yeast extract paste 1% shakes in the bottle cultivation respectively at 30 ℃ and 40 ℃ 220r/min after 24 hours at the 250ml that 20mL is housed, the centrifugal collection thalline of 2000r/min, again the thalline that suspends is being equipped with 20mL, and the 250mL of 20% xylose solution shakes in the bottle.The thalline of 30 ℃ of cultivations continues to continue at 40 ℃ at the thalline of 30 ℃, 40 ℃ cultivations, the cultivation 36-48 of 220r/min hour, uses wood sugar consumption in the high-efficient liquid phase technique assaying reaction liquid and the generation of Xylitol respectively.Maltose candiyeast TIB-Y1020CGMCC No5521 needed 48 hours to run out of wood sugar when transforming for 30 ℃, generated the Xylitol of 160g/L, and only needed 40 hours just to run out of wood sugar when transforming for 40 ℃, produced the Xylitol of about 165g/L.Under the situation that same conversion efficient is arranged substantially, 40 ℃ transform the production time that needs still less, thereby higher production intensity are arranged.
40 ℃ of embodiment 3 maltose candiyeasts (Candida maltosa) TIB-Y1020CGMCC No5521 are shaken bottle and transform the production Xylitol
The 500mL that the 25mL cell culture medium is equipped with in maltose candiyeast TIB-Y1020CGMCC No5521 inoculation shakes in the bottle, and the pH of this substratum is 5.4, contains following component by mass percent: wood sugar 3%, peptone 1%, yeast extract paste 0.5%.At 40 ℃, the cultivation of 220r/min after 16 hours, centrifugal collection thalline, the thalline that suspends again is being equipped with 25mL, and the 250mL that contains the xylose mother liquid of 20% wood sugar shakes in the bottle.Cultivate at 40 ℃ of 220r/min, with the wood sugar consumption in the high-efficient liquid phase technique assaying reaction liquid and the generation of Xylitol.Maltose candiyeast TIB-Y1020CGMCC No5521 can produce the Xylitol that generates 169g/L at 48 hours.Continue centrifugal recovery thalline and carry out 9 and take turns and transform the xylose production Xylitol, Xylitol output can maintain between the 155g/L-165g/L.
The 5L ferment tank of embodiment 4 maltose candiyeasts (Candida maltosa) TIB-Y1020CGMCC No5521 is produced Xylitol
The 500mL that the 50mL cell culture medium is equipped with in maltose candiyeast TIB-Y1020CGMCC No5521 inoculation shakes in the bottle, and the pH of this substratum is 6, contains following component by mass percent: glucose 2%, yeast powder 1%, peptone are 2%., after 40 ℃, the shaking table of 220r/min are cultivated 8 hours, the inoculation of 50mL nutrient solution is had the 7.5L fermentor tank (NBS of 5000mL cell culture medium
115) in, this substratum contains following component by mass percent: wood sugar 5%, soy peptone 2%, yeast extract paste 1%.It is about 5.5 that NaOH by 5M controls pH in real time.At 40 ℃, air flow is to cultivate under 1: 1 the condition after 24 hours, centrifugal collection thalline, the thalline that suspends again is being equipped with 5000mL, contain 15% wood sugar xylose mother liquid 7.5L fermentor tank (NBS
115) in.Air flow is to cultivate under 1: 1 the condition after 36 hours, with the wood sugar consumption in the high-efficient liquid phase technique assaying reaction liquid and the generation of Xylitol.Maltose candiyeast TIB-Y1020CGMCC No5521 can produce the Xylitol that generates 122g/L at 36 hours.
The 30L fermentor tank of embodiment 5 maltose candiyeasts (Candida maltosa) TIB-Y1020CGMCC No5521 transforms produces Xylitol
The 5000mL that the 1000mL cell culture medium is equipped with in maltose candiyeast TIB-Y1020CGMCC No5521 inoculation shakes in the bottle, and the pH of this substratum is 6, contains following component by mass percent: glucose 2%, yeast powder 1%, peptone are 2%.After 40 ℃, the shaking table of 220r/min are cultivated 8 hours, the inoculation of 1000mL nutrient solution is had in the 30L fermentor tank (east, Zhenjiang GUJS-30 type) of 20L cell culture medium, this substratum contains following component by mass percent: wood sugar 5%, soy peptone 2%, yeast extract paste 1%.It is about 5.5 that NaOH by 5M controls pH in real time.At 40 ℃, air flow is to cultivate under 1: 1 the condition after 24 hours, centrifugal collection thalline, and the thalline that suspends again is being equipped with 20L, contains in the 0L fermentor tank (east, Zhenjiang GUJS-30 type) of the xylose mother liquid of 20% wood sugar.Air flow is to cultivate under 1: 1 the condition after 20 hours, with the wood sugar consumption in the high-efficient liquid phase technique assaying reaction liquid and the generation of Xylitol.Maltose candiyeast TIB-Y1020CGMCC No5521 can produce the Xylitol that generates 166g/L at 48 hours.
Claims (11)
1. maltose candiyeast (Candida maltosa) the TIB-Y1020CGMCC No5521 of the product Xylitol that a strain can withstand high temperatures and utilize this bacterium to transform the method for xylose production Xylitol.The method that transforms the xylose production Xylitol is to cultivate maltose candiyeast (Candida maltosa) cell earlier; Yeast cell after the cultivation separates the substrate that the cell transformation that obtains contains wood sugar with substratum; It is disposable or react the generation Xylitol through mode and yeast cell that stream adds to contain the wood sugar substrate; Reacted yeast cell is separated with reaction solution, and cell can repeat repeatedly and contain the wood sugar substrate and react the generation Xylitol.
2. according to right 1 requirement, the substratum of cultivating maltose candiyeast (Candida maltosa) comprises carbon source, nitrogenous source, pH4.5-6.5.
3. according to claim 2, it is characterized in that: described carbon source is wood sugar and/or the biomass by hydrolyzation liquid glucose of 5%-30% for the quality percentage composition.
4. according to claim 2, it is characterized in that: described nitrogenous source is corn steep liquor and/or extractum carnis and/or yeast extract and/or peptone and/or dregs of beans and/or soybean cake powder and/or peanut meal and/or ammonium salt and/or nitrate and/or the urea of 0.1%-5% for the quality percentage composition.
5. according to claim 1,2,3 or 4 described, it is characterized in that: the cell cultures temperature of maltose candiyeast (Candida maltosa) TIB-Y1020CGMCC No5521 is 25-45 ℃.
6. according to claim 1,2,3,4 or 5 described, it is characterized in that: the cell cultures time of maltose candiyeast (Candida maltosa) TIB-Y1020CGMCC No5521 is 8h-48h.
7. method according to claim 1 is characterized in that the used wood sugar substrate that contains is that wood sugar, hemicellulose hydrolysate comprise corn cob, maize straw, wood fragments etc.
8. method according to claim 1, the temperature of reaction that it is characterized in that transforming the xylose production Xylitol is 25-45 ℃.
9. method according to claim 1, the reaction times that it is characterized in that transforming the xylose production Xylitol is 12-120 hour.
10. the stream that contains the wood sugar substrate according to claim 1 adds process, it is characterized in that: the wood sugar substrate gradation that contains with the solid wood sugar or after concentrating adds reaction system, and reaction once finishes.
11. reaction repeated process according to claim 1 is characterized in that: will be from reaction solution isolated cells with contain the wood sugar substrate and carry out conversion reaction after the meeting again.
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|---|---|---|---|---|
| US10759727B2 (en) | 2016-02-19 | 2020-09-01 | Intercontinental Great Brands Llc | Processes to create multiple value streams from biomass sources |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101676403A (en) * | 2008-09-17 | 2010-03-24 | 山东福田药业有限公司 | Screening method of microbial strain for xylitol production |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101676403A (en) * | 2008-09-17 | 2010-03-24 | 山东福田药业有限公司 | Screening method of microbial strain for xylitol production |
Non-Patent Citations (4)
| Title |
|---|
| C.GUO等: "Screening and characterization of yeasts for xylitol production", 《JOURNAL OF APPLIED MICROBIOLOGY》 * |
| 刘畅 等: "耐高温酵母菌的筛选及特性研究", 《酿酒》 * |
| 田沈 等: "一株酵母菌高温驯化与同步糖化发酵研究", 《太阳能学报》 * |
| 耿笑林 等: "适应进化筛选耐高温产木糖醇麦芽糖假丝酵母", 《中国酿造》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10759727B2 (en) | 2016-02-19 | 2020-09-01 | Intercontinental Great Brands Llc | Processes to create multiple value streams from biomass sources |
| US11840500B2 (en) | 2016-02-19 | 2023-12-12 | Intercontinental Great Brands Llc | Processes to create multiple value streams from biomass sources |
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