CN114317639B - Processing method of sugar chain with hypotonic cluster structure - Google Patents
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Abstract
The invention discloses a processing method of a sugar chain with a hypotonic cluster structure, and belongs to the technical field of processing of nutritional food ingredients. The invention takes bulk starch as raw material, and prepares the low-permeability cluster structure sugar chain product by a composite enzymatic catalysis-component grading coupling technology. Weighing a large amount of starch, mixing with a buffer salt solution with the pH value of 5.0-7.0 to prepare a suspension with the mass concentration of 5-30%, heating at 70-95 ℃ for 30-60min, adding glycoside hydrolase, and reacting at the constant temperature of 60-95 ℃ for 2-12h; the hydrolysate of DE5-30 is obtained through fractionation, glycosyltransferase is added and reacts for 2-12 hours at the temperature of 30-70 ℃, and the target product is obtained through heating, enzyme deactivation, centrifugation and drying treatment. The method has simple operation, controllable reaction conditions and continuous green production, and the obtained product can be widely applied to the fields of special medical foods, sports foods, special meal foods, medicines and the like.
Description
Technical Field
The invention belongs to the technical field of processing of nutritional food ingredients, and particularly relates to a processing method of a sugar chain with a hypotonic cluster structure.
Background
Starch is a natural high molecular compound produced by plants through photosynthesis, is the second most renewable resource next to cellulose, and is an important green chemical basic raw material. However, the natural starch has the defects of poor solubility, poor dispersibility, poor film forming property, easy aging, poor peptization stability and the like, and severely restricts the application of the natural starch in various industries. The development, utilization and research of starch resources are very important for various countries in the world for a long time, and the modified starch is particularly used in industries such as food, papermaking, textile, medicine, light industry, building materials, petroleum, bioengineering and the like through modification of starch by a chemical method, a physical method and an enzymatic method.
The total annual output of starch in China exceeds 3000 ten thousand tons, which is second to the United states in the world, but most of the starch is used for manufacturing primary industrial raw materials such as starch sugar, fermented products and the like, and compared with the advanced level of developed countries in the Western world, the starch has key technical bottleneck problems of few varieties of high added value products, low comprehensive utilization level and the like. At present, along with the increasing functional requirements of industrial production on modified starch and the increasing quality requirements, the research and development of modified starch are also increasing, and international across-country companies are suitable for Ruian, jiaji, luo Gaite and the like, and the variety and specification of modified starch by using green clean production technology are more than two thousand, such as new products of resistant dextrin and low-calorie sugar; the domestic enterprises mainly produce and sell traditional products such as acidolysis starch, oxidized starch, acetate starch and the like, and are required to enrich and upgrade the types of the existing products and improve the added value of the products. For the above reasons, in order to widen the application of starch in nutritional health foods, development of a processing method of sugar chains of hypotonic cluster structure is urgently required.
Disclosure of Invention
[ technical problem ]
Domestic starch utilization is mainly limited to primary industrial raw materials, and improvement of added value of starch products is needed.
Technical scheme
In order to solve the problems, the invention provides a processing method of a sugar chain with a hypotonic cluster structure, which takes bulk starch as a raw material and prepares a sugar chain product with the hypotonic cluster structure by a composite enzymatic catalysis-component grading coupling technology.
The first object of the present invention is to provide a method for processing a sugar chain having a hypotonic clustered structure, comprising the steps of:
(1) Weighing a large amount of starch, mixing with a buffer salt solution with the pH value of 5.0-7.0 to prepare a suspension with the mass concentration of 5-30%, heating at 70-95 ℃ for 30-60min, adding glycoside hydrolase, and reacting at the constant temperature of 60-95 ℃ for 2-12h;
(2) The hydrolysate of DE5-30 is obtained through fractionation, glycosyltransferase is added and reacts for 2-12 hours at the temperature of 30-70 ℃, and the target product is obtained through heating, enzyme deactivation, centrifugation and drying treatment.
In one embodiment of the present invention, the bulk starch is crop starch derived from corn, rice, wheat, potato, tapioca, sweet potato, mung bean, oat, buckwheat, sorghum, etc.
In one embodiment of the present invention, the buffer salt solution is one or more of phosphate, acetate, tris salt, barbital salt, and the like.
In one embodiment of the invention, the glycoside hydrolase is one or more of a commercial alpha-amylase, pullulanase, beta-amylase, isoamylase, maltogenic amylase, amylomaltase, etc., derived from GH family 13.
In one embodiment of the invention, the glycoside hydrolase is added in an amount of 50 to 300U/g starch.
In one embodiment of the invention, the glycosyltransferase is one or more of 4-alpha-glucanotransferase, 1, 4-alpha-glucanotransferase, sucrose-4-glucosyltransferase, alpha-4, 6-glucosyltransferase, alpha-4, 3-glucosyltransferase, cyclomaltodextrin enzyme, neopullulanase and the like, derived from archaea or bacteria.
In one embodiment of the invention, the glycosyltransferase is added in an amount of 100 to 800U/g starch.
In one embodiment of the invention, the fractionation is achieved by polyethylene glycol.
The second object of the present invention is a low-permeability cluster-structured sugar chain product produced by the above-mentioned method.
In one embodiment of the present invention, the hypotonic cluster structure sugar chain product has a DE value of 0.1 to 5 and a molecular weight of 1.0 to 8.5X10 5 g/mol, osmotic pressure of 100-150mmol/L.
The third purpose of the invention is to apply the low-permeability cluster structure sugar chain product to the fields of special medical foods, sports foods, special meal foods, medicines and the like.
It is a fourth object of the present invention to provide a specific medical food, sports food, specific dietary food, medicine or the like comprising the aforementioned low-permeability cluster structure sugar chain product.
The invention has the following advantages:
1) The method has the advantages of simple and convenient steps, easy operation, controllable reaction conditions, relatively low cost by adopting an enzyme catalysis process, and no pollution to the environment by adopting a clean production process; meanwhile, the product is developed by fully utilizing a large amount of starch raw materials with rich resources in China and through a modern green production technology, the added value of the product is improved, and a certain basis is provided for comprehensive development and utilization of the product.
2) The DE value of the sugar chain with the cluster structure prepared by the invention is 0.1-5, and the molecular weight is 1.0-8.5X10 5 g/mol, osmotic pressure of 100-150mmol/L, and can be used as a functional ingredient in various fields such as food, medicine and the like, and has good market prospect and wide economic benefit.
Drawings
FIG. 1 shows a molecular weight distribution of sugar chains in a cluster structure.
Detailed Description
The present invention is further described below with reference to examples, but embodiments of the present invention are not limited thereto.
Molecular weight measurement: a high performance liquid phase volume exclusion chromatography, a multi-angle laser light scattering detector and a differential refraction detector combined system are adopted, a Shedx Ohpak SB-805HQ gel chromatographic column is selected, 0.1mol/L sodium nitrate solution is used as a mobile phase, the flow rate is set to 0.7mL/min, and the refractive index is set to dn/dc=0.138.
DE value measurement: starch hydrolysis degree DE (dextrose equivalent), commonly referred to as the percentage of reducing sugar (expressed as glucose) based on dry matter of the syrup, is determined by the Lyn-Ai Nong method
Osmotic pressure measurement: the ElitechGroupWescor5600 dew point osmometer, U.S.A., samples were configured to 10% strength and their molar concentrations were determined.
Glycosyltransferase preparation: the enzyme preparation is obtained by the steps of screening archaea or bacteria from the natural world, carrying out activation culture, fermenting and producing enzyme, collecting thalli, freeze-drying and crushing and the like, and is specifically referred to the literature Biofabric, structure and characterizations of amylopectin-based cyclic glucose.
Example 1:
100g of corn starch and a sodium acetate buffer salt solution with the pH value of 7.0 are weighed and mixed to prepare a suspension with the mass concentration of 10 percent, the suspension is placed in a water bath with the temperature of 85 ℃ for heating treatment for 40min, 100U/g of starch alpha-amylase is added for constant temperature reaction at the temperature of 95 ℃ for 4h, the hydrolysis product of DE16 is obtained by using polyethylene glycol for fractionation, 300U/g of starch 1, 4-alpha-glucan branching enzyme is added for heat preservation reaction for 3h, and the target product is obtained after heating, enzyme deactivation, centrifugation and drying treatment.
The DE value of the sugar chain with the cluster structure is 3.6, and the molecular weight is 6.1X10 5 g/mol, osmotic pressure 137mmol/L.
Example 2:
weighing 100g of potato starch and blending with a phosphate buffer solution with the pH value of 5.0 to prepare a suspension with the mass concentration of 5%, placing the suspension in a water bath with the temperature of 95 ℃ for heating treatment for 30min, and adding 50U/g of starch isoamylase for reacting for 12h at the constant temperature of 60 ℃; the hydrolysis product of DE9 is obtained by using polyethylene glycol for fractionation, 200U/g starch 4-alpha-glucanotransferase is added for reaction for 6 hours, and the target product is obtained by heating, enzyme deactivation, centrifugation and drying treatment.
The DE value of the sugar chain with the cluster structure is 4.4, and the molecular weight is 3.7X10 5 g/mol, osmotic pressure 146mmol/L.
Example 3:
100g of wheat starch and Tris-hydrochloride buffer salt solution with pH of 6.0 are weighed and mixed to prepare suspension with mass concentration of 20%, and after being placed in water bath with the temperature of 70 ℃ for heating treatment for 60min, 300U/g of starch beta-amylase is added for reacting for 5h at the constant temperature of 65 ℃; the hydrolysis product of DE21 is obtained by using polyethylene glycol for fractionation, 100U/g of starch pullulanase is added and the mixture is reacted for 8 hours under the heat preservation, and the target product is obtained by heating, enzyme deactivation, centrifugation and drying treatment.
The DE value of the sugar chain with the cluster structure is 2.5, and the molecular weight is 7.4X10 5 g/mol, osmotic pressure 126mmol/L.
When other starch, buffer salt solution, glycoside hydrolase and glycosyltransferase are used as defined in the summary of the invention, and according to the preparation method defined in the summary of the invention, the obtained cluster structure sugar chain has DE value of 0.1-5 and molecular weight of 1.0-8.5X10 5 g/mol, osmotic pressure of 100-150mmol/L.
Comparative example 1
With reference to example 1, the only difference is that 100U/g of starch alpha-amylase was not added, the other conditions were unchanged, and the corresponding amylase catalytic product was obtained.
The DE value of the sugar chain with the cluster structure is 8.2, and the molecular weight is 3.9X10 7 g/mol, osmotic pressure 432mmol/L.
Comparative example 2
With reference to example 1, the only difference is that 300U/g starch 1,4- α -glucan branching enzyme was not added, and the other conditions were unchanged, giving the corresponding amylase catalyzed product.
The DE value of the sugar chain with the cluster structure is 6.6, and the molecular weight is 6.1X10 7 g/mol, osmotic pressure 475mmol/L.
Comparative example 3
With reference to example 1, the only difference is that the amount of alpha-amylase is replaced by 5U/g substrate and 800U/g substrate from 100U/g starch, respectively, and other conditions are unchanged, so as to obtain the corresponding amylase catalytic product. The performance results of the resulting starch products are shown in Table 1.
TABLE 1 results of amylase catalytic products obtained with varying amounts of alpha-amylase
Comparative example 4
With reference to example 1, the only difference is that the amount of 1,4- α -glucan branching enzyme was replaced by 20U/g substrate and 1000U/g substrate from 300U/g starch, respectively, with the other conditions unchanged, to obtain the corresponding amylase catalytic product. The performance results of the resulting starch products are shown in Table 2.
TABLE 2 results of amylase catalytic products obtained with different amounts of 1, 4-alpha-glucan branching enzyme
Comparative example 5
With reference to example 1, the only difference is that the glycosyltransferase is a commercial cyclodextrin glycosyltransferase (EC 2.4.1.19) from japan field company, the other conditions being unchanged, the corresponding amylase catalytic product being obtained.
The DE value of the sugar chain with the cluster structure is 7.8, and the molecular weight is 5.9X10 7 g/mol, osmotic pressure 452mmol/L.
While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (6)
1. A method for processing a sugar chain with a hypotonic clustered structure, comprising the steps of:
(1) Weighing a large amount of starch, mixing with a buffer salt solution with the pH value of 5.0-7.0 to prepare a suspension with the mass concentration of 5-30%, heating at 70-95 ℃ for 30-60min, and adding glycoside hydrolase for reacting at the constant temperature of 60-95 ℃ for 2-12h; the glycoside hydrolase is selected from one or more of alpha-amylase, beta-amylase and isoamylase; the addition amount of the glycoside hydrolase is 50-300U/g starch; the bulk starch is starch derived from corn, rice, wheat, potato, cassava, sweet potato, mung bean, oat, buckwheat and sorghum;
(2) Grading to obtain hydrolysate with DE value of 5-30, adding glycosyltransferase, reacting at 30-70deg.C for 2-12h, heating to deactivate enzyme, centrifuging, and drying to obtain low-permeability cluster structure sugar chain product; the product has DE value of 0.1-5 and molecular weight of 1.0-8.5X10 5 g/mol, osmotic pressure of 100-150 mmol/L; the glycosyltransferase is one or more of 4-alpha-glucan transferase, 1, 4-alpha-glucan branching enzyme and new pullulanase which are derived from archaea or bacteria; the addition amount of the glycosyltransferase is 100-800U/g starch.
2. The process of claim 1, wherein the buffer salt solution is one or more of phosphate, acetate, tris salt, barbital salt.
3. A low-permeability cluster-structured sugar chain product produced by the processing method according to any one of claims 1 or 2.
4. The hypotonic clustered sugar chain product of claim 3, wherein the hypotonic clustered sugar chain product has a DE value of 0.1 to 5 and a molecular weight of 1.0 to 8.5X10 5 g/mol, osmotic pressure of 100-150mmol/L.
5. A forensic food, sports food, a forensic food or a medicine comprising the hypotonic clustered sugar chain product of claim 3 or 4.
6. Use of the hypotonic clustered sugar chain product of claim 3 or 4 in the preparation of special medical foods, sports foods, special meal foods, and medical products.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010226988A (en) * | 2009-03-26 | 2010-10-14 | Showa Sangyo Co Ltd | Starch decomposition product, and food additive, food and drink and drug containing the starch decomposition product, and method for producing the starch decomposition product |
| TW201731939A (en) * | 2015-12-04 | 2017-09-16 | 林原股份有限公司 | Mixture of alpha-glucans, its production and uses |
| CN110099928A (en) * | 2016-12-27 | 2019-08-06 | 江崎格力高株式会社 | The slow macromolecule glucan of digestion rate |
| CN113679053A (en) * | 2021-08-13 | 2021-11-23 | 江南大学 | Low-glycemic starch derivative and processing method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2010226988A (en) * | 2009-03-26 | 2010-10-14 | Showa Sangyo Co Ltd | Starch decomposition product, and food additive, food and drink and drug containing the starch decomposition product, and method for producing the starch decomposition product |
| TW201731939A (en) * | 2015-12-04 | 2017-09-16 | 林原股份有限公司 | Mixture of alpha-glucans, its production and uses |
| CN110099928A (en) * | 2016-12-27 | 2019-08-06 | 江崎格力高株式会社 | The slow macromolecule glucan of digestion rate |
| CN113679053A (en) * | 2021-08-13 | 2021-11-23 | 江南大学 | Low-glycemic starch derivative and processing method thereof |
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