CN1206303C - Method for preparing hydrolytic gelatin using combined enzyme - Google Patents
Method for preparing hydrolytic gelatin using combined enzyme Download PDFInfo
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- CN1206303C CN1206303C CN 02159288 CN02159288A CN1206303C CN 1206303 C CN1206303 C CN 1206303C CN 02159288 CN02159288 CN 02159288 CN 02159288 A CN02159288 A CN 02159288A CN 1206303 C CN1206303 C CN 1206303C
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Abstract
The present invention relates to a method for preparing hydrolytic gelatin by combined enzymes. Raw gelatin is prepared to be an aqueous solution, and the pH value of the aqueous solution system is regulated by a buffer system of citric acid and phosphate. The aqueous solution is heated to a reaction temperature, enzymes are put in the gelatin solution to be stirred, and the hydrolytic gelatin is obtained by the processes of enzyme elimination through heating, filtration, concentration and drying. The enzymes are the combined enzymes comprising any two kinds of AS1.398 neutral protease, papain, trypsin or bromelin, and the reaction temperature is in the proper temperature range of the single enzymes forming the combined enzymes. The pH value of the system is from 5 to 8.5, reaction time is from 0.5 to 5 hours, and the adding quantity of the single enzymes is 0.01% to 2% of the weight of the gelatin. The gelatin is degraded after all the single enzymes in the combined enzymes are mixed, or is degraded by one kind of single enzymes firstly and is degraded by the other kind of single enzymes secondly. The present invention can decrease the molecular weight of the hydrolytic gelatin, simultaneously improves the stability of the hydrolytic gelatin solution, and eliminates abnormal fishy smell.
Description
Technical field the present invention relates to a kind of method for preparing gelatin hydrolysate with combined protein enzyme liberating gelatin.The present invention can reduce the molecular weight of gelatin hydrolysate, improves the stability of gelatin hydrolysate solution simultaneously, has eliminated different raw meat and has smelt flavor.
Background technology
Gelatin hydrolysate is owing to have identical chemical constitution with gelatin, keeping gelatin institute inherent trophic function, and because its molecular weight is low, its solubleness in water is increased, easier being absorbed by the body, thereby its trophic function is further brought into play, not only in food, medicine, household chemicals field, and also obtained using widely in fields such as weaving, printing and dyeing, coating, papermaking and chemical industry.Gelatinase degraded preparation gelatin hydrolysate has three kinds of methods, i.e. acid system, alkaline process and enzyme process, and wherein acid system and alkali process are simple, cost is low, but in hydrolytic process, the cut randomness of chemical bond is big, therefore reacts wayward, the saltiness height.Enzyme process gelatin DeR is controlled easily, and the degraded product quality that obtains is better.At present, adopt the molecular weight of product of single enzyme liberating gelatin generally about 4000.People such as the yellow wisdom of Beijing University of Chemical Technology have reported that AS1.398 neutral protein enzyme liberating prepares gelatin in " the AS1.398 neutral protease prepares gelatin hydrolysate " (the 71st page 74 pages of chemical industry and engineering the 2nd phases of calendar year 2001).Specific operation process is the aqueous solution that gelatin is mixed with 10% (massfraction), regulation system pH value is 7.0 (with citric acid and phosphate buffer), under 45 ℃ condition, the AS1.398 neutral protease that adds 2% (massfraction), reacted 25 minutes, and killed enzyme, after filtration with the filtrate concentrate drying, promptly obtain the gelatin degraded product, molecular weight is about 4800.At present the gelatin hydrolysate product all in various degree the different raw meat that exists smell flavor.Gelatin hydrolysate is a peptide species, and easily by decomposition such as bacteriums, stability in storage is relatively poor.
Along with the Application and Development of gelatin hydrolysate, require to have the serial gelatin hydrolysate product of different molecular weight, require goods good stability, free from extraneous odour simultaneously.
Summary of the invention the object of the present invention is to provide a kind of method for preparing low-molecular-weight gelatin hydrolysate with the combination enzyme.Its product is that a kind of molecular weight is little, free from extraneous odour, gelatin hydrolysate that aqueous stability is high.
The present invention prepares the method for gelatin hydrolysate with the combination enzyme, with single enzyme prepare the method for gelatin hydrolysate identical be that the raw material gelatin is mixed with the aqueous solution, with citric acid and phosphate buffer regulation system pH value; Be heated to temperature of reaction, enzyme put in the gelatin solution stirred, through adding the heat kill enzyme, filtration, concentrated, dry promptly obtains gelatin hydrolysate.Technical characterictic of the present invention is: the enzyme that is adopted is the combination enzyme, the combination enzyme contains any two kinds in AS1.398 neutral protease, papoid, trypsinase or the bromeline, temperature of reaction is got in single enzyme suitable temp scope of forming the combination enzyme, system pH is 5～8.5, and the reaction times is 0.5～5 hour.The add-on of single enzyme is 0.0 1%～2% of a gelatin weight.The combination enzyme is AS1.398 neutral protease and papoid, and temperature of reaction is 45～65 ℃, and pH is 5～8.The combination enzyme is trypsinase and papoid, and temperature of reaction is 45～65 ℃, and pH is 5～8.5.The combination enzyme is trypsinase and AS1.398 neutral protease, and temperature of reaction is 45～65 ℃, and pH is 7～8.5.The combination enzyme is bromeline and AS1.398 neutral protease, and temperature of reaction is 45～50 ℃, and pH is 7.The combination enzyme is papoid and bromeline, and temperature of reaction is 50～65 ℃, and pH is 5～8.The combination enzyme is trypsinase and bromeline, and temperature of reaction is 45～50 ℃, and pH is 7～8.5.Carry out the gelatin degraded after mixing with whole single enzyme of combination in the enzyme or will wherein a kind of single enzyme add earlier gelatin is degraded, after add another kind of single enzyme again and continue gelatin is degraded.
Enzyme is a biological catalyst, has the height specificity.Be that different proteolytic enzyme acts on different peptide bonds.Therefore with the different proteolytic enzyme combination of specificity gelatin is degraded, can further regulate and control the molecular weight and the molecular weight distribution of gelatin hydrolysate.Just because of the difference of various proteolytic enzyme specificity degree, resulting peptide segment length is different when making the catalyst degradation gelatin with them, i.e. molecular weight and distribution thereof is different, and the end amino acid of its product is also different.The present invention reaches with this principle and obtains the little purpose of degraded product molecular weight.
From prior art as can be known, prepare gelatin hydrolysate with single enzyme, the suitable temp of AS1.398 neutral protease is about 45 ℃, and optimum pH is 7; 65 ℃ of papoid suitable temps, optimum pH are 5～8; 45 ℃ of trypsinase suitable temps, optimum pH are 7.5～8.5; 50 ℃ of bromeline suitable temps, optimum pH are 7.
Raw material gelatin of the present invention comprises skin gelatin and bone gelatin(e).
The selection principle of combination enzyme reaction temperature and the suitable pH of system: the combination enzyme reaction temperature should the suitable temp of each single enzyme and the scope of pH in, that is: be no more than the higher single enzyme suitable temp of suitable temp, also be not less than the lower single enzyme suitable temp of suitable temp.
The present invention Citrate trianion and phosphate buffer soln regulation system pH, the method that the preparation of Citrate trianion and phosphate buffer soln and known single enzyme prepare gelatin hydrolysate is identical.
The test result of the gelatin hydrolysate of the present invention's preparation is listed in the table 1, and the method for relevant test is as follows:
The measuring method of molecular weight: adopt gel filtration chromatography to measure the molecular weight of gelatin hydrolysate.
Gelatin hydrolysate stability of solution measuring method: get the 1.5g gelatin hydrolysate and add water and be mixed with 15% solution, at room temperature place, every 7 days determining molecular weights and transparency.Considerable change is arranged is poor stability for molecular weight and transparency in 7 days; With interior molecular weight and transparency considerable change being arranged at 15 days is less stable; Not have considerable change be good stability for molecular weight and transparency more than 6 months.
Smelling flavor measures: at room temperature, gelatin hydrolysate is added water be mixed with 15% solution, smell its smell.
As can be seen from Table 1, gelatin hydrolysate of the present invention all is as good as raw meat and smells flavor, and the product that Comparative Examples obtains all has different raw meat to smell flavor, illustrates that the combination enzyme has the effect of eliminating peculiar smell.Gelatin hydrolysate aqueous stability of the present invention can be up to 1 year, and the sample that single enzyme liberating gelatin obtains has different fishy smell, deposits about a week muddy rotten.Proved that the combination enzyme is improved the stability of gelatin hydrolysate, has solved this difficult problem of gelatin hydrolysate stability of solution difference, for applying of product had laid a good foundation when eliminating peculiar smell.
Embodiment
Embodiment 1
Gelatin 10 gram being added water 90mL be mixed with the aqueous solution, is 7 with the pH value of Citrate trianion and phosphate buffer soln regulation system; Be warmed up to 50 ℃ of dissolvings, then, add trypsinase 0.01 gram and papoid 0.1 gram, reacted 1 hour, heat 90～100 ℃ and kill enzyme, filter, concentrate drying, promptly get the gelatin degraded product, measure the molecular weight of product, measure the stability of the aqueous solution and smell flavor, the result is as shown in table 1.
Embodiment 2
Gelatin 10 gram being added water 90mL be mixed with the aqueous solution, is 7 with the pH value of Citrate trianion and phosphate buffer soln regulation system; Be warmed up to 45 ℃ of dissolvings, then, add trypsinase 0.05 gram, reacted 2 hours, and heated 90～100 ℃ and kill enzyme, filter, cool to 65 ℃, add papoid 0.1 gram again, reacted 1 hour, heat 90～100 ℃ and kill enzyme, filter, concentrate, drying obtains product, measures the molecular weight of product, measure the stability of the aqueous solution and smell flavor, the result is as shown in table 1.
Embodiment 3
Gelatin 10 gram being added water 90mL be mixed with the aqueous solution, is 8 with the pH value of Citrate trianion and phosphate buffer soln regulation system; Be warmed up to 45 ℃ of dissolvings, then, add trypsinase 0.05 gram, AS1.398 neutral protease 0.2 gram, reacted 3 hours, heat 90～100 ℃ and kill enzyme, filter, concentrate drying, promptly get the gelatin degraded product, measure the molecular weight of product, measure the stability of the aqueous solution and smell flavor, the result is as shown in table 1.
Embodiment 4
Gelatin 10 gram being added water 90mL be mixed with the aqueous solution, is 7.0 with the pH value of Citrate trianion and phosphate buffer soln regulation system; Be warmed up to 50 ℃ of dissolvings, then, add bromeline 0.01 gram, AS1.398 neutral protease 0.2 gram, reacted 0.5 hour, heat 90～100 ℃ and kill enzyme, filter, concentrate drying, promptly get the gelatin degraded product, measure the molecular weight of product, measure the stability of the aqueous solution and smell flavor, the result is as shown in table 1.
Embodiment 5
Gelatin 10 gram being added water 90mL be mixed with the aqueous solution, is 5.0 with the pH value of lemon phosphoric acid and lemon phthalate buffer regulation system; Be warmed up to 65 ℃ of dissolvings, then, add papoid 0.1 gram, AS1.398 neutral protease 0.2 gram, reacted 2 hours, heat 90～100 ℃ and kill enzyme, filter, concentrate drying, promptly get the gelatin degraded product, measure the molecular weight of product, measure the stability of the aqueous solution and smell flavor, the result is as shown in table 1.
Comparative Examples 1
Gelatin 10 gram being added water 90mL be mixed with the aqueous solution, is 7.0 with the pH value of lemon phosphoric acid and lemon phthalate buffer regulation system; Be warmed up to 45 ℃ of dissolvings, then, add AS1.398 neutral protease 0.2 gram, reacted 2 hours, heat 90～100 ℃ and kill enzyme, filter, concentrate drying, promptly get the gelatin degraded product, measure the molecular weight of product, measure the stability of the aqueous solution and smell flavor, the result is as shown in table 1.
Comparative Examples 2
Gelatin 10 gram being added water 90mL be mixed with the aqueous solution, is 7.5 with the pH value of lemon phosphoric acid and lemon phthalate buffer regulation system; Be warmed up to 65 ℃ of dissolvings, then, add papoid 0.1 gram, reacted 5 hours, heat 90～100 ℃ and kill enzyme, filter, concentrate, drying promptly gets the gelatin degraded product, measures the molecular weight of product, measures the stability of the aqueous solution and smells flavor, and the result is as shown in table 1.
Comparative Examples 3
Gelatin 10 gram being added water 90mL be mixed with the aqueous solution, is 7 with the pH value of phosphoric acid salt and citrate buffer regulation system; Be warmed up to 50 ℃ of dissolvings, then, add bromeline 0.05 gram, reacted 2 hours, heat 90～100 ℃ and kill enzyme, filter, concentrate, drying promptly gets the gelatin degraded product, measures the molecular weight of product, measures the stability of the aqueous solution and smells flavor, and the result is as shown in table 1.
Table 1
Sequence number molecular weight Mn smells flavor stability
Embodiment 1 2600 does not have
Embodiment 2 3000 does not have
Embodiment 3 2500 does not have
Embodiment 4 2100 does not have
Embodiment 5 2800 does not have
Comparative Examples 1 4800 different fishy smell are poor
Comparative Examples 2 4000 different fishy smell are poor
Comparative Examples 3 3800 different fishy smell are poor
Claims (8)
1, a kind ofly prepares the method for gelatin hydrolysate, the raw material gelatin is mixed with the aqueous solution, with citric acid and phosphate buffer regulation system pH value with the combination enzyme; Be heated to temperature of reaction, enzyme put in the gelatin solution stir, through adding the heat kill enzyme, filtration, concentrated, dry, promptly obtain gelatin hydrolysate, it is characterized in that: the enzyme that is adopted is the combination enzyme, the combination enzyme contains any two kinds in AS1.398 neutral protease, papoid, trypsinase or the bromeline, temperature of reaction is got in single enzyme suitable temp scope of forming the combination enzyme, system pH is 5～8.5, reaction times is 0.5～5 hour, and the add-on of single enzyme is 0.01%～2% of a gelatin weight.
2, method according to claim 1 is characterized in that: the combination enzyme is AS1.398 neutral protease and papoid, and temperature of reaction is 45～65 ℃, and pH is 5～8.
3, method according to claim 1 is characterized in that: the combination enzyme is trypsinase and papoid, and temperature of reaction is 45～65 ℃, and pH is 5～8.5.
4, method according to claim 1 is characterized in that: the combination enzyme is trypsinase and AS1.398 neutral protease, and temperature of reaction is 45～65 ℃, and pH is 7～8.5.
5, method according to claim 1 is characterized in that: the combination enzyme is bromeline and AS1.398 neutral protease, and temperature of reaction is 45～50 ℃, and pH is 7.
6, method according to claim 1 is characterized in that: the combination enzyme is papoid and bromeline, and temperature of reaction is 50～65 ℃, and pH is 5～8.
7, method according to claim 1 is characterized in that: the combination enzyme is trypsinase and bromeline, and temperature of reaction is 45～50 ℃, and pH is 7～8.5.
8, method according to claim 1 is characterized in that: carry out the gelatin degraded after mixing with whole single enzyme of combination in the enzyme or will wherein a kind of single enzyme add earlier gelatin is degraded, after add another kind of single enzyme again and continue gelatin is degraded.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN 02159288 CN1206303C (en) | 2002-12-30 | 2002-12-30 | Method for preparing hydrolytic gelatin using combined enzyme |
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| CN 02159288 CN1206303C (en) | 2002-12-30 | 2002-12-30 | Method for preparing hydrolytic gelatin using combined enzyme |
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| CN1511910A CN1511910A (en) | 2004-07-14 |
| CN1206303C true CN1206303C (en) | 2005-06-15 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| DE102009033158A1 (en) * | 2009-07-13 | 2011-01-27 | Gelita Ag | Concentrate for the preparation of a cooling and separating agent as well as such cooling and separating agent |
| CN108586692B (en) * | 2018-01-23 | 2020-09-22 | 陕西科技大学 | Preparation method and application of polishable waterborne polyurethane for synthetic leather |
| CN108676502B (en) * | 2018-04-12 | 2020-10-16 | 山东双圆生物科技有限公司 | Method for preparing gelatin by using compound biological organic acid and compound biological enzyme |
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Owner name: BEIJING HUA DA JIERUI BIOTECHNOLOGY CO. Free format text: FORMER OWNER: BEIJING CHEMICAL ENGINEERING UNIV. Effective date: 20080704 |
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Effective date of registration: 20080704 Address after: Beijing City, Haidian District Zizhuyuan Road No. 98 winmore library No. 412 Patentee after: BEIJING HUADA JIERUI BIOTECHNOLOGY Co.,Ltd. Address before: Beijing City, Chaoyang District North Ring Road No. 15 Patentee before: Beijing University of Chemical Technology |
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