CN1088754C - Process for preparing soya-beta amylase - Google Patents
Process for preparing soya-beta amylase Download PDFInfo
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- CN1088754C CN1088754C CN99102506A CN99102506A CN1088754C CN 1088754 C CN1088754 C CN 1088754C CN 99102506 A CN99102506 A CN 99102506A CN 99102506 A CN99102506 A CN 99102506A CN 1088754 C CN1088754 C CN 1088754C
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- concentration
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- ultrafiltration
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Abstract
The present invention provides a preparation technology of a soybean beta amylase with high enzyme activity, which comprises the steps such as ultrafiltration and concentration of defatted soybean powder extract, secondary precipitation and precipitate freeze-thawing treatment. The technology has the advantages of low energy consumption and high production efficiency; the technology is more suitable for industrial production. The present invention also provides a soybean beta amylase having good product performance and produced according to the technology, which is characterized in that the optimum action temperature is 60 DEG C, the action temperature range is from 45 to 65 DEG C, the optimum action pH value is from 5.0 to 6.0, the action pH value range is from 4.0 to 7.0, and the soybean beta amylase does not contain alpha amylases.
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 the hydrolysis meta-bolites has 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, food and fermentation industries 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, filters after 30-40 ℃ of lixiviate through damping fluid, adds the sodium sulfate of 0.2-1.0%, ultrafiltration and concentration, the pH that adjusts concentrated solution is 3.6-5.0, and 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 ℃ refrigeration more than 10 hours, take out precipitation, the adding distil water dissolving is filtered or centrifugal collection clear liquid, 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 of the present invention is based on that following principle draws: 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, only have the beta-amylase vigor of denier residual in the precipitation, and this part sedimentary dry weight account 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, adds 15%-20% sodium sulfate, filter or centrifugal collecting precipitation, crushing screening after 35-40 ℃ of oven dry gets the high enzymatic activity soya-beta amylase, and color and luster is pale yellow, 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 (3)
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 and precipitate, 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, filter 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 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN99102506A CN1088754C (en) | 1999-02-03 | 1999-02-03 | Process for preparing soya-beta amylase |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN99102506A CN1088754C (en) | 1999-02-03 | 1999-02-03 | Process for preparing soya-beta amylase |
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| Publication Number | Publication Date |
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| CN1225943A CN1225943A (en) | 1999-08-18 |
| CN1088754C true 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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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2943686B1 (en) * | 2009-03-30 | 2013-11-01 | Roquette Freres | PROCESS FOR OBTAINING BETA AMYLASE PREPARATION FROM SOLUBLE FRUITS OF STARCH PLANTS |
| 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 |
| FR3022257B1 (en) | 2014-06-16 | 2018-03-30 | Roquette Freres | PROCESS FOR PRODUCING A STABLE AQUEOUS SOLUTION OF BETA-AMYLASE, AQUEOUS SOLUTION OBTAINED AND USES THEREOF |
| 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 |
Citations (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
Patent Citations (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 |
Non-Patent Citations (1)
| Title |
|---|
| 食品与发酵工业,第23卷第6期,第45-49页 1997.1.1 谷海先等,大豆B-淀粉酶的制备及酶学性质的研究 * |
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| CN1225943A (en) | 1999-08-18 |
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