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CN108315374A - A kind of environment-friendly preparation method thereof of superelevation branch converted starch particle - Google Patents

A kind of environment-friendly preparation method thereof of superelevation branch converted starch particle Download PDF

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CN108315374A
CN108315374A CN201810048745.3A CN201810048745A CN108315374A CN 108315374 A CN108315374 A CN 108315374A CN 201810048745 A CN201810048745 A CN 201810048745A CN 108315374 A CN108315374 A CN 108315374A
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CN108315374B (en
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郭丽
崔波
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Qilu University of Technology
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/22Preparation of compounds containing saccharide radicals produced by the action of a beta-amylase, e.g. maltose
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    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/04Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/18Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins

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Abstract

The present invention relates to Starch Production technical fields, particularly disclose a kind of environment-friendly preparation method thereof of superelevation branch converted starch particle.The environment-friendly preparation method thereof of the superelevation branch converted starch particle, it is characterised in that:Using starch from sweet potato as receptor, malto-oligosaccharide is donor, and enzyme hydrolysis and transglucosidation are carried out successively to starch from sweet potato using branching enzyme, beta amylase and glycosyltransferase, obtains the superelevation branch converted starch particle containing higher branch's short chain proportional numbers.Present invention process is simple, short preparation period, need not add special installation, and production cost is low, is suitable for industrialized production, and do not need any additive, safety non-pollution during producing.

Description

一种超高分支变性淀粉颗粒的绿色制备方法A green preparation method of ultra-high branched modified starch granules

(一)技术领域(1) Technical field

本发明涉及淀粉生产技术领域,特别涉及一种超高分支变性淀粉颗粒的绿色制备方法。The invention relates to the technical field of starch production, in particular to a green preparation method of ultra-highly branched modified starch granules.

(二)背景技术(2) Background technology

淀粉作为一种重要的工业原料被广泛应用于食品和工业领域,从而加强改善产品的各种加工性能。淀粉也是人类膳食中的重要组成成分,其被人体消化后转化成葡萄糖以提供人们生命活动所必须的能量。然而,随着食品加工技术的进步及淀粉科学研究的深入,人们发现原淀粉在应用中具有很大的局限性:稳定性比较差,容易老化;溶解性较差;在淀粉凝胶中,淀粉分子的部分链段会发生位移产生粘性流动,导致淀粉凝胶弹性发生改变,硬度大、有粗糙感等。同时,原淀粉在体内消化速率较快而易导致餐后血糖迅速升高,引起高血糖、肥胖病、心血管等疾病。As an important industrial raw material, starch is widely used in food and industrial fields, so as to enhance and improve various processing properties of products. Starch is also an important component of the human diet, which is converted into glucose after being digested by the human body to provide the energy necessary for people's life activities. However, with the advancement of food processing technology and the deepening of starch scientific research, it has been found that native starch has great limitations in application: relatively poor stability, easy aging; poor solubility; in starch gel, starch Partial chain segments of the molecule will be displaced to generate viscous flow, resulting in changes in the elasticity of the starch gel, high hardness, rough feeling, etc. At the same time, the digestion rate of native starch in the body is relatively fast, which can easily lead to a rapid rise in blood sugar after meals, causing hyperglycemia, obesity, cardiovascular and other diseases.

为解决原淀粉所存在的这些问题,目前研究有采用物理、化学、基因或酶法以及复合改性的方法对原淀粉进行改性,从而改善其物化性能和消化性能。其中,化学修饰淀粉已被广泛的应用于食品工业中,但却存在安全性及环境污染等问题,酶法改性淀粉的制备方法环保、作用条件简单温和且有易于被人体消化分解等优点。In order to solve these problems of native starch, the current research has used physical, chemical, genetic or enzymatic methods and compound modification methods to modify native starch, so as to improve its physical and chemical properties and digestibility. Among them, chemically modified starch has been widely used in the food industry, but there are problems such as safety and environmental pollution. The preparation method of enzymatically modified starch is environmentally friendly, the working conditions are simple and mild, and it is easy to be digested and decomposed by the human body.

(三)发明内容(3) Contents of the invention

本发明为了弥补现有技术的不足,提供了一种制备周期短、生产成本低、安全无污染的超高分支变性淀粉颗粒的绿色制备方法。In order to make up for the deficiencies of the prior art, the present invention provides a green preparation method of ultra-highly branched modified starch granules with short preparation period, low production cost, safety and no pollution.

本发明是通过如下技术方案实现的:The present invention is achieved through the following technical solutions:

一种超高分支变性淀粉颗粒的绿色制备方法,其特征在于:以红薯淀粉为受体,麦芽低聚糖为供体,利用分支酶、β-淀粉酶和葡萄糖苷转移酶对红薯淀粉依次进行酶水解和转糖苷作用,得到含有较高分支短链比例数的超高分支变性淀粉颗粒。A green method for preparing ultra-highly branched modified starch granules, characterized in that sweet potato starch is used as an acceptor and maltooligosaccharides as a donor, and the sweet potato starch is sequentially processed by branching enzyme, β-amylase and glucosidyl transferase. Enzymatic hydrolysis and transglycosidation, to obtain ultra-highly branched modified starch granules with a relatively high proportion of branched short chains.

本发明采用复合酶法改性红薯淀粉,首先通过分支酶作用红薯淀粉时,水解淀粉中直链淀粉和支链淀粉直链区的a-1,4-糖苷键产生簇和一些线性的葡萄糖残基,同时催化转移葡萄糖残基连接到其他链上促进a-1,6-糖苷键的形成,产生新的分支,使淀粉分子的分支密度增加。然后利用β-淀粉酶进一步水解使其外部链长缩短,最后利用葡萄糖苷转移酶的转葡萄糖基作用将游离的葡萄糖残基和麦芽低聚糖以a-1,6糖苷键的形式连接到其他链上,形成了较多新的短链侧分支,进一步增加其分支密度,形成较紧密的结构,得到含有较高分支短链比例数的超高分支变性淀粉颗粒,提高了红薯淀粉的分支度,无化学残留。The present invention adopts compound enzyme method to modify sweet potato starch. Firstly, when sweet potato starch is acted on by branching enzymes, the α-1,4-glucosidic bonds in the amylose and amylopectin linear regions of the hydrolyzed starch produce clusters and some linear glucose residues. At the same time, it catalyzes the transfer of glucose residues to connect to other chains to promote the formation of a-1,6-glycosidic bonds, generate new branches, and increase the branch density of starch molecules. Then use β-amylase to further hydrolyze to shorten the external chain length, and finally use glucosyltransferase to connect the free glucose residues and maltooligosaccharides to other amino acids in the form of a-1,6 glycosidic bonds. On the chain, more new short-chain side branches are formed, further increasing its branch density, forming a tighter structure, and obtaining ultra-highly branched modified starch granules with a higher proportion of branched short-chains, which improves the branching degree of sweet potato starch , no chemical residue.

其具体包括如下步骤:It specifically includes the following steps:

(1)将红薯淀粉配置成5(w/v)%的淀粉液,调节淀粉液pH为6.5;(1) Prepare sweet potato starch into 5 (w/v)% starch liquid, and adjust the pH of the starch liquid to 6.5;

(2)向淀粉液中加入分支酶,于水浴中振荡水解,然后灭酶活性,冷却得到酶解液;(2) Add branching enzyme to the starch liquid, shake and hydrolyze in a water bath, then inactivate the enzyme activity, and cool to obtain the enzymatic hydrolysis solution;

(3)调节酶解液pH为5.2,加入β-淀粉酶,于水浴中震荡水解,然后灭酶活性,冷却得到酶解液;(3) Adjust the pH of the enzymatic hydrolysis solution to 5.2, add β-amylase, shake and hydrolyze in a water bath, then inactivate the enzyme activity, and cool to obtain the enzymatic hydrolysis solution;

(4)向酶解液中加入麦芽低聚糖,混合均匀后调节pH至5.0,再加入葡萄糖苷转移酶,置于水浴中振荡酶解,然后灭酶活性,冷却后得到样液;(4) Add maltooligosaccharides to the enzymatic hydrolysis solution, adjust the pH to 5.0 after mixing evenly, then add glucosidyl transferase, place in a water bath for oscillating enzymolysis, then inactivate the enzyme activity, and obtain a sample solution after cooling;

(5)向样液中加入无水乙醇,离心,所得沉淀经冷冻干燥得到产品。(5) Add absolute ethanol to the sample solution, centrifuge, and freeze-dry the obtained precipitate to obtain the product.

本发明的更优技术方案为:The more optimal technical scheme of the present invention is:

步骤(1)中,称取10g红薯淀粉,加入pH为6.5的0.02M醋酸钠缓冲溶液至淀粉浆液体积为200mL。In step (1), 10 g of sweet potato starch was weighed, and 0.02 M sodium acetate buffer solution with a pH of 6.5 was added until the volume of the starch slurry was 200 mL.

步骤(2)中,向淀粉液中加入308U/g的分支酶,置于60℃水浴中振荡20-24h,然后加入2.5mL 1mol/L的NaOH溶液灭酶活性。In step (2), add 308U/g branching enzyme to the starch liquid, place in a 60°C water bath and shake for 20-24h, then add 2.5mL 1mol/L NaOH solution to inactivate the enzyme activity.

步骤(3)中,向酶解液中加入10u/g的β-淀粉酶,置于37℃水浴中振荡4-6h,然后加入2.5mL 1mol/L的NaOH溶液灭酶活性。In step (3), add 10u/g of β-amylase to the enzymolysis solution, place in a 37°C water bath and shake for 4-6h, then add 2.5mL of 1mol/L NaOH solution to inactivate the enzyme activity.

步骤(4)中,向酶解液中加入占其重量比5%的麦芽低聚糖,混合均匀后加入9088U/g的葡萄糖苷转移酶,置于55℃水浴中振荡酶解10-12h,然后加入2.5mL 1mol/L的NaOH溶液灭酶活性;其中,所述麦芽低聚糖由重量配比为20%的麦芽四糖、30%的麦芽五糖、40%的麦芽六糖和10%的麦芽七糖混合组成。In step (4), add maltooligosaccharides accounting for 5% by weight to the enzymolysis solution, mix well, add 9088U/g glucosidyl transferase, place in a 55°C water bath for oscillating enzymolysis for 10-12 hours, Then add 2.5mL of 1mol/L NaOH solution to inactivate the enzyme activity; Composition of maltoheptaose blend.

步骤(5)中,向样液中加入其3倍体积的无水乙醇,于3000r/min的条件下离心10min,得到沉淀。In step (5), 3 times the volume of absolute ethanol was added to the sample liquid, and centrifuged at 3000 r/min for 10 min to obtain a precipitate.

本发明工艺简单,制备周期短,不需要添加特殊设备,生产成本低,适于工业化生产,且生产期间不需要任何添加剂,安全无污染。The invention has the advantages of simple process, short preparation cycle, no need of adding special equipment, low production cost, suitable for industrialized production, no additives during production, and safety and pollution-free.

(四)附图说明(4) Description of drawings

下面结合附图对本发明作进一步的说明。The present invention will be further described below in conjunction with the accompanying drawings.

图1为原红薯淀粉的SEM电镜图;Fig. 1 is the SEM electron micrograph of former sweet potato starch;

图2为本发明变性淀粉颗粒产品的SEM电镜图;Fig. 2 is the SEM electron micrograph of modified starch granule product of the present invention;

图3为原红薯淀粉在偏光显微镜下黑十字观察成像图;Fig. 3 is the black cross observation imaging figure of former sweet potato starch under polarizing microscope;

图4为本发明变性淀粉颗粒在偏光显微镜下黑十字观察成像图。Figure 4 is an image of the modified starch granules of the present invention observed with a black cross under a polarizing microscope.

(五)具体实施方式(5) Specific implementation methods

实施例:一种超高分支变性淀粉颗粒的制备方法Embodiment: A kind of preparation method of ultra-highly branched modified starch granules

具体包括以下步骤:Specifically include the following steps:

(1)红薯淀粉液的配制:准确称取10g红薯淀粉样品配成5%(w/v)淀粉液,调节淀粉溶液pH为6.5;(1) Preparation of sweet potato starch solution: Accurately weigh 10g of sweet potato starch sample to prepare 5% (w/v) starch solution, and adjust the pH of the starch solution to 6.5;

(2)分支酶酶解:分支酶(308U/g),置于60℃水浴振荡24h,然后加入2.5mL 1mol/L的NaOH溶液使酶灭活,冷却;(2) Branching enzyme enzymatic hydrolysis: branching enzyme (308U/g), place in a 60°C water bath and shake for 24 hours, then add 2.5mL 1mol/L NaOH solution to inactivate the enzyme, and cool;

(3)β-淀粉酶水解:调节溶液pH为5.2,β-淀粉酶酶添加量10U/g,酶解时间6h,反应温度37℃,加入2.5mL 1mol/L的NaOH溶液使酶灭活,冷却;(3) β-amylase hydrolysis: adjust the pH of the solution to 5.2, add β-amylase to 10U/g, enzymolysis time 6h, reaction temperature 37°C, add 2.5mL 1mol/L NaOH solution to inactivate the enzyme, cool down;

(4)麦芽低聚糖的添加:称取5g麦芽低聚糖混合物,包括20%麦芽四糖、30%麦芽五糖、40%麦芽六糖、10%麦芽七糖,加入到酶解液中混合均匀;(4) Addition of maltooligosaccharides: Weigh 5g of maltooligosaccharide mixture, including 20% maltotetraose, 30% maltopentaose, 40% maltohexaose, 10% maltoheptaose, and add it to the enzymatic solution well mixed;

(5)葡萄糖苷转移酶酶解:调节溶液pH为5.0,葡萄糖苷转移酶添加量9088U/g,置于55℃水浴振荡酶解12h,然后加入2.5mL 1mol/L的NaOH溶液使酶灭活;(5) Glucosidyl transferase enzymatic hydrolysis: adjust the pH of the solution to 5.0, add glucosidyl transferase to 9088U/g, place in a 55°C water bath for 12 hours, then add 2.5mL 1mol/L NaOH solution to inactivate the enzyme ;

(6)待样液冷却至室温,加入3倍体积的无水乙醇,于3000 r/min下进行10min的离心,所得沉淀冷冻干燥即得到超高分支变性淀粉颗粒。(6) After the sample solution was cooled to room temperature, 3 times the volume of absolute ethanol was added, centrifuged at 3000 r/min for 10 minutes, and the resulting precipitate was freeze-dried to obtain ultra-highly branched modified starch granules.

对本实施例得到的变性淀粉颗粒进行物化及消化特性的检测,经检测可知,与原淀粉相比,本发明的变性淀粉颗粒溶解度从9.86%显著增高到47.65%,淀粉糊粘度增加了43.24%;糊化峰值温度从68.98℃降低到56.94℃;淀粉糊在4℃储藏一周,其老化度从58.94%显著降低到36.57%;缓慢消化淀粉和抗性淀粉含量分别从24.65%和8.68%显著增加到48.34%和26.87%。The physical, chemical and digestive properties of the modified starch granules obtained in this example were tested. It can be seen that compared with the original starch, the solubility of the modified starch granules of the present invention was significantly increased from 9.86% to 47.65%, and the starch paste viscosity increased by 43.24%; The gelatinization peak temperature decreased from 68.98°C to 56.94°C; starch paste was stored at 4°C for one week, and its aging degree was significantly reduced from 58.94% to 36.57%; the contents of slowly digestible starch and resistant starch increased significantly from 24.65% and 8.68% to 48.34% and 26.87%.

将本实施例得到的变性淀粉颗粒进行应用试验:The modified starch granules that present embodiment obtains are carried out application test:

(1)添加在面制品中,延缓老化,改善面制品的口感、黏度稳定性、弹性等;(1) Added to noodle products to delay aging, improve the taste, viscosity stability, elasticity, etc. of noodle products;

(2)应用于化妆品行业,可作为粉底的添加剂;(2) Applied in the cosmetics industry, it can be used as an additive for foundation;

(3)作为一种多功能食品添加剂用于食品加工中,可以方便加工工艺、为食品提供优良的质构, 提高淀粉的增稠、悬浮、保水和稳定能力;(3) As a multifunctional food additive used in food processing, it can facilitate the processing technology, provide food with excellent texture, and improve the thickening, suspension, water retention and stability of starch;

(4)作为功能性成分用于多种食品,显著降低消化速率,从而降低血糖释放量。(4) Used as a functional ingredient in a variety of foods, significantly reducing the rate of digestion, thereby reducing the release of blood sugar.

Claims (8)

1. a kind of environment-friendly preparation method thereof of superelevation branch converted starch particle, it is characterised in that:Using starch from sweet potato as receptor, malt Oligosaccharide is donor, carries out enzyme hydrolysis successively to starch from sweet potato using branching enzyme, beta amylase and glycosyltransferase and turns Glucosides acts on, and obtains the superelevation branch converted starch particle containing higher branch's short chain proportional numbers.
2. the environment-friendly preparation method thereof of superelevation branch converted starch particle according to claim 1, it is characterized in that, it specifically includes Following steps:(1)Starch from sweet potato is configured to 5(w/v)The starch fluid of %, it is 6.5 to adjust starch fluid pH;(2)Add into starch fluid Enter branching enzyme, hydrolysis is vibrated in water-bath, then enzyme deactivation activity, cooling obtains enzymolysis liquid;(3)It is 5.2 to adjust enzymolysis liquid pH, is added Enter beta amylase, hydrolysis is shaken in water-bath, then enzyme deactivation activity, cooling obtains enzymolysis liquid;(4)Malt is added into enzymolysis liquid Oligosaccharide adjusts pH to 5.0, adds glycosyltransferase after mixing, is placed in water-bath and vibrates enzymolysis, then enzyme deactivation Activity obtains sample liquid after cooling;(5)Absolute ethyl alcohol is added into sample liquid, centrifuges, gained precipitation is freeze-dried to obtain product.
3. the environment-friendly preparation method thereof of superelevation branch converted starch particle according to claim 2, it is characterised in that:Step (1)In, 10g starch from sweet potato is weighed, it is 200mL that 0.02M sodium acetate buffers to the farinaceous size volume that pH is 6.5, which is added,.
4. the environment-friendly preparation method thereof of superelevation branch converted starch particle according to claim 2, it is characterised in that:Step (2)In, the branching enzyme of 308U/g is added into starch fluid, is placed in 60 DEG C of water-baths and vibrates 20-24h, 2.5mL is then added The NaOH solution enzyme deactivation activity of 1mol/L.
5. the environment-friendly preparation method thereof of superelevation branch converted starch particle according to claim 2, it is characterised in that:Step (3)In, the beta amylase of 10u/g is added into enzymolysis liquid, is placed in 37 DEG C of water-baths and vibrates 4-6h, 2.5mL is then added The NaOH solution enzyme deactivation activity of 1mol/L.
6. the environment-friendly preparation method thereof of superelevation branch converted starch particle according to claim 2, it is characterised in that:Step (4)In, the malto-oligosaccharide for accounting for its weight ratio 5% is added into enzymolysis liquid, the glucoside of 9088U/g is added after mixing Transferase is placed in oscillation enzymolysis 10-12h in 55 DEG C of water-baths, and the NaOH solution enzyme deactivation activity of 2.5mL 1mol/L is then added.
7. the environment-friendly preparation method thereof of superelevation branch converted starch particle according to claim 2, it is characterised in that:Step (5)In, the absolute ethyl alcohol of its 3 times of volumes is added into sample liquid, 10min is centrifuged under conditions of 3000r/min, is precipitated.
8. the environment-friendly preparation method thereof of the superelevation branch converted starch particle according to claim 2 or 6, it is characterised in that:Institute It is 20% maltotetraose, 30% maltopentaose, 40% maltose and 10% malt that malto-oligosaccharide, which is stated, by weight proportion Seven sugar mixing compositions.
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CN110438183A (en) * 2019-07-25 2019-11-12 哈尔滨商业大学 Combined-enzyme method modified starch prepares the application of the method and low DE value maltodextrin of low DE value maltodextrin in food
CN116766546A (en) * 2023-05-24 2023-09-19 齐鲁工业大学(山东省科学院) Method for preparing starch-based straw by bioenzyme co-extrusion shearing

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