A kind of dregs of beans is through the preparation method of the soybean protein of heat treated
Technical field
A kind of dregs of beans is through the preparation method of the soybean protein of heat treated, and the biochemistry that belongs to processing of farm products and enzyme suppresses the field.
Background technology
Along with the demand in domestic soybean protein market and improving constantly of output, the raising of soybean protein functional character has become an urgent demand of numerous domestic albumen processing factory.With more external famous large-scale soybean protein manufacturers, compare as ADM, only grease, central soybean etc., the no matter kind or the quality of albumen, domestic product all also differs bigger class with it.This gap is little in many production scales, particularly outstanding in the middle-size and small-size soybean protein manufacturing and processing enterprise a little less than the research and development of products ability.Anatomizing influences the functional qualitative factor of soybean protein, mainly contains: 1) soybean varieties, 2) plant natural causes such as regional water and soil, weather, 3) processing mode and the technology of defatted soybean meal, comprise degreasing method (squeezing, lixiviate, CO
2Supercritical extract etc.), the residual oil content of skimming temp, degreasing time and defatted soybean meal etc., 4) processing mode and the technological parameter of soybean protein, as solid-liquid ratio, extraction temperature, extraction time, pH etc.Wherein 1) and 2) belong to the naturality factor, can be by seed selection soybean improved seeds, modes such as enhancement of field management obtain high-quality and process raw material, can be little for the leeway of control for soybean protein processing enterprise; 3) and 4) then belong to the post-production link, can remedy the influence of raw material to a great extent by selecting rational technological process and parameter, and can improve the functional character of soybean protein.
An easy unheeded link is arranged in the soybean protein process, is exactly the Passivation Treatment that defatted soybean meal extracts the soybean protein preferment.Soybean is rich in plurality of enzymes system, and as lipoxygenase, urease, amylase etc., wherein the content of lipoxygenase is the highest, accounts for 1~2% of protein content in the soybean.Because the catalysis of lipoxidase endonuclease capable contains the unrighted acid and the ester class of suitable, suitable-1-4-pentadiene structure, and generate hydroperoxidation lipid, in this process, can produce a large amount of free radicals.The place that this is worth arousing attention in soybean protein processing just, dregs of beans is through ungrease treatment, though most neutral lipid is removed, a spot of polar lipid and remaining neutral lipid can remain.In the protein extraction in later stage, these remaining lipids can interact in the molten step of alkali with the lipoxygenase in the dregs of beans, and the temperature of the environment of leaching liquor meta-alkalescence and 40~50 ℃ is the suitableeest action condition of lipoxygenase especially.Therefore a large amount of free radicals that produce in this process can impel soybean protein to assemble.This bulky, loosely organized aggregation can have a strong impact on the functional character of soybean protein.
As far back as 1966, Roubal and Tappel just studied lipid peroxidation and protein interaction.They think that the free radical that generates can bring out the polymerisation of protein in reaction, and have proposed the mechanism of reaction.What its research was primarily aimed at is the variation that lipid peroxide forms flavor components; But another the important consequence that has also proposed lipid peroxide and protein interaction is that protein function character changes, because protein combines with MDA, the energy state of protein can change, and cause the variation of protein denaturation and thing followed protein solubility, and solubility change and then have influence on the multiple functional character of protein, but do not do further research.
People such as Hettiarachchy find, in the soybean protein isolate leaching process, add the solubility that antioxidant can improve albumen, illustrate that the oxidation of certain composition in the soybean will have influence on the functional character of albumen; Its another research has then disclosed the solubility that snperoxiaized lipid can reduce albumen.People such as Akio obata find the lipoxidase enzymic catalytic reaction initial stage product-hydroperoxides can with soybean protein in-SH group generation-S-S-key ,-SO
2H or-SO
3H, thus make soybean protein form the ability drop of gel.
More than research all in various degree reflection lipid peroxidation the functional character of albumen is affected, but deeply do not excavate its root place.We are the principal element that causes the remaining lipid peroxidation of dregs of beans really by the existence of discovering lipoxygenase, and through after the enzyme processing of going out in various degree, the vigor of lipoxygenase progressively descends in the dregs of beans, and functional characters such as its solubility, gelation are progressively improved.Present material from being grasped, the domestic and international report that does not also have this respect.
Summary of the invention
The objective of the invention is to make the soybean protein functional character that extracts be improved by preliminary treatment to dregs of beans.The existence that confirms remaining lipid of defatted soybean meal and lipoxygenase after deliberation can inevitably cause the functional character of the soybean protein that extracts to suffer damage.For this detrimental effect is reduced to bottom line, adopted the method for several physical heating to come activity of fatty oxygenase in the passivation dregs of beans.After heat treatment, the enzyme in the dregs of beans reaches irreversible thermal denaturation, thereby reaches the purpose of eliminating its adverse effect in the late protein leaching process.
For the effect of eliminating lipoxygenase has two approach available: the one, the substrate of minimizing enzyme effect, because the solvent that is adopted in the dregs of beans degreasing process flow process is generally solvent No. six, its main component is a n-hexane, nonpolar alkane material such as cyclohexane, so the polar lipid in the soybean can not remove substantially, adding the existence of the nonpolar remaining lipid of minute quantity, therefore will reduce the effect substrate of lipoxygenase from the angle that reduces the remaining lipid of dregs of beans, is uneconomical also inconvenient.And second approach is exactly the quantity and the vigor of enzyme in reducing dregs of beans, and the object that this present invention just endeavoured to study improves the functional character that the later stage extracts albumen by the enzyme that goes out.
Technical scheme of the present invention: the enzyme that goes out of defatted soybean meal adopts the physical heating mode.For the xeothermic enzyme that goes out, the water content of dregs of beans is unusual important parameters, because the water content difference can have influence on the moisture activity of lipoxygenase in the dregs of beans, and moisture activity is very huge for the thermal denaturation temperature influence of enzyme.By regulating the dregs of beans water content in advance, it is reached or slightly surpass the moisture critical value of the enzyme denaturation under the heating-up temperature.General dregs of beans water content can be obviously greater than 10% effect, and temperature is 90~110 ℃, heat time heating time 1~30min.For the saturated vapor enzyme that goes out, it is crucial then controlling heat time heating time well, because in hot and humid environment, lipoxygenase is inactivation rapidly no doubt, and also sex change easily of the albumen in the dregs of beans is unfavorable for the later stage extraction.So must get hold of heat time heating time.The saturated vapor temperature is 70~100 ℃, and be 10s~20min heat time heating time.Or use microwave treatment: microwave power is 100~400W, and microwave treatment time is: 5~80s, the microwave treatment amount is 100 gram dregs of beans.
After cooling, air dry, measure the remaining vigor of lipoxygenase through the dregs of beans after the enzyme processing of going out.Linoleic acid is the vigor that the optimum response substrate of lipoxygenase is used to measure enzyme usually.Linoleic acid generates through enzymatic and contains the hydroperoxides of gripping structure altogether, at the 234nm place characteristic absorption is arranged, and its content is directly proportional with the vigor of enzyme, according to this principle, adopts the vigor of spectrophotometry lipoxygenase.Preparation 2.24 * 10
-3The linoleic borate solution (pH9.0) of mol/L, add a certain amount of dregs of beans and extract supernatant, in 30 ℃ of water bath with thermostatic control insulation 3min, add the absolute ethyl alcohol cessation reaction, increasing by 0.001 with the inherent 234nm of 1ml reaction system 1min place's delustring is that a unit calculating enzyme is lived.
Beneficial effect of the present invention: the present invention finds that the existence of lipoxygenase is the principal element that causes the remaining lipid peroxidation of dregs of beans, go out after enzyme handles through heating in various degree, the vigor of lipoxygenase progressively descends in the dregs of beans, and functional characters such as its solubility, gelation improve.Dregs of beans is through after heating the enzyme processing of going out, and functional characters such as the nitrogen soluble index of soybean protein and gel strength all improve a lot.
The specific embodiment
Embodiment 1
Get defatted soybean meal, pulverize the back and regulate water content to 10.2%, the tool plug screw-cap test tube of packing into, the sealing back is in 110 ℃ of heating enzyme 30min that goes out, and ice bath is cooled to room temperature immediately.The vigor of getting this dregs of beans mensuration lipoxygenase is about 400U/mg.Extract protein isolate with this dregs of beans, solid-liquid ratio 1: 10g/ml, 30 ℃ of temperature, at the molten 60min of pH7.0 alkali, the heavy pH4.0 of acid, curdled milk must be separated albumen through readjustment pH7.0 postlyophilization.The nitrogen soluble index (NSI) of measuring protein isolate is 91%.Measure the gel strength of protein isolate again, with the protein solution of this protein isolate preparation 12%, 90 ℃ are heated 15min down, and ice bath is cooled to room temperature, and the gel of formation is measured gel strength with property tester behind 4 ℃ of refrigerator ageing 24h, and its intensity is 206g.
Embodiment 2
Get defatted soybean meal, pulverize the back and regulate water content to 14%, the tool plug screw-cap test tube of packing into, the sealing back is in 90 ℃ of heating enzyme 1min that goes out, and ice bath is cooled to room temperature immediately.The vigor of getting this dregs of beans mensuration lipoxygenase is about 350U/mg.Extract protein isolate with this dregs of beans, solid-liquid ratio 1: 20g/ml, 60 ℃ of temperature, at the molten 30min of pH9.0 alkali, the heavy pH5.0 of acid, curdled milk spray-drying and must separate albumen behind readjustment pH7.0.The nitrogen soluble index (NSI) of measuring protein isolate is 92%.Measure the gel strength of protein isolate again, with the protein solution of this protein isolate preparation 12%, 90 ℃ are heated 15min down, and ice bath is cooled to room temperature, and the gel of formation is measured gel strength with property tester behind 4 ℃ of refrigerator ageing 24h, and its intensity is 220g.
Embodiment 3
Get defatted soybean meal, evenly spread skim in steaming on the frame, the saturation vapour temperature of regulating the constant temperature steam copper is 100 ℃, rapidly steaming is placed in the pot, airtight steam pot cover takes out the steaming frame and is cooled to room temperature behind the 10s, be about 150U/mg through measuring the lipoxidase enzyme activity after the air dry.Extract protein isolate, method is with embodiment 1, and the NSI value of measuring protein isolate is 95%.Measure 12% gel strength (condition is with embodiment 1) and be 315g.
Embodiment 4
Get defatted soybean meal, evenly spread skim in steaming on the frame, the saturation vapour temperature of regulating the constant temperature steam copper is 70 ℃, rapidly steaming is placed in the pot, airtight steam pot cover takes out the steaming frame and is cooled to room temperature behind the 20min, be about 50U/mg through measuring the lipoxidase enzyme activity after the air dry.Extract protein isolate, method is with embodiment 1, and the NSI value of measuring protein isolate is 95%.Measure 12% gel strength (condition is with embodiment 1) and be 356g.
Embodiment 5
Get 100 gram defatted soybean meals, add in the small beaker and seal up preservative film, regulate microwave generator power 100W, control microwave heating time 80s takes out beaker and cools off rapidly, treats that temperature reduces to that to measure the lipoxidase enzyme activity after the room temperature be about 75U/mg.Extract protein isolate, method is with embodiment 1, and the NSI value of measuring protein isolate is 93%.Measure 12% gel strength (condition is with embodiment 1) and be 303g.
Embodiment 6
Get 100 gram defatted soybean meals, add in the small beaker and seal up preservative film, regulate microwave generator power 400W, control microwave heating time 5s takes out beaker and cools off rapidly, treats that temperature reduces to that to measure the lipoxidase enzyme activity after the room temperature be about 80U/mg.Extract protein isolate, method is with embodiment 1, and the NSI value of measuring protein isolate is 89%.Measure 12% gel strength (condition is with embodiment 1) and be 290g.
Embodiment 7
Present embodiment is a reference examples, gets defatted soybean meal, and the vigor of directly measuring lipoxygenase is 4240U/mg.Get this dregs of beans and prepare protein isolate, measure the gel strength of NSI value and 12%, be respectively 83% and 37g with embodiment 1 method.