CN1225943A - Process for preparing soya-beta amylase - Google Patents
Process for preparing soya-beta amylase Download PDFInfo
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- CN1225943A CN1225943A CN 99102506 CN99102506A CN1225943A CN 1225943 A CN1225943 A CN 1225943A CN 99102506 CN99102506 CN 99102506 CN 99102506 A CN99102506 A CN 99102506A CN 1225943 A CN1225943 A CN 1225943A
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Abstract
The preparation process of soybean beta-amylase with high enzyme activity includes the following steps: ultrafiltration and concentration of defatted soybean powder extract, secondary precipitation and precipitate freeze-thawing treatment. Said preparation process is low in energy consumption, high in production efficiency,and more suitable for industrial production. Said invented soybean beta-amylase is characterized by that its optimum action temp. is 60 deg.C, action temp. range is 45-65 deg.C, optimum action pH is 5.0-6.0, action pH range is 4.0-7.0, and it contains no alpha-amylase.
Description
The present invention relates to the beta-amylase production technique in the kind of plant, relate to a kind of preparation technology of high enzymatic activity soya-beta amylase particularly.
Beta-amylase is the circumscribed-type saccharifying enzyme, when it acts on starch, downcuts maltose unit successively from the non reducing end of the sugared sweet key of α-1,4, and hydrolysis thanks to product maltose, trisaccharide maltose and β-boundary dextrin.Beta-amylase is mainly used in foodstuffs industry, as the production of high barley-sugar slurry and superhigh maltose syrup.At present, in the prior art, with the commodity beta-amylase poor heat resistance of Production by Microorganism Fermentation, and cost is higher.Beta-amylase vigor height in the plant, good heat resistance, the action pH scope is wide, but the sweet potato beta-amylase enzyme activity of once producing in the prior art is extremely low, and quality is coarse, eliminates.Soya-beta amylase preparation method according to Gu Haixian, Guo Qingwen report (sees Gu Haixian, Guo Qingwen, 1997, the preparation of soya-beta amylase and the research of zymologic property, instrument and fermentation industry 23 (6) 45-49), comprise dextrin absorption and alcohol precipitation, though the enzyme activity of this method is than sweet potato beta-amylase height, but preparation process consumes a large amount of alcohol, must the supporting alcohol recovery device of setting up, thus make complicated operation, the cost height.Up to now, owing to lack relevant Technology, the high enzymatic activity plant beta-amylase industrial-scale production that still is unrealized.
The objective of the invention is to overcome the above-mentioned shortcoming of prior art, provide that a kind of power consumption is lower, production efficiency is high, be more suitable for industrial high enzymatic activity soya-beta amylase preparation technology.Soya-beta amylase according to the good product performance of this explained hereafter further is provided.
In order to realize above-mentioned purpose of the present invention, the invention provides following technical scheme:
The preparation technology of soya-beta amylase, the ultrafiltration and concentration that comprises the defatted soy flour vat liquor, secondary sedimentation and sedimentary freeze thawing treatment step, particularly, defatted soy flour through damping fluid after 30-40 ℃ of lixiviate, filter, add the sodium sulfate of 0.2-1.0%, ultrafiltration and concentration, the pH that adjusts concentrated solution is 3.6-5.0, adding sodium sulphate to concentration is 24%-31%, filters or centrifugal collecting precipitation, precipitation is placed-10 ℃ to-1 ℃ quick-frozens to frozen state or place 1 ℃ to 5 ℃ to refrigerate more than 10 hours, take out precipitation, the adding distil water dissolving is filtered or centrifugal collection clear liquid, and it is 3.6-5.0 that above-mentioned filtrate is transferred to pH, add 15%-20% sodium sulfate, filter or centrifugal collecting precipitation oven dry, crushing screening.
In the above-mentioned technology, ultrafiltration and concentration can preferably be used the daltonian hollow fiber column of exclusion molecular weight 20K-30K.
In the above-mentioned technology, preferably temperature 35-40 ℃ of oven dry.
Simultaneously, the present invention also provides the soya-beta amylase that the above-mentioned explained hereafter of foundation goes out.Its characteristic is: optimum temperature is 60 ℃, 45-65 ℃ of operative temperature scope, and the suitableeest action pH 5.0-6.0, action pH scope 4.0-7.0 does not contain α-Dian Fenmei.
Excellent effect foot of the present invention draws based on following principle: defatted soy flour is after the damping fluid lixiviate; filter; the sodium sulfate that adds 0.2-1.0% to filtrate; as protein protective agent; carry out ultrafiltration and concentration with the daltonian hollow fiber column of exclusion molecular weight 20K, the pH that adjusts concentrated solution is 3.6-5.5, carries out the precipitation first time; adding sodium sulphate to concentration is 24%-31%, filters or centrifugal collecting precipitation.The enzyme this moment rate of recovery alive is 85%.Precipitation is placed-10 ℃ to-1 ℃ quick-frozens to frozen state or place 1 ℃ to 5 ℃ refrigeration more than 10 hours, take out precipitation, it is molten to add an amount of distillation hydrolysis, filters or centrifugal collection clear liquid, abandons insolubles.Analyzing insolubles is polysaccharose substances such as foreign protein and starch because these material tool cryoprecipitabilities, through degree of depth cooling in addition freezing after, no longer be dissolved in the cold water, and be removed as precipitation; After testing, the precipitation China National Instruments Import ﹠ Export Corporation has the beta-amylase vigor of denier residual, and this part sedimentary dry weight accounts for 1/5th to 1/4th of dried enzyme powder; Therefore, through this purification step, the ratio vigor of enzyme improves 25%-33%.At this moment, the ratio vigor of beta-amylase has reached 30Ku/ml (1u i.e. 1 enzyme activity unit, and it is meant under suitable reaction condition, per hour produces the enzyme amount of 1 milligram of maltose) in the filtrate, adds certain preservatives, can be used as liquid enzymes and directly uses.If will make the dry powder of enzyme, can carry out the precipitation second time to the dissolving clear liquid, being about to above-mentioned filtrate, to transfer to pH be 3.6-5.0, add 15%-20% sodium sulfate, filter or centrifugal collecting precipitation crushing screening after 35-40 ℃ of oven dry, get the high enzymatic activity soya-beta amylase, color and luster is pale yellow, and enzyme activity is up to 430Ku/g, and total enzyme rate of recovery alive is 70%.These indexs all are better than the goods that obtain with prior art.As seen the present invention has overcome the above-mentioned shortcoming of prior art, provides that a kind of power consumption is lower, production efficiency is high, be more suitable for industrial high enzymatic activity soya-beta amylase preparation technology.Soya-beta amylase according to the good product performance of explained hereafter of the present invention also is provided simultaneously.Its characteristic is: optimum temperature is 60 ℃, 45-65 ℃ of operative temperature scope, and the suitableeest action pH 5.0-6.0, action pH scope 4.0-7.0 does not contain α-Dian Fenmei.
Further specify essentiality content of the present invention with embodiment below, but content of the present invention is not limited thereto.
Embodiment 1: take by weighing 1 kilogram of defatted soy flour, add 9 kilograms of damping fluids, 40 ℃ were extracted after 30 minutes, filter, filter residue adds 1 kilogram of distilled water, stirs after-filtration, merging filtrate adds sodium sulfate 45 grams to filtrate, after the dissolving fully, carry out ultrafiltration and concentration with the daltonian hollow fiber column of exclusion molecular weight 20K, the pH that adjusts concentrated solution is 4.5, adds sodium sulphate 560 grams, treat that sodium sulfate crystal dissolves fully after, centrifugal 20 minutes of 6000rpm, collecting precipitation also places-8 ℃ of quick-frozens.Add distilled water 450ml to the precipitation of freezing, centrifugal 25 minutes of dissolving back 6000rpm, supernatant liquor transfers to pH4.5, adds sodium sulfate 104 grams, treat that sodium sulfate crystal dissolves fully after, centrifugal collecting precipitation, it is online that precipitation is dispersed in stainless steel sift, and 40 ℃ of heat were dried by the fire 20 hours, pulverizes the back and cross 120 eye mesh screens, get enzyme powder 39 grams, measuring the beta-amylase vigor is 432Ku/g.
Embodiment 2: take by weighing 2 kilograms of defatted soy flours, add 17 kilograms of damping fluids, 30 ℃ were extracted after 40 minutes, filter, filter residue adds 2 kilograms of distilled water, stirs after-filtration, merging filtrate adds sodium sulfate 88 grams to filtrate, after the dissolving fully, carry out ultrafiltration and concentration with the daltonian hollow fiber column of exclusion molecular weight 30K, the pH that adjusts concentrated solution is 3.8, adds sodium sulphate 990 grams, treat that sodium sulfate crystal dissolves fully after, centrifugal 20 minutes of 6000rpm, collecting precipitation put to refrigerate in the refrigerator and spend the night.Precipitation to refrigeration adds distilled water 900ml, centrifugal 25 minutes of dissolving back 6000rpm, supernatant liquor transfers to pH3.8, adds sodium sulfate 200 grams, treat that sodium sulfate crystal dissolves fully after, centrifugal collecting precipitation, it is online that precipitation is dispersed in stainless steel sift, and 38 ℃ of heat were dried by the fire 24 hours, pulverizes the back and cross 120 eye mesh screens, get enzyme powder 80 grams, measuring the beta-amylase vigor is 424Ku/g.
Embodiment 3: take by weighing 5 kilograms of defatted soy flours, add 45 kilograms of damping fluids, 35 ℃ were extracted after 40 minutes, filter, filter residue adds 5 kilograms of distilled water, stirs after-filtration, merging filtrate adds sodium sulfate 225 grams to filtrate, after the dissolving fully, carry out ultrafiltration and concentration with the daltonian hollow fiber column of exclusion molecular weight 30K, the pH that adjusts concentrated solution is 4.2, adds sodium sulfate 2600 grams, treat that sodium sulfate crystal dissolves fully after, centrifugal 20 minutes of 6000rpm, collecting precipitation put to refrigerate in the refrigerator and spend the night.Precipitation to refrigeration adds distilled water 2250ml, centrifugal 25 minutes of dissolving back 6000rpm, supernatant liquor transfers to pH4.2, adds sodium sulfate 500 grams, treat that sodium sulfate crystal dissolves fully after, centrifugal collecting precipitation, it is online that precipitation is dispersed in stainless steel sift, and 40 ℃ of heat were dried by the fire 20 hours, pulverizes the back and cross 120 eye mesh screens, get enzyme powder 197 grams, measuring the beta-amylase vigor is 430Ku/g.
The soya-beta amylase that the foregoing description 1-3 prepares.Its characteristic is: optimum temperature is 60 ℃, 45-65 ℃ of operative temperature scope, and the suitableeest action pH 5.0-6.0, action pH scope 4.0-7.0 does not contain α-Dian Fenmei.
Claims (4)
1, the preparation technology of soya-beta amylase, the ultrafiltration and concentration that comprises the defatted soy flour vat liquor, secondary sedimentation and sedimentary freeze thawing treatment step, it is characterized in that defatted soy flour through damping fluid after 30-40 ℃ of lixiviate, filter, the sodium sulfate that adds 0.2-1.0%, ultrafiltration and concentration, the pH that adjusts concentrated solution is 3.6-5.0, adding sodium sulphate to concentration is 24%-31%, filter or centrifugal collecting precipitation, precipitation is placed-10 ℃ to-1 ℃ quick-frozens to frozen state or place 1 ℃ to 5 ℃ refrigeration more than 10 hours, take out precipitation, the adding distil water dissolving, filter or centrifugal collection clear liquid, it is 3.6-5.0 that above-mentioned filtrate is transferred to pH, adds 15%-20% sodium sulfate, filters or centrifugal collecting precipitation, oven dry, crushing screening.
2, preparation technology according to claim 1 is characterized in that ultrafiltration and concentration preferably uses the daltonian hollow fiber column of exclusion molecular weight 20K-30K.
3, preparation technology according to claim 1 is characterized in that drying preferred temperature 35-40 ℃.
4, the soya-beta amylase of producing according to claim 1 or 2 or 3 described preparation technologies.
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| CN99102506A CN1088754C (en) | 1999-02-03 | 1999-02-03 | Process for preparing soya-beta amylase |
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| CN99102506A CN1088754C (en) | 1999-02-03 | 1999-02-03 | Process for preparing soya-beta amylase |
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| CN1225943A true CN1225943A (en) | 1999-08-18 |
| CN1088754C CN1088754C (en) | 2002-08-07 |
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| CN99102506A Expired - Fee Related CN1088754C (en) | 1999-02-03 | 1999-02-03 | Process for preparing soya-beta amylase |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102378764A (en) * | 2009-03-30 | 2012-03-14 | 罗盖特兄弟公司 | Method for obtaining a preparation of beta-amylases from the soluble fractions of starch plants |
| CN103232982A (en) * | 2013-05-15 | 2013-08-07 | 常熟市诺科生化工程有限公司 | Preparation method for high-activity beta-amylase |
| WO2015193601A1 (en) | 2014-06-16 | 2015-12-23 | Roquette Freres | Method for manufacturing a stable aqueous solution of β-amylase, aqueous solution obtained and uses thereof |
| CN116286747A (en) * | 2023-05-22 | 2023-06-23 | 烟台麦特尔生物技术有限公司 | Production process for removing alpha-amylase in beta-amylase |
| US11987828B2 (en) | 2015-07-06 | 2024-05-21 | Roquette Freres | Method for producing maltitol, with an increased yield |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1044940A (en) * | 1989-02-14 | 1990-08-29 | 何志存 | Raw material treatment process when fermentative production alcohol and alcohol product before the saccharification |
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1999
- 1999-02-03 CN CN99102506A patent/CN1088754C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102378764A (en) * | 2009-03-30 | 2012-03-14 | 罗盖特兄弟公司 | Method for obtaining a preparation of beta-amylases from the soluble fractions of starch plants |
| CN103232982A (en) * | 2013-05-15 | 2013-08-07 | 常熟市诺科生化工程有限公司 | Preparation method for high-activity beta-amylase |
| CN103232982B (en) * | 2013-05-15 | 2015-03-04 | 常熟市诺科生化工程有限公司 | Preparation method for high-activity beta-amylase |
| WO2015193601A1 (en) | 2014-06-16 | 2015-12-23 | Roquette Freres | Method for manufacturing a stable aqueous solution of β-amylase, aqueous solution obtained and uses thereof |
| US11987828B2 (en) | 2015-07-06 | 2024-05-21 | Roquette Freres | Method for producing maltitol, with an increased yield |
| CN116286747A (en) * | 2023-05-22 | 2023-06-23 | 烟台麦特尔生物技术有限公司 | Production process for removing alpha-amylase in beta-amylase |
| CN116286747B (en) * | 2023-05-22 | 2023-08-08 | 烟台麦特尔生物技术有限公司 | Production process for removing alpha-amylase in beta-amylase |
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| Publication number | Publication date |
|---|---|
| CN1088754C (en) | 2002-08-07 |
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