CN114539332B - Method for preparing xylooligosaccharide by using biomass raw material and application of xylooligosaccharide in preparation of feed additive - Google Patents
Method for preparing xylooligosaccharide by using biomass raw material and application of xylooligosaccharide in preparation of feed additive Download PDFInfo
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Abstract
The invention discloses a method for preparing xylo-oligosaccharide by using a biomass raw material and application thereof in preparing a feed additive, wherein the preparation method comprises the following steps: removing impurities from a biomass raw material containing lignocellulose, crushing, adding water, mixing uniformly, adding into a high-pressure reaction kettle, performing subcritical hydrothermal hydrolysis reaction at 170-220 ℃ under the condition of homogeneous stirring, rapidly cooling to normal temperature after the reaction is finished, performing solid-liquid separation, collecting a hydrolysis mixed solution, and performing aftertreatment. According to the invention, the preparation of the xylo-oligosaccharide can be completed through one-step subcritical hydrolysis reaction, and the yield of the xylo-oligosaccharide can be obviously improved and the generation of byproducts can be effectively reduced by synergistically optimizing the particle size of crushed biomass raw materials and subcritical hydrolysis reaction conditions; the prepared xylo-oligosaccharide is rich in polysaccharide active ingredients, and can effectively promote the utilization of animals on feed; the preparation process avoids the use of auxiliary materials such as maltodextrin, corn starch, beta-cyclodextrin and the like, and can obviously reduce the production cost.
Description
Technical Field
The invention belongs to the technical field of xylo-oligosaccharide preparation, and particularly relates to a method for preparing xylo-oligosaccharide by using a biomass raw material and application of the xylo-oligosaccharide in preparation of a feed additive.
Background
Xylo-oligosaccharides (XOS), which are a mixture of oligosaccharides composed of 2-7 xylose units linked by beta-1,4 glycosidic bonds (GB/T23747-2009). As a super prebiotic, xylooligosaccharide has the effects of low calorie, good stability, inhibition of intestinal inflammatory factors and the like, has the effect of proliferating intestinal probiotics such as bifidobacterium and lactobacillus which is 10 to 20 times of the prebiotics such as fructooligosaccharide, isomaltooligosaccharide and the like, and is an important functional food and feed additive.
At present, the main preparation method of feed-grade xylo-oligosaccharide comprises the processes of crushing, physicochemical pretreatment, acid-base neutralization, enzymolysis, desalination, decolorization, concentration and the like, and has the problems of complex process, long production period, high energy consumption, large reagent dosage and the like.
In addition, in the prior art, the raw material for preparing the feed-grade xylooligosaccharide is mainly corncob, and enterprises for preparing the feed-grade xylooligosaccharide by taking agricultural and forestry wastes such as straw and branches and agricultural processing wastes such as rice husk and bagasse as raw materials exist in part of regions, and there is no report on preparing the feed-grade xylooligosaccharide by using the biomass of reed and triarrhena sacchariflora.
Phragmites australis Trin belongs to the genus Phragmites of the family Gramineae, is a perennial emergent water C3 plant, and is widely distributed in various parts of the world; triarrhena sacchariflora (Miscanthus luteoriparius) belongs to the genus Dirrhena of Gramineae, and is perennial hygrophyte C4.
The content of lignocellulose (including cellulose, hemicellulose and lignin) in the reed is up to more than 70wt%, and the content of lignocellulose in the triarrhena sacchariflora is up to about 90 wt%; and the biomass ash content of the two is very little, the fiber processing quality is excellent, and the two are traditional papermaking raw materials in the middle and lower reaches of Yangtze river all the time. The wetland of the shoal of the Dongting lake region in China has a mixed community of reed and triarrhena sacchariflora of nearly 130 ten thousand acres, but with the comprehensive quit of the traditional paper making industry, the green, novel and high-valued biomass industry of the Dongting lake reed and triarrhena sacchariflora, which can replace the paper making industry, is found, and becomes a major ecological economic problem to be solved urgently at present.
In view of this, the invention is particularly proposed.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a method for preparing xylooligosaccharide by using a biomass raw material and application of the xylooligosaccharide in a feed additive.
In order to realize the purpose, the technical scheme of the invention is as follows:
a method for preparing xylo-oligosaccharide by using biomass raw materials comprises the following steps:
removing impurities from a biomass raw material containing lignocellulose, crushing, adding water, mixing uniformly, adding into a high-pressure reaction kettle, performing subcritical hydrothermal hydrolysis reaction at 170-220 ℃ under the condition of homogeneous stirring, rapidly cooling to normal temperature after the reaction is finished, performing solid-liquid separation, collecting a hydrolysis mixed solution, and performing aftertreatment.
In the technical scheme, the particle size of the biomass raw material after being crushed is 5-100 meshes.
Specifically, in the above technical solution, if the particle size after pulverization is larger than 5 meshes, the self-hydrolysis reaction effect of the xylan component in the biomass in the subcritical hydrothermal reaction process will be significantly affected, and the extraction effect of the xylan component in the biomass will be seriously reduced; if the particle size after crushing is larger than 100 meshes, transitional hydrolysis of xylan components in the biomass in the subcritical hydrothermal reaction process is caused, so that a large amount of monosaccharide and degradation byproducts (formic acid, acetic acid, furfural, 5-HMF and the like) are formed, the concentration of xylooligosaccharide in the product dry powder is remarkably reduced, and even powder preparation cannot be performed.
In the above technical scheme, the stirring speed of the homogenizing stirring is more than 200rpm.
Specifically, in the above technical solution, if the stirring rotation speed is lower than 200rpm, the hydrolysis transition of the xylan component in the biomass raw material in the subcritical hydrothermal reaction process is caused, a hydrolysis mixed solution taking monosaccharides such as xylose and glucose as main components is formed, and certainly, a part of degradation byproducts (formic acid, acetic acid, furfural, 5-HMF and the like) are included, so that the content of xylooligosaccharide is seriously reduced, and the application value of xylooligosaccharide is further influenced.
Further, in the technical scheme, the crushed biomass raw material and water are uniformly mixed according to the solid-liquid ratio of 1 g:10-30 ml to obtain a raw material mixed solution, and the raw material mixed solution is added into a high-pressure reaction kettle with the volume of 1.2-2.5 times of the volume of the raw material mixed solution for subcritical hydrothermal hydrolysis reaction.
Specifically, in the above technical scheme, the pressure of the subcritical hydrolysis reaction is closely related to the reaction product, and when the sample loading amount of the high-pressure reaction kettle is too large, the reaction pressure is too high (greater than 2 Mpa), so that the oligomerization xylose component in the hydrolysis mixed solution is accelerated to be further hydrolyzed into xylose.
Specifically, in the above technical scheme, the time of the subcritical hydrolysis reaction is 10-60min.
Still further, in the above technical scheme, after the subcritical hydrothermal hydrolysis reaction is finished, the temperature is reduced to normal temperature within 24-36 min.
Still further, in the technical scheme, the post-treatment comprises the steps of removing impurities by an ultrafiltration membrane, intercepting by a nanofiltration membrane, distilling, concentrating and drying to prepare powder, so that the xylo-oligosaccharide is obtained.
In a preferred embodiment of the invention, the pore size of the ultrafiltration membrane is 0.22um.
In another preferred embodiment of the present invention, the nanofiltration membrane is a nanofiltration membrane with a molecular weight cut-off of 100 daltons.
In yet another preferred embodiment of the present invention, the concentration factor of the distillation concentration is 5 to 10.
In a particular embodiment of the invention, the biomass feedstock is triarrhena sacchariflora and/or reed.
Specifically, the dry biomass of triarrhena sacchariflora and reed is similar in all components, and specifically comprises the following components: soluble matter, cellulose, hemicellulose, lignin and ash content.
The invention also provides application of the method for preparing xylooligosaccharide by using the biomass raw material in preparation of feed additives.
Specifically, the application is to prepare the feed additive containing the xylo-oligosaccharide after the obtained xylo-oligosaccharide pulp is directly dried.
In an embodiment of a specific application of the present invention, the drying is at least one of freeze drying, low temperature spray drying and vacuum belt drying.
Compared with the prior art, the invention has the following advantages:
(1) According to the invention, the preparation of the xylo-oligosaccharide can be completed through one-step subcritical hydrothermal hydrolysis reaction, and the generation of byproducts such as micromolecular organic acid and furfural is effectively reduced while the yield of the xylo-oligosaccharide is remarkably improved by synergistically optimizing the particle size of the crushed biomass raw material and subcritical hydrothermal hydrolysis reaction conditions;
(2) The xylo-oligosaccharide syrup prepared by the one-step subcritical hydrolysis reaction is rich in polysaccharide active ingredients such as xylose, arabinose and glucose, can effectively promote animals to utilize feeds, effectively avoids the use of auxiliary materials such as maltodextrin, corn starch and beta-cyclodextrin, and obviously reduces the production cost of the animal feeds;
(3) The method for preparing xylo-oligosaccharide by one-step subcritical hydrolysis reaction has the advantages of short production period, low energy consumption, strong operability, no need of adding acid, alkali reagent, xylanase and other biological agents, low carbon, environmental protection and suitability for large-scale production, and the palatability of solid residue obtained after subcritical hydrolysis reaction is obviously improved compared with that of raw materials, so that the method can be further used for processing animal feed, and the full component utilization of reed, triarrhena sacchariflora and other biomass raw materials is realized;
(4) In the method for preparing xylo-oligosaccharide by one-step subcritical hydrolysis, solid impurities such as stone particles, soil and the like can be thoroughly removed by an ultrafiltration membrane, reaction byproducts such as formic acid, acetic acid, hydroxymethyl furfural, inorganic salts and the like are removed by a nanofiltration membrane, the effect of decolouring xylo-oligosaccharide is achieved, and the separation and purification process of xylo-oligosaccharide is effectively simplified;
(5) The biomass raw material for preparing xylo-oligosaccharide by subcritical hydrolysis reaction selected by the invention is the straw of perennial tall grass reed and triarrhena sacchariflora which has high biomass yield and high xylan and is urgently to be utilized in the Dongting lake region at present, has wide source and has great practical significance.
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FIG. 1 is a flow chart of a method for preparing xylo-oligosaccharide by using a biomass raw material in the embodiment of the invention;
FIG. 2 is a high performance liquid chromatography detection result diagram of the solution of the dry powder of triarrhena sacchariflora prepared in example 1 of the present invention;
FIG. 3 is a high performance liquid chromatography detection result chart of the acidolysis solution of the dry powder product of triarrhena sacchariflora prepared in example 1 of the present invention;
FIG. 4 is a diagram of the HPLC detection results of xylobiose, xylotriose and xylotetraose in the solution of the triarrhena sacchariflora dry powder product prepared in example 1 of the present invention;
FIG. 5 is a high performance liquid chromatography detection result chart of the acidolysis solution of the reed dry powder product prepared in example 2 of the present invention
Fig. 6 is a diagram of the detection result of the high performance liquid chromatography of the solution of the reed dry powder product prepared in embodiment 2 of the present invention;
fig. 7 is a high performance liquid chromatography detection result graph of the acidolysis solution of the reed dry powder product prepared in example 2 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments.
It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
In the examples, the means used are conventional in the art unless otherwise specified.
The terms "comprises," "comprising," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.
In addition, the technical features involved in the respective embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
The reeds and triarrhena used in the embodiment of the invention are both taken from Dongting lake areas.
Result detection method
Weighing 0.219g of dry powder, placing the dry powder into a 20mL volumetric flask, adding water to fully dissolve, fixing the volume and carrying out acidolysis.
The method for measuring the content of xylo-oligosaccharide, glucose oligosaccharide, xylose, arabinose, galacturonic acid, formic acid, acetic acid, hydroxymethyl furfural and furfural is as follows:
performing High Performance Liquid Chromatography (HPLC) analysis, specifically referring to GB/T35545-2017 (appendix B) and NREL/TP-510-42623, and calculating by using the difference value of xylose content in a solution before and after dilute sulfuric acid hydrolysis, wherein the system and the operation conditions of a chromatograph are as follows: the Shimadzu LC chromatograph and the workstation adopt Aminex HPX-87H as a chromatographic column and 5mM H as a mobile phase 2 SO 4 Column temperature 65 deg.C, flow rate 0.6mL/min, differential refractometer, sample amount 10uL, external standard method.
The calculation formula of the content of xylooligosaccharide is as follows:
total amount of xylooligosaccharide = (xylose content after acidolysis-xylose content before acidolysis) × 1.1.
The content of xylobiose, xylotriose and xylotetraose is determined as follows:
using High Performance Liquid Chromatography (HPLC) analysis, see in particular GB/T35545-2017 (appendix a), the system and operating conditions of the chromatograph are: shimadzu LC chromatograph and workstation, wherein the chromatographic column is Shodex SUGAR KS-801, the column temperature is 80 ℃, and the measurement is carried out by an external standard method.
Example 1A method for preparing xylooligosaccharide from triarrhena sacchariflora and application of xylooligosaccharide in preparing feed additive
1. The raw materials are used:
the triarrhena sacchariflora biomass contains 35.6wt% of glucan, 20.3 wt% of xylan, 2.5wt% of araban, 25.5wt% of lignin, 3.5wt% of ash and the balance of impurities, wherein the water content is less than 0.001 wt%.
2. Preparation method
As shown in fig. 1, 1.5kg of triarrhena biomass was pulverized to pass through a 5 mesh sieve, and the ratio of 1g (triarrhena biomass powder): mixing 10mL (deionized water) in proportion, adding into a 25L high-pressure reaction kettle, heating to 190 ℃, and keeping for 10min under the condition of stirring rotation speed of 700 rpm.
After the reaction is finished, water is rapidly introduced for cooling, the reaction system is ensured to be cooled to room temperature within 30min, and 12.7L of hydrolysis mixed solution is obtained through solid-liquid separation of filter pressing equipment; then, the obtained hydrolysis mixed solution is filtered and purified by using an ultrafiltration device provided with a 0.22um membrane, and then is intercepted and concentrated in sequence by a nanofiltration device provided with a 100d membrane and a thin film evaporator to obtain 1.05L of xylo-oligosaccharide syrup; finally, the xylo-oligosaccharide syrup was dried and powdered by a freeze dryer to obtain 275.4g of dry powder.
3. The result of the detection
The water content is 4.2wt%;
ash content 9.7wt%;
meets the requirements of GB/T23747-2009 on the contents of water (less than or equal to 8 wt%) and ash (less than or equal to 15 wt%) in the xylo-oligosaccharide feed additive.
From FIGS. 2-3, it can be calculated that:
and formic acid, acetic acid, hydroxymethyl furfural and furfural are not detected in the byproducts.
In conclusion, the total amount of the active polysaccharide reaches 57.9wt%; wherein the content of the reduced absolute dry xylo-oligosaccharide is 40.3wt%, which is far higher than the requirement of GB/T23747-2009 on the content of the component in the xylo-oligosaccharide feed additive (absolute dry is more than or equal to 35%).
From fig. 4, it can be calculated that:
7.5wt% of xylobiose;
xylotriose 9.1wt%;
6.6wt% of xylotetraose;
the total content of the three xylo-oligosaccharides accounts for 59.9wt% of the total xylo-oligosaccharide content, which is much higher than that of the XOS in the food grade XOS-70 type xylo-oligosaccharides in GB/T35545-2017 2-4 The content requirement (absolute dry content is more than or equal to 50wt percent).
1. The raw materials are used:
the reed biomass contains water content of less than 0.001wt% and components of 37.8wt% of glucan, 18.0 wt% of xylan, 1.2wt% of araban, 26.2wt% of lignin, 5.1wt% of ash and the balance of impurities.
2. Preparation method
1.5kg of reed biomass was crushed to pass through a 5 mesh sieve, and the weight ratio of 1g (reed biomass powder): 10mL (deionized water) are mixed and then added into a 25L high-pressure reaction kettle, the temperature is raised to 190 ℃, and the mixture is kept for 10min under the condition of the stirring rotating speed of 700 rpm.
After the reaction is finished, water is rapidly introduced for cooling, the reaction system is ensured to be cooled to room temperature within 30min, and 12.6L of hydrolysis mixed liquid is obtained through solid-liquid separation of filter pressing equipment; then, the obtained hydrolysis mixed solution is filtered and purified by using an ultrafiltration device provided with a 0.22um membrane, and then is intercepted and concentrated in sequence by a nanofiltration device provided with a 100d membrane and a thin film evaporator to obtain 1.2L of xylo-oligosaccharide syrup; finally, drying the xylooligosaccharide syrup by a freeze dryer to prepare powder, so as to obtain 161.8 g of product dry powder.
3. The result of the detection
The water content is 6.4wt%;
ash content 13.7wt%;
meets the requirements of GB/T23747-2009 on the content of water (less than or equal to 8 wt%) and ash (less than or equal to 15 wt%) in the xylo-oligosaccharide feed additive.
From fig. 5-6, it can be calculated that:
and formic acid, acetic acid, hydroxymethyl furfural and furfural are not detected in the byproducts.
In conclusion, the total amount of the active polysaccharide reaches 48.9wt%; wherein the content of the xylo-oligosaccharide in the oven dry is 32.1wt percent and is slightly lower than the requirement of GB/T23747-2009 on the content of the xylo-oligosaccharide feed additive (the oven dry is more than or equal to 35 percent).
From fig. 7, it can be calculated that:
xylobiose 3.1wt%;
xylotriose 4.0wt%;
5.8wt% of xylotetraose;
the total content of the three xylo-oligosaccharides accounts for 43.2wt% of the total content of the xylo-oligosaccharides, which is far higher than that of XOS in food grade XOS-35 type xylo-oligosaccharides of GB/T35545-2017 2-4 And (4) the requirement of content.
Comparative example 1
The invention provides a method for preparing xylooligosaccharide by utilizing triarrhena sacchariflora and application of the xylooligosaccharide in preparing feed additives.
Compared with example 1, the only difference is that:
in the preparation method, the grain diameter of the crushed and sieved triarrhena sacchariflora is 300 meshes, the reaction stirring speed is 120rpm, and the rest is the same as that of the embodiment 1.
The results show that the total amount of active polysaccharide in the product is 30.1wt%; wherein the content of the reduced absolute dry xylo-oligosaccharide is 12.9wt%; the sum of the three xylo-oligosaccharides accounts for 36.1wt% of the total xylo-oligosaccharide content.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is specific and detailed, but not to be understood as limiting the scope of the invention.
It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (3)
1. A method for preparing xylo-oligosaccharide by using biomass raw materials is characterized in that,
the method comprises the following steps: removing impurities from a biomass raw material containing lignocellulose, crushing the biomass raw material containing lignocellulose to obtain particles with the particle size of 5-100 meshes, adding water, mixing uniformly, adding the mixture into a high-pressure reaction kettle, performing subcritical hydrothermal hydrolysis reaction at 170-220 ℃ under the condition of homogeneous stirring, rapidly cooling to normal temperature within 24-36min after the reaction is finished, performing solid-liquid separation, collecting a hydrolysis mixed solution, and performing post-treatment to obtain xylo-oligosaccharide dry powder;
the biomass raw material is triarrhena sacchariflora;
the solid-liquid ratio of the crushed biomass raw material to water is 1 g: uniformly mixing 10-30ml of the raw materials to obtain a raw material mixed solution, and adding the raw material mixed solution into a high-pressure reaction kettle with the volume 1.2-2.5 times of the volume of the raw material mixed solution for subcritical hydrothermal hydrolysis reaction;
the stirring speed of the homogenizing stirring is more than 200 rpm;
the post-treatment comprises the steps of sequentially removing impurities by an ultrafiltration membrane, intercepting by a nanofiltration membrane, distilling, concentrating and drying to prepare powder;
the aperture of the ultrafiltration membrane is 0.22 um; the nanofiltration membrane is a nanofiltration membrane with the molecular weight cutoff of 100 daltons.
2. The method for preparing xylooligosaccharide from biomass feedstock according to claim 1, wherein,
the time of the subcritical hydrolysis reaction is 10-60min.
3. The method for preparing xylooligosaccharide from biomass feedstock according to claim 1, wherein,
the concentration multiple of the distillation concentration is 5-10 times.
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