CN115651950A - Preparation method of long chain amylopectin - Google Patents
Preparation method of long chain amylopectin Download PDFInfo
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- CN115651950A CN115651950A CN202211406986.3A CN202211406986A CN115651950A CN 115651950 A CN115651950 A CN 115651950A CN 202211406986 A CN202211406986 A CN 202211406986A CN 115651950 A CN115651950 A CN 115651950A
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- starch
- long chain
- amylopectin
- chain amylopectin
- debranching
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- 229920000945 Amylopectin Polymers 0.000 title claims abstract description 63
- 238000002360 preparation method Methods 0.000 title abstract description 14
- 229920002472 Starch Polymers 0.000 claims abstract description 117
- 235000019698 starch Nutrition 0.000 claims abstract description 115
- 239000008107 starch Substances 0.000 claims abstract description 114
- 238000000034 method Methods 0.000 claims abstract description 34
- 235000013336 milk Nutrition 0.000 claims abstract description 25
- 239000008267 milk Substances 0.000 claims abstract description 25
- 210000004080 milk Anatomy 0.000 claims abstract description 25
- 238000010438 heat treatment Methods 0.000 claims abstract description 17
- 108090000790 Enzymes Proteins 0.000 claims abstract description 16
- 102000004190 Enzymes Human genes 0.000 claims abstract description 16
- 239000012634 fragment Substances 0.000 claims abstract description 12
- 238000001035 drying Methods 0.000 claims abstract description 11
- 239000002244 precipitate Substances 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 108090000637 alpha-Amylases Proteins 0.000 claims description 8
- 108010028688 Isoamylase Proteins 0.000 claims description 4
- 230000003301 hydrolyzing effect Effects 0.000 claims description 2
- 238000012545 processing Methods 0.000 abstract description 4
- 229920002261 Corn starch Polymers 0.000 description 17
- 239000008120 corn starch Substances 0.000 description 17
- 238000006116 polymerization reaction Methods 0.000 description 14
- 230000002255 enzymatic effect Effects 0.000 description 9
- 238000001816 cooling Methods 0.000 description 8
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 8
- 229920001592 potato starch Polymers 0.000 description 8
- 238000005406 washing Methods 0.000 description 8
- 239000002994 raw material Substances 0.000 description 6
- 240000003183 Manihot esculenta Species 0.000 description 5
- 235000016735 Manihot esculenta subsp esculenta Nutrition 0.000 description 5
- 108010033764 Amylosucrase Proteins 0.000 description 4
- 229930006000 Sucrose Natural products 0.000 description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 4
- 239000005720 sucrose Substances 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 3
- 235000019621 digestibility Nutrition 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 229920000856 Amylose Polymers 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 241000588660 Neisseria polysaccharea Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 229940100486 rice starch Drugs 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 229940100445 wheat starch Drugs 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- IPCSVZSSVZVIGE-UHFFFAOYSA-M hexadecanoate Chemical compound CCCCCCCCCCCCCCCC([O-])=O IPCSVZSSVZVIGE-UHFFFAOYSA-M 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
Landscapes
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of starch deep processing, and discloses a preparation method of long chain amylopectin, which comprises the following steps: (1) Preparing starch into starch milk, and performing heat treatment in a water bath environment at a temperature lower than the gelatinization termination temperature of the starch; (2) Carrying out enzymolysis debranching treatment on the starch after heat treatment by using debranching enzyme; (3) And centrifuging the enzyme-hydrolyzed starch solution after debranching treatment, collecting centrifugal starch precipitate, adding ethanol solution to passivate debranching enzyme, drying and crushing to obtain starch fragments rich in long chain amylopectin. The method provided by the invention can effectively enrich long chain amylopectin, and then obviously improve the average chain length of the amylopectin.
Description
Technical Field
The invention belongs to the technical field of starch deep processing, and particularly relates to a preparation method of long chain amylopectin.
Background
Starch is the only carbohydrate in nature in particulate form, a macromolecular glucan stored mainly in the cells of seeds and subterranean stems of higher plants. The starch is heteropolysaccharide composed of amylose and amylopectin, and the amylose is mainly composed of glucose molecules connected by alpha-1,4 glycosidic bonds and is in a linear chain; amylopectin is glycosidically linked at the branches by α -1,6, with the linear portion also being α -1,4. The fine structure of starch directly affects the functional properties of starch (such as paste properties, heat stability and digestibility); among these, the content of long chain amylopectin significantly affects the starch paste properties and its digestibility in the human body. Compared with short amylopectin, long amylopectin has higher possibility of self-aggregation or forming ordered aggregates by interaction with hydrophobic guest molecules (such as lipid, palmitate and the like), and the regulation and control on starch paste properties and digestibility are more remarkable. Therefore, finding a way with mild preparation conditions and simple operation to obtain long chain amylopectin becomes one of the research contents concerned in the field of starch deep processing at home and abroad.
In recent years, the preparation of long chain amylopectin mainly utilizes the catalytic action of amylosucrase (from Neisseria polysaccharea) on starch and sucrose to transfer glucose in sucrose to starch molecular chains, thereby realizing the extension of the starch molecular chains. However, at present, the amylosucrase is mainly derived from neisseria polysaccharea, and the isolation process is complex and has not been popularized and applied commercially, so that the development of screening and preparation of long chain amylopectin based on amylosucrase catalysis is limited. In particular, until now, no other separation methods have been available to efficiently synthesize or isolate long chain amylopectin. Therefore, the development of a preparation method of the long chain amylopectin, which is simple to operate, green and environment-friendly and does not need special raw materials and equipment, is a key problem to be solved for industrial large-scale production of the long chain amylopectin.
Disclosure of Invention
Aiming at the problem of application limitation of the prior art, particularly the preparation of long chain amylopectin by using amylosucrase, the invention provides the long chain amylopectin and the preparation method thereof, and the screening and the preparation of the starch fragment rich in the long chain amylopectin can be realized by three simple procedures of starch milk preheating, debranching enzyme enzymolysis and centrifugal separation.
The technical scheme of the invention is as follows:
one of the purposes of the invention is a preparation method of long chain amylopectin, which comprises the following steps:
(1) Preparing starch into starch milk, and performing heat treatment in a water bath environment at a temperature lower than the gelatinization termination temperature of the starch;
(2) Carrying out enzymolysis debranching treatment on the starch subjected to heat treatment by using debranching enzyme;
(3) And centrifuging the debranched enzymatic starch, collecting centrifugal starch precipitate, adding ethanol solution to passivate debranching enzyme, drying and crushing to obtain starch fragments rich in long chain amylopectin.
Further, the starch is a large commercial starch, including corn starch, potato starch, tapioca starch, rice starch, wheat starch, and the like.
Further, the mass concentration of the starch milk prepared and formed in the step (1) is 1-15%.
Further, the temperature for carrying out heat treatment on the starch milk in the step (1) is 58-80 ℃; preferably, 3-10 ℃ below the starch gelatinization termination temperature; the heat treatment time is 10-60min; preferably, 20-40min
Further, the debranching enzyme is one or more of pullulanase or isoamylase.
Further, the adding amount of the debranching enzyme in the step (2) is 20-1000U/g starch; preferably, it is 200 to 600U/g starch.
Further, the temperature of the debranching enzyme for enzymolysis of starch in the step (2) is 55-65 ℃, preferably 58-62 ℃; the time for enzymolysis of starch is 2-48h, preferably 6-24h.
Further, the pH of the solution of debranching enzyme for hydrolyzing starch in the step (2) is 4.5-6.5, preferably 5.0-6.0.
Further, the centrifugal rotating speed of the starch enzymolysis liquid in the step (3) is 500-8000r/min, preferably 2000-5000r/min.
The second object of the present invention is to provide a long chain amylopectin starch separated according to the above-mentioned preparation method.
Compared with the prior art, the invention has the beneficial effects that:
1. the method provided by the invention can obtain the long chain amylopectin by carrying out simple heat treatment, enzymolysis and centrifugal separation on the starch milk, and has the advantages of simple operation and environmental protection; most importantly, the raw materials required in the invention are simple in source and do not need any special raw materials and equipment, and the invention is beneficial to the industrial production of the long chain amylopectin.
2. The method only regulates and controls the structure of the starch, does not relate to the use of raw materials such as sucrose in the preparation of the long chain amylopectin by the traditional starch sucrose enzyme method, saves the raw material investment and the production cost, and can be widely applied to the field of fine and deep processing of starch resources.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments; all other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the invention, all the percentages are weight units, and the adopted equipment, raw materials or materials and the like can be purchased from the market or are commonly used in the field; the methods in the following examples are conventional in the art unless otherwise specified.
Example 1
A method for preparing long chain amylopectin starch from waxy corn starch comprises the following steps:
(1) Preparing waxy corn starch into starch milk with concentration of 10%, and heating in water bath environment at 70 deg.C for 20min;
(2) Cooling the starch milk to 58 ℃, adjusting the pH value of the solution to 5.8, then adding 200U/g pullulanase, and carrying out enzymolysis debranching treatment for 24 hours;
(3) Centrifuging the debranched enzymatic starch at a centrifugal speed of 3000r/min, washing the obtained precipitate for 2 times by using 70% ethanol water solution, drying and crushing to obtain starch fragments rich in long chain amylopectin;
the waxy corn starch without any treatment in this example had an average degree of polymerization of 20.86 and a long chain amylopectin (degree of polymerization. Gtoreq.37) content of 11.25%; the average degree of polymerization and the content of amylopectin in the starch samples separated according to the method of this example were higher, 22.21 and 13.55%, respectively.
Example 2
A method for preparing long chain amylopectin starch from waxy corn starch comprises the following steps:
(1) Preparing waxy corn starch into starch milk with concentration of 15%, and heating in water bath environment at 80 deg.C for 20min;
(2) Cooling the starch milk to 60 ℃, adjusting the pH value of the solution to 5.5, then adding 1000U/g starch isoamylase, and carrying out enzymolysis for 2h;
(3) Centrifuging the debranched enzymatic starch at a centrifugal speed of 1000r/min, washing the obtained precipitate for 2 times by using 70% ethanol water solution, drying and crushing to obtain the starch fragment rich in long chain amylopectin.
The starch samples isolated according to the method of this example had an average degree of polymerization and a content of amylopectin higher than the waxy corn starch without any treatment, 28.09 and 25.19%, respectively.
Example 3
A method for preparing long chain amylopectin starch from waxy corn starch comprises the following steps:
(1) Preparing waxy corn starch into starch milk with concentration of 1%, and heating in water bath environment at 75 deg.C for 60min;
(2) Cooling the starch milk to 55 ℃, adjusting the pH value of the solution to 4.5, then adding 1000U/g pullulanase, and carrying out enzymolysis for 12h;
(3) Centrifuging the debranched enzymatic starch at a centrifugal speed of 500r/min, washing the obtained precipitate for 2 times by using 70% ethanol water solution, drying and crushing to obtain the starch fragment rich in long chain amylopectin.
The starch samples isolated according to the method of this example had an average degree of polymerization and a content of amylopectin higher, 23.61 and 14.83% respectively, than the waxy corn starch without any treatment.
Example 4
A method for preparing long chain amylopectin starch from waxy corn starch comprises the following steps:
(1) Preparing waxy corn starch into starch milk with concentration of 10%, and heating in 78 deg.C water bath environment for 30min;
(2) Cooling the starch milk to 65 ℃, adjusting the pH value of the solution to 5.2, then adding pullulanase of 500U/g starch, and carrying out enzymolysis for 48h;
(3) Centrifuging the debranched enzymatic starch at a centrifugation speed of 1000r/min, washing the obtained precipitate for 2 times by using 70% ethanol water solution, drying and crushing to obtain the starch fragment rich in long chain amylopectin.
The starch samples isolated according to the method of this example had an average degree of polymerization and a content of amylopectin higher than the waxy corn starch without any treatment, 25.63 and 19.68%, respectively.
Example 5
A method for preparing long chain amylopectin starch from waxy corn starch comprises the following steps:
(1) Preparing waxy corn starch into 10% starch milk, and heating in 78 deg.C water bath environment for 60min;
(2) Cooling the starch milk to 55 ℃, adjusting the pH value of the solution to 5.0, then adding 400U/g pullulanase, and carrying out enzymolysis for 36h;
(3) Centrifuging the debranched enzymatic starch at a centrifugal speed of 2000r/min, washing the obtained precipitate for 2 times by using 70% ethanol water solution, drying and crushing to obtain the starch fragment rich in long chain amylopectin.
The starch samples isolated according to the method of this example had an average degree of polymerization and a higher content of amylopectin, 29.81 and 29.64% respectively, compared to the waxy corn starch without any treatment.
Example 6
A method for preparing long chain amylopectin from common potato starch comprises the following steps:
(1) Preparing common potato starch into starch milk with the concentration of 5%, and heating in water bath environment at 60 deg.C for 10min;
(2) Cooling the starch milk to 58 ℃, adjusting the pH value of the solution to 5.2, then adding 200U/g starch isoamylase, and carrying out enzymolysis for 24 hours;
(3) Centrifuging the debranched enzymatic starch at a centrifugal speed of 3000r/min, washing the obtained precipitate for 2 times by using 70% ethanol water solution, drying and crushing to obtain the starch fragment rich in long chain amylopectin.
Ordinary potato starch without any treatment in this example had an average degree of polymerization of amylopectin of 22.72 and a long-chain amylopectin (degree of polymerization. Gtoreq.37) content of 15.72%; the average degree of polymerization and the content of amylopectin of the starch samples isolated according to the method of this example were higher, 24.87 and 20.68%, respectively.
Example 7
A method for preparing long chain amylopectin from common potato starch comprises the following steps:
(1) Preparing common potato starch into starch milk with concentration of 6%, and heating in water bath environment at 65 deg.C for 30min;
(2) Cooling the starch milk to 60 ℃, adjusting the pH value of the solution to 5.2, then adding pullulanase of 500U/g starch, and carrying out enzymolysis for 36 hours;
(3) Centrifuging the debranched enzymatic starch at a centrifugation speed of 1000r/min, washing the obtained precipitate for 2 times by using 70% ethanol water solution, drying and crushing to obtain the starch fragment rich in long chain amylopectin.
The starch samples isolated according to the method of this example had an average degree of polymerization and a higher content of amylopectin, 29.17 and 30.49% respectively, compared to normal potato starch without any treatment.
Example 8
A method for preparing long chain amylopectin from common tapioca starch comprises the following steps:
(1) Preparing common cassava starch into 9% starch milk, and heating in 75 deg.C water bath environment for 30min;
(2) Cooling the starch milk to 60 ℃, adjusting the pH value of the solution to 5.0, then adding 1000U/g pullulanase of starch, and carrying out enzymolysis for 6 hours;
(3) Centrifuging the debranched enzymatic starch at a centrifugation speed of 1000r/min, washing the obtained precipitate for 2 times by using 70% ethanol water solution, drying and crushing to obtain the starch fragment rich in long chain amylopectin.
The average polymerization degree of amylopectin of the common tapioca starch without any treatment in the embodiment is 20.65, and the content of long chain amylopectin (polymerization degree is more than or equal to 37) is 13.92%; the starch samples isolated according to the method of this example had higher average degrees of polymerization and amylopectin contents, 27.82 and 25.37%, respectively.
In addition to the waxy corn starch, normal potato starch and normal tapioca starch used in the above examples, other commercially available starches, such as rice starch or wheat starch, may also be used in the present invention.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (10)
1. A method for preparing long chain amylopectin is characterized by comprising the following steps:
(1) Preparing starch into starch milk, and performing heat treatment in a water bath environment at a temperature lower than the gelatinization termination temperature of the starch;
(2) Carrying out enzymolysis debranching treatment on the starch subjected to heat treatment by using debranching enzyme;
(3) And centrifuging the enzyme-hydrolyzed starch solution after debranching treatment, collecting centrifugal starch precipitate, adding ethanol solution to passivate debranching enzyme, drying and crushing to obtain starch fragments rich in long chain amylopectin.
2. The method of claim 1, wherein the starch is a bulk commercial starch.
3. The method for preparing long chain amylopectin according to claim 1, wherein the starch milk prepared in the step (1) has a mass concentration of 1-15%.
4. The method for preparing long chain amylopectin according to claim 1, wherein the temperature for heat-treating the starch milk in step (1) is 58-80 ℃ and the time for heat-treating is 10-60min.
5. The method according to claim 1, wherein the debranching enzyme is one or more of pullulanase and isoamylase.
6. The method according to claim 1, wherein the debranching enzyme is added in an amount of 20 to 1000U/g starch in step (2).
7. The method for preparing long chain amylopectin according to claim 1, wherein the temperature for enzymolysis of starch by debranching enzyme in the step (2) is 55-65 ℃ and the time for enzymolysis of starch is 2-48h.
8. The method according to claim 1, wherein the pH of the solution of debranching enzyme for hydrolyzing starch in step (2) is 4.5 to 6.5.
9. The method for preparing long chain amylopectin according to claim 1, wherein the centrifugal rotation speed of the starch enzymolysis solution in the step (3) is 500-8000r/min.
10. A long chain amylopectin isolated by the method according to any one of claims 1 to 7.
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| CN202211406986.3A CN115651950A (en) | 2022-11-10 | 2022-11-10 | Preparation method of long chain amylopectin |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1615317A (en) * | 2001-10-17 | 2005-05-11 | 巴斯福种植科学有限公司 | Starch |
| US20080016598A1 (en) * | 2002-03-27 | 2008-01-17 | Broglie Karen E | Maize starch containing elevated amounts of actual amylose |
| CN112921059A (en) * | 2021-02-05 | 2021-06-08 | 王学勇 | Preparation method of canna RS3 resistant starch and application of canna RS3 resistant starch in functional food and anti-Parkinsonian drugs |
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- 2022-11-10 CN CN202211406986.3A patent/CN115651950A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1615317A (en) * | 2001-10-17 | 2005-05-11 | 巴斯福种植科学有限公司 | Starch |
| US20080016598A1 (en) * | 2002-03-27 | 2008-01-17 | Broglie Karen E | Maize starch containing elevated amounts of actual amylose |
| CN112921059A (en) * | 2021-02-05 | 2021-06-08 | 王学勇 | Preparation method of canna RS3 resistant starch and application of canna RS3 resistant starch in functional food and anti-Parkinsonian drugs |
Non-Patent Citations (2)
| Title |
|---|
| 徐兵等: ""普鲁兰酶脱支对淀粉结构及热稳定性的影响"", 《食品科技》, vol. 45, no. 7, 31 July 2020 (2020-07-31), pages 255 - 261 * |
| 雷鸣: """部分糊化及酶脱支处理对淀粉物化特性影响研究""", 《全国优秀硕士论文集》, vol. 2022, no. 10, 15 October 2022 (2022-10-15), pages 1 - 81 * |
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