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 PDFInfo
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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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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
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.
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.
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. method of agricultural wastes being produced the functional food additives xylo-oligosaccharide through enzyme liberating, it is characterized in that, with the agricultural wastes that are rich in hemicellulose through ultrasonication is substrate, used waste is a particulate state, be specially the corn cob meal particle, the bean straw powder particle, cotton seed hulls powder particles or bagasse powder particle, producing bacterial strain with the novel zytase of a strain---the endo-xylanase that the branch streptomycete produces is degraded to this substrate, steps of the method are: (1) is put in the agricultural wastes particulate material in the special container of ultrasonic equipment, add sig water, solid-to-liquid ratio is scattered in the NaOH dilute alkaline soln of 10% (w/v) at 1: 10, in 50 ℃-80 ℃, supersound process 30min-35min under the 120-200W condition; (2) material that step (1) is produced filters, and discards filter residue and removes solid substance, collects filtrate, and regulates the pH value to subacidity with acidic buffer, obtains hemicellulose solution; (3) branch streptomycete (Streptomyces rameus) internal cutting type xylanase behind the adding purifying carries out hydrolysis of hemicellulose in above-mentioned hemicellulose solution, and wherein enzyme concentration is the 50U/g substrate, and temperature of reaction is 50 ℃, vibrating at constant temperature for enzymolysis 24h; (4) to the above-mentioned solution enzyme that goes out, centrifugal, separate, collect supernatant liquor, again through decolouring, ion-exchange, concentration process refiningly obtain xylo-bioses, xylotriose is main xylo-oligosaccharide product, the polymerization degree that obtains this xylo-oligosaccharide product is 2 to 5.
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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 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2000333692A (en) * | 1999-03-23 | 2000-12-05 | Oji Paper Co Ltd | Production of xylooligosaccharide |
| CN101182559A (en) * | 2007-11-16 | 2008-05-21 | 江南大学 | Method for preparing low polyxylose through squeezing-assisted wheat bran enzymolysis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2000333692A (en) * | 1999-03-23 | 2000-12-05 | Oji Paper Co Ltd | Production of xylooligosaccharide |
| CN101182559A (en) * | 2007-11-16 | 2008-05-21 | 江南大学 | Method for preparing low polyxylose through squeezing-assisted wheat bran enzymolysis |
Non-Patent Citations (2)
| Title |
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| 张海波.小麦麸皮制备低聚木糖的研究.《江南大学硕士学位论文》.2009,全文. * |
| 杨健等.超声波法提取玉米芯木聚糖的研究.《北京化工大学学报》.2005,第32卷(第3期),107-110. * |
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