WO2021082440A1 - Method for producing xylooligosaccharide under catalysis of xylonic acid - Google Patents

Method for producing xylooligosaccharide under catalysis of xylonic acid Download PDF

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WO2021082440A1
WO2021082440A1 PCT/CN2020/093889 CN2020093889W WO2021082440A1 WO 2021082440 A1 WO2021082440 A1 WO 2021082440A1 CN 2020093889 W CN2020093889 W CN 2020093889W WO 2021082440 A1 WO2021082440 A1 WO 2021082440A1
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xylan
xylonic acid
oligosaccharides
reaction
xylo
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PCT/CN2020/093889
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French (fr)
Chinese (zh)
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徐勇
周鑫
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南京林业大学
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Publication of WO2021082440A1 publication Critical patent/WO2021082440A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/02—Monosaccharides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/12—Disaccharides

Definitions

  • the invention relates to the technical fields of food engineering and chemical engineering, in particular to a method for catalyzing the production of xylo-oligosaccharides by xylonic acid.
  • Xylooligosaccharides also known as xylo-oligosaccharides, are mainly derived from the hydrolysis of xylan in lignocellulosic fibers. As a functional food or feed additive, they cannot be absorbed by the digestive system, but they can selectively proliferate bifidobacteria in the intestinal tract. Activate a variety of immune cell activities; therefore, driven by the rapid development of human health and micro-ecology, food safety and green animal breeding, ecological agriculture and other industries, xylo-oligosaccharide products derived from wood fiber are used as a kind of " The development prospects of "Super Prebiotics" are very broad.
  • xylo-oligosaccharides mainly uses endo-xylanase preparations to catalyze the hydrolysis of alkali-extracted xylan, but this method relies on high-priced biological enzymes, high cost, long cycle, and the alkali extraction treatment process is complicated and difficult;
  • the ordinary strong acid hydrolysis method can be used to prepare xylo-oligosaccharides.
  • the yield of xylo-oligosaccharides is low and there are many by-products and the product quality is not high.
  • the present invention overcomes the deficiencies in the prior art and provides a method for catalyzing the production of xylo-oligosaccharides by xylonic acid.
  • the present invention provides the following technical solutions: a method for producing xylo-oligosaccharides from xylonic acid, which includes mixing xylan raw materials and xylonic acid, heating and stirring the reaction; calculated in parts by mass
  • the xylan raw material is 1 part, and the xylonic acid is 5-12 parts.
  • the xylan raw material is xylan and/or xylan-containing lignocellulosic raw material.
  • the stirring rate of the heating and stirring reaction is 30-100 rpm
  • the temperature is 130-170° C.
  • the time is 0.25-2.0 h.
  • the fermentation liquid is placed in a bipolar membrane electrodialysis salt chamber, and the acid chamber and the alkali chamber are added to the salt chamber respectively.
  • Ionized water is used to drive the electrodialysis reaction by applying a direct current power supply, and the xylonic acid solution in the acid chamber is added to the xylan raw material; the electrodialysis reaction is performed until the conductivity of the salt chamber is stable.
  • the bacteria are Xylose Oxidizing Bacillus, which is 0.01 to 0.1 parts.
  • the pH is adjusted after cooling down to room temperature and then adjusted to weak acidity, and the temperature of the low-temperature stirring is 25-35°C, The stirring rate is 100 ⁇ 200rmp.
  • the present invention overcomes the deficiencies in the prior art and provides a method for fermentatively catalyzed production of xylooligosaccharides, which includes mixing xylose with bacterial cells to obtain a mixed liquid; adjusting the mixed liquid pH, stirring at low temperature; adding xylan raw materials, heating and stirring, to obtain xylo-oligosaccharides.
  • the method for producing xylo-oligosaccharides by fermentation and catalysis wherein: after the biological oxidation, it also includes placing the mixed solution in a bipolar membrane electrodialysis salt chamber, and adding deionization to the acid chamber and the alkali chamber respectively.
  • Water is used to drive the electrodialysis reaction by applying a direct current power supply, and the xylonic acid solution in the acid chamber is added to the xylan raw material; the electrodialysis reaction is performed until the conductivity of the salt chamber is stable.
  • the bacterial cell is xylosoxidizing bacteria
  • the xylan raw material is xylan and/or xylan-containing xylan Fiber raw materials
  • the xylose is 1 part
  • the bacteria is 0.01 to 0.1 parts
  • the xylan raw material is 1 to 5 parts.
  • the pH of the mixed liquid is adjusted to be weakly acidic; the temperature of the low-temperature stirring is 25-35°C, The stirring rate is 100 to 200 rpm, the stirring rate of the heating and stirring reaction is 30 to 100 rpm, the temperature is 130 to 170° C., and the time is 0.25 to 2.0 h.
  • the present invention uses xylonic acid as a catalyst, with high yield and less by-product xylose furfural; the present invention chooses bio-oxidation and electrodialysis coupling technology to convert xylose into xylonic acid, wherein xylonic acid can be recycled and low as a self-supply catalyst. Xylose products are more pure.
  • Figure 1 is a high-efficiency anion exchange chromatographic analysis chart in Example 1;
  • Figure 2 is a flow chart of the production process in Example 6;
  • FIG. 3 is a schematic diagram of the electrodialysis reaction in Example 6.
  • FIG. 3 is a schematic diagram of the electrodialysis reaction in Example 6.
  • the "one embodiment” or “embodiment” referred to herein refers to a specific feature, structure, or characteristic that can be included in at least one implementation of the present invention.
  • the appearances of "in one embodiment” in different places in this specification do not all refer to the same embodiment, nor are they separate or selectively mutually exclusive embodiments with other embodiments.
  • Thermo Fisher ICS5000 ion chromatography equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column
  • the temperature is 30°C
  • the sample volume is 10 ⁇ L
  • the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
  • Xylonic acid (XA) and xylose (Xylose), xylobiose (X2), xylotriose (X3), xylotetraose (X4), xylopentaose (X5), Xylose (X6), xyloseptaose (X7), and xylooctaose (X8) can be detected at the same time.
  • the main components are xylose to xylooctaose, and the yields are 27.6%, 19.2%, 13.1%, 8.8%, 5.9%, 2.5%, 1.6% and 1.3%, a total of 80%, of which xylo-oligosaccharides total 52.4%; in addition, the furfural yield is 0.05%.
  • Thermo Fisher ICS5000 ion chromatography equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column
  • the temperature is 30°C
  • the sample volume is 10 ⁇ L
  • the mobile phase is 100mmol/L sodium hydroxide and 500mmol/L sodium acetate for binary gradient elution
  • the flow rate is 0.3mL/min.
  • the analysis chart shows that xylonic acid (XA) and xylose (Xylose), xylobiose (X2), xylotriose (X3), xylotetraose (X4), xylopentaose (X5), xylohexaose (X6) ), Xylose (X7), Xylose (X8) can be detected at the same time.
  • the main components are xylose to xylooctaose, and the yields are respectively 30.5%, 17.2%, 13.2%, 9.1%, 6.8%, 3.9%, 2.6% and 1.7%, totaling 86.2%, of which xylooligosaccharides totaling 55.7 %; In addition, the furfural yield is 0.07%.
  • the xylan hydrolysate was neutralized with sodium hydroxide to pH 5.5 and 2 g/L of Gluconobacter oxydans was added to a 2L bioreactor at the same time to carry out the biological oxidation reaction.
  • the conditions of the biological oxidation reaction were: temperature 30°C, stirring speed It is 150rpm, the air volume is 0.5vvm, the reaction time is 24h, 98% of the xylose is converted into xylonic acid; after the reaction, the bacteria and the fermentation broth are separated by centrifugation, the centrifugal condition: 5000rpm, 5min; the fermentation broth is mainly The ingredients are xylonic acid and xylo-oligosaccharides; the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, 500mL deionized water is added to the acid chamber and the alkali chamber respectively, and the electrodialysis reaction is driven by an external DC power supply.
  • the conductivity of the salt chamber is Refer to the detection and separation reaction process. After 1 hour of reaction, the conductivity of the salt chamber is stable and the reaction ends. At this time, the recovery of xylonic acid in the acid chamber is 96.8%, and the recovery rate of xylo-oligosaccharide in the salt chamber is 100%.
  • Thermo Fisher ICS5000 ion chromatography equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column
  • the temperature is 30°C
  • the sample volume is 10 ⁇ L
  • the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
  • xylonic acid XA
  • xylose Xylose
  • X2 xylobiose
  • X3 xylotriose
  • X4 xylotetraose
  • X5 xylohexaose
  • Xylose Xylose
  • X8 Xylose
  • the main components are xylose to xylooctaose, and the yields are 16.2%, 15.8%, 10.2%, 9.3%, 6.1%, 4.8%, 2.5%, and 1.8%, a total of 66.7%, of which a total of 50 xylo-oligosaccharides %; In addition, the furfural yield is 0.03%.
  • the xylan hydrolysate is neutralized with sodium hydroxide to pH 5.5 and 8g/L Gluconobacter oxydans (American Type Strain Collection ATCC 621H) is simultaneously added to a 2L bioreactor for biological oxidation reaction, biological oxidation reaction
  • the conditions are: the temperature is 30°C, the stirring speed is 150rpm, the air volume is 0.5vvm, the reaction time is 12h, and 99% of the xylose is converted into xylonic acid; after the reaction, the bacteria and the fermentation broth are separated by centrifugation.
  • the main components of the fermentation broth are xylonic acid and xylo-oligosaccharides;
  • the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, 500mL deionized water is added to the acid chamber and the alkali chamber, and the electricity is driven by an external DC power supply.
  • the conductivity of the salt chamber is used as a reference to detect the separation reaction process. After 1 hour of reaction, the conductivity of the salt chamber is stable and the reaction ends. At this time, the recovery of xylonic acid in the acid chamber is 96.3%, and the recovery rate of xylo-oligosaccharides in the salt chamber is 100%. .
  • Thermo Fisher ICS5000 ion chromatography equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column
  • the temperature is 30°C
  • the sample volume is 10 ⁇ L
  • the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
  • xylonic acid (XA) and xylose (Xylose), xylobiose (X2), xylotriose (X3), xylotetraose (X4), xylopentaose (X5), xylohexaose (X6), wood Heptaose (X7) and Xylose (X8) can be detected at the same time.
  • the main components are xylose to xylooctaose, and the yields are respectively 35.2%, 16.5%, 12.1%, 8.2%, 6.3%, 4.4%, 3.6%, 1.9% and 1.6%, a total of 89.8%, of which oligomeric wood
  • the total sugar is 54.6%; in addition, the furfural yield is 0.08%.
  • the xylan hydrolysate is neutralized with sodium hydroxide to pH 5.5 and 2g/L of Gluconobacter Fortorii (American Type Strain Collection ATCC IFO 3264) is simultaneously added to a 2L bioreactor for biological oxidation reaction.
  • the conditions of the biological oxidation reaction are: temperature 30°C, stirring speed 150rpm, air flow rate 0.5vvm, reaction time 12h, 94% xylose is converted into xylonic acid; after the reaction, the bacteria and fermentation are separated by centrifugation Centrifugation conditions: 5000rpm, 5min; the main components of the fermentation broth are xylonic acid and xylo-oligosaccharides; the fermentation broth is applied to the bipolar membrane electrodialysis salt chamber, and 500mL of deionized water is added to the acid chamber and the alkali chamber, respectively, through external direct current The electrodialysis reaction is driven by the power supply, and the separation reaction process is detected with the conductivity of the salt chamber as a reference.
  • Thermo Fisher ICS5000 ion chromatography equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column
  • the temperature is 30°C
  • the sample volume is 10 ⁇ L
  • the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
  • xylonic acid XA
  • xylose Xylose
  • X2 xylobiose
  • X3 xylotriose
  • X4 xylotetraose
  • X5 xylohexaose
  • Xylose Xylose
  • X8 Xylose
  • the main components xylose to xylooctaose were 19.1%, 17.6%, 11.2%, 8.4%, 6.8%, 5.1%, 2.5%, and 1.3%, a total of 72%, of which xylo-oligosaccharides total 52.9%; in addition, the furfural yield is 0.06%.
  • the production process flow chart is shown in Figure 2.
  • a 1L mechanically stirred stainless steel high-pressure reaction tank add 50g of corncob base to extract xylan and 500mL of 10% (mass fraction) xylonic acid solution. After sealing, turn on the stirring ( 60rpm), and heat to 150°C for 45min; after the reaction, after the reaction tank is lowered to room temperature, put the reacted solid-liquid mixture into a vacuum washer, and squeeze and filter to separate the solids that have not been hydrolyzed With xylan hydrolysate (the hydrolysate is mainly a mixture of xylose, xylonic acid and xylo-oligosaccharides).
  • the obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography.
  • the chromatographic conditions Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10 ⁇ L; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
  • the xylan hydrolysate was neutralized with sodium hydroxide to pH 5.5 and 5 g/L of Gluconobacter oxydans was added to a 2L bioreactor at the same time to carry out the biological oxidation reaction.
  • the conditions of the biological oxidation reaction were: temperature 30°C, stirring speed At 150rpm, the air volume is 0.5vvm, the reaction time is 12h, 98% of the xylose is converted into xylonic acid; after the reaction, the bacteria and the fermentation broth are separated by centrifugation, the centrifugal condition: 5000rpm, 5min; the fermentation broth is mainly The components are xylonic acid and xylo-oligosaccharides; the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, and the electrodialysis reaction principle diagram is shown in Figure 3. The acid chamber and the alkali chamber are respectively added with 500mL deionized water, and a DC power supply is added.
  • the electrodialysis reaction is driven, and the separation reaction process is detected with the conductivity of the salt chamber as a reference. After 1 hour of reaction, the conductivity of the salt chamber is stable and the reaction ends. At this time, the recovery rate of xylonic acid in the acid chamber is 97.9%, and the mass concentration is about 11%. The recovery rate of xylo-oligosaccharides in the salt chamber is 100%.
  • the hydrolysate is mainly a mixture of xylose, xylonic acid and xylo-oligosaccharides
  • the yields are 29.0%, 16.1%, 13.1%, 9.6%, 6.1%, 3.4 %, 1.9% and 0.9%, a total of 80.1%, of which xylo-oligosaccharides are 51.1%; in addition, the furfural yield is 0.04%.
  • Thermo Fisher ICS5000 ion chromatography equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column
  • the temperature is 30°C
  • the sample volume is 10 ⁇ L
  • the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
  • xylose to xylooctaose The main components of xylose to xylooctaose, the yields are 72.6%, 8.2%, 6.1%, 3.2%, 0.9%, 0.1%, 0.1% and 0.02%, a total of 91.2%, of which xylo-oligosaccharides are 18.6% ; In addition, the furfural yield is 0.8%. Because under high-intensity (high acid, high temperature) conditions, xylonic acid makes the hydrolysis of xylan more complete, but the xylose content will be higher.
  • Thermo Fisher ICS5000 ion chromatography equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column
  • the temperature is 30°C
  • the sample volume is 10 ⁇ L
  • the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
  • the main components of xylose to xylooctaose are 65.2%, 9.2%, 7.1%, 4.2%, 3.0%, 1.6%, 0.5% and 0.1%, totaling 90.9%, of which xylooligosaccharides totaling 25.7% ;
  • the furfural yield is 0.4%.
  • Thermo Fisher ICS5000 ion chromatography equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column
  • the temperature is 30°C
  • the sample volume is 10 ⁇ L
  • the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
  • the main components of xylose to xylooctaose are 11.6%, 8.8%, 8.2%, 7.3%, 5.0%, 3.9%, 2.1% and 1.5% respectively, totaling 48.4%, of which xylooligosaccharides totaling 36.8% ;
  • the furfural yield is 0.04%.
  • the obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography.
  • the chromatographic conditions Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10 ⁇ L; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
  • xylonic acid (XA) and xylose (Xylose), xylobiose (X2), xylotriose (X3), xylotetraose (X4), xylopentaose (X5), xylohexaose (X6) ), Xylose (X7), Xylose (X8) can be detected at the same time.
  • the main components of xylose to xylooctaose, the yields were 10.1%, 9.8%, 9.2%, 8.1%, 6.9%, 4.8%, 3.7%, and 3.3%, totaling 55.9%. 45.8%; in addition, the furfural yield is 0.02%.
  • the xylan hydrolysate was neutralized with sodium hydroxide to pH 5.5 and 4g/L of Gluconobacter oxydans was added to a 2L bioreactor at the same time to carry out the biological oxidation reaction.
  • the conditions of the biological oxidation reaction were: temperature 30°C, stirring speed It is 150rpm, the air volume is 0.5vvm, the reaction time is 6h, 98% of the xylose is converted into xylonic acid; after the reaction, the bacteria and the fermentation broth are separated by centrifugation, the centrifugal condition: 5000rpm, 5min; the fermentation broth is mainly
  • the ingredients are xylonic acid and xylo-oligosaccharides; the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, 500mL deionized water is added to the acid chamber and the alkali chamber respectively, and the electrodialysis reaction is driven by an external DC power supply.
  • the conductivity of the salt chamber is Refer to the detection and separation reaction process. After 1 hour of reaction, the conductivity of the salt chamber is stable and the reaction ends. At this time, the recovery rate of xylonic acid in the acid chamber is 96.5%, the mass concentration is about 5.5%, and the recovery rate of xylo-oligosaccharides in the salt chamber is 100%. .
  • the hydrolysate is mainly a mixture of xylose, xylonic acid and xylo-oligosaccharides), among which xylose to xylooctaose, the yields (calculated based on the initial raw material 50g xylan) are 8.2%, 7.2%, 6.3 %, 4.1%, 3.2%, 2.4%, 0.9% and 0.7%, a total of 33%, of which xylo-oligosaccharides total 24.8%; in addition, the furfural yield is 0.04%.
  • Thermo Fisher ICS5000 ion chromatography equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column
  • the temperature is 30°C
  • the sample volume is 10 ⁇ L
  • the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
  • the main ingredients xylose to xylooctaose are 21.6%, 18.7%, 14.9%, 13.8%, 9.6%, 5.7%, 2.9% and 1.5%, a total of 88.7%, of which 67.1% of xylo-oligosaccharides ;
  • the furfural yield is 0.05%.
  • the xylan hydrolysate was neutralized with sodium hydroxide to pH 5.5 and 5 g/L of Gluconobacter oxydans was added to a 2L bioreactor at the same time to carry out the biological oxidation reaction.
  • the conditions of the biological oxidation reaction were: temperature 30°C, stirring speed At 150rpm, the air volume is 0.5vvm, the reaction time is 12h, 98% of the xylose is converted into xylonic acid; after the reaction, the bacteria and the fermentation broth are separated by centrifugation, the centrifugal condition: 5000rpm, 5min; the fermentation broth is mainly The ingredients are xylonic acid and xylo-oligosaccharides; the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, 500mL deionized water is added to the acid chamber and the alkali chamber respectively, and the electrodialysis reaction is driven by an external DC power supply.
  • the conductivity of the salt chamber is Refer to the detection and separation reaction process.
  • the conductivity of the salt chamber is stable and the reaction ends.
  • the recovery rate of xylonic acid in the acid chamber is 97.9%
  • the mass concentration is about 11%
  • the recovery rate of xylooligosaccharides in the salt chamber is 100%.
  • the hydrolysate is mainly Is a mixture of xylose, xylonic acid and xylo-oligosaccharides
  • the yields are respectively 22.0%, 19.2%, 15.3%, 11.2%, 8.0%, 4.9%, 1.8% and 0.5%, a total of 82.9%, of which xylo-oligosaccharides totaled 60.9%; in addition, the furfural yield was 0.04%.
  • a 3L bioreactor add 1.5L of 100g/L xylose solution and 9g of Gluconobacter oxydans to perform xylose biooxidation reaction.
  • the conditions of the biooxidation reaction are: temperature 30°C, stirring speed 150rpm, and air
  • the amount is 1.0vvm
  • the reaction time is 24h
  • the pH during the fermentation process is controlled by sodium hydroxide
  • 98% of the xylose is converted into xylonic acid at the end of the reaction
  • the bacteria and the fermentation broth are separated by centrifugation at the end of the reaction, and the centrifugal conditions: 5000rpm, 5min
  • the main component of the fermentation broth is xylonic acid
  • the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, and 1.5L of deionized water is added to the acid chamber and the alkali chamber respectively, and the electrodialysis reaction is driven by the external DC power supply.
  • the conductivity is the reference detection and separation reaction process. After 1
  • Thermo Fisher ICS5000 ion chromatography equipped with CarboPacTM PA200 (3mm ⁇ 250mm) chromatographic column, PAD integral amperometric detector, column
  • the temperature is 30°C
  • the sample volume is 10 ⁇ L
  • the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
  • the yields are 35.2%, 16.5%, 11.7%, 9.1%, 5.9%, 4.6%, 3.7% and 1.4%, totaling 88.1%, of which xylooligosaccharides totaling 52.9% ;
  • the furfural yield is 0.08%.
  • the invention utilizes the reaction of microbial whole-cell biological oxidation of xylose to xylose, and uses the produced xylonic acid as a catalyst. Compared with acetic acid and other inorganic acids, the produced polysaccharides are not easily degraded excessively, and the yield is high and the by-product xylose There is little furfural; this method has technical universality and can be used for a variety of lignocellulosic raw materials (alkali extraction of xylan, straw, corncob, bagasse, etc.); in the present invention, the amount of xylonic acid, time and temperature need to be strictly controlled.
  • the present invention selects biological oxidation and electrodialysis coupling technology to convert xylose into xylonic acid, wherein the xylose acid can be recovered as a self-supplied catalyst and the xylo-oligosaccharide product is more pure.

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Abstract

Disclosed in the present invention is a method for producing xylooligosaccharide under catalysis of a xylonic acid. The method for producing xylooligosaccharide using a xylonic acid comprises: mixing a xylan raw material and the xylonic acid, and performing heating and stirring to undergo a reaction to obtain xylooligosaccharide, in parts by mass, the xylan raw material being 1 part, and the xylonic acid being 5-12 parts. The method for producing xylooligosaccharide by fermentation under catalysis comprises: mixing xylose and bacteria to obtain a mixed liquid; adjusting the pH of the mixed liquid; stirring at a low temperature to undergo a biological oxidation reaction; and adding a xylan raw material, and performing heating and stirring to obtain xylooligosaccharide. The present invention utilizes the reaction of microbial whole cell biological oxidation of the xylose to the xylonic acid, and uses the produced xylonic acid as a catalyst; compared with acetic acid and other inorganic acids, the produced glycan is not easily excessively degraded, and has a high yield and less by-product xylose furfural.

Description

一种木糖酸催化生产低聚木糖的方法Method for producing xylo-oligosaccharides catalyzed by xylonic acid
本申请要求于2019年10月28日提交中国专利局、申请号为201911032990.6、发明名称为“一种木糖酸催化生产低聚木糖的方法”的中国专利申请的优先权，其全部内容通过引用结合在本申请中。This application claims the priority of a Chinese patent application filed with the Chinese Patent Office on October 28, 2019, the application number is 201911032990.6, and the invention title is "a method for the production of xylo-oligosaccharides catalyzed by xylonic acid", the entire content of which is approved The reference is incorporated in this application.
技术领域Technical field
本发明涉及食品工程和化学工程技术领域，尤其涉及一种木糖酸催化生产低聚木糖的方法。The invention relates to the technical fields of food engineering and chemical engineering, in particular to a method for catalyzing the production of xylo-oligosaccharides by xylonic acid.
背景技术Background technique
低聚木糖又称木寡糖，主要来源于木质纤维中的木聚糖水解，其作为功能性食品或饲料添加剂，不能被消化系统吸收，但能够选择性增殖肠道内的双歧杆菌，同时激活多种免疫细胞活性；因此，在人类大健康与微生态、食品安全与绿色动物养殖、生态农业等产业高速发展的牵引和驱动下，源于木质纤维的低聚木糖产品作为一种“超强益生元”发展前景十分广阔。当下低聚木糖生产主要采用内切型木聚糖酶制剂催化水解碱提取的木聚糖，但该方法依赖于高价格生物酶，成本高、周期长，且碱提取处理工艺复杂困难；另可采用普通的强酸水解法制备低聚木糖，低聚木糖收率低副产品多，产品品质不高。Xylooligosaccharides, also known as xylo-oligosaccharides, are mainly derived from the hydrolysis of xylan in lignocellulosic fibers. As a functional food or feed additive, they cannot be absorbed by the digestive system, but they can selectively proliferate bifidobacteria in the intestinal tract. Activate a variety of immune cell activities; therefore, driven by the rapid development of human health and micro-ecology, food safety and green animal breeding, ecological agriculture and other industries, xylo-oligosaccharide products derived from wood fiber are used as a kind of " The development prospects of "Super Prebiotics" are very broad. The current production of xylo-oligosaccharides mainly uses endo-xylanase preparations to catalyze the hydrolysis of alkali-extracted xylan, but this method relies on high-priced biological enzymes, high cost, long cycle, and the alkali extraction treatment process is complicated and difficult; The ordinary strong acid hydrolysis method can be used to prepare xylo-oligosaccharides. The yield of xylo-oligosaccharides is low and there are many by-products and the product quality is not high.
发明内容Summary of the invention
本部分的目的在于概述本发明的实施例的一些方面以及简要介绍一些较佳实施例。在本部分以及本申请的说明书摘要和发明名称中可能会做些简化或省略以避免使本部分、说明书摘要和发明名称的目的模糊，而这种简化或省略不能用于限制本发明的范围。The purpose of this section is to outline some aspects of the embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this part, the description abstract and the title of the invention in this application to avoid obscuring the purpose of this part, the description abstract and the title of the invention, and such simplifications or omissions cannot be used to limit the scope of the present invention.
鉴于上述的技术缺陷，提出了本发明。In view of the above technical defects, the present invention is proposed.
因此，作为本发明其中一个方面，本发明克服现有技术中存在的不足，提供一种木糖酸催化生产低聚木糖的方法。Therefore, as one of the aspects of the present invention, the present invention overcomes the deficiencies in the prior art and provides a method for catalyzing the production of xylo-oligosaccharides by xylonic acid.
为解决上述技术问题，本发明提供了如下技术方案：一种木糖酸生产低聚木糖的方法，其包括，将木聚糖原料和木糖酸混合，加热搅拌反应；按质量份数计，所述木聚糖原料为1份，所述木糖酸为5～12份。In order to solve the above-mentioned technical problems, the present invention provides the following technical solutions: a method for producing xylo-oligosaccharides from xylonic acid, which includes mixing xylan raw materials and xylonic acid, heating and stirring the reaction; calculated in parts by mass The xylan raw material is 1 part, and the xylonic acid is 5-12 parts.
作为本发明所述的木糖酸生产低聚木糖的方法的优选方案，其中：所述木聚糖原料为木聚糖和/或含有木聚糖的木质纤维原料。As a preferred solution of the method for producing xylo-oligosaccharides with xylonic acid according to the present invention, the xylan raw material is xylan and/or xylan-containing lignocellulosic raw material.
作为本发明所述的木糖酸生产低聚木糖的方法的优选方案，其中：所述加热搅拌反应的搅拌速率为30～100rmp，温度为130～170℃，时间为0.25～2.0h。As a preferred solution of the method for producing xylooligosaccharides with xylonic acid according to the present invention, the stirring rate of the heating and stirring reaction is 30-100 rpm, the temperature is 130-170° C., and the time is 0.25-2.0 h.
作为本发明所述的木糖酸生产低聚木糖的方法的优选方案，其中：加热搅拌反应后还包括，降温后调节pH值，加入菌体，低温搅拌进行生物氧化，得到发酵液；在发酵液中加入木聚糖原料，加热搅拌。As a preferred solution of the method for producing xylo-oligosaccharides with xylonic acid according to the present invention, wherein: after heating and stirring the reaction, it also includes adjusting the pH value after cooling down, adding bacteria, and stirring at low temperature for biological oxidation to obtain a fermentation broth; Add xylan raw materials to the fermentation broth, heat and stir.
作为本发明所述的木糖酸生产低聚木糖的方法的优选方案，其中：所述生物氧化后还包括将发酵液置于双极膜电渗析盐室，酸室和碱室分别添加去离子水，通过外加直流电源驱动电渗析反应，取酸室中的木糖酸溶液加入所述木聚糖原料；所述电渗析反应至盐室电导率稳定。As a preferred solution of the method for producing xylo-oligosaccharides with xylonic acid according to the present invention, wherein: after the biological oxidation, the fermentation liquid is placed in a bipolar membrane electrodialysis salt chamber, and the acid chamber and the alkali chamber are added to the salt chamber respectively. Ionized water is used to drive the electrodialysis reaction by applying a direct current power supply, and the xylonic acid solution in the acid chamber is added to the xylan raw material; the electrodialysis reaction is performed until the conductivity of the salt chamber is stable.
作为本发明所述的木糖酸生产低聚木糖的方法的优选方案，其中：所述菌体为木糖氧化杆菌，其为0.01～0.1份。As a preferred solution of the method for producing xylo-oligosaccharides with xylonic acid according to the present invention, the bacteria are Xylose Oxidizing Bacillus, which is 0.01 to 0.1 parts.
作为本发明所述的木糖酸生产低聚木糖的方法的优选方案，其中：所述降温后调节pH为降至室温后调节至弱酸性，所述低温搅拌的温度为25～35℃，搅拌速率为100～200rmp。As a preferred solution of the method for producing xylo-oligosaccharides with xylonic acid according to the present invention, wherein: the pH is adjusted after cooling down to room temperature and then adjusted to weak acidity, and the temperature of the low-temperature stirring is 25-35°C, The stirring rate is 100～200rmp.
作为本发明所述的木糖酸生产低聚木糖的方法的优选方案，其中：所述将木聚糖原料和木糖酸混合与所述加入木聚糖原料中的所述木聚糖原料不相同。As a preferred solution of the method for producing xylo-oligosaccharides with xylonic acid according to the present invention, wherein: the xylan raw material and xylanic acid are mixed with the xylan raw material added to the xylan raw material Are not the same.
作为本发明另一个方面，本发明克服现有技术中存在的不足，提供一种发酵催化生产低聚木糖的方法，其包括，将木糖与菌体混合，得到混合液；调节混合液的pH，低温搅拌；加入木聚糖原料，加热搅拌，得到低聚木糖。As another aspect of the present invention, the present invention overcomes the deficiencies in the prior art and provides a method for fermentatively catalyzed production of xylooligosaccharides, which includes mixing xylose with bacterial cells to obtain a mixed liquid; adjusting the mixed liquid pH, stirring at low temperature; adding xylan raw materials, heating and stirring, to obtain xylo-oligosaccharides.
作为本发明所述的发酵催化生产低聚木糖的方法的优选方案，其中：所述生物氧化后还包括将混合液置于双极膜电渗析盐室，酸室和碱室分别添加去离子水，通过外加直流电源驱动电渗析反应，取酸室中的木糖酸溶液加入所述木聚糖原料；所述电渗析反应至盐室电导率稳定。As a preferred solution of the method for producing xylo-oligosaccharides by fermentation and catalysis according to the present invention, wherein: after the biological oxidation, it also includes placing the mixed solution in a bipolar membrane electrodialysis salt chamber, and adding deionization to the acid chamber and the alkali chamber respectively. Water is used to drive the electrodialysis reaction by applying a direct current power supply, and the xylonic acid solution in the acid chamber is added to the xylan raw material; the electrodialysis reaction is performed until the conductivity of the salt chamber is stable.
作为本发明所述的发酵催化生产低聚木糖的方法的优选方案，其中：所述菌体为木糖氧化杆菌；所述木聚糖原料为木聚糖和/或含有木聚糖的木质纤维原料；按质量份数计，所述木糖为1份，所述菌体为0.01～0.1份，所述木聚糖原料为1～5份。As a preferred solution of the method for producing xylo-oligosaccharides by fermentation and catalysis according to the present invention, wherein: the bacterial cell is xylosoxidizing bacteria; the xylan raw material is xylan and/or xylan-containing xylan Fiber raw materials; in parts by mass, the xylose is 1 part, the bacteria is 0.01 to 0.1 parts, and the xylan raw material is 1 to 5 parts.
作为本发明所述的发酵催化生产低聚木糖的方法的优选方案，其中：所述调节混合液的pH为调节混合液的pH至弱酸性；所述低温搅拌的温度为25～35℃，搅拌速率为100～200rmp，所述加热搅拌反应的搅拌速率为30～100rmp，温度为130～170℃，时间为0.25～2.0h。As a preferred solution of the method for fermentation and catalytic production of xylo-oligosaccharides according to the present invention, wherein: the pH of the mixed liquid is adjusted to be weakly acidic; the temperature of the low-temperature stirring is 25-35°C, The stirring rate is 100 to 200 rpm, the stirring rate of the heating and stirring reaction is 30 to 100 rpm, the temperature is 130 to 170° C., and the time is 0.25 to 2.0 h.
本发明的有益效果：The beneficial effects of the present invention:
本发明采用木糖酸作为催化剂，得率高且副产品木糖糠醛少；本发明选择生物氧化、电渗析耦合技术以转化木糖为木糖酸，其中木糖酸作为自供给催化剂可回收且低聚木糖产品更纯。The present invention uses xylonic acid as a catalyst, with high yield and less by-product xylose furfural; the present invention chooses bio-oxidation and electrodialysis coupling technology to convert xylose into xylonic acid, wherein xylonic acid can be recycled and low as a self-supply catalyst. Xylose products are more pure.
附图说明Description of the drawings
为了更清楚地说明本发明实施例的技术方案，下面将对实施例描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动性的前提下，还可以根据这些附图获得其它的附图。其中：In order to explain the technical solutions of the embodiments of the present invention more clearly, the following will briefly introduce the drawings used in the description of the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative labor, other drawings can be obtained from these drawings. among them:
图1为实施例1中高效阴离子交换色谱分析图谱；Figure 1 is a high-efficiency anion exchange chromatographic analysis chart in Example 1;
图2为实施例6中的生产工艺流程图；Figure 2 is a flow chart of the production process in Example 6;
图3为实施例6中的电渗析反应原理图。FIG. 3 is a schematic diagram of the electrodialysis reaction in Example 6. FIG.
具体实施方式Detailed ways
为使本发明的上述目的、特征和优点能够更加明显易懂，下面结合具体实施例对本发明的具体实施方式做详细的说明。In order to make the above objectives, features, and advantages of the present invention more obvious and understandable, the specific embodiments of the present invention will be described in detail below in conjunction with specific embodiments.
在下面的描述中阐述了很多具体细节以便于充分理解本发明，但是本发明还可以采用其他不同于在此描述的其它方式来实施，本领域技术人员可以在不违背本发明内涵的情况下做类似推广，因此本发明不受下面公开的具体实施例的限制。In the following description, many specific details are explained in order to fully understand the present invention, but the present invention can also be implemented in other ways different from those described here, and those skilled in the art can do so without departing from the connotation of the present invention. Similar promotion, therefore, the present invention is not limited by the specific embodiments disclosed below.
其次，此处所称的“一个实施例”或“实施例”是指可包含于本发明至少一个实现方式中的特定特征、结构或特性。在本说明书中不同地方出现的“在一个实施例中”并非均指同一个实施例，也不是单独的或选择性的与其他实施例互相排斥的实施例。Secondly, the "one embodiment" or "embodiment" referred to herein refers to a specific feature, structure, or characteristic that can be included in at least one implementation of the present invention. The appearances of "in one embodiment" in different places in this specification do not all refer to the same embodiment, nor are they separate or selectively mutually exclusive embodiments with other embodiments.
实施例1Example 1
在1L机械搅拌式不锈钢高压反应罐中，加入50g玉米芯碱抽提木聚糖和5％(质量分数)的木糖酸溶液500mL，密封后开启搅拌(50rpm)，并加 热至150℃保温75min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。In a 1L mechanically agitated stainless steel high-pressure reaction tank, add 50g corncob base to extract xylan and 5% (mass fraction) xylonic acid solution 500mL, after sealing, turn on the stirring (50rpm), and heat to 150℃ for 75min After the reaction is over, after the reaction tank is lowered to room temperature, the solid-liquid mixture after the reaction is loaded into a vacuum washing machine, and the unhydrolyzed solids are separated from the xylan hydrolysate (the hydrolysate is mainly It is a mixture of xylose, xylonic acid and xylo-oligosaccharides). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min.
其分析图谱如图1所示，木糖酸(XA)与木糖(Xylose)、木二糖(X2)、木三糖(X3)、木四糖(X4)、木五糖(X5)、木六糖(X6)、木七糖(X7)、木八糖(X8)可以同时检测。其中主要成分为木糖至木八糖，其得率分别为27.6％、19.2％、13.1％、8.8％、5.9％、2.5％、1.6％和1.3％，共计80％，其中低聚木糖共52.4％；此外糠醛得率0.05％。The analysis chart is shown in Figure 1. Xylonic acid (XA) and xylose (Xylose), xylobiose (X2), xylotriose (X3), xylotetraose (X4), xylopentaose (X5), Xylose (X6), xyloseptaose (X7), and xylooctaose (X8) can be detected at the same time. Among them, the main components are xylose to xylooctaose, and the yields are 27.6%, 19.2%, 13.1%, 8.8%, 5.9%, 2.5%, 1.6% and 1.3%, a total of 80%, of which xylo-oligosaccharides total 52.4%; in addition, the furfural yield is 0.05%.
此外，以100mmol/L氢氧化钠(NaOH)与500mmol/L的醋酸钠(NaAc)为流动相进行二元梯度淋洗的具体操作见下表：In addition, the specific operation of binary gradient elution with 100mmol/L sodium hydroxide (NaOH) and 500mmol/L sodium acetate (NaAc) as mobile phases is shown in the following table:
时间(min)Time (min) 100mmol/LNaOH(％)100mmol/LNaOH(%) 500mmol/LNaAc(％)500mmol/LNaAc(%)
00 100100 00
99 100100 00
2626 9292 88
2626 5050 5050
4040 5050 5050
4040 100100 00
5050 100100 00
实施例2Example 2
在1L机械搅拌式不锈钢高压反应罐中，加入50g玉米芯碱抽提木聚糖和10％(质量分数)的木糖酸溶液500mL，密封后开启搅拌(50rpm)，并加热至160℃保温45min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L 氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。其分析图谱显示木糖酸(XA)与木糖(Xylose)、木二糖(X2)、木三糖(X3)、木四糖(X4)、木五糖(X5)、木六糖(X6)、木七糖(X7)、木八糖(X8)可以同时检测。主要成分为木糖至木八糖，其得率分别为30.5％、17.2％、13.2％、9.1％、6.8％、3.9％、2.6％和1.7％，共计86.2％，其中低聚木糖共55.7％；此外糠醛得率0.07％。木聚糖水解液用氢氧化钠中和至pH5.5后和2g/L的氧化葡萄糖酸杆菌同时加入容量2L的生物反应器进行生物氧化反应，生物氧化反应条件为：温度30℃，搅拌转速为150rpm，空气通入量为0.5vvm，反应时间24h，98％的木糖被转化为木糖酸；反应结束通过离心机离心分离菌体与发酵液，离心条件：5000rpm，5min；发酵液主要成分为木糖酸与低聚木糖；将发酵液至于双极膜电渗析盐室，酸室和碱室分别添加500mL去离子水，通过外加直流电源驱动电渗析反应，以盐室电导率为参考检测分离反应过程，反应1h后盐室电导率稳定反应结束，此时，酸室中木糖酸回收96.8％，盐室中低聚木糖回收率100％。In a 1L mechanically agitated stainless steel high-pressure reaction tank, add 50g corncob base to extract xylan and 10% (mass fraction) xylonic acid solution 500mL, seal and turn on the stirring (50rpm), and heat to 160℃ for 45min After the reaction is over, after the reaction tank is lowered to room temperature, the solid-liquid mixture after the reaction is loaded into a vacuum washing machine, and the unhydrolyzed solids are separated from the xylan hydrolysate (the hydrolysate is mainly It is a mixture of xylose, xylonic acid and xylo-oligosaccharides). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the mobile phase is 100mmol/L sodium hydroxide and 500mmol/L sodium acetate for binary gradient elution, and the flow rate is 0.3mL/min. The analysis chart shows that xylonic acid (XA) and xylose (Xylose), xylobiose (X2), xylotriose (X3), xylotetraose (X4), xylopentaose (X5), xylohexaose (X6) ), Xylose (X7), Xylose (X8) can be detected at the same time. The main components are xylose to xylooctaose, and the yields are respectively 30.5%, 17.2%, 13.2%, 9.1%, 6.8%, 3.9%, 2.6% and 1.7%, totaling 86.2%, of which xylooligosaccharides totaling 55.7 %; In addition, the furfural yield is 0.07%. The xylan hydrolysate was neutralized with sodium hydroxide to pH 5.5 and 2 g/L of Gluconobacter oxydans was added to a 2L bioreactor at the same time to carry out the biological oxidation reaction. The conditions of the biological oxidation reaction were: temperature 30℃, stirring speed It is 150rpm, the air volume is 0.5vvm, the reaction time is 24h, 98% of the xylose is converted into xylonic acid; after the reaction, the bacteria and the fermentation broth are separated by centrifugation, the centrifugal condition: 5000rpm, 5min; the fermentation broth is mainly The ingredients are xylonic acid and xylo-oligosaccharides; the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, 500mL deionized water is added to the acid chamber and the alkali chamber respectively, and the electrodialysis reaction is driven by an external DC power supply. The conductivity of the salt chamber is Refer to the detection and separation reaction process. After 1 hour of reaction, the conductivity of the salt chamber is stable and the reaction ends. At this time, the recovery of xylonic acid in the acid chamber is 96.8%, and the recovery rate of xylo-oligosaccharide in the salt chamber is 100%.
实施例3Example 3
在1L机械搅拌式不锈钢高压反应罐中，加入50g玉米芯和5％(质量分数)的木糖酸溶液500mL，密封后开启搅拌(50rpm)，并加热至170℃保温15min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。其分析图谱显示木糖酸(XA)与木糖(Xylose)、木二糖(X2)、木三糖(X3)、木四糖(X4)、木五糖(X5)、木六糖(X6)、木七糖(X7)、木八糖(X8)可以同时检测。主要成分为木糖至木八糖，其得率分别为16.2％、15.8％、10.2％、9.3％、6.1％、4.8％、2.5％和1.8％，共计66.7％，其中低聚木糖共50％；此外糠醛得率0.03％。木聚糖水解液用氢氧化钠中和至pH5.5后和8g/L的氧化葡萄糖酸杆菌(美国典型菌株保藏中心ATCC 621H)同时加入容量2L的生 物反应器进行生物氧化反应，生物氧化反应条件为：温度30℃，搅拌转速为150rpm，空气通入量为0.5vvm，反应时间12h，99％的木糖被转化为木糖酸；反应结束通过离心机离心分离菌体与发酵液，离心条件：5000rpm，5min；发酵液主要成分为木糖酸与低聚木糖；将发酵液至于双极膜电渗析盐室，酸室和碱室分别添加500mL去离子水，通过外加直流电源驱动电渗析反应，以盐室电导率为参考检测分离反应过程，反应1h后盐室电导率稳定反应结束，此时，酸室中木糖酸回收96.3％，盐室中低聚木糖回收率100％。In a 1L mechanically agitated stainless steel high-pressure reaction tank, add 50g corn cob and 5% (mass fraction) xylonic acid solution 500mL, turn on the stirring (50rpm) after sealing, and heat to 170℃ for 15min; after the reaction, wait for the reaction After the body tank is lowered to room temperature, the reacted solid-liquid mixture is loaded into a vacuum washer, and the unhydrolyzed solids are separated from the xylan hydrolysate (the hydrolysate is mainly xylose and xylose) by squeezing and filtering. Acid and xylo-oligosaccharide mixture). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min. Its analysis chart shows that xylonic acid (XA) and xylose (Xylose), xylobiose (X2), xylotriose (X3), xylotetraose (X4), xylopentaose (X5), xylohexaose (X6) ), Xylose (X7), Xylose (X8) can be detected at the same time. The main components are xylose to xylooctaose, and the yields are 16.2%, 15.8%, 10.2%, 9.3%, 6.1%, 4.8%, 2.5%, and 1.8%, a total of 66.7%, of which a total of 50 xylo-oligosaccharides %; In addition, the furfural yield is 0.03%. The xylan hydrolysate is neutralized with sodium hydroxide to pH 5.5 and 8g/L Gluconobacter oxydans (American Type Strain Collection ATCC 621H) is simultaneously added to a 2L bioreactor for biological oxidation reaction, biological oxidation reaction The conditions are: the temperature is 30℃, the stirring speed is 150rpm, the air volume is 0.5vvm, the reaction time is 12h, and 99% of the xylose is converted into xylonic acid; after the reaction, the bacteria and the fermentation broth are separated by centrifugation. Conditions: 5000rpm, 5min; the main components of the fermentation broth are xylonic acid and xylo-oligosaccharides; the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, 500mL deionized water is added to the acid chamber and the alkali chamber, and the electricity is driven by an external DC power supply. In the dialysis reaction, the conductivity of the salt chamber is used as a reference to detect the separation reaction process. After 1 hour of reaction, the conductivity of the salt chamber is stable and the reaction ends. At this time, the recovery of xylonic acid in the acid chamber is 96.3%, and the recovery rate of xylo-oligosaccharides in the salt chamber is 100%. .
实施例4Example 4
在1L机械搅拌式不锈钢高压反应罐中，加入50g甘蔗渣干粉和10％(质量分数)的木糖酸溶液500mL，密封后开启搅拌(50rpm)，并加热至150℃保温60min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。其中木糖酸(XA)与木糖(Xylose)、木二糖(X2)、木三糖(X3)、木四糖(X4)、木五糖(X5)、木六糖(X6)、木七糖(X7)、木八糖(X8)可以同时检测。主要成分为木糖至木八糖，其得率分别为35.2％、16.5％、12.1％、8.2％、6.3％、4.4％、3.6％、1.9％和1.6％，共计89.8％，其中低聚木糖共54.6％；此外糠醛得率0.08％。木聚糖水解液用氢氧化钠中和至pH5.5后和2g/L的弗托式葡萄糖酸杆菌(美国典型菌株保藏中心ATCC IFO 3264)同时加入容量2L的生物反应器进行生物氧化反应，生物氧化反应条件为：温度30℃，搅拌转速为150rpm，空气通入量为0.5vvm，反应时间12h，94％的木糖被转化为木糖酸；反应结束通过离心机离心分离菌体与发酵液，离心条件：5000rpm，5min；发酵液主要成分为木糖酸与低聚木糖；将发酵液至于双极膜电渗析盐室，酸室和碱室分别添加500mL去离子水，通过外加直流电源驱动电渗析反应，以盐室电导率为参考检测分离反应过程，反应1h 后盐室电导率稳定反应结束，此时，酸室中木糖酸回收97.1％，盐室中低聚木糖回收率100％。In a 1L mechanically agitated stainless steel high-pressure reaction tank, add 50g bagasse dry powder and 10% (mass fraction) xylonic acid solution 500mL, turn on the stirring (50rpm) after sealing, and heat to 150℃ for 60min; after the reaction, wait After the reaction tank is lowered to room temperature, the reacted solid-liquid mixture is loaded into a vacuum washer, and the unhydrolyzed solids are separated from the xylan hydrolysate by squeezing and filtering (the hydrolysing solution is mainly xylose, wood Sugar acid and xylo-oligosaccharide mixture). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min. Among them, xylonic acid (XA) and xylose (Xylose), xylobiose (X2), xylotriose (X3), xylotetraose (X4), xylopentaose (X5), xylohexaose (X6), wood Heptaose (X7) and Xylose (X8) can be detected at the same time. The main components are xylose to xylooctaose, and the yields are respectively 35.2%, 16.5%, 12.1%, 8.2%, 6.3%, 4.4%, 3.6%, 1.9% and 1.6%, a total of 89.8%, of which oligomeric wood The total sugar is 54.6%; in addition, the furfural yield is 0.08%. The xylan hydrolysate is neutralized with sodium hydroxide to pH 5.5 and 2g/L of Gluconobacter Fortorii (American Type Strain Collection ATCC IFO 3264) is simultaneously added to a 2L bioreactor for biological oxidation reaction. The conditions of the biological oxidation reaction are: temperature 30℃, stirring speed 150rpm, air flow rate 0.5vvm, reaction time 12h, 94% xylose is converted into xylonic acid; after the reaction, the bacteria and fermentation are separated by centrifugation Centrifugation conditions: 5000rpm, 5min; the main components of the fermentation broth are xylonic acid and xylo-oligosaccharides; the fermentation broth is applied to the bipolar membrane electrodialysis salt chamber, and 500mL of deionized water is added to the acid chamber and the alkali chamber, respectively, through external direct current The electrodialysis reaction is driven by the power supply, and the separation reaction process is detected with the conductivity of the salt chamber as a reference. After 1 hour of reaction, the conductivity of the salt chamber is stable and the reaction ends. At this time, 97.1% of xylonic acid in the acid chamber is recovered, and xylo-oligosaccharides in the salt chamber are recovered The rate is 100%.
实施例5Example 5
在1L机械搅拌式不锈钢高压反应罐中，加入50g玉米芯干粉和5％(质量分数)的木糖酸溶液500mL，密封后开启搅拌(50rpm)，并加热至150℃保温70min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。其分析图谱显示木糖酸(XA)与木糖(Xylose)、木二糖(X2)、木三糖(X3)、木四糖(X4)、木五糖(X5)、木六糖(X6)、木七糖(X7)、木八糖(X8)可以同时检测。其中主要成分木糖至木八糖，其得率分别为19.1％、17.6％、11.2％、8.4％、6.8％、5.1％、2.5％、和1.3％，共计72％，其中低聚木糖共52.9％；此外糠醛得率0.06％。In a 1L mechanically agitated stainless steel high-pressure reaction tank, add 50g corncob dry powder and 5% (mass fraction) xylonic acid solution 500mL, turn on the stirring (50rpm) after sealing, and heat to 150℃ for 70min; wait after the reaction is over After the reaction tank is lowered to room temperature, the reacted solid-liquid mixture is loaded into a vacuum washer, and the unhydrolyzed solids are separated from the xylan hydrolysate by squeezing and filtering (the hydrolysing solution is mainly xylose, wood Sugar acid and xylo-oligosaccharide mixture). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min. Its analysis chart shows that xylonic acid (XA) and xylose (Xylose), xylobiose (X2), xylotriose (X3), xylotetraose (X4), xylopentaose (X5), xylohexaose (X6) ), Xylose (X7), Xylose (X8) can be detected at the same time. Among them, the main components xylose to xylooctaose, the yields were 19.1%, 17.6%, 11.2%, 8.4%, 6.8%, 5.1%, 2.5%, and 1.3%, a total of 72%, of which xylo-oligosaccharides total 52.9%; in addition, the furfural yield is 0.06%.
实施例6Example 6
生产工艺流程图如图2所示，在1L机械搅拌式不锈钢高压反应罐中，加入50g玉米芯碱抽提木聚糖和10％(质量分数)的木糖酸溶液500mL，密封后开启搅拌(60rpm)，并加热至150℃保温45min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。主要成分木糖至木八糖，其得率分别为25.6％、15.8％、12.2％、10.1％、5.5％、4.2％、2.8％和1.7％，共计77.9％，其中低聚木糖共52.3％；此外糠醛得率0.04％。木聚糖水解 液用氢氧化钠中和至pH5.5后和5g/L的氧化葡萄糖酸杆菌同时加入容量2L的生物反应器进行生物氧化反应，生物氧化反应条件为：温度30℃，搅拌转速为150rpm，空气通入量为0.5vvm，反应时间12h，98％的木糖被转化为木糖酸；反应结束通过离心机离心分离菌体与发酵液，离心条件：5000rpm，5min；发酵液主要成分为木糖酸与低聚木糖；将发酵液至于双极膜电渗析盐室，电渗析反应原理图如图3所示，酸室和碱室分别添加500mL去离子水，通过外加直流电源驱动电渗析反应，以盐室电导率为参考检测分离反应过程，反应1h后盐室电导率稳定反应结束，此时，酸室中木糖酸回收率为97.9％，质量浓度约为11％，盐室中低聚木糖回收率100％。The production process flow chart is shown in Figure 2. In a 1L mechanically stirred stainless steel high-pressure reaction tank, add 50g of corncob base to extract xylan and 500mL of 10% (mass fraction) xylonic acid solution. After sealing, turn on the stirring ( 60rpm), and heat to 150°C for 45min; after the reaction, after the reaction tank is lowered to room temperature, put the reacted solid-liquid mixture into a vacuum washer, and squeeze and filter to separate the solids that have not been hydrolyzed With xylan hydrolysate (the hydrolysate is mainly a mixture of xylose, xylonic acid and xylo-oligosaccharides). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min. The main components of xylose to xylooctaose, the yields are 25.6%, 15.8%, 12.2%, 10.1%, 5.5%, 4.2%, 2.8% and 1.7%, totaling 77.9%, of which xylooligosaccharides totaling 52.3% ; In addition, the furfural yield is 0.04%. The xylan hydrolysate was neutralized with sodium hydroxide to pH 5.5 and 5 g/L of Gluconobacter oxydans was added to a 2L bioreactor at the same time to carry out the biological oxidation reaction. The conditions of the biological oxidation reaction were: temperature 30°C, stirring speed At 150rpm, the air volume is 0.5vvm, the reaction time is 12h, 98% of the xylose is converted into xylonic acid; after the reaction, the bacteria and the fermentation broth are separated by centrifugation, the centrifugal condition: 5000rpm, 5min; the fermentation broth is mainly The components are xylonic acid and xylo-oligosaccharides; the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, and the electrodialysis reaction principle diagram is shown in Figure 3. The acid chamber and the alkali chamber are respectively added with 500mL deionized water, and a DC power supply is added. The electrodialysis reaction is driven, and the separation reaction process is detected with the conductivity of the salt chamber as a reference. After 1 hour of reaction, the conductivity of the salt chamber is stable and the reaction ends. At this time, the recovery rate of xylonic acid in the acid chamber is 97.9%, and the mass concentration is about 11%. The recovery rate of xylo-oligosaccharides in the salt chamber is 100%.
将酸室回收的11％木糖酸溶液稀释至10％后取500mL与50g玉米芯碱抽提木聚糖混合于1L机械搅拌式不锈钢高压反应罐中，密封后开启搅拌(60rpm)，并加热至155℃保温45min，反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)，其中木糖至木八糖，其得率分别为29.0％、16.1％、13.1％、9.6％、6.1％、3.4％、1.9％和0.9％，共计80.1％，其中低聚木糖共51.1％；此外糠醛得率0.04％。Dilute the 11% xylonic acid solution recovered in the acid chamber to 10%, then take 500mL and 50g of corncob base to extract the xylan and mix it in a 1L mechanically stirred stainless steel high-pressure reaction tank. After sealing, turn on the stirring (60rpm) and heat it. Keep the temperature at 155°C for 45 minutes. After the reaction, after the reaction tank is lowered to room temperature, put the reacted solid-liquid mixture into a vacuum washer, and squeeze and filter to separate the unhydrolyzed solids and xylan hydrolysis Liquid (the hydrolysate is mainly a mixture of xylose, xylonic acid and xylo-oligosaccharides), among which xylose to xylooctaose, the yields are 29.0%, 16.1%, 13.1%, 9.6%, 6.1%, 3.4 %, 1.9% and 0.9%, a total of 80.1%, of which xylo-oligosaccharides are 51.1%; in addition, the furfural yield is 0.04%.
实施例7Example 7
在1L机械搅拌式不锈钢高压反应罐中，加入50g玉米芯干粉和10％(质量分数)的木糖酸溶液500mL，密封后开启搅拌(50rpm)，并加热至170℃保温50min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。主要成分木糖至木八糖，其得率分别为72.6％、8.2％、6.1％、3.2％、0.9％、0.1％、0.1％和0.02％，共计91.2％，其中低聚木糖共18.6％；此外糠醛得 率为0.8％。因为在高强度(高酸，高温)条件下木糖酸使木聚糖水解更彻底但木糖含量会偏高。In a 1L mechanically stirred stainless steel high-pressure reaction tank, add 50g corn cob dry powder and 10% (mass fraction) xylonic acid solution 500mL, turn on the stirring (50rpm) after sealing, and heat to 170℃ for 50min; wait after the reaction is over After the reaction tank is lowered to room temperature, the reacted solid-liquid mixture is loaded into a vacuum washer, and the unhydrolyzed solids are separated from the xylan hydrolysate by squeezing and filtering (the hydrolysing solution is mainly xylose, wood Sugar acid and xylo-oligosaccharide mixture). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min. The main components of xylose to xylooctaose, the yields are 72.6%, 8.2%, 6.1%, 3.2%, 0.9%, 0.1%, 0.1% and 0.02%, a total of 91.2%, of which xylo-oligosaccharides are 18.6% ; In addition, the furfural yield is 0.8%. Because under high-intensity (high acid, high temperature) conditions, xylonic acid makes the hydrolysis of xylan more complete, but the xylose content will be higher.
实施例8Example 8
在1L机械搅拌式不锈钢高压反应罐中，加入50g玉米芯干粉和10％(质量分数)的酒石酸溶液500mL，密封后开启搅拌(50rpm)，并加热至150℃保温45min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。主要成分木糖至木八糖，其得率分别为65.2％、9.2％、7.1％、4.2％、3.0％、1.6％、0.5％和0.1％，共计90.9％，其中低聚木糖共25.7％；此外糠醛得率为0.4％。In a 1L mechanically agitated stainless steel high-pressure reaction tank, add 50g corncob dry powder and 10% (mass fraction) tartaric acid solution 500mL, turn on the stirring (50rpm) after sealing, and heat to 150℃ for 45min; after the reaction, the reaction body After the tank is lowered to room temperature, the reacted solid-liquid mixture is loaded into a vacuum washer, and the unhydrolyzed solids are separated from the xylan hydrolyzate by squeezing and filtering (the hydrolyzed liquid is mainly xylose and xylonic acid). Mixed with xylo-oligosaccharides). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min. The main components of xylose to xylooctaose, the yields are 65.2%, 9.2%, 7.1%, 4.2%, 3.0%, 1.6%, 0.5% and 0.1%, totaling 90.9%, of which xylooligosaccharides totaling 25.7% ; In addition, the furfural yield is 0.4%.
实施例9Example 9
在1L机械搅拌式不锈钢高压反应罐中，加入50g杨木粉和10％(质量分数)的木糖酸溶液500mL，密封后开启搅拌(50rpm)，并加热至150℃保温45min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。主要成分木糖至木八糖，其得率分别为11.6％、8.8％、8.2％、7.3％、5.0％、3.9％、2.1％和1.5％，共计48.4％，其中低聚木糖共36.8％；此外糠醛得率0.04％。In a 1L mechanically stirred stainless steel high-pressure reaction tank, add 50g of poplar wood powder and 500mL of 10% (mass fraction) xylonic acid solution. After sealing, turn on the stirring (50rpm), and heat to 150℃ for 45 minutes; After the reaction tank is lowered to room temperature, the reacted solid-liquid mixture is loaded into a vacuum washer, and the unhydrolyzed solids are separated from the xylan hydrolysate by squeezing and filtering (the hydrolysing solution is mainly xylose, wood Sugar acid and xylo-oligosaccharide mixture). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min. The main components of xylose to xylooctaose, the yields are 11.6%, 8.8%, 8.2%, 7.3%, 5.0%, 3.9%, 2.1% and 1.5% respectively, totaling 48.4%, of which xylooligosaccharides totaling 36.8% ; In addition, the furfural yield is 0.04%.
实施例10Example 10
在1L机械搅拌式不锈钢高压反应罐中，加入50g玉米芯碱抽提木聚 糖和5％(质量分数)的木糖酸溶液500mL，密封后开启搅拌(50rpm)，并加热至170℃保温15min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物废渣与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。其分析图谱显示木糖酸(XA)与木糖(Xylose)、木二糖(X2)、木三糖(X3)、木四糖(X4)、木五糖(X5)、木六糖(X6)、木七糖(X7)、木八糖(X8)可以同时检测。其中主要成分木糖至木八糖，其得率分别为10.1％、9.8％、9.2％、8.1％、6.9％、4.8％、3.7％、和3.3％，共计55.9％，其中低聚木糖共45.8％；此外糠醛得率0.02％。In a 1L mechanically agitated stainless steel high-pressure reaction tank, add 50g corncob base to extract xylan and 5% (mass fraction) xylonic acid solution 500mL, after sealing, turn on the stirring (50rpm), and heat to 170℃ for 15min After the reaction, after the reaction tank is lowered to room temperature, the solid-liquid mixture after the reaction is loaded into the vacuum washing machine, and the unhydrolyzed solid waste residue and the xylan hydrolyzate (hydrolyzed liquid) are separated by squeezing and filtering. Mainly a mixture of xylose, xylonic acid and xylo-oligosaccharides). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min. Its analysis chart shows that xylonic acid (XA) and xylose (Xylose), xylobiose (X2), xylotriose (X3), xylotetraose (X4), xylopentaose (X5), xylohexaose (X6) ), Xylose (X7), Xylose (X8) can be detected at the same time. Among them, the main components of xylose to xylooctaose, the yields were 10.1%, 9.8%, 9.2%, 8.1%, 6.9%, 4.8%, 3.7%, and 3.3%, totaling 55.9%. 45.8%; in addition, the furfural yield is 0.02%.
木聚糖水解液用氢氧化钠中和至pH5.5后和4g/L的氧化葡萄糖酸杆菌同时加入容量2L的生物反应器进行生物氧化反应，生物氧化反应条件为：温度30℃，搅拌转速为150rpm，空气通入量为0.5vvm，反应时间6h，98％的木糖被转化为木糖酸；反应结束通过离心机离心分离菌体与发酵液，离心条件：5000rpm，5min；发酵液主要成分为木糖酸与低聚木糖；将发酵液至于双极膜电渗析盐室，酸室和碱室分别添加500mL去离子水，通过外加直流电源驱动电渗析反应，以盐室电导率为参考检测分离反应过程，反应1h后盐室电导率稳定反应结束，此时，酸室中木糖酸回收率为96.5％，质量浓度约为5.5％，盐室中低聚木糖回收率100％。The xylan hydrolysate was neutralized with sodium hydroxide to pH 5.5 and 4g/L of Gluconobacter oxydans was added to a 2L bioreactor at the same time to carry out the biological oxidation reaction. The conditions of the biological oxidation reaction were: temperature 30℃, stirring speed It is 150rpm, the air volume is 0.5vvm, the reaction time is 6h, 98% of the xylose is converted into xylonic acid; after the reaction, the bacteria and the fermentation broth are separated by centrifugation, the centrifugal condition: 5000rpm, 5min; the fermentation broth is mainly The ingredients are xylonic acid and xylo-oligosaccharides; the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, 500mL deionized water is added to the acid chamber and the alkali chamber respectively, and the electrodialysis reaction is driven by an external DC power supply. The conductivity of the salt chamber is Refer to the detection and separation reaction process. After 1 hour of reaction, the conductivity of the salt chamber is stable and the reaction ends. At this time, the recovery rate of xylonic acid in the acid chamber is 96.5%, the mass concentration is about 5.5%, and the recovery rate of xylo-oligosaccharides in the salt chamber is 100%. .
将酸室回收的5.5％木糖酸溶液与上述反应后的玉米芯碱抽提木聚糖固体废渣混合于1L机械搅拌式不锈钢高压反应罐中，密封后开启搅拌(60rpm)，并加热至170℃保温15min，反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)，其中木糖至木八糖，其得率(针对于初始原料50g木聚糖计算)分别为8.2％、7.2％、6.3％、4.1％、3.2％、2.4％、0.9％和0.7％，共计33％，其中低聚木糖共24.8％；此外糠醛得率0.04％。Mix the 5.5% xylonic acid solution recovered from the acid chamber with the corncob alkali extraction xylan solid waste residue after the above reaction in a 1L mechanically agitated stainless steel high-pressure reaction tank. After sealing, turn on the stirring (60rpm) and heat to 170 Keep the temperature at ℃ for 15min. After the reaction, after the reaction tank is lowered to room temperature, put the reacted solid-liquid mixture into a vacuum washer, and squeeze and filter to separate the unhydrolyzed solids and the xylan hydrolysate ( The hydrolysate is mainly a mixture of xylose, xylonic acid and xylo-oligosaccharides), among which xylose to xylooctaose, the yields (calculated based on the initial raw material 50g xylan) are 8.2%, 7.2%, 6.3 %, 4.1%, 3.2%, 2.4%, 0.9% and 0.7%, a total of 33%, of which xylo-oligosaccharides total 24.8%; in addition, the furfural yield is 0.04%.
实施例11Example 11
在1L机械搅拌式不锈钢高压反应罐中，加入50g玉米芯碱抽提木聚糖和5％(质量分数)的木糖酸溶液500mL，密封后开启搅拌(60rpm)，并加热至170℃保温30min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。主要成分木糖至木八糖，其得率分别为21.6％、18.7％、14.9％、13.8％、9.6％、5.7％、2.9％和1.5％，共计88.7％，其中低聚木糖共67.1％；此外糠醛得率0.05％。木聚糖水解液用氢氧化钠中和至pH5.5后和5g/L的氧化葡萄糖酸杆菌同时加入容量2L的生物反应器进行生物氧化反应，生物氧化反应条件为：温度30℃，搅拌转速为150rpm，空气通入量为0.5vvm，反应时间12h，98％的木糖被转化为木糖酸；反应结束通过离心机离心分离菌体与发酵液，离心条件：5000rpm，5min；发酵液主要成分为木糖酸与低聚木糖；将发酵液至于双极膜电渗析盐室，酸室和碱室分别添加500mL去离子水，通过外加直流电源驱动电渗析反应，以盐室电导率为参考检测分离反应过程，反应1h后盐室电导率稳定反应结束，此时，酸室中木糖酸回收率为97.9％，质量浓度约为11％，盐室中低聚木糖回收率100％。In a 1L mechanically agitated stainless steel high-pressure reaction tank, add 50g corncob base to extract xylan and 5% (mass fraction) xylonic acid solution 500mL, seal and turn on the stirring (60rpm), and heat to 170℃ for 30min After the reaction is over, after the reaction tank is lowered to room temperature, the solid-liquid mixture after the reaction is loaded into a vacuum washing machine, and the unhydrolyzed solids are separated from the xylan hydrolysate (the hydrolysate is mainly It is a mixture of xylose, xylonic acid and xylo-oligosaccharides). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min. The main ingredients xylose to xylooctaose, the yields are 21.6%, 18.7%, 14.9%, 13.8%, 9.6%, 5.7%, 2.9% and 1.5%, a total of 88.7%, of which 67.1% of xylo-oligosaccharides ; In addition, the furfural yield is 0.05%. The xylan hydrolysate was neutralized with sodium hydroxide to pH 5.5 and 5 g/L of Gluconobacter oxydans was added to a 2L bioreactor at the same time to carry out the biological oxidation reaction. The conditions of the biological oxidation reaction were: temperature 30°C, stirring speed At 150rpm, the air volume is 0.5vvm, the reaction time is 12h, 98% of the xylose is converted into xylonic acid; after the reaction, the bacteria and the fermentation broth are separated by centrifugation, the centrifugal condition: 5000rpm, 5min; the fermentation broth is mainly The ingredients are xylonic acid and xylo-oligosaccharides; the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, 500mL deionized water is added to the acid chamber and the alkali chamber respectively, and the electrodialysis reaction is driven by an external DC power supply. The conductivity of the salt chamber is Refer to the detection and separation reaction process. After 1h of reaction, the conductivity of the salt chamber is stable and the reaction ends. At this time, the recovery rate of xylonic acid in the acid chamber is 97.9%, the mass concentration is about 11%, and the recovery rate of xylooligosaccharides in the salt chamber is 100%. .
将酸室回收的5.5％木糖酸溶液稀释至5％后取500mL与50g甘蔗渣干粉混合于1L机械搅拌式不锈钢高压反应罐中，密封后开启搅拌(60rpm)，并加热至170℃保温30min，反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)，其中木糖至木八糖，其得率分别为22.0％、19.2％、15.3％、11.2％、8.0％、4.9％、1.8％和0.5％，共计82.9％，其中低聚木糖共60.9％；此外糠醛得率0.04％。Dilute the 5.5% xylonic acid solution recovered in the acid chamber to 5%, then take 500mL and 50g of bagasse dry powder and mix them in a 1L mechanically stirred stainless steel high-pressure reaction tank. After sealing, turn on the stirring (60rpm), and heat to 170℃ for 30min. After the reaction, after the reaction tank is lowered to room temperature, the reacted solid-liquid mixture is loaded into a vacuum washer, and the unhydrolyzed solids are separated from the xylan hydrolysate by squeezing and filtering (the hydrolysate is mainly Is a mixture of xylose, xylonic acid and xylo-oligosaccharides), in which xylose to xylooctaose, the yields are respectively 22.0%, 19.2%, 15.3%, 11.2%, 8.0%, 4.9%, 1.8% and 0.5%, a total of 82.9%, of which xylo-oligosaccharides totaled 60.9%; in addition, the furfural yield was 0.04%.
实施例12Example 12
在3L的生物反应器内加入1.5L的100g/L的木糖溶液与9g的氧化葡萄糖酸杆菌进行木糖生物氧化反应，生物氧化反应条件为：温度30℃，搅拌转速为150rpm，空气通入量为1.0vvm，反应时间24h，以氢氧化钠控制发酵过程中的pH，反应结束98％的木糖被转化为木糖酸；反应结束通过离心机离心分离菌体与发酵液，离心条件：5000rpm，5min；发酵液主要成分为木糖酸；将发酵液至于双极膜电渗析盐室，酸室和碱室分别添加1.5L去离子水，通过外加直流电源驱动电渗析反应，以盐室电导率为参考检测分离反应过程，反应1h后盐室电导率稳定反应结束，此时，酸室中木糖酸回收率为97.0％，质量浓度约为10％。In a 3L bioreactor, add 1.5L of 100g/L xylose solution and 9g of Gluconobacter oxydans to perform xylose biooxidation reaction. The conditions of the biooxidation reaction are: temperature 30℃, stirring speed 150rpm, and air The amount is 1.0vvm, the reaction time is 24h, the pH during the fermentation process is controlled by sodium hydroxide, 98% of the xylose is converted into xylonic acid at the end of the reaction; the bacteria and the fermentation broth are separated by centrifugation at the end of the reaction, and the centrifugal conditions: 5000rpm, 5min; the main component of the fermentation broth is xylonic acid; the fermentation broth is placed in the bipolar membrane electrodialysis salt chamber, and 1.5L of deionized water is added to the acid chamber and the alkali chamber respectively, and the electrodialysis reaction is driven by the external DC power supply. The conductivity is the reference detection and separation reaction process. After 1 hour of reaction, the conductivity of the salt chamber is stable and the reaction ends. At this time, the recovery rate of xylonic acid in the acid chamber is 97.0%, and the mass concentration is about 10%.
在1L机械搅拌式不锈钢高压反应罐中，加入50g玉米芯干粉和上述制备的10％的木糖酸溶液500mL，密封后开启搅拌(60rpm)，并加热至155℃保温60min；反应结束后待反应体罐降至室温后将反应后的固液混合物装入真空洗浆机中，通过挤压、过滤以分离未被水解的固形物与木聚糖水解液(水解液主要为木糖、木糖酸与低聚木糖混合液)。所获得的木聚糖水解液样品采用高效阴离子交换色谱分析其糖组分，色谱条件：美国赛默飞ICS5000型离子色谱，配置CarboPacTM PA200(3mm×250mm)色谱柱，PAD积分安培检测器，柱温30℃，进样体积10μL；以100mmol/L氢氧化钠与500mmol/L的醋酸钠为流动相进行二元梯度淋洗，流速0.3mL/min。主要成分木糖至木八糖，其得率分别为35.2％、16.5％、11.7％、9.1％、5.9％、4.6％、3.7％和1.4％，共计88.1％，其中低聚木糖共52.9％；此外糠醛得率0.08％。In a 1L mechanically stirred stainless steel high-pressure reaction tank, add 50g corncob dry powder and 500mL of the 10% xylonic acid solution prepared above, turn on the stirring (60rpm) after sealing, and heat to 155℃ for 60min; after the reaction, wait for the reaction After the body tank is lowered to room temperature, the reacted solid-liquid mixture is loaded into a vacuum washer, and the unhydrolyzed solids are separated from the xylan hydrolysate (the hydrolysate is mainly xylose and xylose) by squeezing and filtering. Acid and xylo-oligosaccharide mixture). The obtained xylan hydrolysate sample was analyzed by high-performance anion exchange chromatography. The chromatographic conditions: Thermo Fisher ICS5000 ion chromatography, equipped with CarboPacTM PA200 (3mm×250mm) chromatographic column, PAD integral amperometric detector, column The temperature is 30°C, the sample volume is 10μL; the binary gradient elution is performed with 100mmol/L sodium hydroxide and 500mmol/L sodium acetate as the mobile phase, and the flow rate is 0.3mL/min. The main components of xylose to xylooctaose, the yields are 35.2%, 16.5%, 11.7%, 9.1%, 5.9%, 4.6%, 3.7% and 1.4%, totaling 88.1%, of which xylooligosaccharides totaling 52.9% ; In addition, the furfural yield is 0.08%.
本发明利用微生物全细胞生物氧化木糖至木糖酸的反应，将产生的木糖酸作为催化剂，与醋酸和其他无机酸相比，产生的聚糖不易过度降解，得率高且副产品木糖糠醛少；此法具有技术普遍性可用于多种木质纤维原料(碱抽提木聚糖、秸秆、玉米芯，甘蔗渣等)；本发明中木糖酸用量，时间以及温度需严格控制。酸浓度过低处理时间长能耗高；温度和时间过高过长产品容易发生过度降解产率下降；需合适条件与比例。本发明选择生物氧化、电渗析耦合技术以转化木糖为木糖酸，其中木糖酸作为自供给催化剂可回收且低聚木糖产品更纯。The invention utilizes the reaction of microbial whole-cell biological oxidation of xylose to xylose, and uses the produced xylonic acid as a catalyst. Compared with acetic acid and other inorganic acids, the produced polysaccharides are not easily degraded excessively, and the yield is high and the by-product xylose There is little furfural; this method has technical universality and can be used for a variety of lignocellulosic raw materials (alkali extraction of xylan, straw, corncob, bagasse, etc.); in the present invention, the amount of xylonic acid, time and temperature need to be strictly controlled. If the acid concentration is too low, the treatment time is long and the energy consumption is high; the temperature and the time are too high and the product is prone to excessive degradation and the yield is reduced; suitable conditions and proportions are required. The present invention selects biological oxidation and electrodialysis coupling technology to convert xylose into xylonic acid, wherein the xylose acid can be recovered as a self-supplied catalyst and the xylo-oligosaccharide product is more pure.
应说明的是，以上实施例仅用以说明本发明的技术方案而非限制，尽 管参照较佳实施例对本发明进行了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的精神和范围，其均应涵盖在本发明的权利要求范围当中。It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be carried out. Modifications or equivalent replacements without departing from the spirit and scope of the technical solutions of the present invention should be covered by the scope of the claims of the present invention.

Claims (12)

  1. 一种木糖酸生产低聚木糖的方法，其特征在于：包括，A method for producing xylo-oligosaccharides with xylonic acid, which is characterized in that it comprises:
    将木聚糖原料和木糖酸混合，加热搅拌反应，得到低聚木糖；Mix xylan raw materials and xylonic acid, heat and stir to react to obtain xylo-oligosaccharides;
    按质量份数计，所述木聚糖原料为1份，所述木糖酸为5～12份。In terms of parts by mass, the xylan raw material is 1 part, and the xylonic acid is 5-12 parts.
  2. 如权利要求1所述的木糖酸生产低聚木糖的方法，其特征在于：所述木聚糖原料为木聚糖和/或含有木聚糖的木质纤维原料。The method for producing xylo-oligosaccharides with xylonic acid according to claim 1, wherein the xylan raw material is xylan and/or xylan-containing lignocellulosic raw material.
  3. 如权利要求1所述的木糖酸生产低聚木糖的方法，其特征在于：所述加热搅拌反应的搅拌速率为30～100rmp，温度为130～170℃，时间为0.25～2.0h。The method for producing xylo-oligosaccharides with xylonic acid according to claim 1, wherein the stirring rate of the heating and stirring reaction is 30-100 rpm, the temperature is 130-170°C, and the time is 0.25-2.0 h.
  4. 如权利要求1～3任一所述的木糖酸生产低聚木糖的方法，其特征在于：加热搅拌反应后还包括，降温后调节pH值，加入菌体，低温搅拌进行生物氧化，得到发酵液；The method for producing xylo-oligosaccharides with xylonic acid according to any one of claims 1 to 3, characterized in that: after the heating and stirring reaction, the method further comprises: adjusting the pH value after cooling down, adding bacteria, and stirring at low temperature for biological oxidation to obtain Fermentation broth
    在发酵液中加入木聚糖原料，加热搅拌。Add xylan raw materials to the fermentation broth, heat and stir.
  5. 如权利要求4所述的木糖酸生产低聚木糖的方法，其特征在于：所述生物氧化后还包括将发酵液置于双极膜电渗析盐室，酸室和碱室分别添加去离子水，通过外加直流电源驱动电渗析反应，取酸室中的木糖酸溶液加入所述木聚糖原料；The method for producing xylo-oligosaccharides with xylonic acid according to claim 4, characterized in that: after the biological oxidation, it further comprises placing the fermentation broth in a bipolar membrane electrodialysis salt chamber, and the acid chamber and the alkali chamber are added separately Ionized water, driven by an external DC power supply to drive the electrodialysis reaction, and add the xylonic acid solution in the acid chamber to the xylan raw material;
    所述电渗析反应至盐室电导率稳定。The electrodialysis reacts until the conductivity of the salt chamber becomes stable.
  6. 如权利要求4或5所述的木糖酸生产低聚木糖的方法，其特征在于：所述菌体为木糖氧化杆菌，其为0.01～0.1份。The method for producing xylo-oligosaccharides with xylonic acid according to claim 4 or 5, wherein the bacteria are Xylose Oxidizing Bacillus, which is 0.01 to 0.1 parts.
  7. 如权利要求4或5所述的木糖酸生产低聚木糖的方法，其特征在于：所述降温后调节pH为降至室温后调节至弱酸性，所述低温搅拌的温度为25～35℃，搅拌速率为100～200rmp。The method for producing xylo-oligosaccharides with xylonic acid according to claim 4 or 5, characterized in that: the pH is adjusted after cooling down to room temperature and then adjusted to weak acidity, and the temperature of the low-temperature stirring is 25-35 ℃, the stirring rate is 100～200rmp.
  8. 如权利要求4或5所述的木糖酸生产低聚木糖的方法，其特征在于：所述将木聚糖原料和木糖酸混合与所述加入木聚糖原料中的所述木聚糖原料不相同。The method for producing xylo-oligosaccharides with xylonic acid according to claim 4 or 5, characterized in that: the xylan raw material and xylonic acid are mixed with the xylan raw material added to the xylan raw material. Sugar raw materials are not the same.
  9. 一种发酵催化生产低聚木糖的方法，其特征在于：包括，A method for producing xylo-oligosaccharides by fermentation and catalysis, which is characterized in that it comprises:
    将木糖与菌体混合，得到混合液；调节混合液的pH，低温搅拌进行生物氧化反应，加入木聚糖原料，加热搅拌，得到低聚木糖。The xylose is mixed with the bacterial cells to obtain a mixed liquid; the pH of the mixed liquid is adjusted, low-temperature stirring is carried out for biological oxidation reaction, xylan raw materials are added, and heated and stirred to obtain xylooligosaccharides.
  10. 如权利要求9所述的发酵催化生产低聚木糖的方法，其特征在于：所述生物氧化后还包括将混合液置于双极膜电渗析盐室，酸室和碱室分别添加去离子水，通过外加直流电源驱动电渗析反应，取酸室中的木糖酸溶液加入所述木聚糖原料；The method for producing xylo-oligosaccharides by fermentation and catalysis according to claim 9, characterized in that: after the biological oxidation, it further comprises placing the mixed solution in a bipolar membrane electrodialysis salt chamber, and adding deionization to the acid chamber and the alkali chamber respectively. Water, the electrodialysis reaction is driven by an external DC power supply, and the xylonic acid solution in the acid chamber is added to the xylan raw material;
    所述电渗析反应至盐室电导率稳定。The electrodialysis reacts until the conductivity of the salt chamber becomes stable.
  11. 如权利要求9或10所述的发酵催化生产低聚木糖的方法，其特征在于：所述菌体为木糖氧化杆菌；所述木聚糖原料为木聚糖和/或含有木聚糖的木质纤维原料；The method for fermentation and catalytic production of xylo-oligosaccharides according to claim 9 or 10, characterized in that: the bacteria are xylosoxidizing bacteria; the xylan raw material is xylan and/or contains xylan Of lignocellulosic raw materials;
    按质量份数计，所述木糖为1份，所述菌体为0.01～0.1份，所述木聚糖原料为1～5份。In terms of parts by mass, the xylose is 1 part, the fungus is 0.01 to 0.1 parts, and the xylan raw material is 1 to 5 parts.
  12. 如权利要求9～11任意一项所述的发酵催化生产低聚木糖的方法，其特征在于：所述调节混合液的pH为调节混合液的pH至弱酸性；所述低温搅拌的温度为25～35℃，搅拌速率为100～200rmp；The method for fermentation and catalytic production of xylo-oligosaccharides according to any one of claims 9 to 11, wherein the pH of the mixed liquid is adjusted to be weakly acidic; the temperature of the low-temperature stirring is 25～35℃, stirring rate is 100～200rmp;
    所述加热搅拌的搅拌速率为30～100rmp，温度为130～170℃，时间为0.25～2.0h。The stirring rate of the heating and stirring is 30-100 rpm, the temperature is 130-170° C., and the time is 0.25-2.0 h.
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