CN101381713A - Preparation technology of high temperature resistant feeding cellulase - Google Patents
Preparation technology of high temperature resistant feeding cellulase Download PDFInfo
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- CN101381713A CN101381713A CNA2007100186513A CN200710018651A CN101381713A CN 101381713 A CN101381713 A CN 101381713A CN A2007100186513 A CNA2007100186513 A CN A2007100186513A CN 200710018651 A CN200710018651 A CN 200710018651A CN 101381713 A CN101381713 A CN 101381713A
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Abstract
The invention relates to a process for preparing cellulase used for heat resisting feeds. The process comprises the following steps: screening out heat resisting strains for preservation by performing heat resisting domestication first on the strains for the cellulase used for feed production, and performing transfer of culture on the preserved strains; and then fermenting the screened heat resisting strains to prepare into the mixed enzyme liquid, and concentrating the mixed enzyme liquid to certain concentration to perform microcapsule coating. The process can improve the tolerance temperature of the cellulase used for the feeds from 65 DEG C keeping for 10 minutes to 85 DEG C keeping for 10 minutes, and basically solve the difficult problem that the cellulase used for the feeds does not resist high temperature, thereby laying the technical foundation for the wide application.
Description
Technical field
The present invention relates to a kind of preparation technology of high temperature resistant feeding cellulase
Background technology
Feeding cellulase is a kind of as fodder enzyme preparation, can increase substantially the utilization ratio of non-starch polysaccharide in the feed, saves feed, promotes growth of animal and improves its production performance, and its effect is confirmed by vast Feed Enterprise and plant.But some defective effects of zymin inherent its to the more popularization of wide spectrum, feeding cellulase is no exception.In many influence factors (such as: bacterial classification source, leavening temperature, humidity, potential of hydrogen, time, complete processing, condition of storage or the like) in, feed processing technology is very big to the influence of feeding cellulase action effect, and especially some feed (as: granulated feed) needs " high temperature granulating " process (being generally 80-90 ℃, 10 minutes).And the optimum temperuture that feeding cellulase plays a role is 40 ℃, can cause most of zymoprotein inactivation more than 80 ℃, active decline 50-80%, even to no effect.This has had a strong impact on its application in granulated feed.Granulated feed generally simply or is not as yet grown at digestive tubes such as aquatic animal, livestock and poultry younglings and is improved animal specific, the existence of feeding cellulase can help these animals to perfect the digestive ferment system, quickens digestion, reduces chyme viscosity, reduce disease, makes animal health and saves feed.The feeding cellulase optimum temperuture is 40 ℃ in the prior art; But the highest tolerable temperature has only 65 ℃, this shows, the tolerance Wen Wendu that how to improve feeding cellulase is a problem that urgency is to be solved.
Summary of the invention
The technical problem that the present invention solves: the preparation technology that a kind of high temperature resistant feeding cellulase is provided, by carrying out double-layer microcapsule bag quilt, make the high-temperature stability of feeding cellulase bring up to 85 ℃, 10 minutes by 65 ℃, 10 minutes to the high temperature resistant domestication of feeding cellulase bacterial strain with to feeding cellulase.
Technical solution of the present invention:
A kind of preparation technology of high temperature resistant feeding cellulase:
1. the bacterial strain that produces feeding cellulase is carried out filtering out the high-temperature resistant strain preservation after the high temperature resistant domestication, and the bacterial classification of preservation is gone down to posterity;
2. mixed enzyme solution is made in fermentation to the high-temperature resistant strain that filters out, and carries out microcapsule bag quilt after mixed enzyme solution is concentrated into finite concentration.
Described high temperature resistant domestication process is:
The first step, earlier the certain density cellulase solution that described bacterial strain metabolism is produced place respectively 45 ℃, 50 ℃, 55 ℃, 60 ℃, 65 ℃, 70 ℃ water-baths heat 5 ', 10 ', 15 ', surveying enzyme respectively with the DNS method lives, draw the enzyme live data of this bacterial strain after differing temps, different time are handled before and after the domestication, the bacterial strain of selecting the high temperature resistant relative long period is for just sending out bacterial strain;
Second step, adopt ultraviolet irradiation to carry out mutagenesis to just sending out bacterial strain, described bacterial strain is exposed to shines 30 under the ultraviolet lamp respectively ', 60 ', 90 ', the bacterial strain of getting different irradiation times carries out multiplication culture, dilution separates, obtain single bacterium colony, choosing colony is done strain identification then, the bacterial strain that meets proterties is produced high temperature resistant feeding cellulase ability with the DNS method measure;
The 3rd step, with enzyme work under same concentrations, the 40 ℃ of conditions is the relatively the first step, enzyme in second step rate of loss of living of benchmark, filter out the bacterial strain of high temperature resistant relative long period, and repeat second process, until filtering out required high-temperature resistant strain, but and to verify that the high-temperature resistant strain that filters out has good hereditary potency.
Described microcapsule bag by process is: the strain fermentation that filters out after the high temperature resistant domestication is made mixed enzyme solution, will be concentrated into certain density mixed enzyme solution and live with DNS method survey enzyme; Prepare 4.5%～5.5% gum arabic aqueous solution and 9%～11% aqueous gelatin solution respectively, 45 ℃ of insulations are standby; With mixed enzyme solution and 4.5%～5.5% gum arabic aqueous solution vibration mixing rapidly after 1.8～2.2:1 mixed by volume, transfer pH value to 4.2～4.6, and be cooled to 19.5～20.5 ℃ rapidly and make mixed solution, 1.8～2.2: 1 ratio adds 9%～11% aqueous gelatin solution by volume in mixed solution, continue to stir, and be cooled to rapidly below 4 ℃, transfer pH value to 2.0 back to continue to stir 30min, dry getting final product.
The present invention can make the tolerable temperature of feeding cellulase bring up to 85 ℃, 10 minutes by 65 ℃, 10 minutes, solves the difficult problem of feeding cellulase non-refractory substantially, for technical foundation has been established in its widespread use.
Embodiment
1, produce the high temperature resistant domestication of feeding cellulase bacterial strain:
The first step, earlier the concentration that described bacterial strain metabolism is produced be 80% cellulase solution place respectively 45 ℃, 50 ℃, 55 ℃, 60 ℃, 65 ℃, 70 ℃ water-baths heat 5 ', 10 ', 15 ', surveying enzyme respectively with the DNS method lives, draw the enzyme live data of this bacterial strain after differing temps, different time are handled before and after the domestication, the bacterial strain of selecting the high temperature resistant relative long period is for just sending out bacterial strain;
Second step, adopt ultraviolet irradiation to carry out mutagenesis, described bacterial strain is exposed to shines 30 under the ultraviolet lamp respectively ', 60 ', 90 ', the bacterial strain of getting different irradiation times carries out multiplication culture, dilution separates, obtain single bacterium colony, choosing colony is done strain identification then, the bacterial strain that meets proterties is produced high temperature resistant feeding cellulase ability with the DNS method measure;
The 3rd step, with enzyme work under same concentrations, the 40 ℃ of conditions is the relatively the first step, enzyme in second step rate of loss of living of benchmark, filter out the bacterial strain of high temperature resistant relative long period, and repeat second process, until filtering out required high-temperature resistant strain, but and to verify that the high-temperature resistant strain that filters out has good hereditary potency.
2, with microcapsule with enzyme liquid bag quilt: the strain fermentation that filters out after the high temperature resistant domestication is made mixed enzyme solution, is that 80% mixed enzyme solution is surveyed enzyme with the DNS method and lived with being concentrated into concentration; Prepare the 5% gum arabic aqueous solution and 10% aqueous gelatin solution respectively, 45 ℃ of insulations are standby; With enzyme liquid and 5% gum arabic aqueous solution vibration mixing rapidly after 1.8～2.2 mixed by volume, transfer pH value to 4.6, and be cooled to 20 ℃ rapidly and make mixed solution, 1.8～2.2 ratios add 10% aqueous gelatin solution by volume in mixed solution, continue to stir, and be cooled to rapidly below 4 ℃, transfer pH value to 2.0 back to continue to stir 30min, Air drying.
3, DNS method survey enzyme is lived.Place 65 ℃, 70 ℃, 75 ℃, 80 ℃, 85 ℃, 90 ℃, 95 ℃ water-baths to be incubated 10 minutes respectively in above-mentioned different batches sample, survey enzyme with the DNS method and live, relatively enzyme rate of loss alive.The tolerable temperature that shows feeding cellulase reaches by 85 ℃, 10 minutes, solves the difficult problem of feeding cellulase non-refractory substantially, for technical foundation has been established in its widespread use.
4, " in-vitro simulated stomach ENVIRONMENTAL LAW " bilayered microcapsule broken wall test.
(1) preparation of the external shell-broken liquid of simulation stomach environment.
No. 1 liquid: it is an amount of that dilute hydrochloric acid solution pH is adjusted to 2.0+ aspartic protease (our company's self-control);
No. 2 liquid: get No. 1 liquid 750ml and add the 250ml0.2mol/L metabisulfite solution, transfer pH to 6.8;
(2) bilayered microcapsule broken wall test.
Accurately a certain amount of dry sample of weighing is put in No. 1 liquid of certain volume, and 37 ℃ of water bath heat preservation 30min transfer pH to 6.8, add isopyknic No. 2 liquid, 37 ℃ of insulation 30min.Centrifugal, taking precipitate, the microscopy sporoderm-broken rate reaches more than 90%.
Claims (3)
1, a kind of preparation technology of high temperature resistant feeding cellulase is characterized in that:
1. the bacterial strain that produces feeding cellulase is carried out filtering out the high-temperature resistant strain preservation after the high temperature resistant domestication, and the bacterial classification of preservation is gone down to posterity;
2. mixed enzyme solution is made in fermentation to the high-temperature resistant strain that filters out, and carries out microcapsule bag quilt after mixed enzyme solution is concentrated into finite concentration.
2, the preparation technology of a kind of high temperature resistant feeding cellulase according to claim 1 is characterized in that described high temperature resistant domestication process is:
The first step, earlier the certain density cellulase solution that described bacterial strain metabolism is produced place respectively 45 ℃, 50 ℃, 55 ℃, 60 ℃, 65 ℃, 70 ℃ water-baths heat 5 ', 10 ', 15 ', surveying enzyme respectively with the DNS method lives, draw the enzyme live data of this bacterial strain after differing temps, different time are handled before and after the domestication, the bacterial strain of selecting the high temperature resistant relative long period is for just sending out bacterial strain;
Second step, adopt ultraviolet irradiation to carry out mutagenesis to just sending out bacterial strain, described bacterial strain is exposed to shines 30 under the ultraviolet lamp respectively ', 60 ', 90 ', the bacterial strain of getting different irradiation times carries out multiplication culture, dilution separates, obtain single bacterium colony, choosing colony is done strain identification then, the bacterial strain that meets proterties is produced high temperature resistant feeding cellulase ability with the DNS method measure;
The 3rd step, with enzyme work under same concentrations, the 40 ℃ of conditions is the relatively the first step, enzyme in second step rate of loss of living of benchmark, filter out the bacterial strain of high temperature resistant relative long period, and repeat second process, until filtering out required high-temperature resistant strain, but and to verify that the high-temperature resistant strain that filters out has good hereditary potency.
3, the preparation technology of a kind of high temperature resistant feeding cellulase according to claim 1, it is characterized in that described microcapsule bag by process is: the strain fermentation that filters out after the high temperature resistant domestication is made mixed enzyme solution, will be concentrated into certain density mixed enzyme solution and live with DNS method survey enzyme; Prepare 4.5%～5.5% gum arabic aqueous solution and 9%～11% aqueous gelatin solution respectively, 45 ℃ of insulations are standby; With mixed enzyme solution and 4.5%～5.5% gum arabic aqueous solution vibration mixing rapidly after 1.8～2.2:1 mixed by volume, transfer pH value to 4.2～4.6, and be cooled to 19.5～20.5 ℃ rapidly and make mixed solution, 1.8～2.2: 1 ratio adds 9%～11% aqueous gelatin solution by volume in mixed solution, continue to stir, and be cooled to rapidly below 4 ℃, transfer pH value to 2.0 back to continue to stir 30min, dry getting final product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100186513A CN101381713A (en) | 2007-09-08 | 2007-09-08 | Preparation technology of high temperature resistant feeding cellulase |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100186513A CN101381713A (en) | 2007-09-08 | 2007-09-08 | Preparation technology of high temperature resistant feeding cellulase |
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| CN101381713A true CN101381713A (en) | 2009-03-11 |
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| CNA2007100186513A Pending CN101381713A (en) | 2007-09-08 | 2007-09-08 | Preparation technology of high temperature resistant feeding cellulase |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102771630A (en) * | 2012-06-07 | 2012-11-14 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and cellulose |
| CN103355502A (en) * | 2012-06-07 | 2013-10-23 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and compound enzyme |
| CN107173639A (en) * | 2017-06-27 | 2017-09-19 | 句容市华南水产养殖专业合作社 | A kind of production technology of Big Salangid micro-granulated feed |
| CN113424884A (en) * | 2021-05-08 | 2021-09-24 | 上海共得健康科技集团有限公司 | Method for preparing monascus tartary buckwheat tea by using high-temperature monascus |
| CN114958809A (en) * | 2022-06-14 | 2022-08-30 | 中农华威生物制药(湖北)有限公司 | Construction method of endo-beta-glucanase suitable for high-temperature granulation of feed |
| CN117296994A (en) * | 2023-11-30 | 2023-12-29 | 潍坊新希望六和饲料科技有限公司 | Application of chicken stomach microorganism source cellulase in preparation of chicken feed additive |
-
2007
- 2007-09-08 CN CNA2007100186513A patent/CN101381713A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102771630A (en) * | 2012-06-07 | 2012-11-14 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and cellulose |
| CN102771630B (en) * | 2012-06-07 | 2013-10-16 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and cellulose |
| CN103355502A (en) * | 2012-06-07 | 2013-10-23 | 宁夏伊品生物科技股份有限公司 | Fermentation and coating of lysine and compound enzyme |
| CN103444983A (en) * | 2012-06-07 | 2013-12-18 | 宁夏伊品生物科技股份有限公司 | Lysine and cellulase fermented and coated feed |
| CN103355502B (en) * | 2012-06-07 | 2015-11-18 | 宁夏伊品生物科技股份有限公司 | The fermentation of Methionin, prozyme and bag quilt |
| CN103444983B (en) * | 2012-06-07 | 2015-11-18 | 宁夏伊品生物科技股份有限公司 | The fermentation of Methionin, cellulase and bag are by feed |
| CN107173639A (en) * | 2017-06-27 | 2017-09-19 | 句容市华南水产养殖专业合作社 | A kind of production technology of Big Salangid micro-granulated feed |
| CN113424884A (en) * | 2021-05-08 | 2021-09-24 | 上海共得健康科技集团有限公司 | Method for preparing monascus tartary buckwheat tea by using high-temperature monascus |
| CN114958809A (en) * | 2022-06-14 | 2022-08-30 | 中农华威生物制药(湖北)有限公司 | Construction method of endo-beta-glucanase suitable for high-temperature granulation of feed |
| CN117296994A (en) * | 2023-11-30 | 2023-12-29 | 潍坊新希望六和饲料科技有限公司 | Application of chicken stomach microorganism source cellulase in preparation of chicken feed additive |
| CN117296994B (en) * | 2023-11-30 | 2024-03-22 | 潍坊新希望六和饲料科技有限公司 | Application of chicken stomach microorganism source cellulase in preparation of chicken feed additive |
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Open date: 20090311 |