CN102533689A - Preparation method for vacuum drying of transglutaminase - Google Patents
Preparation method for vacuum drying of transglutaminase Download PDFInfo
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- CN102533689A CN102533689A CN201110452678XA CN201110452678A CN102533689A CN 102533689 A CN102533689 A CN 102533689A CN 201110452678X A CN201110452678X A CN 201110452678XA CN 201110452678 A CN201110452678 A CN 201110452678A CN 102533689 A CN102533689 A CN 102533689A
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Abstract
The invention discloses a preparation method for vacuum drying of transglutaminase. The preparation method comprises the following steps of: adding a heat protectant in fermentation broth of the transglutaminase; standing the mixed solution at room temperature; carrying out high-speed freezing and centrifuging on the mixed solution to obtain supernate; precipitating the fermentation broth with 95 percent ethanol; carrying out high-speed freezing and centrifuging on the precipitated solution to obtain a precipitate; adding an antioxidant protectant; and carrying out uniform mixing and vacuum drying to obtain an transglutaminase preparation. The transglutaminase obtained by using the preparation method disclosed by the invention has greatly-improved stability.
Description
Technical field
The present invention relates to a kind of production of enzyme preparation technology; Relate to the vacuum drying preparation method of a kind of Transglutaminase EC2.3.2.13 particularly; Add making Transglutaminase EC2.3.2.13 enzyme stable heat-resisting, anti-oxidant protective agent, use boulton process and come the dried microorganism fermented liquid to produce Transglutaminase EC2.3.2.13.
Background technology
Transglutaminase EC2.3.2.13 is a kind of microbial enzyme of catalyzing acyl shift reaction, and catalysis is present in peptide intrachain L-glutamic acid Gln residue γ-carboxy and amide groups and Methionin Lys residue epsilon-amino generation crosslinked action; In that intramolecularly or intermolecular generation ε-(the bridge formation cohesive action of γ-Glu)-Lys forms crosslinked protein structure, thereby improves protein function character; Its safety and superpower crosslinking feature; Receive domestic and international food science and technology worker extensive concern, the laudatory title of " super tackiness agent of 21st century " is arranged, often be used to the meat that bonds as modifying agent; Fish is rotten, in the bread.
Transglutaminase EC2.3.2.13 is to be a kind of mercaptan enzyme of active site residue with the halfcystine, and oxidation-resistance is poor, is prone to inactivation under the hot conditions, is difficult for preserving.The method of producing glutamine transaminage at present mainly is lyophilize, and cost is high, and cost is big, is unfavorable for large-scale industrial production.
The present invention proposes a kind of method that improves Transglutaminase EC2.3.2.13 stability, through adding the thermostability of glycitols protective material raising Transglutaminase EC2.3.2.13, through adding the antioxidative stabilizer of bioactive peptide raising Transglutaminase EC2.3.2.13.Can utilize process of vacuum drying to take the dried microorganism fermented liquid like this, solve the problem that to produce Transglutaminase EC2.3.2.13 in the industrial production with lyophilize, improve the stability of Transglutaminase EC2.3.2.13, practice thrift production cost.
Summary of the invention
The present invention proposes the vacuum drying preparation method of a kind of Transglutaminase EC2.3.2.13, in glutamine transferred amine enzyme fermentation broth, add thermal protecting agent, room temperature leaves standstill; Obtain supernatant through the high speed frozen centrifugation; With 95% ethanol said supernatant is precipitated, obtain throw out, add anti-oxidant protective agent through the high speed frozen centrifugation; Through mix, vacuum-drying, obtain the Transglutaminase EC2.3.2.13 zymin; Wherein, said thermal protecting agent is the glycitols material, comprises Sorbitol Powder and maltose alcohol, and said anti-oxidant protective agent comprises bioactive peptide.
The enzyme work of Transglutaminase EC2.3.2.13 is 10 ~ 50u/ml in the described fermented liquid.
The addition of said Sorbitol Powder is 0.5% ~ 5%w/v, and preferred addition is 0.5% ~ 2%w/v; The addition of said maltose alcohol is 1% ~ 10%w/v, and preferred addition is 2% ~ 4%w/v.
Said room temperature time of repose is 0.5h-1h.
The volume ratio of said alcoholic acid addition and supernatant is 0.5:1 ~ 1.5:1, and preferred proportion is 0.5:1.
Said anti-oxidant protective agent is preferably the wheat protein peptide.
The addition of said anti-oxidant protective agent and sedimentary mass ratio are 0.5:1 ~ 1:1.5, and preferred proportion is 0.75:1.
Under 35 ℃-40 ℃, carry out vacuum-drying, the preferred time is 3h-5h.
Among the present invention; Adopt the high speed frozen centrifugation twice; Behind fermented liquid adding thermal protecting agent, get supernatant for the first time and obtain fermented liquid supernatant liquid, carry out post precipitation through the throw out of high speed frozen centrifugation acquisition for the second time through adding ethanol, and add inhibitor through the high speed frozen centrifugation.Among the present invention, adopt twice high speed frozen centrifugation to reach the technique effect of highly purified Transglutaminase EC2.3.2.13.
Transglutaminase EC2.3.2.13 is interior and intermolecular generation covalent cross-linking through the catalytic proteins peptide molecule, thereby improves proteinic 26S Proteasome Structure and Function.But the poor stability of Transglutaminase EC2.3.2.13 self, unsuitable standing storage, thus influence the application of Transglutaminase EC2.3.2.13 at field of food.Thereby the present invention relates to a kind of technology that boulton process is produced Transglutaminase EC2.3.2.13 of using, and in glutamine transferred amine enzyme fermentation broth, add thermal protecting agent, room temperature is placed for some time; Select for use 95% ethanol that said fermented liquid is precipitated, the high speed frozen centrifugation is abandoned supernatant; In the throw out that obtains, add anti-oxidant protective agent; Mix, vacuum-drying obtains the Transglutaminase EC2.3.2.13 zymin.
Among the present invention, utilize vacuum-drying to replace lyophilize to produce glutamine transaminage, solved the influence of high temperature Transglutaminase EC2.3.2.13 stability.Thermal protecting agent glycitols material among the present invention is the aldehyde radical of monose molecule or the compound that ketone group is reduced into alcohol radical; Make sugar change polyvalent alcohol into; Under thermal environment, advantages of higher stability is arranged, this is because contain great amount of hydroxy group in the sugar alcohol, can change proteinic hydration structure in the solution; Promote the hydrophobic interaction between protein molecule, thereby improved the stability when protein is heated.The glycitols that is suitable among the present invention comprises Sorbitol Powder, mannitol, erythrose alcohol, maltose alcohol, Saccharum lactis, Xylitol etc.Different sugar alcohols is discrepant to proteinic provide protection, and this possibly be because cause different with the position of quantity of oh group.It is ideal to proteinic heat protection effect that the present invention proposes in the sugar alcohol Sorbitol Powder and maltose alcohol, secondly is Saccharum lactis and Xylitol.
Adopt bioactive peptide as anti-oxidant protective agent among the present invention.Bioactive peptide is the linearity from the dipeptides to the complicacy that 20 natural amino acids constitute with different compositions and arrangement mode in the protein, the general name of the different peptides of ring-type result; According to the category division of raw material, the bioactive peptide that the inventive method is suitable for comprises wheat peptide, soybean peptides, corn peptide, white of an egg peptide, silk-protein peptide and compound peptide etc.Bioactive peptide has certain nutrition effect and physiological function.The existence form of glutaminic acid residue in protein is the Stimulina of its amide form, and Stimulina is the precursor substance of extremely important inhibitor-reduced glutathion in the compound body, thereby the present invention adopts bioactive peptide as anti-oxidant protective agent.Glutamic is that content is the highest in all vegetable-proteins in the wheat protein peptide that the present invention adopts, and accounts for 35% of Tot Prot, therefore, adds the wheat protein peptide and can significantly improve the Transglutaminase EC2.3.2.13 resistance of oxidation.Except that the wheat protein peptide, most of protein peptide also can improve the resistance of oxidation of Transglutaminase EC2.3.2.13, but effect is not as the wheat protein peptide.With other protective materials (like amino acids, unit price salt etc.) effect of Transglutaminase EC2.3.2.13 is compared; The antioxygenation that bioactive peptide is mainly brought into play is the most remarkable; The quality of the Transglutaminase EC2.3.2.13 that obtains is maximum, and in prolonged preservation, the decline alive of total enzyme is minimum; And the bioactive peptide cost is lower, is suitable for being widely used in suitability for industrialized production.
Innovative point of the present invention is; In general production Transglutaminase EC2.3.2.13 process, mainly add thermal protecting agent and stablize Transglutaminase EC2.3.2.13, among the present invention; Add thermal protecting agent and anti-oxidant protective agent simultaneously to fermented liquid; The antioxygenation effect is obvious, and can significantly improve enzyme and live, and better with the enzyme stability of this method production.The present invention selects secondary high speed frozen centrifugation for use, the zymin that obtains than frozen centrifugation obtain zymin, purity is higher.Beneficial effect of the present invention is: thermal protecting agent can improve the thermostability of Transglutaminase EC2.3.2.13 when being heated; Contain multiple amino acids in anti-oxidant protective agent such as the wheat protein peptide, can improve the oxidation-resistance of enzyme in the long-term put procedure.The present invention efficiently solves the problem of Transglutaminase EC2.3.2.13 poor stability, and has greatly reduced production cost.
The glutamine transaminage normal temperature of the present invention's preparation is placed every enzyme activity determination that carries out at a distance from January.Detected result shows that the glutamine transaminage that obtains according to preparation method of the present invention has good enzyme stability.
Description of drawings
Shown in Figure 1 is the stable time dependent synoptic diagram of the prepared Transglutaminase EC2.3.2.13 zymin of embodiment 1.
Shown in Figure 2 is the stable time dependent synoptic diagram of the prepared Transglutaminase EC2.3.2.13 zymin of embodiment 2.
Shown in Figure 3 is the stable time dependent synoptic diagram of the prepared Transglutaminase EC2.3.2.13 zymin of embodiment 3.
Shown in Figure 4 is the stable time dependent synoptic diagram of the prepared Transglutaminase EC2.3.2.13 zymin of embodiment 4.
Shown in Figure 5 is the stable time dependent synoptic diagram of the prepared Transglutaminase EC2.3.2.13 zymin of embodiment 5.
Shown in Figure 6 is the stable time dependent synoptic diagram of the prepared Transglutaminase EC2.3.2.13 zymin of embodiment 6.
Shown in Figure 7 is the stable time dependent synoptic diagram of the prepared Transglutaminase EC2.3.2.13 zymin of embodiment 7.
Embodiment
In conjunction with following specific embodiment and accompanying drawing, the present invention is done further detailed description, protection content of the present invention is not limited to following examples.Under spirit that does not deviate from inventive concept and scope, variation and advantage that those skilled in the art can expect all are included among the present invention, and are protection domain with the appending claims.
The vacuum drying preparation method of Transglutaminase EC2.3.2.13 of the present invention may further comprise the steps: in glutamine transferred amine enzyme fermentation broth, add thermal protecting agent, room temperature leaves standstill; Obtain supernatant through the high speed frozen centrifugation; With 95% ethanol supernatant is precipitated, obtain throw out, add anti-oxidant protective agent through the high speed frozen centrifugation; Through mix, vacuum-drying, obtain the Transglutaminase EC2.3.2.13 zymin; Wherein, thermal protecting agent is the glycitols material, comprises Sorbitol Powder and maltose alcohol, and anti-oxidant protective agent comprises bioactive peptide.
Wherein, the enzyme work of Transglutaminase EC2.3.2.13 is 10 ~ 50u/ml in the glutamine transferred amine enzyme fermentation broth.
Among the present invention, the Sorbitol Powder addition is 0.5% ~ 5%w/v, and preferred addition is 0.5% ~ 2%w/v.The maltose alcohol addition is 1% ~ 10%w/v, and preferred addition is 2% ~ 4%w/v.Wheat protein peptide addition with the throw out quality than preferred 0.75:1 (w/w).
Among the present invention, the fermented liquid that adds thermal protecting agent is abandoned throw out through the high speed frozen centrifugation, obtains supernatant.Consumption of ethanol and adding thermal protecting agent secondary fermentation liquid are 0.5:1 (v/v) through the volume ratio of the supernatant that the high speed frozen centrifugation obtains.
After fermented liquid added thermal protecting agent, room temperature was placed 0-2h, and be 0.5-1h preferred storage period.
Add the alcoholic acid fermented liquid and obtain throw out, add anti-oxidant protective agent through the high speed frozen centrifugation, after abandoning supernatant.
Wherein, vacuum-drying is carried out under 35 ℃ of-45 ℃ of vacuum environments, and be 3-5h preferred time of drying.
Thermal protecting agent is the glycitols material among the present invention, comprises maltose alcohol, Xylitol, Saccharum lactis, Sorbitol Powder, mannitol, erythritol etc.Anti-oxidant protective agent is a bioactive peptide among the present invention, comprises newborn peptide, wheat peptide, soybean peptides, corn peptide, white of an egg peptide, silk-protein peptide and compound peptide etc.
Xiang Maoyuan wheel Streptothrix (
Streptoverticillium mobaraense) among the glutamine transferred amine enzyme fermentation broth 100ml that produced with the ethanol of 0.5:1: fermentating liquid volume precipitates enzyme than adding ethanol; High speed freezing centrifuge is set: 6000r/min, 4 ℃, centrifugal 20min obtain throw out; Throw out is put into vacuum drying oven; 40 ℃ of dry 3h obtain the Transglutaminase EC2.3.2.13 zymin.Seal with valve bag, and the results of regular determination enzymic activity.
Xiang Maoyuan wheel Streptothrix (
Streptoverticillium mobaraense) add 1% (w/v) Sorbitol Powder among the glutamine transferred amine enzyme fermentation broth 100ml that produced; Stir the back room temperature and place 30min; Ethanol with 0.5:1: the volume ratio of fermented liquid is added ethanol enzyme is precipitated, and high speed freezing centrifuge is set: 6000r/min, 4 ℃, centrifugal 20min obtain throw out, and throw out is put into vacuum drying oven; 40 ℃ of dry 3h obtain the Transglutaminase EC2.3.2.13 zymin.Seal the results of regular determination enzymic activity with valve bag.
Xiang Maoyuan wheel Streptothrix (
Streptoverticillium mobaraense) add 3% (w/v) maltose alcohol among the glutamine transferred amine enzyme fermentation broth 100ml that produced; Stir the back room temperature and place 30min; Ethanol with 0.5:1: the volume ratio of fermented liquid is added ethanol enzyme is precipitated, and high speed freezing centrifuge is set: 6000r/min, 4 ℃, centrifugal 20min obtain throw out, and throw out is put into vacuum drying oven; 40 ℃ of dry 3h obtain the Transglutaminase EC2.3.2.13 zymin.Seal the results of regular determination enzymic activity with valve bag.
Xiang Maoyuan wheel Streptothrix (
Streptoverticillium mobaraense) add 3% (w/v) maltose alcohol, 1% Sorbitol Powder among the glutamine transferred amine enzyme fermentation broth 100ml that produced; Stir the back room temperature and place 30min; Ethanol with 0.5:1: the volume ratio of fermented liquid is added ethanol enzyme is precipitated, and high speed freezing centrifuge is set: 6000r/min, 4 ℃, centrifugal 20min obtain throw out, and throw out is put into vacuum drying oven; 40 ℃ of dry 3h obtain the Transglutaminase EC2.3.2.13 zymin.Seal the results of regular determination enzymic activity with valve bag.
Xiang Maoyuan wheel Streptothrix (
Streptoverticillium mobaraense) among the glutamine transferred amine enzyme fermentation broth 100ml that produced with the ethanol of 0.5:1: fermentating liquid volume precipitates enzyme than adding ethanol; High speed freezing centrifuge is set: 6000r/min, 4 ℃, centrifugal 20min obtain throw out; With the wheat protein peptide: throw out is that the mass ratio of 0.75:1 adds the wheat protein peptide; Put into vacuum drying oven after mixing, dry 3h obtains the Transglutaminase EC2.3.2.13 zymin.Seal the results of regular determination enzymic activity with valve bag.
Xiang Maoyuan wheel Streptothrix (
Streptoverticillium mobaraense) add 1% (w/v) Sorbitol Powder among the glutamine transferred amine enzyme fermentation broth 100ml that produced, stir the back room temperature and place 30min, with the ethanol of 0.5:1: the volume ratio of fermented liquid is added ethanol enzyme is precipitated; High speed freezing centrifuge is set: 6000r/min, 4 ℃, centrifugal 20min; In the throw out that obtains, add the wheat protein peptide, addition and sedimentary mass ratio are 0.75:1, put into vacuum drying oven; Dry 3h obtains the Transglutaminase EC2.3.2.13 zymin.Seal the results of regular determination enzymic activity with valve bag.
Embodiment 7
Xiang Maoyuan wheel Streptothrix (
Streptoverticillium mobaraense) add 1% (w/v) Sorbitol Powder and 3% (w/v) maltose alcohol among the glutamine transferred amine enzyme fermentation broth 100ml that produced; Stir the back room temperature and place 30min; Ethanol with 0.5:1: the volume ratio of fermented liquid is added ethanol enzyme is precipitated, and high speed freezing centrifuge is set: 6000r/min, 4 ℃, centrifugal 20min add the wheat protein peptide in the throw out that obtains; Addition and sedimentary mass ratio are 0.75:1; Put into vacuum drying oven, dry 3h obtains the Transglutaminase EC2.3.2.13 zymin.Seal the results of regular determination enzymic activity with valve bag.
Embodiment 8
Transglutaminase EC2.3.2.13 zymin through standard hydroxamate assay method prepares embodiment 1-7 is carried out enzyme biopsy survey, the steps include: to get respectively 2ml reagent A liquid and reagent B liquid and adds in the rub oral examination tube, and 37 ℃ are incubated 15 minutes; Add 0.2ml enzyme liquid, reacted 10 minutes, the test tube that adds A liquid adds 2mlB liquid termination reaction; The test tube that adds B liquid adds 2ml A liquid; Filter, as blank, the 525nm place measures the OD value with second test-tube reaction liquid.Do typical curve with L-L-glutamic acid-γ-single hydroxamic acid.
Reagent A (reaction solution): contain 0.2 M Tris-HCl, 0.1M oxammonium hydrochloride, 0.01 M reduced glutathion, 0.03 M Na-CBZ-Gln-Gly, pH 6.0.
Reagent B (stop buffer): following three kinds of compositions are the mixed of 1:1:1 by volume.3 mol/L HCl, 12 % trichoroacetic acid(TCA)s (W/V), 5 % iron trichlorides (W/V).
It is as shown in Figure 1 that the result is surveyed in the Transglutaminase EC2.3.2.13 zymin enzyme biopsy of embodiment 1 preparation, do not add any protectant Transglutaminase EC2.3.2.13, and total enzyme is lived low; 2098 (U) only, and As time goes on, total enzyme live descend more; Drop to 1132U from 2098U; Fall reaches more than 50%, in the visible prolonged preservation process, and less stable.
It is as shown in Figure 2 that the result is surveyed in the Transglutaminase EC2.3.2.13 zymin enzyme biopsy of embodiment 2 preparation, with Fig. 1 relatively, add Sorbitol Powder after, total enzyme work of the Transglutaminase EC2.3.2.13 that obtains is significantly higher than the total enzyme that does not add the Transglutaminase EC2.3.2.13 that protective material obtains and lives.Total enzyme is lived and is 2743U, improves 700U, and As time goes on, enzyme is lived and descended seldom, drops to 2400U from 2743U.
It is as shown in Figure 3 that the result is surveyed in the Transglutaminase EC2.3.2.13 zymin enzyme biopsy of embodiment 3 preparations; With Fig. 1 relatively, add maltose alcohol after, total enzyme work of the Transglutaminase EC2.3.2.13 that obtains is significantly higher than the total enzyme that does not add the Transglutaminase EC2.3.2.13 that protective material obtains and lives; Total enzyme is lived and is 2631U; And As time goes on, enzyme is lived and is descended seldom, drops to 2389U from 2631U.
It is as shown in Figure 4 that the result is surveyed in the Transglutaminase EC2.3.2.13 zymin enzyme biopsy of embodiment 4 preparations, compares with Fig. 1, behind adding Sorbitol Powder and the maltose alcohol; The total enzyme work of the Transglutaminase EC2.3.2.13 that obtains is significantly higher than the total enzyme that does not add the Transglutaminase EC2.3.2.13 that protective material obtains and lives; Also be higher than total enzyme work that independent interpolation Sorbitol Powder and maltose alcohol (embodiment 2-3) obtain, total enzyme is lived and is 3023U, As time goes on; Enzyme is lived in descending and is lacked, and drops to 2632U from 3023U.
It is as shown in Figure 5 that the result is surveyed in the Transglutaminase EC2.3.2.13 zymin enzyme biopsy of embodiment 5 preparations; With Fig. 1 relatively, add the wheat protein peptide after, total enzyme work of the Transglutaminase EC2.3.2.13 that obtains is significantly higher than the total enzyme that does not add the Transglutaminase EC2.3.2.13 that protective material obtains and lives; Total enzyme work reaches 2754U; And As time goes on, enzyme is lived and is descended seldom, drops to 2471U from 2754U.
It is as shown in Figure 6 that the result is surveyed in the Transglutaminase EC2.3.2.13 zymin enzyme biopsy of embodiment 6 preparations, compares with Fig. 1, behind adding Sorbitol Powder and the wheat protein peptide; Total enzyme work of the Transglutaminase EC2.3.2.13 that obtains is significantly higher than the total enzyme that does not add the Transglutaminase EC2.3.2.13 that protective material obtains and lives; The total enzyme that also is higher than the Transglutaminase EC2.3.2.13 that obtains through embodiment 2-4 is lived, and total enzyme is lived and is 2980U, and As time goes on; Enzyme is lived and is descended seldom, drops to 2687U from 2980U.
It is as shown in Figure 7 that the result is surveyed in the Transglutaminase EC2.3.2.13 zymin enzyme biopsy of embodiment 7 preparations; With embodiment 1-6 contrast, according to the preparation method of Transglutaminase EC2.3.2.13 of the present invention, behind adding Sorbitol Powder, maltose alcohol and the wheat protein peptide; Total enzyme of the Transglutaminase EC2.3.2.13 that obtains is lived the highest; Total enzyme is lived and is 3543U, and passes the fall minimum that enzyme is lived in time, drops to 3331U from 3543U.It is thus clear that the wheat protein peptide of the Sorbitol Powder of interpolation 0.5%-2% (w/v), the maltose alcohol of 2%-4% (w/v) and certain mass can improve the thermostability and the oxidation-resistance of Transglutaminase EC2.3.2.13, in prolonged preservation, has brought into play effect.
Embodiment 9
In the present embodiment with Saccharum lactis and Xylitol as thermal protecting agent, all the other working method and embodiment 4 are basic identical.Through detecting, the total enzyme work of the Transglutaminase EC2.3.2.13 that obtains is higher than the total enzyme that does not add any protectant Transglutaminase EC2.3.2.13 lives, but alive not as adding total enzyme that Transglutaminase EC2.3.2.13 that Sorbitol Powder and maltose alcohol obtain obtains.Therefore, the present invention proposes Sorbitol Powder and maltose alcohol as thermal protecting agent, and its effect that protein heat is protected is ideal.
Embodiment 10
In the present embodiment with soybean protein peptide as anti-oxidant protective agent, all the other working method and embodiment 5 are basic identical.Through detecting, the Transglutaminase EC2.3.2.13 that obtains is higher than the total enzyme that does not add any protectant Transglutaminase EC2.3.2.13 lives, but total enzyme that the Transglutaminase EC2.3.2.13 that obtains not as interpolation wheat protein peptide obtains is lived.Therefore, as anti-oxidant protective agent, its effect to the protein anti-oxidation protection is ideal with the aleuronat peptide for the present invention's proposition.
Claims (8)
1. the vacuum drying preparation method of Transglutaminase EC2.3.2.13 is characterized in that, said method comprises adds thermal protecting agent in glutamine transferred amine enzyme fermentation broth; Room temperature leaves standstill, and obtains supernatant through the high speed frozen centrifugation, with 95% ethanol said supernatant is precipitated; Obtain throw out through the high speed frozen centrifugation; Add anti-oxidant protective agent, through mix, vacuum-drying, obtain the Transglutaminase EC2.3.2.13 zymin; Wherein, said thermal protecting agent is the glycitols material, comprises Sorbitol Powder and maltose alcohol, and said anti-oxidant protective agent comprises bioactive peptide.
2. the vacuum drying preparation method of Transglutaminase EC2.3.2.13 according to claim 1 is characterized in that, the enzyme work of Transglutaminase EC2.3.2.13 is 10 ~ 50u/ml in the described fermented liquid.
3. the vacuum drying preparation method of Transglutaminase EC2.3.2.13 according to claim 1 is characterized in that the addition of said Sorbitol Powder is 0.5% ~ 5%w/v, and preferred addition is 0.5% ~ 2%w/v; The addition of said maltose alcohol is 1% ~ 10%w/v, and preferred addition is 2% ~ 4%w/v.
4. the vacuum drying preparation method of Transglutaminase EC2.3.2.13 according to claim 1 is characterized in that said room temperature time of repose is 0.5h-1h.
5. the vacuum drying preparation method of Transglutaminase EC2.3.2.13 according to claim 1 is characterized in that the volume ratio of said alcoholic acid addition and supernatant is 0.5:1 ~ 1.5:1, and preferred proportion is 0.5:1.
6. the vacuum drying preparation method of Transglutaminase EC2.3.2.13 according to claim 1 is characterized in that said anti-oxidant protective agent is preferably the wheat protein peptide.
7. the vacuum drying preparation method of Transglutaminase EC2.3.2.13 according to claim 1 is characterized in that the addition of said anti-oxidant protective agent and sedimentary mass ratio are 0.5:1 ~ 1:1.5, and preferred proportion is 0.75:1.
8. the vacuum drying preparation method of Transglutaminase EC2.3.2.13 according to claim 1 is characterized in that, under 35 ℃-40 ℃, carries out vacuum-drying, and the preferred time is 3h-5h.
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| CN108018279A (en) * | 2018-01-24 | 2018-05-11 | 江南大学 | A kind of glutamine transaminage built agent and its application |
| CN108624573A (en) * | 2018-05-17 | 2018-10-09 | 江苏鸣生物股份有限公司 | A kind of method that industry improves glutamine transaminage storage-stable |
| CN110373406A (en) * | 2019-07-15 | 2019-10-25 | 泰兴市东圣生物科技有限公司 | A kind of preparation method of immobilization glutamine transaminage |
| CN110623249A (en) * | 2019-11-05 | 2019-12-31 | 泰兴市东圣生物科技有限公司 | Instant enzyme preparation and preparation method thereof |
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| CN107531747A (en) * | 2015-04-03 | 2018-01-02 | 罗盖特公司 | The stabilization of protein |
| CN108018279A (en) * | 2018-01-24 | 2018-05-11 | 江南大学 | A kind of glutamine transaminage built agent and its application |
| CN108018279B (en) * | 2018-01-24 | 2020-09-04 | 江南大学 | Glutamine transaminase compound agent and application thereof |
| CN108624573A (en) * | 2018-05-17 | 2018-10-09 | 江苏鸣生物股份有限公司 | A kind of method that industry improves glutamine transaminage storage-stable |
| CN110373406A (en) * | 2019-07-15 | 2019-10-25 | 泰兴市东圣生物科技有限公司 | A kind of preparation method of immobilization glutamine transaminage |
| CN110623249A (en) * | 2019-11-05 | 2019-12-31 | 泰兴市东圣生物科技有限公司 | Instant enzyme preparation and preparation method thereof |
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