CN116035196B - Polysaccharide-protein complex, and preparation method and application thereof - Google Patents
Polysaccharide-protein complex, and preparation method and application thereof Download PDFInfo
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/10—Foods or foodstuffs containing additives; Preparation or treatment thereof containing emulsifiers
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- General Preparation And Processing Of Foods (AREA)
Abstract
The invention provides a polysaccharide-protein complex, which is obtained by covalent crosslinking of polysaccharide and protein through oxidoreductase; the weight average molecular weight of the polysaccharide is 300-600 kDa; the weight average molecular weight of the protein is 9-800 kDa; the hydration radius of the polysaccharide-protein complex is 5-40 nm. Compared with the prior art, the polysaccharide-protein compound is obtained by selecting the polysaccharide and protein with the weight average molecular weight in a specific range and covalently crosslinking the polysaccharide-protein compound by an enzyme method, so that the compound has stronger emulsifying capacity and emulsifying stability, can be used in fat-containing products, can effectively prevent the problems of floating, layering and the like of fat without adding a chemical emulsifier, can not introduce the peculiar smell such as sour taste, astringent taste and the like, and achieves the purposes of stabilizing a liquid or solid fat-containing food system, ensuring the pure taste of the food and ensuring the nature of label ingredients.
Description
Technical Field
The invention belongs to the technical field of emulsifying agents, and particularly relates to a polysaccharide-protein compound, a preparation method and application thereof.
Background
Emulsifiers, which are an important class of food additives, play an important role in the food industry, and are now an important component of the food industry, where they are required in amounts of about 50% of the additives. Based on the interaction of the surface activity property and food components, the emulsifier not only plays roles of emulsification, dispersion, lubrication, stabilization and the like in a series of processing procedures of mixing, fusing and the like of various raw materials, but also can improve and enhance the quality and stability of food.
Conventional emulsifiers are largely classified into two types depending on the source: the emulsifier is chemically synthesized, such as tween, span, diacetyl tartaric acid mono-glyceride, diglycerol ester and the like, has heavy chemical smell, can bring bad flavor when being applied to food, has thin film formed by emulsifying and wrapping oil drops because the emulsifier is mostly small molecules, is easy to aggregate in long-term storage, has poor emulsion stability and often needs to be compounded for use to improve the emulsion stability; the other is a natural emulsifier, such as sodium caseinate, which is made of natural raw materials and has high safety, but the natural emulsifier is not closely arranged on the surface of oil drops and has general emulsifying capacity, so that the natural emulsifier needs to be modified to improve the emulsifying capacity.
The published materials mostly utilize polysaccharide and protein combinations to modify natural emulsifiers, and the presently disclosed compositions are largely divided into three categories: the two are simply mixed, namely, a composition is formed by electrostatic combination, but the combination is not firm enough, and the emulsion performance is poor due to the fact that the composition is easily influenced by factors such as pH, temperature and the like of a system when the composition is applied (for example, chinese patent with publication number CN 102227170B); the second is covalent bonding of the two through Maillard reaction, although the bonding is firm and not easily affected by the system, the reaction process is complex and difficult to control directionally, the odor such as burnt smell and the like and the darker color are often carried out, the application is limited, and the emulsifying performance is general because the polysaccharide and the protein raw materials are not specially selected (Chinese patent with publication number of CN 101181068A); the third category is that polysaccharide and protein are covalently crosslinked through an enzymatic method (Chinese patent publication No. CN 104304947A), but the method has no requirement on the weight average molecular weight, hydration radius and the like of raw materials, and the protein is only bovine serum albumin, so that the emulsifying effect of the prepared composition is far less than that of an emulsifying agent, the use amount of the emulsifying agent needs to be greatly increased to improve the emulsifying property of an application system, and the derivative problem is that too much protein and polysaccharide are introduced into the system, and the processing property and palatability are affected by too large viscosity of the emulsion.
Disclosure of Invention
In view of the above, the technical problem to be solved by the present invention is to provide a polysaccharide-protein complex with high emulsifying capacity and emulsion stability, and a preparation method and application thereof.
The invention provides a polysaccharide-protein complex, which is obtained by covalent crosslinking of polysaccharide and protein through oxidoreductase; the weight average molecular weight of the polysaccharide is 300-600 kDa; the weight average molecular weight of the protein is 9-800 kDa; the hydration radius of the polysaccharide-protein complex is 5-40 nm.
Preferably, the hydration radius of the polysaccharide-protein complex is 15-25 nm; and/or the weight average molecular weight of the polysaccharide is 400-500 kDa; and/or the weight average molecular weight of the protein is 10-700 kDa.
Preferably, the polysaccharide is selected from dietary fiber and/or starch; and/or the protein is selected from milk proteins and/or vegetable proteins;
Preferably, the oxidoreductase is selected from one or more of laccase, catalase and peroxidase.
Preferably, the mass ratio of the polysaccharide to the protein is 1: (0.2-3).
The invention also provides a preparation method of the polysaccharide-protein complex, which comprises the following steps:
S1) mixing polysaccharide, protein and water, and heating to obtain a mixed solution;
s2) mixing the mixed solution with oxidoreductase, heating for reaction, and then inactivating enzyme to obtain the polysaccharide-protein complex.
Preferably, the mass of the oxidoreductase is 0.0001-1% of the mass of the mixed solution in terms of 10 ten thousand U/mL of enzyme activity; and/or the mass of the polysaccharide is 0.01-6% of the total mass of the polysaccharide and water.
Preferably, the temperature of the heating treatment in the step S1) is 40-80 ℃; the heating treatment time is 5-30 min;
The temperature of the heating reaction in the step S2) is 40-80 ℃; the heating reaction time is 1-10 h;
the temperature of the enzyme deactivation treatment is 80-90 ℃; the enzyme deactivation treatment time is 5-40 min.
The invention also provides an application of the polysaccharide-protein complex as an emulsifier; or as a stabilizing system for liquid and/or solid fat-containing foods.
The invention also provides a food containing the polysaccharide-protein complex according to any one of claims 1 to 5 or the polysaccharide-protein complex prepared by the preparation method according to claims 6 to 8; preferably, the food product comprises at least one of a beverage, yogurt, modified milk, and ice cream.
The invention provides a polysaccharide-protein complex, which is obtained by covalent crosslinking of polysaccharide and protein through oxidoreductase; the weight average molecular weight of the polysaccharide is 300-600 kDa; the weight average molecular weight of the protein is 9-800 kDa; the hydration radius of the polysaccharide-protein complex is 5-40 nm. Compared with the prior art, the polysaccharide-protein compound with the specific hydration radius range is obtained by selecting the polysaccharide with the specific weight average molecular weight and the protein to carry out covalent crosslinking through an enzymatic method, the compound is tightly arranged on the surface of oil drops, the wrapped oil drops are compact, the solubility is higher, and meanwhile, the film formed by the wrapped oil drops is thicker, so that the aggregation of the oil drops can be effectively blocked, the compound has stronger emulsifying capacity and emulsifying stability, can be used in fat-containing products, can effectively prevent the problems of fat floating, layering and the like under the condition of not adding a chemical emulsifying agent, can not introduce peculiar smell such as sour taste, astringent taste and the like, and achieves the purposes of stabilizing a liquid or solid fat-containing food system, and guaranteeing the pure taste of the food and the natural ingredients of the label.
Drawings
FIG. 1 is a photograph showing oil and water separation after the emulsion stability of the samples obtained in examples 1 to 5 and comparative examples 1 to 6 of the present invention was measured;
FIG. 2 is a chart showing the stability analysis test of turbiscan of the modified milk prepared by the sample of comparative example 4 of the present invention;
FIG. 3 is a chart showing the stability analysis test of turbiscan of the modified milk prepared as the sample in example 1 of the present invention;
FIG. 4 is a graph depicting QDA for quantitative analysis of the modified milk obtained in the application example of the present invention.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The invention provides a polysaccharide-protein complex, which is obtained by covalent crosslinking of polysaccharide and protein through oxidoreductase; the weight average molecular weight of the polysaccharide is 300-600 kDa; the weight average molecular weight of the protein is 9-800 kDa.
The weight average molecular weight of a polysaccharide or protein is an important factor affecting the emulsifying capacity and the emulsion stability of the polysaccharide-protein complex. The weight average molecular weight is the average molecular weight counted by weight, and the sum of the weight fractions occupied by the molecules with different molecular weights and the products of the corresponding molecular weights can be measured by adopting a dynamic laser light scattering instrument. The polysaccharide or protein has too large weight average molecular weight, the composition is not tightly arranged on the surface of oil drops, the oil drops are not tightly packed, the emulsifying capacity is poor, and the solubility is low; the weight average molecular weight of the polysaccharide or protein is too small, the film formed by coating oil drops by the composition is too thin, the oil drops are easy to aggregate during long-term storage, and the emulsion stability is poor, so the polysaccharide and protein with the weight average molecular weight in a specific range are selected. In the present invention, it is further preferable that the weight average molecular weight of the polysaccharide is 400 to 500kDa; in the examples provided herein, the polysaccharide has a weight average molecular weight of, in particular, 300kDa, 600kDa, 500kDa, 400kDa or 450kDa; further preferably, the weight average molecular weight of the protein is 10 to 700kDa; in the examples provided herein, the weight average molecular weight of the protein is specifically 800kDa, 9kDa, 90kDa, 50kDa or 400kDa.
The polysaccharide is preferably dietary fiber and/or starch; the dietary fiber and the starch are preferably derived from cereal; further preferably, the dietary fiber is one or more of corn fiber, oat fiber, wheat fiber and Arabic; the source treatment process of the polysaccharide is not particularly limited, and the polysaccharide can be obtained by treating raw materials through one or more of fractional precipitation, column chromatography and gel chromatography molecules.
The protein is preferably a milk protein and/or a vegetable protein; further preferably, the milk protein comprises at least one of isolated whey protein and casein, and even more preferably one or more of isolated whey protein, casein and milk protein; the vegetable protein is preferably one or more of soybean protein, pea protein, rice protein and wheat protein; further preferably, the soy protein comprises one or more of soy protein isolate, soy protein concentrate and soy whey protein; the method for preparing the protein is not particularly limited, and the protein can be obtained by one or more combined treatment processes of salting out, dialysis, gel treatment and ion exchange chromatography.
Covalent crosslinking of polysaccharide and protein by oxidoreductase to obtain polysaccharide-protein complex; the mass ratio of the polysaccharide to the protein is preferably 1: (0.2-3); in the embodiment provided by the invention, the mass ratio of the polysaccharide to the protein is specifically 1: 3. 1:0.2, 1: 1. 1:0.5 or 1:2.5; the oxidoreductase is preferably one or more of laccase, catalase and peroxidase.
Another important factor affecting the emulsifying capacity and the emulsifying stability of the polysaccharide-protein complex provided by the invention is the hydration radius of the polysaccharide-protein complex, wherein the hydration radius is the radius of the space occupied by a moving molecule in a solution, is a numerical value obtained by integrating the shape characteristics of the molecule and the interaction characteristics of the molecule and a solvent, and can be calculated by adopting a curve measured by a dynamic laser light scattering instrument. The hydration radius of the polysaccharide-protein complex is too large, the migration rate is slow, the combination rate with oil drops in the complex emulsifying process is slow, and the emulsifying capacity is poor; the composition has too small hydration radius, and the barrier formed by the compound is difficult to effectively block aggregation of oil drops during long-term storage, so that the emulsion stability is poor. Only if the weight average molecular weight of the raw materials and the hydration radius of the compound meet certain conditions, the polysaccharide-protein compound has higher emulsifying capacity and emulsifying stability. In the present invention, the hydration radius of the polysaccharide-protein complex is preferably 5 to 40nm, more preferably 10 to 35nm, still more preferably 15 to 25nm; in the examples provided herein, the radius of hydration of the polysaccharide-protein complex is specifically 40nm, 5nm, 17nm, 20nm or 35nm.
According to the invention, the polysaccharide-protein compound with the specific hydration radius range is obtained by selecting the polysaccharide with the specific range of weight average molecular weight and the protein to be subjected to covalent crosslinking through an enzymatic method, the compound is tightly arranged on the surface of oil drops, the wrapped oil drops are compact, the solubility is higher, meanwhile, the film formed by wrapping the oil drops is thicker, and the aggregation of the oil drops can be effectively blocked, so that the compound has stronger emulsifying capacity and emulsifying stability, can be used in fat-containing products, can effectively prevent the problems of fat floating, layering and the like under the condition of not adding a chemical emulsifier, can not introduce peculiar smell such as sour taste, astringent taste and the like, and achieves the purposes of stabilizing a liquid or solid fat-containing food system, ensuring the pure taste of the food and natural tag ingredients.
The invention also provides a preparation method of the polysaccharide-protein complex, which comprises the following steps: s1) mixing polysaccharide, protein and water, and heating to obtain a mixed solution; s2) mixing the mixed solution with oxidoreductase, heating for reaction, and then inactivating enzyme to obtain the polysaccharide-protein complex.
The source of all the raw materials is not particularly limited, and the raw materials are commercially available. The types and proportions of the polysaccharide, protein and oxidoreductase are as described above, and are not described in detail herein.
Mixing polysaccharide and protein with water; in the present invention, it is preferable to mix the polysaccharide with water to form a polysaccharide solution and then add the protein; the mass concentration of the polysaccharide in the polysaccharide solution is preferably 0.01% -6%, more preferably 0.05% -5%, still more preferably 0.1% -4%, still more preferably 0.5% -3%, and most preferably 1% -2%.
After mixing, heating to obtain a mixed solution; the temperature of the heating treatment is preferably 40-80 ℃, and in the embodiment provided by the invention, the temperature of the heating treatment is specifically 80 ℃,40 ℃,50 ℃, 60 ℃ or 65 ℃; the heating treatment time is preferably 5 to 30 minutes; the heating treatment time in the embodiment provided by the invention is specifically 5min, 30min, 10min, 20min or 15min; suitable conditions for the enzymatic crosslinking reaction can be provided by heat treatment.
Mixing the mixed solution with oxidoreductase, and performing a heating reaction, namely performing an enzyme crosslinking reaction; the mass of the oxidoreductase is preferably 0.0001-1% of the total mass of the mixed solution according to the enzyme activity of 10 ten thousand U/mL; in the embodiment provided by the invention, the mass of the oxidoreductase is specifically 1%, 0.0001%, 0.03%, 0.005% or 0.8% of the total mass of the mixed solution in terms of 10 ten thousand U/mL of enzyme activity; the temperature of the heating reaction is preferably 40-80 ℃; in the embodiment provided by the invention, the temperature of the heating reaction is specifically 80 ℃,40 ℃, 50 ℃, 60 ℃ or 65 ℃; the heating reaction time is preferably 1 to 10 hours; in the embodiment provided by the invention, the heating reaction time is specifically 1h, 10h, 5h, 4h or 8h.
After the heating reaction is finished, inactivating enzyme, stopping enzyme crosslinking reaction, and ensuring the stability of the product; the temperature of the enzyme deactivation treatment is preferably 80-90 ℃; the time of the enzyme deactivation treatment is preferably 5 to 40 minutes.
After enzyme deactivation treatment, the powder polysaccharide-protein compound or the paste polysaccharide-protein compound can be obtained through drying or concentration; the drying process can be freeze drying, spray drying or hot air drying; the process of concentration may be rotary evaporation, membrane concentration or vacuum concentration.
The invention also provides an application of the polysaccharide-protein complex as an emulsifier.
The invention also provides application of the polysaccharide-protein complex as a stabilizing system of liquid and/or solid fat-containing foods; the mass of the polysaccharide-protein complex is preferably 0.05 to 0.5 percent, more preferably 0.1 to 0.5 percent, and even more preferably 0.2 to 0.3 percent of the mass of the liquid and/or solid fat-containing food; the liquid fat-containing food is preferably a modified milk, a milk-containing beverage, a vegetable protein beverage or a cereal beverage; the solid fat food is preferably milk powder.
Taking polysaccharide-protein complex as a stabilizing system for preparing milk, the preparation process is as follows: mixing raw milk, anhydrous butter and polysaccharide-protein complex, heating for emulsification, homogenizing, and sterilizing to obtain concocted milk; the mass of the polysaccharide-protein complex is preferably 0.1-0.5% of the mass of the prepared milk, more preferably 0.2-0.3%; the temperature of the emulsification is preferably 70-75 ℃; the time of the emulsification is preferably 10 to 40min, more preferably 20 to 30min; the homogenization temperature is preferably 65-70 ℃; the secondary pressure of the homogenization is preferably 30 to 50bar, more preferably 40bar; the primary pressure for homogenization is preferably 150 to 250bar, more preferably 150 to 200bar, still more preferably 180bar.
The invention also provides a food containing the polysaccharide-protein complex; preferably, the food product comprises at least one of a beverage, yogurt, modified milk, and ice cream.
In order to further illustrate the present invention, the following describes in detail a polysaccharide-protein complex, a preparation method and application thereof provided in the present invention with reference to examples.
The reagents used in the examples below are all commercially available; the catalase was purchased from Shandong Bioengineering Co., ltd; laccase was purchased from Ningxia-Shang Utility Co., ltd; milk proteins were purchased from Shanghai purkinje International trade company Limited; casein was purchased from a constant natural group; pea proteins were purchased from jiaji; the isolated soy protein is purchased from jaboticaba; isolated whey protein was purchased from Shanghai purkinje International trade company Limited; rice proteins were purchased from tin-free gold biotechnology limited; wheat proteins were purchased from sitlesh agrobiotechnology limited; acacia gum is purchased from yiruian; corn fiber was purchased from western safety australian biotechnology limited; oat fiber and wheat fiber were purchased from reddenmel (JRS).
Comparative example 1
Selecting acacia having weight average molecular weight of 300kDa, preparing 2% acacia solution, selecting milk protein having weight average molecular weight of 9kDa, and mixing acacia and milk protein according to ratio 1:0.2 adding milk protein into acacia solution, and preserving heat at 40 ℃ for 5min; adding laccase (enzyme activity is 10 ten thousand U/mL) according to 0.0001% of total mass of acacia and milk protein solution, stirring, and maintaining at 40deg.C for 1 hr. After the reaction is finished, heating to 80 ℃ and maintaining for 40min for enzyme deactivation, and then freeze-drying to obtain a powder finished product. The radius of hydration of the finished product is 4nm.
Comparative example 2
Selecting corn fiber with weight average molecular weight of 500kDa, preparing 2% corn fiber solution, selecting casein with weight average molecular weight of 20kDa, and mixing the corn fiber and casein according to the proportion of 1:0.5 adding casein into the corn fiber solution, and preserving the temperature at 40 ℃ for 5min; catalase (enzyme activity is 10 ten thousand U/mL) is added according to 1% of the total mass of the corn fiber and casein solution, and the mixture is stirred and incubated for 10 hours at 40 ℃. After the reaction is finished, heating to 80 ℃ and maintaining for 40min for enzyme deactivation, and then spray drying to obtain a powder finished product. The radius of hydration of the finished product is 42nm.
Comparative example 3
Oat fiber with weight average molecular weight of 100kDa is selected, 2% oat fiber solution is prepared, pea protein with weight average molecular weight of 60kDa is selected, and the ratio of oat fiber to pea protein is 1:3 adding pea protein into the oat fiber solution, and preserving the temperature at 80 ℃ for 30min; catalase (enzyme activity is 10 ten thousand U/mL) is added according to 0.1% of the total mass of the oat fiber and pea protein solution, and the mixture is stirred and kept at 80 ℃ for 10 hours. After the reaction is finished, heating to 90 ℃ and keeping for 5min for enzyme deactivation, and then carrying out vacuum concentration to obtain a pasty finished product. The radius of hydration of the finished product is 25nm.
Comparative example 4
Selecting wheat fiber with weight average molecular weight of 800kDa, preparing 2% wheat fiber solution, selecting pea protein with weight average molecular weight of 100kDa, and mixing the wheat fiber and pea protein according to the proportion of 1:2 adding pea protein into the wheat fiber solution, and preserving the temperature at 70 ℃ for 30min; catalase (enzyme activity is 10 ten thousand U/mL) was added according to 0.01% of the total mass of the wheat fiber and pea protein solution, stirred, and incubated at 70℃for 5h. After the reaction is finished, heating to 90 ℃ and keeping for 5min for enzyme deactivation, and then performing rotary evaporation to obtain a pasty finished product. The radius of hydration of the finished product is 20nm.
Comparative example 5
Selecting acacia having weight average molecular weight of 300kDa, preparing 2% acacia solution, selecting milk protein having weight average molecular weight of 7kDa, and mixing acacia and milk protein according to ratio 1:1, adding milk protein into acacia solution, and preserving heat at 50 ℃ for 15min; laccase (enzyme activity is 10 ten thousand U/mL) is added according to 0.5% of the total mass of the acacia and milk protein solution, and the mixture is stirred and kept at 50 ℃ for 3 hours. After the reaction is finished, heating to 85 ℃ and keeping for 20min for enzyme deactivation, and then freeze-drying to obtain a powder finished product. The radius of hydration of the finished product is 14nm.
Comparative example 6
Oat fiber with weight average molecular weight of 500kDa is selected, 2% oat fiber solution is prepared, pea protein with weight average molecular weight of 900kDa is selected, and the ratio of oat fiber to pea protein is 1:0.7 adding pea protein into the oat fiber solution, and preserving the temperature at 60 ℃ for 30min; catalase (enzyme activity is 10 ten thousand U/mL) is added according to 0.03 percent of the total mass of the oat fiber and pea protein solution, and the mixture is stirred and is kept at 60 ℃ for 8 hours. After the reaction is finished, heating to 90 ℃ and keeping for 5min for enzyme deactivation, and then carrying out vacuum concentration to obtain a pasty finished product. The radius of hydration of the finished product is 30nm.
Example 1
Selecting a wheat fiber with a weight average molecular weight of 300kDa, preparing a 2% wheat fiber solution, selecting a soybean protein isolate with a weight average molecular weight of 800kDa, and mixing the wheat fiber and the soybean protein isolate according to a ratio of 1:3 adding the isolated soy protein into the wheat fiber solution, and preserving the temperature at 80 ℃ for 5min; catalase (enzyme activity is 10 ten thousand U/mL) is added according to 1% of the total mass of the wheat fiber and soybean protein isolate solution, and the mixture is stirred and incubated at 80 ℃ for 1h. After the reaction is finished, heating to 90 ℃ and keeping for 5min for enzyme deactivation, and then carrying out vacuum concentration to obtain a pasty finished product. The radius of hydration of the finished product is 40nm.
Example 2
Acacia having a weight average molecular weight of 600kDa was selected, a 2% acacia solution was prepared, casein having a weight average molecular weight of 9kDa was selected, and the ratio of acacia to casein was 1:0.2 adding casein into acacia solution, and keeping the temperature at 40 ℃ for 30min; catalase (enzyme activity was 10 ten thousand U/mL) was added at 0.0001% of the total mass of the gum arabic and casein solution, stirred, and incubated at 40℃for 10 hours. After the reaction is finished, heating to 80 ℃ and keeping for 40min for enzyme deactivation, and then performing rotary evaporation and concentration to obtain a pasty finished product. The radius of hydration of the finished product is 5nm.
Example 3
Selecting corn fiber with weight average molecular weight of 500kDa, preparing 2% corn fiber solution, selecting separated whey protein with weight average molecular weight of 90kDa, and mixing corn fiber and separated whey protein according to the ratio of 1:1 adding the separated whey protein into corn fiber solution, and preserving the temperature at 50 ℃ for 10min; laccase (enzyme activity is 10 ten thousand U/mL) is added according to 0.03% of the total mass of the corn fiber and the separated whey protein solution, and the mixture is stirred and kept at 50 ℃ for 5 hours. After the reaction is finished, heating to 85 ℃ and keeping for 20min for enzyme deactivation, and then spray drying to obtain a powdery finished product. The radius of hydration of the finished product is 17nm.
Example 4
Selecting oat fiber with weight average molecular weight of 400kDa, preparing 2% oat fiber solution, selecting rice protein with weight average molecular weight of 50kDa, and mixing the oat fiber and the rice protein according to the proportion of 1:0.5 adding rice protein into oat fiber solution, and preserving heat at 60 ℃ for 20min; adding catalase (enzyme activity is 10 ten thousand U/mL) according to 0.005% of the total mass of the oat fiber and rice protein solution, stirring, and preserving at 60 ℃ for 4 hours. After the reaction is finished, heating to 90 ℃ and keeping for 5min for enzyme deactivation, and then freeze-drying to obtain a powdery finished product. The radius of hydration of the finished product is 20nm.
Example 5
Selecting corn fiber with weight average molecular weight of 450kDa, preparing 2% corn fiber solution, selecting wheat protein with weight average molecular weight of 400kDa, and mixing corn fiber and wheat protein according to a ratio of 1:2.5 adding wheat protein into the corn fiber solution, and preserving the temperature at 65 ℃ for 15min; catalase (enzyme activity is 10 ten thousand U/mL) is added according to 0.8 percent of the total mass of the corn fiber and the wheat protein solution, and the mixture is stirred and is kept at 65 ℃ for 8 hours. After the reaction is finished, heating to 85 ℃ and keeping for 15min for enzyme deactivation, and then spray drying to obtain a powdery finished product. The radius of hydration of the finished product is 35nm.
The results of the measurements of the composite products obtained in comparative examples 1 to 6 and examples 1 to 5 were shown in Table 1, based on the following water-oil model measurements.
Emulsification capacity measurement: preparing 0.2% emulsifier solution, shearing with a shearing machine at a shearing speed of 3000r/min, slowly dripping vegetable oil, adjusting shearing speed to 6000r/min, when the oil drops are dripped to a certain degree, increasing emulsion viscosity, yellowing emulsion color, immediately stopping oiling, and recording total dripping amount of oil. The total amount of vegetable oil that can be emulsified per 1g of emulsifier is the emulsifying capacity.
Emulsion stability determination: the emulsion is placed in a centrifuge tube, and centrifuged for 10min at 4000r/min at 25 ℃, and the oil and water separation condition is observed, so that the oil and water separation photographs are shown in figure 1, wherein 1-6 in figure 1 are respectively comparative examples 1-6, and 7-11 are respectively examples 1-5. As can be seen from FIG. 1, oil-water separation is evident in comparative examples 1, 3 and 5, oil-floating is present on the surfaces of comparative examples 2, 4 and 6, examples 1 to 5 are more uniform, oil-water separation is not present, and oil-floating is not present on the surfaces.
Table 1 results of measurement of the emulsifying properties of the compound
| Sample of | Emulsifying capacity (g/g) | Emulsion stability |
| Comparative example 1 | 15.0 | Oil-water separation |
| Comparative example 2 | 12.3 | The surface is provided with floating oil |
| Comparative example 3 | 15.7 | Oil-water separation |
| Comparative example 4 | 13.1 | The surface is provided with floating oil |
| Comparative example 5 | 16.4 | Oil-water separation |
| Comparative example 6 | 12.8 | The surface is provided with floating oil |
| Example 1 | 18.1 | No oil slick and no oil-water separation on the surface |
| Example 2 | 20.3 | No oil slick and no oil-water separation on the surface |
| Example 3 | 19.2 | No oil slick and no oil-water separation on the surface |
| Example 4 | 17.7 | No oil slick and no oil-water separation on the surface |
| Example 5 | 19.5 | No oil slick and no oil-water separation on the surface |
Application examples
The formula of the modified milk comprises the following components: raw milk 98.9%, anhydrous cream 0.9%, and the sample obtained in comparative example 4 or example 1 0.2%.
The process comprises the following steps: emulsifying for 30min at 70-75 ℃, homogenizing at 65-70 ℃ for one time, wherein the homogenizing pressure is 40bar (secondary pressure)/180 bar (primary pressure), and the UHT tube sterilization is carried out under 138 ℃ for 4s.
Detecting the performance of the prepared milk, and detecting the particle size by using a laser particle sizer; emulsion stability (12 h at 25 ℃) was measured using turbiscan stability analyzer to obtain TSI value, light intensity variation value, top peak height (25 ℃). The results are shown in Table 2, FIG. 2 and FIG. 3. Wherein FIG. 2 is a chart of a turbiscan stability analysis test of the prepared milk from the sample of comparative example 4; FIG. 3 is a chart of a turbiscan stability analysis test of the modified milk prepared from the sample of example 1; TSI represents overall stability, with smaller values being more stable; the light intensity change value indicates particle agglomeration, and the larger the absolute value is, the more serious the particle agglomeration is; the peak height at the top indicates that the grease floats up, and the larger the value, the more serious the grease floats up. The particle agglomeration, top oil lifting and overall stability of example 1 were all better than comparative example 4, indicating that example 1 has better emulsion stability than comparative example 4.
Table 2 test results of milk properties
Sensory testing of the modified milk: quantitative analysis and description QDA graphs are obtained through professional sensory team evaluation, and are shown in FIG. 4. Evaluation criteria: panelist n=6, flavor evaluation used 10 minutes, specific criteria: less than or equal to 2 minutes: none; 3-4 minutes: identifiable, not very strong; 5-6 minutes: moderately strong; 7-8 minutes: strong; 9-10 minutes: is very strong.
As can be seen from fig. 4, the samples of comparative example 4 and example 1 each contained no off-flavors such as sour taste, bitter taste, and astringency; the samples containing comparative example 4 had a lower smoothness than example 1 and a higher thickness than example 1, since comparative example 4 had a larger weight average molecular weight, resulting in the application product tasting sticky and not smooth enough; the front flavor score of the milk, cream, and condensed milk containing the sample of comparative example 4 was slightly lower than that of example 1, because of the higher weight average molecular weight of comparative example 4, the product was thick and the front flavor was masked.
Claims (10)
1. A polysaccharide-protein complex is characterized in that the polysaccharide-protein complex is obtained by covalent crosslinking of polysaccharide and protein through oxidoreductase; the weight average molecular weight of the polysaccharide is 300-600 kDa; the weight average molecular weight of the protein is 9-800 kDa; the hydration radius of the polysaccharide-protein complex is 5-40 nm;
the oxidoreductase is selected from laccase and/or catalase.
2. The polysaccharide-protein complex of claim 1, wherein the hydration radius of the polysaccharide-protein complex is 15-25 nm; and/or the weight average molecular weight of the polysaccharide is 400-500 kDa; and/or the weight average molecular weight of the protein is 10-700 kDa.
3. The polysaccharide-protein complex of claim 1, wherein the polysaccharide is selected from dietary fiber and/or starch; and/or the number of the groups of groups,
The protein is selected from one or more of milk protein and/or vegetable protein.
4. The polysaccharide-protein complex of claim 1, wherein the mass ratio of polysaccharide to protein is 1: (0.2-3).
5. A method of preparing the polysaccharide-protein complex of claim 1, comprising:
s1) mixing polysaccharide, protein and water, and heating to obtain a mixed solution;
S2) mixing the mixed solution with oxidoreductase, heating for reaction, and then inactivating enzyme to obtain the polysaccharide-protein complex.
6. The preparation method according to claim 5, wherein the mass of the oxidoreductase is 0.0001% -1% of the mass of the mixed solution in terms of 10 ten thousand U/mL of enzyme activity; and/or the mass of the polysaccharide is 0.01% -6% of the total mass of the polysaccharide and water.
7. The method according to claim 5, wherein the temperature of the heating treatment in the step S1) is 40 ℃ to 80 ℃; the heating treatment time is 5-30 min;
the temperature of the heating reaction in the step S2) is 40-80 ℃; the heating reaction time is 1-10 h;
The temperature of the enzyme deactivation treatment is 80-90 ℃; the enzyme deactivation treatment time is 5-40 min.
8. The use of the polysaccharide-protein complex of any one of claims 1 to 4 or the polysaccharide-protein complex prepared by the preparation method of any one of claims 5 to 7 as an emulsifier; or as a stabilizing system for liquid and/or solid fat-containing foods.
9. A food product comprising the polysaccharide-protein complex of any one of claims 1 to 4 or the polysaccharide-protein complex prepared by the preparation method of any one of claims 5 to 7.
10. The food product of claim 9, wherein the food product comprises at least one of a beverage, yogurt, modified milk, and ice cream.
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