CN102660530A - Alkali protease composition with enhanced activity, and application thereof - Google Patents
Alkali protease composition with enhanced activity, and application thereof Download PDFInfo
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- CN102660530A CN102660530A CN2012101516634A CN201210151663A CN102660530A CN 102660530 A CN102660530 A CN 102660530A CN 2012101516634 A CN2012101516634 A CN 2012101516634A CN 201210151663 A CN201210151663 A CN 201210151663A CN 102660530 A CN102660530 A CN 102660530A
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- xylooligosaccharides
- sumizyme
- enhanced activity
- enzyme composition
- basic protein
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Abstract
The invention relates to an alkali protease composition with enhanced activity, and the alkali protease composition comprises alkali protease and xylo-oligosaccharide, wherein the content of the xylo-oligosaccharide in every 1000mu of alkali protease is less than 100g. The invention also relates to a product with a proteolysis function, and the product comprises the alkali protease composition with enhanced activity; and the product is an enzyme-containing detergent, a fiber modifying agent, a leather treating agent, make-up cosmetic, food or the like.
Description
Technical field
The present invention relates to a kind of basic protein enzyme composition, be specifically related to a kind of basic protein enzyme composition with enhanced activity.
Background technology
Sumizyme MP is through submerged fermentation, extracts and the refining a kind of proteolytic ferment that forms.It can generate polypeptide or amino acid by protein hydrolysate molecule peptide chain, has the ability of strong decomposing protein.75% is proteolytic ferment in the present industrial enzymes, and proteolytic enzyme is to occupy the maximum enzyme of ratio in the industrial enzymes, accounts for about 60% of the annual total sales volume in the whole world greatly.
Utilize its proteolyze characteristic, Sumizyme MP in food-processing, brewage, numerous industries such as silk, process hides and washing composition all are widely used:
Sumizyme MP is a popular washing additive in the market, can increase substantially the washing soil removability, especially protein dirts such as bloodstain, sweat stain, milk stain, oil stain is had unique washing effect.
Sumizyme MP is used for handling protein fibres such as hair, silk; Can play polishing action to wool product; The wool product glossiness that makes is high, and the lovely luster of low temperature dyeing is after softening; Fabric has soft, smooth performance, has nursing functions such as machine washable, anti, fluffing; Can also effectively eliminate organochlorine in addition, and human body is not had injury, also can be used for coming unstuck of silk, harmless fiber.
Sumizyme MP is when being used to soak leather (animal skin), but the aquation leather makes the fresh hide backwater, is used for dry-salted hide or wet salted hide, can avoid between the shelf lives leather to rot, and this soaking enzyme can be saved the about 4-6 of soak time hour, save energy.
Sumizyme MP can be made into perfumed soap, soap, cream frost, cleansing milk, washing composition, washing powder, Liquid soap, body wash, cosmetics and skincare product etc.Can act on the aging cutin layer on the human body skin; Impel its degeneration, decomposition, removal, promote skin metabolism, reduce the dark phenomenon of the colour of skin that causes because of Exposure to Sunlight; Make skin care present good delicate state; Excellent effect with unique whitening and softening skin, the health care of U.S. face, removing beverage dirt, and soil removability is strong, degerming sterilization, relieved safely.
In food-processing: Sumizyme MP can be applicable to the hydrolysis of food service industry plant animal protein, generates polypeptide or amino acid, forms the protein hydrolyzate with peculiar flavour, is widely used in the middle of sauce and the health products trade.Comprise: utilize basic protein enzyme extraction rice X 1000.Rice protein is formed with its reasonable amino acid, and characteristics such as higher bioavailability and distinctive hypoallergenic are acknowledged as quality plant albumen.Compare with traditional alkali extraction and acid precipitation, Enzymatic Extraction is to utilize degraded and the modification of proteolytic enzyme to rice protein, makes it become the peptide of solubility and is extracted out.The Enzymatic Extraction reaction conditions is gentle, and protein and peptide chain hydrolyzable is a short peptide chain, has improved proteinic solvability; The liquid-solid ratio of its reaction simultaneously is little; Improved the solid content in the extracting solution, reduced the energy expenditure that is used to remove extracting solution moisture, for industrial production has been created favourable condition.The application of Sumizyme MP in making the bread process: flour is the main raw material of bread making process.It contains undissolved gluten, and this has determined the quality of bread.Can modify wheat gluten through restricted degraded through Sumizyme MP.The dough that enzyme is handled can increase its toughness and thereby physical strength increases its range of application.Excessive use proteolytic enzyme can reduce the mixing time and the output that increases bread of dough.Use bacteria protease can increase the ductility of dough.
Because the production process of Sumizyme MP is complicated, just can improve utilization ratio if can improve its activity to it, reduce cost indirectly.
(xylooligosaccharide XOS), is called wood oligose again to xylooligosaccharides; Be by the functional polymerization sugar of 2-7 wood sugar molecule with β-1,4 glycosidic link be combined into, its natural being present in bamboo root, fruit, vegetables, cow's milk and the honey; Wide material sources, it is low to obtain cost.Xylooligosaccharides has great DEVELOPMENT PROSPECT with its significant bifidus bacillus proliferation function and other effects and widespread use in food, medicine and healthcare products becomes a kind of functional foodstuff that latest developments are got up.
Yet, understand according to the contriver, at present as yet not relevant for the report of xylooligosaccharides to the alkaline protease activity enhancement.
Summary of the invention
The purpose of this invention is to provide a kind of basic protein enzyme composition with enhanced activity, with and use.
For this reason, one aspect of the present invention provides a kind of basic protein enzyme composition with enhanced activity, comprises Sumizyme MP and xylooligosaccharides; The xylooligosaccharides content of wherein per 1000 u Sumizyme MPs correspondence is less than 100 g, as being 0.01-100 g, 1-100 g; 80-100 g, or be 100 g.
The present invention provides a kind of product with protein decomposition function on the other hand; Comprise said basic protein enzyme composition with enhanced activity, said product can be enzyme-containing detergent, fibre modifier, leather treatment, makeup, food, medical supplies etc.
Beneficial effect of the present invention is: utilize the enhancement of xylooligosaccharides to alkaline protease activity, improved the utilization ratio of Sumizyme MP, reduced the cost of the product of decomposing protein; Utilize the multi-functional of xylooligosaccharides self, like hypoglycemic, hypotensive, bifidus bacillus cultivation effect etc., make product functionality be able to expansion, Application Areas is more extensive.
Description of drawings
Fig. 1 illustrates the figure that concerns between alkaline protease activity enhancing rate and the xylooligosaccharides relative content.
Embodiment
Below in conjunction with accompanying drawing and specific embodiment the present invention is done further explain.
Folin's methods according in " the general TP of QB/T 1803-1993 industrial enzyme preparation " is measured protease activities.
Material and reagent
Folin reagent: in 2000 mL ground refluxes, add sodium wolframate (Na
2WO
4-2H
2O) 100 g, Sodium orthomolybdate (Na
2MoO
4-2H
2O) 25 g, water 700 mL, 85% phosphoric acid, 50 mL, concentrated hydrochloric acid 100 mL, little fiery boiling reflux 10 h take off reflux cooler, in stink cupboard, add Lithium Sulphate (Li
2SO
4) 50 g, water 50 mL and several dense bromine waters (99%), little again 15 min that boil are to remove unnecessary bromine, and cooling adds water and is settled to 1000 mL, and mixing filters.Be stored in the brown bottle.
Mix with two parts of water with a Folin reagent during use, shake up.
Sodium carbonate solution: 0.4 mol/L
Trichoroacetic acid(TCA): 0. 4 mol/L
Sodium hydroxide solution: 0. 5 mol/L
Buffered soln: borate buffer solution (pH=10.5) is applicable to Sumizyme MP
First liquid: take by weighing Sodium Tetraborate (borax) 19.08 g, be dissolved in water and be settled to 1000 mL;
Second liquid: weighing sodium hydroxide 4.0 g are dissolved in water and are settled to 1000 mL.
In the use, get first liquid 500 mL, second liquid 400 mL mixings, be diluted with water to 1000 mL, and proofread and correct with the pH agent.
Xylooligosaccharides: Suzhou is wealthy bio tech ltd product earlier.
Sumizyme MP: available from source, Shanghai leaf bio tech ltd (1000 U/g), dilution is 600 times during use.
Casein:, be made into 10g/L during use available from source, Shanghai leaf bio tech ltd.
Alkaline protease activity test control experiment:
(1). the xylooligosaccharides solution of the various concentration of the Sumizyme MP of adding 1mL and 1mL in reagent bottle (comprising that concentration is zero, does not promptly contain xylooligosaccharides) leaves standstill 2min in 40 ℃ ± 0.2 ℃ environment;
(2). in the mentioned reagent bottle, add the trichoroacetic acid(TCA) of 2.00mL again, shake up, in 40 ℃ ± 0.2 ℃ environment, leave standstill 10min;
(3). in the mentioned reagent bottle, add the 1.00mL casein again, mix;
(4). the 10min after-filtration is left standstill in the taking-up of mentioned reagent bottle;
(5). get 1.00mL filtrating with measuring cup, in measuring cup, add the 5.00mL sodium carbonate solution again;
(6). in above-mentioned measuring cup, add the 1.00mL Folin reagent and use solution, mix;
(7). final solution in the step (6) is placed 40 ℃ ± 0.2 ℃, 20 min that develop the color, and, use the 10mm cuvette in the 680nm wavelength, choosing xylooligosaccharides concentration is that zero sample hose is established the background pipe, and zeroing.
Sample test:
(1). the xylooligosaccharides solution of the various concentration of the Sumizyme MP of adding 1mL and 1mL in reagent bottle (comprising that concentration is zero, does not promptly contain xylooligosaccharides) leaves standstill 2min in 40 ℃ ± 0.2 ℃ environment;
(2). in the mentioned reagent bottle, add 1.00 mL caseins again, mix, in 40 ℃ ± 0.2 ℃ environment, leave standstill 10min;
(3). in the mentioned reagent bottle, add the trichoroacetic acid(TCA) of 2.00 mL again, shake up;
(4). 10 min after-filtration are left standstill in the taking-up of mentioned reagent bottle;
(5). get 1.00mL filtrating with measuring cup, in measuring cup, add the 5.00mL sodium carbonate solution again;
(6). in above-mentioned measuring cup, add the 1.00mL Folin reagent again and use solution, mix;
(7). final solution in the step (6) is placed 40 ℃ ± 0.2 ℃, 20 min that develop the color, and in the 680nm wavelength, with its absorbancy of 10mm cuvette test.
Typical curve:
Draw the tyrosine typical curve contain the different content xylooligosaccharides accordingly, calculate the extinction constant K whether in specialized range.Calculate K=98.2895 ± 0.0013.Be that the K value is basic identical.The xylooligosaccharides of testing said concentration does not have obvious absorbancy in the 680nm wavelength.
X?=?5/10?×?A?×?K?×?n
The enzyme activity of X-----sample, u/g (u/mL)
A-----sample absorbancy;
K------extinction constant;
The TV of 5------reaction reagent, mL;
10----reaction times 10min is in l min;
The n-----extension rate.
This shows that because the K value is basic identical, the variation that enzyme is lived can be calculated by the variation of A, embodiment provided by the invention characterizes the enzymic activity velocity of variation of sample based on this principle through calculation sample absorbance A velocity of variation.
The raising rate is according to computes:
Xylooligosaccharides is to enhancement=(the sample absorbancy-sample absorbancy of Sumizyme MP when not adding xylooligosaccharides of Sumizyme MP behind the adding xylooligosaccharides)/sample absorbancy * 100% of Sumizyme MP when not adding xylooligosaccharides of Sumizyme MP.
The result is shown in the following table 1, and the final absorbancy of Sumizyme MP sample that does not contain xylooligosaccharides is 0.0364:
| Embodiment | The xylooligosaccharides amount (g) that per 1000 u Sumizyme MPs are corresponding | Absorbancy | Alkaline protease activity enhancing rate (%) |
| 1 | 0.01 | 0.0407 | 11.81 |
| 2 | 1 | 0.0470 | 29.12 |
| 3 | 20 | 0.0490 | 34.62 |
| 4 | 50 | 0.0500 | 37.36 |
| 5 | 80 | 0.0507 | 39.29 |
| 6 | 100 | 0.0510 | 40.11 |
Fig. 1 illustrates among the embodiment 1-6, the relation of alkaline protease activity enhancing rate and xylooligosaccharides relative content.Data in the associative list 1, visible from Fig. 1, xylooligosaccharides increases along with the increase of xylooligosaccharides concentration the enhancement of alkaline protease activity.This increase trend is the most obvious when 1 g (embodiment 2) is following at per 1000 u Sumizyme MPs in the xylooligosaccharides relative concentration; And after the xylooligosaccharides relative concentration is increased to 80 g; Alkaline protease activity enhancing rate almost no longer include remarkable increase (39.29% of embodiment 5, to embodiment 6 40.11%).Therefore, the inventor thinks the xylooligosaccharides content of per 1000 u Sumizyme MPs when 100 g are following, and the alkaline protease activity reinforced effects is better.Do not considering under the condition of cost that the relative content of xylooligosaccharides can be 0.01-100 g in the compsn of the present invention, or 1-100 g, or 80-100 g, or 100 g.Should be understood that those skilled in the art putting into practice when of the present invention, can weigh as required between the alkaline protease activity reinforced effects of cost and xylooligosaccharides, to realize best embodiment.
Embodiment 7:
The application of compound basic protein enzyme composition on leather is made
In leather industry, when utilizing the basic protein enzyme composition to be used to soak leather (animal skin), but the aquation leather makes the fresh hide backwater, is used for dry-salted hide or wet salted hide, can avoid between the shelf lives leather to rot.Because xylooligosaccharides can improve alkaline protease activity, therefore adopt compound basic protein enzyme composition to replace traditional Sumizyme MP, its effectiveness is better; Two of advantage can reduce the consumption of Sumizyme MP, reduces manufacturing cost greatly.
Embodiment 8:
The application of compound basic protein enzyme composition on fibre modification
Sumizyme MP is used for handling protein fibres such as hair, silk, can play polishing action to wool product, and the wool product glossiness that makes is high, can also effectively eliminate organochlorine in addition, and human body is not had injury, also can be used for coming unstuck of silk, harmless fiber.Because xylooligosaccharides can improve alkaline protease activity, therefore adopt compound basic protein enzyme composition to replace traditional Sumizyme MP, effect is more obvious; Two of advantage can reduce the consumption of Sumizyme MP, and burden greatly reduces cost.
Embodiment 9:
The application of compound basic protein enzyme composition on food
In food-processing, Sumizyme MP has many purposes.As can be applicable to the hydrolysis of food service industry plant animal protein, and generate polypeptide or amino acid, form protein hydrolyzate with peculiar flavour, be widely used in the middle of sauce and the health products trade.It is medium simultaneously can also to apply to bread.Use compound basic protein enzyme composition to replace simple Sumizyme MP, because xylooligosaccharides has improved alkaline protease activity, so effect improves; Two of advantage, cost can be than original reduction; Three of advantage, xylooligosaccharides itself just are widely used in the food, and be painted like improved products, reduces water loss etc., therefore can improve food in many-side by compound Sumizyme MP; Four of advantage, xylooligosaccharides have splendid health care and are worth, like propagation probiotic bacterium, lowering blood glucose, multinomial function such as reducing cholesterol.Therefore, the use of compound basic protein enzyme composition has increased many high added values to food, has improved the value of product.
The application that a kind of compound basic protein enzyme composition is in the suds.
In the washing powder prescription, add an amount of Sumizyme MP, can the protein spot be decomposed into material soluble in water.So not only can obviously improve detergency power of laundry powder, and can reduce amount of surfactant, reduce the harm of tensio-active agent environment.Adopt the xylooligosaccharides and the Sumizyme MP composition forms of specified proportion to add in the articles for washing; Can improve the activity of Sumizyme MP greatly; Reduce enzyme dosage; Further reduce the usage quantity of tensio-active agent, strengthen the soil removability of articles for washing, especially remove the ability of protein spot.
Embodiment 11
The application of a kind of compound basic protein enzyme composition in beauty treatment
Compound Sumizyme MP can promote skin metabolism, reduces the dark phenomenon of the colour of skin that causes because of Exposure to Sunlight, makes skin care present good delicate state, has the excellent effect of unique whitening and softening skin, the health care of U.S. face, removing beverage dirt.And because xylooligosaccharides has promoted the activity of Sumizyme MP greatly, its effect is used Sumizyme MP merely than tradition, and tangible lifting is arranged.
The above embodiment of the present invention does not constitute the qualification to protection domain of the present invention.Various other corresponding changes and distortion that any technical conceive according to the present invention has been done all should be included in the protection domain of claim of the present invention.
Claims (7)
1. the basic protein enzyme composition with enhanced activity comprises Sumizyme MP and xylooligosaccharides, and the xylooligosaccharides content of wherein per 1000 u Sumizyme MPs correspondence is less than 100 g.
2. the described basic protein enzyme composition with enhanced activity of claim 1 is characterized in that, the xylooligosaccharides content that per 1000 u Sumizyme MPs are corresponding is 0.01-100 g.
3. the described basic protein enzyme composition with enhanced activity of claim 2 is characterized in that, the xylooligosaccharides content that per 1000 u Sumizyme MPs are corresponding is 1-100 g.
4. the described basic protein enzyme composition with enhanced activity of claim 3 is characterized in that, the xylooligosaccharides content that per 1000 u Sumizyme MPs are corresponding is 80-100 g.
5. the described basic protein enzyme composition with enhanced activity of claim 4 is characterized in that, the xylooligosaccharides content that per 1000 u Sumizyme MPs are corresponding is 100 g.
6. product with protein decomposition function comprises each described basic protein enzyme composition with enhanced activity of claim 1-5.
7. the said product with protein decomposition function of claim 6, said product is enzyme-containing detergent, fibre modifier, leather treatment, cosmetics, food.
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| CN1332787A (en) * | 1998-10-23 | 2002-01-23 | 宝洁公司 | Fabrid care composition and method |
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| WO2007064356A1 (en) * | 2005-11-30 | 2007-06-07 | Knowles Electronics, Llc | Balanced armature bone conduction shaker |
| CN101182559A (en) * | 2007-11-16 | 2008-05-21 | 江南大学 | Method for preparing low polyxylose through squeezing-assisted wheat bran enzymolysis |
| CN101524150A (en) * | 2009-04-10 | 2009-09-09 | 广东省微生物研究所 | Microbial ecological agent and preparation method thereof |
| CN101695372A (en) * | 2009-10-30 | 2010-04-21 | 太原纠偏食品开发有限公司 | Food additive for expelling toxin and nourishing skin and preparation method thereof |
-
2012
- 2012-05-16 CN CN2012101516634A patent/CN102660530A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1332787A (en) * | 1998-10-23 | 2002-01-23 | 宝洁公司 | Fabrid care composition and method |
| CN1570057A (en) * | 2003-07-21 | 2005-01-26 | 梁浩东 | Natural highly effective environmental protection type cleaning agent and its making method |
| WO2007064356A1 (en) * | 2005-11-30 | 2007-06-07 | Knowles Electronics, Llc | Balanced armature bone conduction shaker |
| CN101182559A (en) * | 2007-11-16 | 2008-05-21 | 江南大学 | Method for preparing low polyxylose through squeezing-assisted wheat bran enzymolysis |
| CN101524150A (en) * | 2009-04-10 | 2009-09-09 | 广东省微生物研究所 | Microbial ecological agent and preparation method thereof |
| CN101695372A (en) * | 2009-10-30 | 2010-04-21 | 太原纠偏食品开发有限公司 | Food additive for expelling toxin and nourishing skin and preparation method thereof |
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| Title |
|---|
| 吴京平: "多羟基化合物对碱性蛋白酶热失活半寿期的影响", 《中国食品添加剂》, no. 2, 15 April 2011 (2011-04-15), pages 187 - 190 * |
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