CN113930414A - Method for preparing complex enzyme for beef tallow enzymolysis and complex enzyme prepared by method - Google Patents

Method for preparing complex enzyme for beef tallow enzymolysis and complex enzyme prepared by method Download PDF

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CN113930414A
CN113930414A CN202111203017.3A CN202111203017A CN113930414A CN 113930414 A CN113930414 A CN 113930414A CN 202111203017 A CN202111203017 A CN 202111203017A CN 113930414 A CN113930414 A CN 113930414A
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enzymolysis
protease
beef tallow
standing
compound enzyme
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王俏君
杨礼学
赵谋明
苏国万
马国丽
王冲
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Guanghan Maidele Food Co ltd
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
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Abstract

The invention relates to the technical field of compound enzyme preparation, in particular to a preparation method of compound enzyme for beef tallow enzymolysis and the compound enzyme, which comprises the following steps: s1, preprocessing; s2, inactivating; s3, modifying and standing; s4, temperature control culture; s5, circulating the steps S1-S4 until the production operation is finished, adopting a preparation method completely different from the prior art, adding the steps of inactivation, modification and temperature control culture, and ensuring the full utilization of the activity of the protease to the maximum extent to obtain more reliable active protease.

Description

Method for preparing complex enzyme for beef tallow enzymolysis and complex enzyme prepared by method
Technical Field
The invention relates to the technical field of compound enzyme preparation, in particular to a preparation method of compound enzyme for beef tallow enzymolysis and the compound enzyme.
Background
The protease has the capability of hydrolyzing a protein peptide chain into small-molecule peptides and amino acids, and is mainly used for modifying proteins in food, hydrolyzing and tenderizing meat, changing the characteristics of the proteins in baked flour products and improving the flavor of the baked flour products, degrading the proteins in the food into the small-molecule peptides or amino acids so as to improve the stability and the flavor of the product, accelerate the maturation of fermented foods, improve the quality of brewed seasonings and the like in the aspect of food.
In the prior art, no relevant patent exists for the application of enzyme in the beef tallow and the influence of the enzyme on the components and the content of aromatic substances after the beef tallow is smelted into the beef tallow. Therefore, research and development of an enzymolysis technology are urgently needed in the market, so that the outflow of the beef fat is accelerated, the production efficiency is improved, the yield of the grease is improved, the loss of nutrient substances in the beef fat is reduced, and the extraction of the grease is improved.
Disclosure of Invention
The invention aims to provide a method for preparing a complex enzyme for beef tallow enzymolysis and the complex enzyme, and solves the problem of poor beef tallow enzymolysis effect in the prior art.
The purpose of the invention is realized by the following technical scheme, which comprises the following steps: s1, preprocessing, namely adding a plurality of proteases into raw beef tallow according to a specific proportion, mixing and grinding, wherein the enzyme activity is added according to 50000U, standing for enzymolysis for 2-4 h to obtain an intermediate enzymolysis reactant; s2, inactivation, namely performing boiling water perfusion treatment on the plurality of proteases subjected to enzymolysis, and continuously performing perfusion treatment for 5-15 min to obtain the inactivated proteases; s3, modifying and standing, namely standing the inactivated protease with the specific pH of 6.0-8.5 to obtain active protease; s4, temperature-controlled culture, namely culturing and standing the obtained active protease at different temperatures, wherein the temperature setting range is 45-60 ℃, and the duration time is 1-2.5 hours; s5, circulating the steps S1-S4 until the production operation is finished.
It should be noted that, the preparation method adopted in the application is completely different from the prior art, and the steps of inactivation, modification and temperature-controlled culture are added, so that the activity of the protease can be fully utilized to the greatest extent, and more reliable active protease can be obtained.
The S1 includes the following: the protease is papain, flavourzyme, neutral protease, bromelain and trypsin, and the specific proportion addition amount is 0.2-3.0/g.
It should be noted that, after a lot of experiments, the applicant determines the addition amount of a specific column to be between 0.2/g and 3.0/g, and overcomes more uncertain factors such as temperature, humidity, culture time and the like in the middle, so that the technical scheme of the application has higher activity guarantee in practical use.
The S2 includes the following: the addition amount of boiling water in the perfusion treatment process is 20-33L per minute.
It should be noted that the inactivation can be effectively performed by adding boiling water, the prior art is usually adopted only when a certain active ingredient is eliminated in the link, and the application of the protease to the preparation process finds that the protease which is firstly subjected to primary inactivation is more active in subsequent culture.
The S3 includes the following: the pH value is adjusted once at an interval of 10min in the standing treatment process, and the adjustment range is 6.0-8.5.
It should be noted that the pH adjustment can help the whole protease to be in a better recovery environment.
The S4 includes the following: the active protease is cultured at the temperature of 45-60 ℃, and the mixing reaction is carried out by filling quantitative original protease every 10 minutes in the culture process.
It should be noted that, in a better pH recovery environment, the original protease is added, and then the combination of the activity of the new protease and the activity of the old protease can be performed by combining with a proper temperature, so that the overall activity operation effect is improved.
The complex enzyme prepared by the method of the invention is as follows: the compound enzyme comprises papain, flavourzyme and bromelain, wherein the papain, the bromelain and the flavourzyme are prepared from the following components in percentage by weight: bromelain: the flavourzyme is 1:3: 2.
It should be noted that the above proportion is a better proportion obtained by the applicant in practical experiments, and the speed and efficiency of beef tallow enzymolysis can be improved to the maximum extent.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. the preparation method is completely different from the prior art, and the steps of inactivation, modification and temperature-controlled culture are added, so that the activity of the protease can be fully utilized to the greatest extent, and more reliable active protease can be obtained;
2. the protease is applied to the preparation process, and the protease subjected to primary inactivation is more active in subsequent culture;
3. in a better pH recovery environment, the original protease is added, and then the combination of the activity of the new protease and the activity of the old protease can be carried out by combining with a proper temperature, so that the overall activity operation effect is improved.
Drawings
FIG. 1 is a schematic process flow diagram of the present invention.
Detailed Description
Referring to the accompanying drawing 1, the embodiment provides a method for preparing a complex enzyme for beef tallow enzymolysis and the complex enzyme, and the method for preparing the complex enzyme for beef tallow enzymolysis and the complex enzyme are mainly used for solving the problem that beef tallow enzymolysis effect is poor in the prior art, and the method and the complex enzyme are already in the actual use stage.
It should be noted that the components of the complex enzyme in the present application are all selected from the group consisting of Novoxin (China) Biotechnology Ltd.
Example 1
The present example was carried out according to the following steps: s1, preprocessing, namely adding a plurality of proteases into raw beef tallow according to a specific proportion, mixing and grinding, wherein the enzyme activity is added according to 50000U, standing for enzymolysis for 2 hours, and obtaining an intermediate enzymolysis reactant; s2, inactivation, namely performing boiling water perfusion treatment on the plurality of proteases subjected to enzymolysis, and continuously performing perfusion treatment for 5min to obtain the inactivated proteases; s3, modifying and standing, namely standing the inactivated protease with the specific pH of 6.0 to obtain active protease; s4, temperature-controlled culture, namely culturing and standing the obtained active protease at different temperatures, wherein the temperature setting range is 45 ℃, and the duration time is 1 h; s5, circulating the steps S1-S4 until the production operation is finished.
The protease is papain, flavourzyme, neutral protease, bromelain and trypsin, the adding amount of the specific proportion is 0.2/g, the adding amount of boiling water in the perfusion treatment process is 20L per minute, the pH value is adjusted once at an interval of 10min in the standing treatment process, the temperature-controlled culture of 45 ℃ is carried out on the active protease, and the mixing reaction is carried out by perfusing quantitative original protease at an interval of 10min in the culture process.
Example 2
The technical features in this embodiment are substantially the same as those described in the first embodiment, and the same technical features and technical solutions are not described again, and only the differences between the second embodiment and the first embodiment are described here, and this embodiment is performed according to the following steps: s1, preprocessing, namely adding a plurality of proteases into raw beef tallow according to a specific proportion, mixing and grinding, wherein the enzyme activity is added according to 50000U, and standing for enzymolysis for 3 hours to obtain an intermediate enzymolysis reactant; s2, inactivation, namely performing boiling water perfusion treatment on the plurality of proteases subjected to enzymolysis, and continuously performing perfusion treatment for 10min to obtain the inactivated proteases; s3, modifying and standing, namely standing the inactivated protease with the specific pH of 7.0 to obtain active protease; s4, temperature-controlled culture, namely culturing and standing the obtained active protease at different temperatures, wherein the temperature setting range is 55 ℃, and the duration time is 2 hours; s5, circulating the steps S1-S4 until the production operation is finished.
The protease is papain, flavourzyme, neutral protease, bromelain and trypsin, the adding amount of the specific proportion is 1.5/g, the adding amount of boiling water in the perfusion treatment process is 25L per minute, the pH value is adjusted once at an interval of 10min in the standing treatment process, the temperature-controlled culture of 55 ℃ is carried out on the active protease, and the mixing reaction is carried out by perfusing quantitative original protease at an interval of 10min in the culture process.
Example 3
The technical features in this embodiment are substantially the same as those described in embodiment two, and the same technical features and technical solutions are not described again, and only the differences between embodiment three and embodiment two are described here, and this embodiment is performed according to the following steps: s1, preprocessing, namely adding a plurality of proteases into raw beef tallow according to a specific proportion, mixing and grinding, wherein the enzyme activity is added according to 50000U, and standing for enzymolysis for 4 hours to obtain an intermediate enzymolysis reactant; s2, inactivation, namely performing boiling water perfusion treatment on the plurality of proteases subjected to enzymolysis, and continuously performing perfusion treatment for 15min to obtain the inactivated proteases; s3, modifying and standing, namely standing the inactivated protease with the specific pH of 8.5 to obtain active protease; s4, temperature-controlled culture, namely culturing and standing the obtained active protease at different temperatures, wherein the temperature setting range is 60 ℃, and the duration time is 2.5 hours; s5, circulating the steps S1-S4 until the production operation is finished.
The protease is papain, flavourzyme, neutral protease, bromelain and trypsin, the adding amount of the specific proportion is 3.0/g, the adding amount of boiling water in the perfusion treatment process is 20L per minute, the pH value is adjusted once at an interval of 10min in the standing treatment process, the temperature-controlled culture of 60 ℃ is carried out on the active protease, and the mixing reaction is carried out by perfusing quantitative original protease at an interval of 10min in the culture process.
Example 4
The technical features in this embodiment are substantially the same as those described in embodiment two, and the same technical features and technical solutions are not described again, and only the differences between embodiment three and embodiment two are described here, and this embodiment is performed according to the following steps: s1, preprocessing, namely adding a plurality of proteases into raw beef tallow according to a specific proportion, mixing and grinding, wherein the enzyme activity is added according to 50000U, and standing for enzymolysis for 3 hours to obtain an intermediate enzymolysis reactant; s2, inactivation, namely performing boiling water perfusion treatment on the plurality of proteases subjected to enzymolysis, and continuously performing perfusion treatment for 10min to obtain the inactivated proteases; s3, modifying and standing, namely standing the inactivated protease with the specific pH of 6.2 to obtain active protease; s4, temperature-controlled culture, namely culturing and standing the obtained active protease at different temperatures, wherein the temperature setting range is 50 ℃, and the duration time is 2.5 hours; s5, circulating the steps S1-S4 until the production operation is finished.
The protease is papain, flavourzyme, neutral protease, bromelain and trypsin, the adding amount of the specific proportion is 0.8/g, the adding amount of boiling water in the perfusion treatment process is 30L per minute, the pH value is adjusted once at an interval of 10min in the standing treatment process, the temperature-controlled culture of 50 ℃ is carried out on the active protease, and the mixing reaction is carried out by perfusing quantitative original protease at an interval of 10min in the culture process.
Based on the above examples, the applicant carried out comparative experiments on the relevant parameters, giving the data shown in table 1 below:
class of enzyme pH value Temperature of Time (h) Enzyme addition amount/g Proteolysis Rate/%)
Papain 6.5 50 3 0.5 5.06±0.15
Neutral protease 7.5 55 3 0.33 3.11±0.27
Flavourzyme protease 6.0 50 3 0.5 7.51±0.30
Trypsin 8.3 50 3 2.5 2.09±0.11
Bromelain 6.2 55 3 0.5 6.47±0.38
TABLE 1 Experimental data
As can be seen from table 1, under the condition that the relevant experimental parameters are changed, the three beef tallow enzymolysis products including papain, flavourzyme and bromelain have high proteolysis rates, so that the papain, flavourzyme and bromelain are selected as the enzyme preparations for beef tallow enzymolysis.
After obtaining the related active protease based on the above method, the applicant performed the determination experiment of the component ratio, and performed the experiment of multiple sets of various factors including but not limited to temperature, concentration, pH value and feed-liquid ratio on each active protease, to obtain the following data table 2:
Figure BDA0003305718960000051
Figure BDA0003305718960000061
table 2 experimental results of factors
As can be seen from Table 2, the proteolytic rate of the enzyme-digested beef oil is highest in all groups when the ratio of papain to bromelain to flavourzyme in group 3 is 1:3: 2.
The complex enzyme therefore includes papain, flavourzyme and bromelain, and papain: bromelain: when the flavor protease is 1:3:2, the beef fat enzymolysis effect is optimal, and the beef fat enzymolysis efficiency is the best.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.

Claims (6)

1. A method for preparing a complex enzyme for beef tallow enzymolysis is characterized by comprising the following steps:
s1, preprocessing, namely adding a plurality of proteases into raw beef tallow according to a specific proportion, mixing and grinding, wherein the enzyme activity is added according to 50000U, standing for enzymolysis for 2-4 h to obtain an intermediate enzymolysis reactant;
s2, inactivation, namely performing boiling water perfusion treatment on the plurality of proteases subjected to enzymolysis, and continuously performing perfusion treatment for 5-15 min to obtain the inactivated proteases;
s3, modifying and standing, namely standing the inactivated protease with the specific pH of 6.0-8.5 to obtain active protease;
s4, temperature-controlled culture, namely culturing and standing the obtained active protease at different temperatures, wherein the temperature setting range is 45-60 ℃, and the duration time is 1-2.5 hours;
s5, circulating the steps S1-S4 until the production operation is finished.
2. The method for preparing compound enzyme for beef tallow enzymolysis according to claim 1, wherein S1 comprises the following contents: the protease is papain, flavourzyme, neutral protease, bromelain and trypsin, and the specific proportion addition amount is 0.2-3.0/g.
3. The method for preparing compound enzyme for beef tallow enzymolysis according to claim 1, wherein S2 comprises the following contents: the addition amount of boiling water in the perfusion treatment process is 20-33L per minute.
4. The method for preparing compound enzyme for beef tallow enzymolysis according to claim 1, wherein S3 comprises the following contents: the pH value is adjusted once at an interval of 10min in the standing treatment process, and the adjustment range is 6.0-8.5.
5. The method for preparing compound enzyme for beef tallow enzymolysis according to claim 1, wherein S4 comprises the following contents: the active protease is cultured at the temperature of 45-60 ℃, and the mixing reaction is carried out by filling quantitative original protease every 10 minutes in the culture process.
6. The compound enzyme for the preparation method of the compound enzyme for beef tallow enzymolysis according to any one of claims 1-5, wherein the compound enzyme comprises papain, flavourzyme and bromelain, and the papain, bromelain and flavourzyme are prepared from the following components in proportion: bromelain: the flavourzyme is 1:3: 2.
CN202111203017.3A 2021-10-15 2021-10-15 Method for preparing complex enzyme for beef tallow enzymolysis and complex enzyme prepared by method Pending CN113930414A (en)

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Citations (6)

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Publication number Priority date Publication date Assignee Title
US20150173389A1 (en) * 2013-12-23 2015-06-25 Jiangnan University Method for Producing High Quality Animal Oil with Low Cholesterol Levels
CN107594503A (en) * 2017-09-11 2018-01-19 广东省农业科学院蚕业与农产品加工研究所 A kind of application of complex enzyme nutrition emulsion and preparation method thereof and complex enzyme
CN108782783A (en) * 2018-06-22 2018-11-13 山东农业大学 Edible butter and preparation method thereof
CN110894516A (en) * 2019-11-22 2020-03-20 四川航佳生物科技有限公司 Method for preparing functional beef tallow by biological method
CN110973564A (en) * 2019-12-06 2020-04-10 李茜 Sichuan beef tallow hotpot condiment
CN112640962A (en) * 2020-12-03 2021-04-13 东莞波顿香料有限公司 Butter base material and preparation method thereof

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CN107594503A (en) * 2017-09-11 2018-01-19 广东省农业科学院蚕业与农产品加工研究所 A kind of application of complex enzyme nutrition emulsion and preparation method thereof and complex enzyme
CN108782783A (en) * 2018-06-22 2018-11-13 山东农业大学 Edible butter and preparation method thereof
CN110894516A (en) * 2019-11-22 2020-03-20 四川航佳生物科技有限公司 Method for preparing functional beef tallow by biological method
CN110973564A (en) * 2019-12-06 2020-04-10 李茜 Sichuan beef tallow hotpot condiment
CN112640962A (en) * 2020-12-03 2021-04-13 东莞波顿香料有限公司 Butter base material and preparation method thereof

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