CN115176960A - Thermostable type blue pigment rich in phycocyanin and preparation method thereof - Google Patents
Thermostable type blue pigment rich in phycocyanin and preparation method thereof Download PDFInfo
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- CN115176960A CN115176960A CN202210785455.3A CN202210785455A CN115176960A CN 115176960 A CN115176960 A CN 115176960A CN 202210785455 A CN202210785455 A CN 202210785455A CN 115176960 A CN115176960 A CN 115176960A
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- phycocyanin
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- polylysine
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Images
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- 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
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/40—Colouring or decolouring of foods
- A23L5/42—Addition of dyes or pigments, e.g. in combination with optical brighteners
- A23L5/46—Addition of dyes or pigments, e.g. in combination with optical brighteners using dyes or pigments of microbial or algal origin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/20—Proteins from microorganisms or unicellular algae
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/347—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of proteins from microorganisms or unicellular algae
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- 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
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3526—Organic compounds containing nitrogen
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
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- 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
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Abstract
The invention relates to the technical field of pigment processing, in particular to a thermostable blue pigment rich in phycocyanin and a preparation method thereof. The blue pigment consists of phycocyanin, glutamine transaminase, polylysine and sulfated polysaccharide, wherein the content of the phycocyanin is not lower than 20%, the thermostable blue pigment is easy to dissolve in water, has the same ultraviolet-visible absorption spectrum and fluorescence spectrum as the phycocyanin, and can keep bright blue hue after being heated for 3-5min at 100-130 ℃. The invention solves the defect that the phycocyanin is easy to change and fade when being heated in the production and processing process on one hand, and solves the technical problems of low efficiency, low safety, complex process, difficult standardization and large-scale production for improving the thermal stability of the phycocyanin on the other hand.
Description
Technical Field
The invention relates to the technical field of pigment processing, in particular to a thermostable blue pigment rich in phycocyanin and a preparation method thereof.
Background
Blue is one of the three primary colors, but the blue pigment has few types and is scarce in resources. Phycocyanin (PC) is a natural pigment extracted from blue algae, presents itself bright blue, and is the only natural blue food material approved by the Food and Drug Administration (FDA). Compared with other blue pigments, the blue pigment has the advantages of high safety, wide color generation range and pleasant and bright blue color tone. In addition, phycocyanin also has fluorescent properties, antioxidant, anti-inflammatory, anticancer and liver protecting effects, and is a functional food with high nutritional value. However, phycocyanin is sensitive to light, heat, acid and alkali environments, poor in stability, easy to lose color and the like, so that phycocyanin is only used for low-temperature processed products such as ice cream and instant cold drinks at present, and the application range of phycocyanin is severely limited. The technical processes of mixing, sterilizing and the like in the processing process of foods and medicines, and links of long-term storage and transportation of products and the like all put higher requirements on the heat stability of the phycocyanin. Physical embedding is a common means for improving stability, chinese patent CN 112190697A utilizes tannin and other substances to embed phycocyanin, retention rate of phycocyanin is still over 80% after heating in 45 ℃ water bath for 12h, and the method has the advantages of simple process and large-scale production, but cannot solve the problem of high-temperature fading. In addition, researchers select stabilizers such as preservatives and polysaccharides to improve the heat stability of phycocyanin, the improvement effect is not obvious, and the quality of products is affected due to the fact that a large amount of the stabilizers are added. Li et al published in 2006 "Journal of Biotechnology" 121 of Chemical stabilization of the phytocyanin from cyanobacteria Spirolina, and the publication discloses that 2.5% (v/v) formaldehyde is effective in preventing thermal denaturation of phycocyanin, and remains stable even when heated at 100 ℃ for 10min, and has the advantages of high efficiency and simplicity. However, formaldehyde is an organic reagent, has a large potential safety hazard and is harmful to organisms. Therefore, it is necessary to develop a safe, green and efficient preparation method to improve the thermal stability of phycocyanin.
Disclosure of Invention
The invention aims to solve the technical problems that phycocyanin is sensitive to light, heat and acid-base environments, poor in stability, easy to denature and fade and the like, so that phycocyanin is only used for low-temperature processing products such as ice cream and instant cold drinks at present, and the application range of phycocyanin is severely limited. The technical processes of mixing, sterilizing and the like in the processing process of foods and medicines, and links of long-term storage and transportation of products and the like all put higher requirements on the heat stability of phycocyanin.
In order to solve the problems, the invention provides a thermostable type blue pigment rich in phycocyanin and a green, efficient and simple preparation method, which solve the defect that phycocyanin is easy to change and fade when being heated in the production and processing process on one hand, and solve the technical problems of low efficiency, low safety, complex process, difficult standardization and large-scale production for improving the thermal stability of phycocyanin on the other hand.
In order to achieve the above object, the present invention is specifically realized by the following technical scheme, and a preparation method of a thermostable type blue pigment rich in phycocyanin comprises the following steps:
(1) Weighing phycocyanin, adding distilled water, stirring to dissolve completely to obtain phycocyanin solution with concentration range of 0.5-10 g/L;
(2) Weighing glutamine transaminase, and dissolving with distilled water to obtain activated glutamine transaminase solution with concentration of 5-50 g/L; the catalytic action of glutaminase is utilized to regulate and control the space configuration of phycocyanin to maintain a relatively stretched state, and protein aggregation caused by thermal denaturation is avoided.
(3) Slowly adding the activated glutamine transaminase solution obtained in the step (2) into the phycocyanin solution obtained in the step (1) according to the proportion of adding 200-400U of active enzyme into phycocyanin per unit mass, fully mixing, and standing for 10-120min in a dark place at the reaction temperature of 4-50 ℃ to obtain a reaction solution I; in the step, the addition amount of glutamine transaminase obviously influences the space structure of phycocyanin, when the addition amount is too large, the space structure of phycocyanin can form a large amount of alpha helices and beta turns, beta folding of the stretched polypeptide chain is reduced, protein tends to aggregate, and the high catalytic efficiency of a large amount of glutamine transaminase is not beneficial to controlling the reaction rate of a system.
(4) Weighing polylysine with a certain mass, adding distilled water, stirring at room temperature, and dissolving to obtain a polylysine solution with a concentration range of 1-5 g/L; the polylysine is a linear chain polymer of lysine, lysine residues are positively charged and can interact with negatively charged amino acid residues on the surface of the phycocyanin, the polylysine is a linear molecule with 25-30 polymerization degrees, and the chain length of the polylysine can ensure that polylysine molecules are fully fixed around the outer structural domain of the phycocyanin to limit the depolymerization or the allosteric polymerization of phycocyanin monomers. Meanwhile, polylysine with the molecular weight range of 3600-4300 daltons is the best food grade preservative, the polymerization degree of the selected polylysine is 25-30, the polylysine has excellent sterilization and corrosion resistance, and the addition of the polylysine is favorable for improving the storage quality and shelf life of the phycocyanin.
Weighing a certain mass of sulfated polysaccharide, adding distilled water, stirring at room temperature, and dissolving to obtain a sulfated polysaccharide solution with a concentration range of 1-5 g/L; dextran sulfate is anionic polysaccharide, has negative charges, can be further combined with polylysine to form a stable embedding shell layer, and the formed micro-nano particles are not only favorable for maintaining the structural stability of phycocyanin, but also can remarkably reduce the fluid viscosity of high-molecular concentrated solution and are favorable for subsequent spray drying.
(5) Adding phycocyanin: polylysine: the mass ratio of sulfated polysaccharide is 1: (0.01-1): (0.02-2) sequentially and slowly adding the polylysine solution and the sulfated polysaccharide solution obtained in the step (4) to obtain a reaction solution II; the importance of the step lies in that the non-covalent interaction between protein and polysaccharide molecules is utilized, the product of the reaction solution I is embedded by utilizing steric hindrance to limit the deformation space of the phycocyanin structure domain while the excessive catalytic reaction of glutamine transaminase is hindered, and the phycocyanin is further stabilized, wherein the material adding proportion and the adding sequence directly influence the microstructure of the product.
(6) Drying and dehydrating the reaction solution II to obtain the thermostable type blue pigment rich in phycocyanin.
Further, the dissolving temperature of the step (1) and the step (2) is 4-30 ℃; the temperature of the magnetic stirring in the step (5) is 4-30 ℃, and the loss of phycocyanin in the preparation process is reduced to the maximum extent.
Further, in order to improve the catalytic efficiency of the transglutaminase, the transglutaminase of step (2) is activated at 30-50 ℃ for 15-60min.
Further, in order to achieve the effects of full mixing and complete reaction and avoid the problems of high stirring speed and protein loss caused by excessive shearing force, the step (3) and the step (5) are mixed under the magnetic stirring at the rotating speed of 400-800 rpm.
Further, the slow addition in the steps (3) and (5) is a flow rate of 10-200 ml/min. The flow-adding speed is too high, the phycocyanin and the glutamine transaminase are not mixed well, the glutamine transaminase only catalyzes a part of phycocyanin to be excessively crosslinked, and the flow-adding speed is too high, so that the polylysine and the dextran sulfate of the packing material are not mixed well, and the embedding effect is poor.
Further, the sulfated polysaccharide in step (4) is a type of polysaccharide containing sulfate groups, preferably dextran sulfate, chondroitin sulfate, fucoidan sulfate.
Further, the drying dehydration in the step (6) is spray drying or freeze drying, wherein the inlet air temperature of the spray drying is not higher than 170 ℃, the feeding flow rate is controlled to be 10-20ml/min, the amount of dry substances in the feed liquid is not lower than 0.5%, and the spray drying treatment of the material is ensured to be in a dry powder state.
The thermostable type blue pigment rich in phycocyanin prepared by the method comprises phycocyanin, glutamine transaminase, polylysine and sulfated polysaccharide, wherein the content of phycocyanin is not less than 20%, the blue color of protein powder is prevented from being too light due to too low phycocyanin content, the thermostable type blue pigment is easy to dissolve in water, an ultraviolet-visible absorption spectrum and a fluorescence spectrum of the thermostable type blue pigment have the same characteristic peaks as those of the phycocyanin, and bright blue color tone can be kept after the thermostable type blue pigment is heated for 3-5min at 100-130 ℃.
Furthermore, the phycocyanin is phycocyanin from spirulina platensis and spirulina maxima, and the purity is not lower than 4.0 according to the detection of the A620/A280 method.
Furthermore, the glutamine transaminase is independent of calcium ions, namely can catalyze crosslinking reaction under the condition of no calcium ions, and has enzyme activity of more than 100U/g, preferably 1000U/g, of microbial sources, preferably the glutamine transaminase derived from streptoverticillium mobaraense.
The invention has the beneficial effects that:
(1) The product of the invention is composed of phycocyanin, sulfated polysaccharide, polylysine and glutamine transaminase, wherein the phycocyanin accounts for more than 20% of the total solid content, the reaction process is mild, no organic solvent participates in the reaction, the prepared product is blue peculiar to the phycocyanin, the color difference value delta E between the product and the phycocyanin is less than 30, the product can be dissolved in aqueous solution with the temperature of more than 4 ℃ and the pH value of 7-8, the dissolved solution is in brilliant blue color, the maximum absorption peak is 620nm through ultraviolet-visible spectrum detection, and after the solution is respectively heated and treated for 5 and 3min at the temperature of 100 ℃ and 130 ℃, the retention rate of the solution is respectively improved by 48.88% and 62.99% compared with that of the simple phycocyanin. The higher the purity of the phycocyanin raw material is, the better the thermal stability of the prepared product is; compared with the pure phycocyanin, the heat stability of the same batch of products at higher temperature is more obvious.
(2) The product of the invention still maintains the advantages of strong coloring power and strong functional activity of the phycocyanin, the used raw materials have no toxic or harmful reagents and are food-grade raw materials, the preparation method is green and mild, no special processing equipment such as high temperature and high pressure is needed, and the expanded production is easy to realize.
(3) The preparation method has safe raw materials, simple process and easy amplification production, and does not need special equipment such as high temperature, high pressure, corrosion resistance and the like. The method not only solves the defect that the phycocyanin is easy to deform and fade when being heated in the production and processing process, but also solves the technical problems of low action efficiency, low safety and difficult large-scale production in the aspect of improving the heat stability of the phycocyanin.
Drawings
FIG. 1 is a photograph of lyophilized powder and reconstituted solution of the product (A) and the control phycocyanin (B) in example 1.
FIG. 2 is a UV spectrum of the phycocyanin solution of the product of example 2 and a control group.
FIG. 3 is a graph showing the change in retention of phycocyanin solutions of the product of example 3 and the control group after heat treatment at 100 ℃ for various periods of time.
FIG. 4 is a graph showing the change in retention of phycocyanin solutions of the product of example 4 and the control group at different heat treatment periods at 130 ℃.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the embodiments described are only a few embodiments of the present invention, and not all embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
Example 1:
accurately weighing 100mg phycocyanin, adding 25ml distilled water, and magnetically stirring at 500r/min for 5min to dissolve phycocyanin completely, wherein the whole process is in a dark condition. Activating 8mg/ml glutamine transaminase aqueous solution at 40 deg.C for 30min, adding 3.75ml glutamine transaminase aqueous solution into the phycocyanin solution to make the addition amount of glutamine transaminase be 300U/g, shaking, mixing, and standing at 25 deg.C in dark for 60min. Under the magnetic stirring condition of 550rpm, 4.5mg/ml polylysine solution is added at the flow rate of 25ml/min, after stirring for 5min, 5mg/ml dextran sulfate is added at the same flow rate, and finally phycocyanin: glutamine transaminase: polylysine: the mass ratio of the dextran sulfate is 1:0.3:0.8:1.6.
example 2:
accurately weighing 1g phycocyanin, adding 0.5L distilled water, and magnetically stirring at 500r/min for 15min to dissolve phycocyanin completely, wherein the whole process is in a dark condition. Activating 20g/L of transglutaminase aqueous solution at 50 deg.C for 15min, adding 20ml of transglutaminase aqueous solution into the phycocyanin solution to make the addition amount of transglutaminase 400U/g, shaking and mixing, and standing at 50 deg.C in dark for 15min. Then under the condition of magnetic stirring with the rotation speed of 400rpm, adding 3g/L polylysine solution into the mixture at the flow rate of 200ml/min, stirring for 5min, then adding 2g/L dextran sulfate at the same flow rate, and finally adding phycocyanin: transglutaminase: polylysine: the mass ratio of the dextran sulfate is 1:0.4:1:2.
example 3:
accurately weighing 10mg phycocyanin, adding 20ml distilled water, and magnetically stirring at 500r/min for 5min to dissolve phycocyanin completely, wherein the whole process is in a dark condition. Activating 10mg/ml transglutaminase aqueous solution at 40 deg.C for 30min, adding 0.3ml transglutaminase aqueous solution into the phycocyanin solution to make the addition amount of transglutaminase 300U/g, shaking and mixing, and standing at 37 deg.C in dark for 10min. Adding 3mg/ml polylysine solution at the flow rate of 50ml/min under the magnetic stirring condition of 800rpm, stirring for 5min, adding 2mg/ml dextran sulfate at the same flow rate, and finally adding phycocyanin: transglutaminase: polylysine: the mass ratio of the dextran sulfate is 1:0.3:0.8:1.2.
example 4:
accurately weighing 5g phycocyanin, adding 1L distilled water, and magnetically stirring at 500r/min for 20min to dissolve phycocyanin completely, wherein the whole process is in a dark condition. Activating 50g/L glutamine transaminase aqueous solution at 30 deg.C for 60min, adding 20ml glutamine transaminase aqueous solution into the phycocyanin solution to make the addition of glutamine transaminase 200U/g, shaking, mixing, and standing at 37 deg.C in dark for 30min. Under the magnetic stirring condition of 600rpm, adding 1mg/ml polylysine solution at the flow rate of 10ml/min, stirring for 5min, adding 1mg/ml chondroitin sulfate at the same flow rate, and finally adding phycocyanin: glutamine transaminase: polylysine: the mass ratio of the chondroitin sulfate is 1:0.2:0.01:0.02.
example 5:
the products prepared in examples 1-4 were frozen at-80 ℃ overnight and then freeze-dried for 48h to obtain the lyophilized powders of the products prepared in examples 1-4. Taking 1g of freeze-dried powder sample, dissolving in water, diluting to 100mL, diluting by 20 times, measuring the light absorption values at 620nm, 280nm and 618nm by using an ultraviolet-visible spectrophotometer through a 1cm cuvette, and calculating the phycocyanin content, purity and color value according to the formula (1), the formula (2) and the formula (3). The solutions containing the same phycocyanin mass concentration as in examples 1 to 4 were prepared, and colorimetric values L, a, b were measured using a spectrocolorimeter, and Δ E was calculated according to the formula (4), and the obtained results are shown in table 1.
Phycocyanin content (%) = M PC /M formula (1)
Purity = a 620 /A 280 Formula (2)
In the formula, M PC And M represents the mass of phycocyanin and the total mass of the sample, respectively; a. The 620 、A 280 And A 618 Respectively represent light absorption values at wavelengths of 620nm, 280nm and 618 nm; m represents the mass of the sample; f represents the dilution factor; the values of L, a and b represent the chromatic values of the products prepared in examples 1 to 4; l is a radical of an alcohol 0 、a 0 、b 0 Representing the colorimetric value of phycocyanin.
Table 1: phycocyanin content, purity, color number and Δ E of phycocyanin of products and controls in examples 1-4
As can be seen from the above table, the purity of the products in examples 1-4 is 0.96, 1.00, 0.96 and 2.15, which are all greater than 0.9, and the products are food grade phycocyanin. The phycocyanin contents of the products of examples 1-4 were 27%, 22.7%, 30.3% and 81.3%, respectively, while the phycocyanin-rich color numbers of 11.48-29.02, which are lower than phycocyanin, are probably due to the low phycocyanin content. The Δ E values for the products of examples 1-4 were 20.52, 21.48, 19.68 and 1.33, respectively, which may be the result of occlusion masking.
Example 6:
the product prepared in example 1 was lyophilized to obtain phycocyanin-rich blue pigment powder, 3.5mg of the lyophilized powder was weighed and added to 2ml of water for reconstitution, and then photographed, with phycocyanin as a blank control, and the obtained results are shown in fig. 1. Compared with the phycocyanin control group, the blue color depth of the phycocyanin-rich lyophilized powder and the complex solution prepared in example 1 is reduced, and is consistent with the results of the color value and the delta E in Table 1.
Example 7:
the blue color of phycocyanin originates from chromophores, which are closely related to the absorption spectrum. The light absorption of the product prepared in example 2 in the wavelength range of 200-800nm was measured by an ultraviolet spectrophotometer, and the result obtained using phycocyanin solution as a blank is shown in fig. 2. The phycocyanin in the control group has a characteristic absorption peak at 620nm, and the product in the example 2 also has a maximum absorption peak at 620nm, which shows that the thermostable type blue pigment rich in phycocyanin prepared by the invention does not change the space conformation of the chromophoric group of the phycocyanin and keeps the blue color of the phycocyanin.
Example 8:
the product prepared in example 3 was used for 100 ℃ thermal stability testing. The product prepared in example 3 was heated in a water bath at 100 ℃ for 20min, sampled every 5min, rapidly cooled at 4 ℃, measured for absorbance at 620nm and 652nm, and the retention of phycocyanin was calculated according to equations (5) and (6) using phycocyanin solution as a blank, and the results are shown in FIG. 3.
Phycocyanin concentration (mg/ml) = (A) 620 -0.474×A 652 ) /5.34 equation (5)
Retention (%) = C t /C 0 X 100% formula (6)
Wherein A is 620 And A 652 Represents the absorbance of the sample at 620nm and 652 nm; c 0 And C t Represents the initial and residual concentrations of phycocyanin in g/L.
The retention rate of phycocyanin of the phycocyanin-rich pigment prepared in example 3 after being subjected to water bath at 100 ℃ for 5min is 69.25%, which is 48.88% higher than that of the phycocyanin control group. Meanwhile, the retention rate of the phycocyanin in the control group is only 13.80% even if the phycocyanin is heated at 100 ℃ for 20min, which shows that the phycocyanin rich in the phycocyanin prepared by the method has strong tolerance to high-temperature environment.
Example 9:
the product prepared in example 4 was used for the 130 ℃ thermal stability test. The product prepared in example 4 was heat-treated at 130 ℃ for 8min, sampled every 1min and rapidly cooled in a 4 ℃ refrigerator, and the absorbance values at 620nm and 652nm were measured, and the retention of phycocyanin was calculated according to the formulas (5) and (6), and the result obtained using the phycocyanin solution as a blank was shown in FIG. 4. The retention rate of the product prepared in example 4 after being heated at 130 ℃ for 3min is 85.43%, while the retention rate of phycocyanin in the control group is only 22.44%, which is 62.99% higher than that in the control group, which shows that the phycocyanin-rich blue pigment prepared by the invention not only has good high-temperature stability, but also has better improvement effect on the stability of phycocyanin when the temperature is higher.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and any person skilled in the art may modify or modify the technical details disclosed above into equivalent embodiments with equivalent variations. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention are within the protection scope of the technical solution of the present invention.
Claims (10)
1. A preparation method of a thermostable type blue pigment rich in phycocyanin is characterized by comprising the following steps:
(1) Weighing phycocyanin, adding distilled water, stirring to dissolve completely to obtain phycocyanin solution with concentration range of 0.5-10 g/L;
(2) Weighing glutamine transaminase, adding distilled water for dissolving to obtain activated glutamine transaminase solution with concentration range of 5-50 g/L;
(3) Slowly adding the activated glutamine transaminase solution obtained in the step (2) into the phycocyanin solution obtained in the step (1) according to the proportion of adding 200-400U of active enzyme into phycocyanin per unit mass, fully mixing, and standing for 10-120min in a dark place at the reaction temperature of 4-50 ℃ to obtain a reaction solution I;
(4) Weighing polylysine with a certain mass, adding distilled water, stirring at room temperature, and dissolving to obtain a polylysine solution with a concentration range of 1-5 g/L;
weighing a certain mass of sulfated polysaccharide, adding distilled water, stirring at room temperature, and dissolving to obtain a sulfated polysaccharide solution with a concentration range of 1-5 g/L;
(5) Adding phycocyanin: polylysine: the mass ratio of the dextran sulfate is 1: (0.01-1): (0.02-2) sequentially and slowly adding the polylysine solution and the sulfated polysaccharide solution obtained in the step (4) to obtain a reaction solution II;
(6) And drying and dehydrating the reaction solution II to obtain the thermostable type blue pigment rich in phycocyanin.
2. The method of claim 1, wherein: the dissolving temperature of the step (1) and the step (2) is 4-30 ℃; the temperature of the magnetic stirring in the step (5) is 4-30 ℃.
3. The method of claim 1, wherein: activating the glutamine transaminase obtained in step (2) at 30-50 deg.C for 15-60min.
4. The method of claim 1, wherein: and (5) mixing the step (3) and the step (5) under the magnetic stirring at the rotating speed of 400-800 rpm.
5. The method of claim 1, wherein: the slow addition in steps (3) and (5) is a flow rate of 10-200 ml/min.
6. The method of claim 1, wherein: the sulfated polysaccharide in the step (4) is polysaccharide containing sulfate groups.
7. The method of claim 1, wherein: the drying dehydration in the step (6) is spray drying or freeze drying.
8. The method of claim 7, wherein: the air inlet temperature of spray drying is not higher than 170 ℃, the feeding flow rate is controlled to be 10-20ml/min, and the amount of dry substances in the feed liquid is not lower than 0.5%.
9. A phycocyanin-rich, thermostable blue pigment prepared according to any one of claims 1 to 8, characterized by: consists of phycocyanin, glutamine transaminase, polylysine and sulfated polysaccharide, wherein the content of phycocyanin is not less than 20%, and the bright blue color tone can be maintained after short-time heating at 100-130 ℃ for 3-5 min.
10. The thermostable blue pigment according to claim 9, wherein: the phycocyanin is phycocyanin from Spirulina platensis and Spirulina maxima, and the purity is not lower than 4.0 according to A620/A280 method.
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| US20190254302A1 (en) * | 2018-01-23 | 2019-08-22 | Cornell University | Systems and methods for controlling the release from enzyme-responsive microcapsules with a smart natural shell |
| CN111481675A (en) * | 2020-04-26 | 2020-08-04 | 大连工业大学 | Preparation method of phycocyanin-sodium alginate/lysozyme complex coacervate |
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| US20090004278A1 (en) * | 2006-01-30 | 2009-01-01 | Fujifilm Corporation | Enzymatically Crosslinked Protein Nanoparticles |
| US20190254302A1 (en) * | 2018-01-23 | 2019-08-22 | Cornell University | Systems and methods for controlling the release from enzyme-responsive microcapsules with a smart natural shell |
| CN111481675A (en) * | 2020-04-26 | 2020-08-04 | 大连工业大学 | Preparation method of phycocyanin-sodium alginate/lysozyme complex coacervate |
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