CN101597629B - Method for preparing xyio-oligosacchandes by using ultrasonic to treat agricultural wastes - Google Patents

Method for preparing xyio-oligosacchandes by using ultrasonic to treat agricultural wastes Download PDF

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CN101597629B
CN101597629B CN2009101579913A CN200910157991A CN101597629B CN 101597629 B CN101597629 B CN 101597629B CN 2009101579913 A CN2009101579913 A CN 2009101579913A CN 200910157991 A CN200910157991 A CN 200910157991A CN 101597629 B CN101597629 B CN 101597629B
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hemicellulose
solution
xylo
agricultural wastes
powder particles
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CN101597629A (en
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李秀婷
吕跃钢
曹雁平
马家津
宋焕禄
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Beijing Technology and Business University
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Beijing Technology and Business University
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Abstract

The invention discloses a method for preparing functional xyio-oligosacchandes by taking agricultural wastes as raw materials and using ultrasonic treatment and bio-enzyme degradation. The method comprises the following steps: hydrolyzing the agricultural wastes after the ultrasonic treatment by using endo xylanase generated by Streptomyces rameus, converting hemicellulose in the agricultural wastes into the high-quality xyio-oligosacchandes, and preparing a xyio-oligosacchandes product having low xylose content and the polymerization degree between 2 and 5.

Description

A kind of method of utilizing the ultrasonication agricultural wastes to prepare xylo-oligosaccharide
Technical field
The present invention relates to a kind of method for preparing xylo-oligosaccharide, the method that adopts ultrasonication, bio-enzyme degradation agricultural wastes (as corn cob, stalk, rice husk, beans bar, bagasse etc.) to produce xylo-oligosaccharide.
Background technology
Xylo-oligosaccharide (also claiming wood oligose xylooligosaccharides), be with the xylan be substrate, by the endo-xylanase hydrolyzed xylan form by β-1, the small molecules oligosaccharide mixture of the 2-7 of 4 glycosidic link be combined intos D-wood sugar, its main component are xylo-bioses and xylotriose.At present studies confirm that already that the major physiological function of xylo-oligosaccharide comprised the following aspects:
(1) provides lower energy, satisfy the requirement that happiness food sweets is worried the person of getting fat again, also can supply diabetics, adiposis patient and hypertensive patient edible;
(2) bifidus bacillus and promote its propagation in the activation enteron aisle suppresses pathogenic bacteria, prevents diarrhoea;
(3) prevent constipation;
(4) reduce cholesterol in serum content, bring high blood pressure down, generate nutritive substance, enhancing body immunizing power and opposing tumour;
(5) can not cause carious tooth, help oral Health;
(6) remove enterogenous endotoxin.
As the lignocellulose of preparation xylo-oligosaccharide raw material, aboundresources is extensive, comprises agricultural wastes such as corn cob, cotton seed hulls, bagasse.With China is example, and only agricultural crop straw, cot are one, and annual output just reaches more than 700,000,000 ton.Annual nearly 40%~50% agricultural crop straw is burned, and a large amount of organic substances is run off owing to burning, and it burns the flue dust that is produced and has also caused topsoil simultaneously.With above-mentioned agricultural wastes is the xylo-oligosaccharide that feedstock production has high added value, not only promotes Chinese oligose industrial expansion, can also make full use of agricultural wastes, turns waste into wealth the protection environment.Therefore, the research and development to xylo-oligosaccharide production have very great economic benefit, social benefit.
At present, method for preparing lower polyxylose mainly contains chemical degradation method (as acid-hydrolysis method), mechanical degradation method (as hot-water extraction, microwave method etc.), and biological degradation method or enzyme process.Yet adopt chemistry, mechanical degradation legal system to be equipped with xylo-oligosaccharide, it is low that its preparation process is difficult to control, complex process, yield, is not suitable for suitability for industrialized production.The technology of therefore researching and developing a kind of preparation xylo-oligosaccharide of new high-efficiency and low-pollution has great economic worth and social effect.The ultrasonication technology is the new technology that development in recent years is got up, and the applied research on foodstuffs industry, pharmaceutical industry and chemical industry shows, compares with common process, and its processing speed, processing efficiency and product quality all have significantly raising.
Adopt enzyme process to prepare that xylo-oligosaccharide has efficiently, the advantage of environmental protection, its key is to select high vigor and has the zytase that reasonable enzyme is.Zytase can the degradation of xylan class, and extensively be present in nature, and the bacterial strain of zytase that can degradation of xylan class system, mould and the aspergillus as wood, zytase enzyme that it produced system forms complicated, and often because have the β-D-xylosidase of high vigor, the polymerization degree is that 2~5 xylo-oligosaccharide content is not high in the product, and the too high levels of wood sugar has influenced the quality and the result of use of xylo-oligosaccharide.
Purpose of the present invention is just at the problems referred to above and a kind of high-level efficiency of special development, oligosaprobic ultrasonication agricultural wastes prepare the method for xylo-oligosaccharide, this method has overcome traditional method and has prepared shortcomings such as high pollution in the xylo-oligosaccharide process, expensive and time be long, and use special zytase, reduce the zytase consumption on the one hand, reduced raw materials cost; Product is mainly xylo-bioses and xylotriose on the other hand, and is purer and yield is high, compares with present production xylo-oligosaccharide method, and target product content height has reduced subsequent product purified difficulty, has reduced production cost, thereby has had bright development prospect.
Summary of the invention
Technical problem solved by the invention is hemicellulose in the agricultural wastes to be converted into high-quality xylo-oligosaccharide, particularly a kind of processing method for preparing the xylo-oligosaccharide product.
The method that ultrasonication agricultural wastes of the present invention prepare xylo-oligosaccharide comprises following processing step:
1. ultrasonication agricultural wastes
The agricultural wastes particulate material is put in the ultrasonic wave special container, adds sig water, in ultrasonic equipment, handle.
Wherein, the ultrasonication technology of agricultural wastes is as follows: raw material is scattered in according to solid-to-liquid ratio in the NaOH solution of 10% (w/v) through pulverizing at 1: 10, in 50-80 ℃, and supersound process under the 120-200W condition;
2. filter, discard filter residue, regulate the pH value
Filter and remove solid substance, collect filtrate, and regulate the pH value to subacidity, obtain hemicellulose solution with acidic buffer;
3. zytase carries out enzymolysis
Branch streptomycete (Streptomyces rameus) internal cutting type xylanase in above-mentioned hemicellulose solution behind the adding purifying carries out hydrolysis of hemicellulose, and wherein enzyme concentration is the 50U/g substrate, and temperature of reaction is 50 ℃, vibrating at constant temperature for enzymolysis 24h.
4. xylo-oligosaccharide preparation
To the above-mentioned solution enzyme that goes out, centrifugal, separate, collect supernatant liquor, again through decolouring, ion-exchange, process such as concentrate refiningly obtain xylo-bioses, xylotriose is main xylo-oligosaccharide product.
With this understanding gained xylan enzymolysis liquid has been carried out the thin-layer chromatography stratographic analysis and shown (seeing accompanying drawing 1), main component is xylo-bioses and xylotriose in the enzymolysis solution.
Description of drawings
Fig. 1 is agricultural wastes corn cob enzymolysis product thin-layer chromatogram (row X n, wood sugar is to the mixture of wooden pentasaccharides.)
Embodiment
The present invention is described further below in conjunction with specific examples, but is not restriction the present invention.
Embodiment 1
Step 1: the corn cob meal particle is scattered in by solid-to-liquid ratio in the NaOH solution of 10% (w/v) at 1: 10, in 45-60 ℃, supersound process 30-40min under the 110-130W condition; The best is 50 ℃, supersound process 35min under the 120W condition;
Step 2: filter and remove solid substance, collect filtrate, and regulate the pH value to subacidity, obtain hemicellulose solution with acidic buffer;
Step 3: branch streptomycete (Streptomyces rameus) internal cutting type xylanase in above-mentioned hemicellulose solution behind the adding purifying carries out hydrolysis of hemicellulose, wherein enzyme concentration is the 50U/g substrate, temperature of reaction is 45-55 ℃, and the best is 50 ℃, vibrating at constant temperature for enzymolysis 24h;
Step 4: above-mentioned solution is boiled the 10min enzyme that goes out in boiling water is abundant, centrifugal, separate, collect supernatant liquor, again through decolouring, ion-exchange, process such as concentrate refiningly obtain xylo-bioses, xylotriose is main xylo-oligosaccharide product.
Embodiment 2
Step 1: the bean straw powder particle is scattered in by solid-to-liquid ratio in the NaOH solution of 10% (w/v) at 1: 10, in 60-80 ℃, supersound process 30-40min under the 180-200W condition; The best is 70 ℃, supersound process 35min under the 190W condition;
Step 2: filter and remove solid substance, collect filtrate, and regulate the pH value to subacidity, obtain hemicellulose solution with acidic buffer;
Step 3: branch streptomycete (Streptomyces rameus) internal cutting type xylanase in above-mentioned hemicellulose solution behind the adding purifying carries out hydrolysis of hemicellulose, wherein enzyme concentration is the 50U/g substrate, temperature of reaction is 45-55 ℃, and the best is 50 ℃, vibrating at constant temperature for enzymolysis 24h;
Step 4: above-mentioned solution is boiled the 10min enzyme that goes out in boiling water is abundant, centrifugal, separate, collect supernatant liquor, again through decolouring, ion-exchange, process such as concentrate refiningly obtain xylo-bioses, xylotriose is main xylo-oligosaccharide product.
Embodiment 3
Step 1: the cotton seed hulls powder particles is scattered in by solid-to-liquid ratio in the NaOH solution of 10% (w/v) at 1: 10, in 60-80 ℃, supersound process 30-40min under the 180-200W condition; The best is 70 ℃, supersound process 35min under the 190W condition;
Step 2: filter and remove solid substance, collect filtrate, and regulate the pH value to subacidity, obtain hemicellulose solution with acidic buffer;
Step 3: branch streptomycete (Streptomyces rameus) internal cutting type xylanase in above-mentioned hemicellulose solution behind the adding purifying carries out hydrolysis of hemicellulose, wherein enzyme concentration is the 50U/g substrate, temperature of reaction is 45-55 ℃, and the best is 50 ℃, vibrating at constant temperature for enzymolysis 24h;
Step 4: above-mentioned solution is boiled the 10min enzyme that goes out in boiling water is abundant, centrifugal, separate, collect supernatant liquor, again through decolouring, ion-exchange, process such as concentrate refiningly obtain xylo-bioses, xylotriose is main xylo-oligosaccharide product.
Embodiment 4
Step 1: the bagasse powder particle is scattered in by solid-to-liquid ratio in the NaOH solution of 10% (w/v) at 1: 10, in 45-60 ℃, supersound process 25-35min under the 180-200W condition; The best is 50 ℃, supersound process 30min under the 190W condition;
Step 2: filter and remove solid substance, collect filtrate, and regulate the pH value to subacidity, obtain hemicellulose solution with acidic buffer;
Step 3: branch streptomycete (Streptomyces rameus) internal cutting type xylanase in above-mentioned hemicellulose solution behind the adding purifying carries out hydrolysis of hemicellulose, wherein enzyme concentration is the 50U/g substrate, temperature of reaction is 45-55 ℃, and the best is 50 ℃, vibrating at constant temperature for enzymolysis 24h.
Step 4: above-mentioned solution is boiled the 10min enzyme that goes out in boiling water is abundant, centrifugal, separate, collect supernatant liquor, again through decolouring, ion-exchange, process such as concentrate refiningly obtain xylo-bioses, xylotriose is main xylo-oligosaccharide product.

Claims (1)

1.一种将农业废弃物经酶降解制取功能性食品添加剂低聚木糖的方法,其特征在于,以经超声波处理富含半纤维素的农业废弃物为底物,所用废弃物为颗粒状,具体为玉米芯粉颗粒、豆杆粉颗粒、棉籽壳粉颗粒或者甘蔗渣粉颗粒,以一株新型木聚糖酶产生菌株——枝链霉菌产生的内切木聚糖酶对该底物进行降解,该方法的步骤为:(1)将农业废弃物颗粒料放于超声波设备的专用容器中,加入稀碱液,固液比1∶10分散于10%(w/v)的NaOH稀碱溶液中,于50℃-80℃,120-200W条件下超声处理30min-35min;(2)对步骤(1)产生的物料进行过滤,弃去滤渣去除固形物,收集滤液,并用酸性缓冲液调节pH值至微酸性,得到半纤维素溶液;(3)在上述半纤维素溶液中加入纯化后的枝链霉菌(Streptomyces rameus)内切型木聚糖酶进行半纤维素水解,其中加酶量为50U/g底物,反应温度为50℃,恒温振荡酶解24h;(4)对上述溶液灭酶,离心,分离,收集上清液,再经脱色、离子交换、浓缩过程精制得到木二糖、木三糖为主的低聚木糖产品,得到该低聚木糖产品的聚合度为2至5。 1. A method for producing functional food additive xylooligosaccharides by enzymatic degradation of agricultural waste, characterized in that, the ultrasonic treatment of agricultural waste rich in hemicellulose is a substrate, and the waste used is a particle shape, specifically corn cob powder particles, bean stalk powder particles, cottonseed hull powder particles or bagasse powder particles. The steps of the method are: (1) put the agricultural waste granules in a special container of ultrasonic equipment, add dilute lye, and disperse in 10% (w/v) NaOH with a solid-liquid ratio of 1:10 In a dilute alkaline solution, ultrasonically treat for 30min-35min at 50°C-80°C, 120-200W; (2) Filter the material produced in step (1), discard the filter residue to remove solids, collect the filtrate, and buffer with acid solution to adjust the pH value to slightly acidic to obtain a hemicellulose solution; (3) adding purified Streptomyces rameus endo-xylanase to the above hemicellulose solution to hydrolyze hemicellulose, wherein The amount of enzyme is 50U/g substrate, the reaction temperature is 50°C, and the enzyme is hydrolyzed at constant temperature for 24 hours; (4) the above solution is deenzyme, centrifuged, separated, and the supernatant is collected, and then refined through decolorization, ion exchange, and concentration processes to obtain A xylooligosaccharide product based on xylobiose and xylotriose, the obtained xylooligosaccharide product has a degree of polymerization of 2 to 5. the
CN2009101579913A 2009-07-21 2009-07-21 Method for preparing xyio-oligosacchandes by using ultrasonic to treat agricultural wastes Expired - Fee Related CN101597629B (en)

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CN101792495A (en) * 2010-02-11 2010-08-04 浙江理工大学 Method for ultrasonically catalyzing and extracting hemicellulose, cellulose and lignin from plant straws
CN102206691B (en) * 2011-03-26 2012-05-23 北海田野食品有限公司 Method for producing xylose oligomer from bagasse
CN102320921B (en) * 2011-05-12 2014-04-02 中科院广州化学有限公司 Method for preparing biomass polylol by using ultrasonic waves and application thereof
CN102887965A (en) * 2012-10-22 2013-01-23 中国药科大学 Preparation method for obtaining uniform xylan from corn cob
CN103614435B (en) * 2013-12-02 2016-05-04 广西壮族自治区农业科学院农产品加工研究所 A kind of method of preparing xylo-oligosaccharide from bagasse
CN105440165B (en) * 2015-10-20 2018-09-04 石河子大学 The method that microwave radiation technology lye extracts the xylan in cotton seed hulls
CN105695533A (en) * 2016-03-09 2016-06-22 孟繁志 Additive for preparation of cellulose and preparation method of cellulose
CN115678946A (en) * 2022-11-02 2023-02-03 无锡秋可生物科技有限公司 A method for the efficient preparation of xylooligosaccharides by ultrasonic-assisted composite ionic liquid pretreatment
CN115707709A (en) * 2022-12-07 2023-02-21 河南科技大学 Preparation method of corncob xylooligosaccharide

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