Background technology
Papoid (Papain) is called for short papain, is called Fructus Chaenomelis ferment again.Be the milk that utilizes in immature papaya (Carica papaya) fruit, adopt modern biotechnology to refine the pure-natural biological enzyme prepn that forms.It is to dredge base (SH) endopeptidase has the activity of proteolytic enzyme and esterase, and specificity is more widely arranged a kind of containing; Animal/vegetable protein, polypeptide, ester, acid amides etc. there is stronger hydrolysis ability; Simultaneously, also have complex functionality, can synthesize proteinoid to proteolysate.Its applied industry is more extensive, comprising: medicine industry, foodstuffs industry, cosmetics industry, fodder industry, leather industry, textile industry etc.In the medicine industry, contain the medicine of papoid, can play anticancer, tumour, lymphoid leukemia, former bacterium and parasite, tubercule bacillus etc., but anti-inflammatory, cholagogic, pain relieving, aid digestion.Treatment gynecopathy, glaucoma, hyperosteogeny, the healing of rifle knife wound mouth, blood group discriminating, sting etc.In foodstuffs industry, enzymatic reaction capable of using makes the macromolecular proteolysis of food become small-molecular peptides or amino acid.Be used for hydrolysising animal-plant protein, process tenderizer, hydrolysis sheep placental extract, hydrolyzed soy, the agent of biscuit pineization, noodles stablizer, beer beverage finings, senior oral liquid, protective foods, soy sauce brewing and drinks starter etc.Effectively the utilization of transforming protein matter improves food value greatly, reduces cost, and helps the digestion and the absorption of human body.In cosmetics industry; Papoid joined contain in protein and the greasy makeup; Have unique whitening and softening skin, the health care of U.S. face, removing beverage dirt, stimulate circulation, improve effect such as skin, the raising product specification also can be made into diet tea, cosmetics and skincare product etc.In the industry of daily use chemicals article, the perfumed soap of interpolation papoid, soap, washing composition, washing powder, Liquid soap etc., strong detergency, degerming sterilization, relieved safely.In fodder industry, papoid is used for fodder additives, and the exploitation protein source is beneficial to absorption, improves the utilization ratio of feed, practices thrift cost, helps livestock digestion, accelerating growth.As in feeds such as pig, ox, sheep, chicken, duck, goose, fish, shrimp, adding.Can also be as the additive of vegetable and fruit high-grade compound fertilizer of compound material.In leather industry, utilize papoid to process trichogen, tan leather has the pore exquisiteness, and the dermatoglyph brightness effect can be used for also handling that wool, silkworm chrysalis come unstuck, the silk refining, can reach effects such as soft, comfortable, anti-contraction, tensile strength.
Because the Application Areas that papoid relates to is extensive, people are also more deep to its understanding, and its Application Areas is also increasing with value.Some enterprise begins to produce papoid now, and still, most of product is relatively low in enzymic activity, and Application Areas is limited.
Papoid is normally produced with the raw product form, and main source is from the fruit of pawpaw tree, to extract the dried food and nuts of processing after the emulsion (latex).Emulsion is to collect through the neck that cuts fruit, or directly air-dry on fruit, or splashes in the container.Emulsion after air-dry is exactly the raw product of papoid.If need it is carried out further purifying to remove impurity, will dissolve raw product earlier, and purify with the process of a standard.Papoid behind the purifying can be made as dry powder or liquid.
(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 is because of its stability, and multiple physiological is functional, is used widely in various fields such as food, medicine, health care, feeds.Along with going deep into of research, people find gradually that also xylooligosaccharides also has much more more other functions, and as a kind of oligose of tool market potential, xylooligosaccharides also remains further exploitation at the potential of more Application Areass.Recently, also there is the people that xylooligosaccharides is used for the production of bread, reduces the hardness of bread, make the water-retentivity of bread better, obtain the quality guaranteed period that prolongs.
Therefore, it is particularly important to seek the product of the high papoid of a kind of enzymic activity.
Embodiment
Below in conjunction with specific embodiment compound pawpaw protease composition is further specified.
A kind of compound pawpaw protease composition comprises xylooligosaccharides and papoid, and wherein, the ratio of said xylooligosaccharides quality (g) and papoid active (u) is more than or equal to 0.002:1000.
Further, said xylooligosaccharides quality (g) is 0:002:1000~200:1000 with papoid active (u) ratio.
Further, said xylooligosaccharides quality (g) is 0.01:1000 with papoid active (u) ratio.
Further, said xylooligosaccharides quality (g) is 0.02:1000 with papoid active (u) ratio.
Further, said xylooligosaccharides quality (g) is 1:1000 with papoid active (u) ratio.
Further, said xylooligosaccharides quality (g) is 50:1000 with papoid active (u) ratio.
Further, said xylooligosaccharides quality (g) is 200:1000 with papoid active (u) ratio.
Beneficial effect of the present invention is: compound pawpaw protease composition provided by the invention has added xylooligosaccharides in traditional papoid; Improved the activity of papoid; Reduce with respect to preparation cost like this, make that the use range of papoid is more extensive with identical active papoid.
In addition,, effectively utilize the physiological function of xylooligosaccharides, make the function of papoid obtain expansion owing to added xylooligosaccharides at papoid.
The present invention also provides the application of this compound pawpaw protease composition in medicine industry, foodstuffs industry, beauty treatment industry, the industry of daily use chemicals article, fodder industry and leather industry; With less compound pawpaw protease composition addition; Realize active significant lifting of papoid, reduced production cost.
Below be the specific embodiment part:
Adopt the method for measuring protease activity in " the general TP of QBT1803-1993 industrial enzyme preparation " to experimentize.
1. experiment reagent
(1). the preparation of Folin reagent: in 2000mL ground reflux, add 100g sodium wolframate (Na
2WO
4-2H
2O), 25g Sodium orthomolybdate (Na
2MoO
4-2H
2O), 700mL water, 50mL 85% phosphoric acid, 100mL concentrated hydrochloric acid, place little fire boiling reflux 10h, take off reflux, in stink cupboard, add 50g Lithium Sulphate (Li
2SO
4), 50mL water and several dense bromine waters (99%), little again 15min that boils, to remove unnecessary bromine, cooling adds water and is settled to 1000mL. mixing, filtration, is stored in the brown bottle;
(2). use solution: a Folin reagent mixes with two parts of water, shakes up;
(3) .0.4mol/L sodium carbonate solution (Na
2CO
3);
(4) .0.4mol/L trichoroacetic acid(TCA);
(5) .0.5mol/L sodium hydroxide solution (NaOH);
(6) .l mol/L hydrochloric acid soln (HCl) and 0.lmol/L hydrochloric acid soln (HCl);
(7). buffered soln: preparation pH=7.5 phosphoric acid buffer takes by weighing 6.02g Sodium phosphate, dibasic (Na
2HPO12H
2O) and 0.5g SODIUM PHOSPHATE, MONOBASIC (NaH
2PO2H
2O), be dissolved in water and be settled to 1000mL, proofread and correct to 7.5 with the PH agent;
(8). papoid: available from source, Shanghai leaf bio tech ltd (12000U/g), dilution is 6000 times during use.
(9). casein:, be made into 10g/L during use available from source, Shanghai leaf bio tech ltd.
2. experimental technique
Control experiment:
(1). the xylooligosaccharides solution of the various concentration of the papoid 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 ℃, colour developing 20min, 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:
Concrete technology is following:
(1). the xylooligosaccharides solution of the various concentration of the papoid 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 1.00mL casein again, mix, in 40 ℃ ± 0.2 ℃ environment, leave standstill 10min;
(3). in the mentioned reagent bottle, add the trichoroacetic acid(TCA) of 2.00mL again, shake up;
(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 again and use solution, mix;
(7). final solution in the step (6) is placed 40 ℃ ± 0.2 ℃, colour developing 20min, 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 lmin;
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 the velocity of variation of calculation sample absorbance A.
The raising rate is according to computes:
Xylooligosaccharides is to enhancement=(the sample absorbancy-sample absorbancy of papoid when not adding xylooligosaccharides of papoid behind the adding xylooligosaccharides)/sample absorbancy * 100% of papoid when not adding xylooligosaccharides of papoid.
Comparative Examples:
Add when not adding xylooligosaccharides, according to the described experimental technique of above-mentioned test, when promptly the papoid activity was not influenced by xylooligosaccharides, the test absorbancy was 0.0406.
Embodiment 1:
Add xylooligosaccharides quality (g): papoid active (u) is 0.002:1000; According to the described experimental technique of above-mentioned official test; Testing its absorbancy is 0.0428; Adopt above-mentioned raising rate calculation formula to draw, the papoid that present embodiment provides active (u) improves 5.42% with respect to Comparative Examples papoid active (u).
Embodiment 2:
Add xylooligosaccharides quality (g): papoid active (u) is 0.01:1000; Experimental technique according to above-mentioned sample test; Testing its absorbancy is 0.0438; Adopt above-mentioned raising rate calculation formula to draw, the papoid that present embodiment provides active (u) improves 7.88% with respect to Comparative Examples papoid active (u).
Embodiment 3:
Add xylooligosaccharides quality (g): papoid active (u) is 0.02:1000; Experimental technique according to above-mentioned sample test; Testing its absorbancy is 0.0479; Adopt above-mentioned raising rate calculation formula to draw, the papoid that present embodiment provides active (u) improves 17.98% with respect to Comparative Examples papoid active (u).
Embodiment 4:
Add xylooligosaccharides quality (g): papoid active (u) is 1:1000; Experimental technique according to above-mentioned sample test; Testing its absorbancy is 0.0531; Adopt above-mentioned raising rate calculation formula to draw, the papoid that present embodiment provides active (u) improves 30.79% with respect to Comparative Examples papoid active (u).
Embodiment 5:
Add xylooligosaccharides quality (g): papoid active (u) is 50:1000; Experimental technique according to above-mentioned sample test; Testing its absorbancy is 0.0544; Adopt above-mentioned raising rate calculation formula to draw, the papoid that present embodiment provides active (u) improves 33.99% with respect to Comparative Examples papoid active (u).
Embodiment 6:
Add xylooligosaccharides quality (g): papoid active (u) is 200:1000; Experimental technique according to above-mentioned sample test; Testing its absorbancy is 0.0555; Adopt above-mentioned raising rate calculation formula to draw, the papoid that present embodiment provides active (u) improves 36.70% with respect to Comparative Examples papoid active (u).
Table 1 is absorbancy and the enzymic activity raising rate that papoid provided by the invention adds the xylooligosaccharides of different mass.
The various papoid compsns of table 1-are active
From table 1, draw: xylooligosaccharides has the enhanced effect to the enzymic activity of papoid, and the also corresponding increase of per-cent that improves along with the enzymic activity of the increase papoid of the xylooligosaccharides quality of adding.
Compound pawpaw protease composition provided by the invention has added xylooligosaccharides in traditional papoid; Improved the activity of papoid; Reduce with respect to preparation cost like this, make that the use range of papoid is more extensive with identical active papoid.
The compound pawpaw protease composition that the foregoing description improves can be in the application in medicine and health care industry, foodstuffs industry, beauty treatment industry, the industry of daily use chemicals article, fodder industry and the leather industry owing to having good enzymic activity.
Embodiment 7:
The application of compound pawpaw protease composition on leather is made
In leather industry, utilize the papoid compsn to process trichogen, tan leather; Has the pore exquisiteness; The dermatoglyph brightness effect can be used for also handling that wool, silkworm chrysalis come unstuck, the silk refining, can reach effects such as soft, comfortable, anti-contraction, tensile strength.Because xylooligosaccharides can improve the papoid activity, therefore adopt compound pawpaw protease composition to replace traditional papoid, its effectiveness is better, and the fur that produces is evaluated through many people, and is higher than original quality; Two of advantage can reduce the consumption of papoid, reduces manufacturing cost.
Embodiment 8:
Compound pawpaw protease composition is in Application in Food Processing
1 compound pawpaw protease composition also can be used in the tenderization.The method of tenderization has physics, chemistry and biological method.Add proteolytic enzyme and be and utilize proteolytic enzyme to destroy collagen protein and elastin in the meat reticular tissue, make the soft tenderization of muscle, do not have coarse striped, advantage such as it is few, simple to operate to have a used device, and effect is obvious.Papoid more than 84%, is that ability is the strongest in numerous proteolytic enzyme to the meat proteins percent hydrolysis.One of advantage is because therefore the enzymic activity of compound pawpaw protease composition has improved efficient greatly, and can reduce cost greater than the simple papoid that uses; Two 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 pawpaw protease composition has increased many high added values to food, has improved the local flavor and the value of product.No matter experiment showed, the meat that uses the tenderization of compound pawpaw protease composition through subjective appreciation, only use the meat of papoid than tradition, be flavor taste, or outward appearance, all is significantly improved.
2. bakery
Like biscuit, cake, bread etc., papoid can improve and cure effect in bakery, make it to paint more easily, and wheat is aromatic strongly fragrant, and excellent taste, it is good to eat that biscuit is crisp, and the cake finished product is moistening tasty and refreshing, and bread dough is even, the taste improvement.Use compound pawpaw protease composition to replace simple papoid,, therefore render a service better because xylooligosaccharides has improved the papoid activity; Two of advantage, cost can be than original reduction; Three of advantage, xylooligosaccharides itself just are widely used in the bakery, and be painted like improved products, reduces water loss etc., and therefore compound papoid can be improved food in many-side; Four of advantage, xylooligosaccharides have splendid health care and are worth, like propagation probiotic bacterium, lowering blood glucose, reducing cholesterol etc.Therefore, the use of compound pawpaw protease composition has increased many high added values to food, has improved the value of product.Through the evaluation of many people organoleptic test, the bakery that uses compound papoid to process uses independent papoid to compare with tradition, and everyway has improvement greatly.
3. papoid can be used for egg-white food processing.Compound pawpaw protease composition can be used to process animal proteinum raw materials such as meat, becomes various natural meat additives, and performance is good, aromatic flavour.Simultaneously can process all kinds of amino-acid nutrition liquids by hydrolyzed vegetable protein, beverage made of fruits or vegetables or food flavouring.
4. clarify beer
Compound pawpaw protease composition is in making beer technology; Can improve the saccharification wort composition, promote the degraded of fermentation di-acetyl, the free protein in the beer of degrading effectively; Improve the non-biostability of beer; The clarification that improves beer is antifreeze, stops cold turbid formation in the cold storage procedure, prolongs the maintenance phase of beer.One of advantage, because xylooligosaccharides has improved the papoid activity, so effect is better.Two of advantage can reduce cost.Three 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 pawpaw protease composition, the additional health care that has improved beer is worth.
In a word, the higher enzymatic reaction of compound pawpaw protease composition vigor capable of using makes the macromolecular proteolysis of food become small-molecular peptides or amino acid.Be used for hydrolysising animal-plant protein, process tenderizer, hydrolysis sheep placental extract, hydrolyzed soy, the agent of biscuit pineization, noodles stablizer, beer beverage finings, senior oral liquid, beverage made of fruits or vegetables, soy sauce brewing and drinks starter etc.Effectively the utilization of transforming protein matter improves food value greatly, helps the digestion and the absorption of human body, can also reduce cost.
Embodiment 9:
The application of compound pawpaw protease composition in industry of daily use chemicals article and beauty treatment
Papoid can be made into the white skin perfumed soap of pawpaw, soap, cream frost, cleansing milk, washing composition, washing powder, Liquid soap, body wash, diet tea, cosmetics and skincare product etc.
Papoid joined contain in protein and the greasy makeup, have unique whitening and softening skin, the health care of U.S. face, removing beverage dirt, stimulate circulation, improve effect such as skin, raising product specification.
And adopt compound pawpaw protease composition to replace traditional independent use papoid, owing to enzymic activity is improved, therefore better than original effect.The application of xylooligosaccharides has simultaneously also increased many value addeds of product.As:
A kind of beauty and make-up water that contains compound pawpaw protease composition comprises:
Xylooligosaccharides 0.3%-1%
Papoid (50,000 u/g): 2%-8%
Vitamins C: 0.5%
Vitamin E: 0.5%
Surplus is a water.
Above-mentioned beauty and make-up water good hand touch.Except that the characteristic of general cleaning skin, but skin care also, the nutrition body.Xylooligosaccharides-papoid compsn can act on the aging cutin layer on the human body skin, impels its degeneration, decomposition, removal, promotes skin metabolism, reduces the skin color depth phenomenon that causes because of Exposure to Sunlight.Xylooligosaccharides is removed the effect of radical in addition, slows down skin aging, makes skin care present good delicate state.
Embodiment 10:
The application of papoid preparation in feed
Papoid is added feed formulation or is blended directly in the feed, can improve proteinic utilization ratio and transformation efficiency greatly, and can develop wider protein source, thereby reduce feed cost.Can be extensively with the fodder additives of raising pig, ox, sheep, chicken, duck, goose, shrimp, eel, milk cow etc.
Xylooligosaccharides has improved the activity of papoid, and therefore the effect of compound pawpaw protease composition is better than simple papoid.And xylooligosaccharides itself can reduce antibiotic application as a kind of fodder additives widespread use, improves the meat quality of domestic birds and animals, improves its intestinal microflora, thereby improves its resistance against diseases, has high using value.Therefore, the application of compound pawpaw protease composition, its value is far longer than the papoid of independent use.
Embodiment 11:
Compound pawpaw protease composition is in the application of medicine, healthcare products industry
In the application of medicine, healthcare products; Add papoid and can play the effect of anticancer, antitumor, antiangiogenic property white blood disease, anti-microbial pathogen and parasite, tubercule bacillus; And can prevent and treat hypertension, ephritis, help help digestion, relax bowel, toxin expelling, anti-treating constipation and the aid digestion effect of controlling stomach trouble.Can be made into pawpaw throat lozenge, pawpaw enteric coated tablet or capsule, product has anti-inflammatory analgetic, cholagogic, aid digestion, improves functions such as immunizing power.Because it is active that xylooligosaccharides has increased papoid, adopt xylooligosaccharides and papoid compsn to be applied in the Medicines and Health Product, effect is better than independent papoid.And xylooligosaccharides itself has good health care and is worth, like the propagation beneficial bacteria of intestinal tract, and anti-treating constipation, anticancer, prevent and treat hypertension etc.Therefore, compound pawpaw protease composition, its functional effect and range of application all are far longer than papoid, have broad application prospects.
The invention provides the application of this compound pawpaw protease composition in medicine industry, foodstuffs industry, beauty treatment industry, the industry of daily use chemicals article, fodder industry and leather industry; With less compound pawpaw protease composition addition; Realize active significant lifting of papoid, reduced production cost.
The above embodiment has only expressed several kinds of embodiments of the present invention, and it describes comparatively concrete and detailed, but can not therefore be interpreted as the restriction to claim of the present invention.Should be pointed out that for the person of ordinary skill of the art under the prerequisite that does not break away from the present invention's design, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with accompanying claims.