CN102020705A - Method for preparing spirulina protein isolate - Google Patents
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- CN102020705A CN102020705A CN2009100920991A CN200910092099A CN102020705A CN 102020705 A CN102020705 A CN 102020705A CN 2009100920991 A CN2009100920991 A CN 2009100920991A CN 200910092099 A CN200910092099 A CN 200910092099A CN 102020705 A CN102020705 A CN 102020705A
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Abstract
The invention relates to a method for preparing spirulina protein isolate, and belongs to the technical field of food science and engineering. The method is characterized by comprising the following steps of: (1) preparing a spirulina cell disrupted, decolorized, deodorized and degreased material; (2) extracting protein by using dilute solution of salt and centrifuging; (3) adjusting the pH until the solution is acidic in a gradient and sectional way, performing acid precipitation to obtain the protein, and adjusting the pH until the solution is neutral; and (4) performing ultra-high temperature short-time sterilization and spray drying. The spirulina protein isolate produced by the method has the advantages of light color, light fishy odor, high protein content, high dissolubility and easiness of digestion, and can be used as a high-quality full-value protein nutritional supplement or additive.
Description
Technical field:
The present invention relates to a kind of preparation method of spirulina protein isolate, belong to the Food Science and Engineering technical field.
Background technology:
Spirulina (Spirulina) is called cyanobacteria again, belongs to Cyanophyta, Cyanophyceae, and Oscillatoriales, Oscillariaceae, Spirullina or artrospira spirulina platymiscium are the thread blue-green algaes of a kind of many cells, are made up of the short cylindrical shape cell, are the filament spirrillum.Spirulina originates in Africa and North America, is a kind of low waterplant of waiting, and existing known helical Trentepohlia has 36 kinds, is distributed widely in ocean, lake, the hot spring of the torrid zone, subtropics and warm temperate zone, particularly the salt alkali lake.The spirulina that batch production is both at home and abroad at present produced mainly is meant bigger spirulina plalensis of the bodily form (Spirulina platensis) and spirulina maxim (Spirulina maxima).
The biochemical composition of spirulina is because of kind difference difference to some extent, containing extremely, rich in protein is one of prominent feature of nutritive component of spirulina, the protein of spirulina powder is about 50%～70%%, and the amino acid of hydrolysis of protein spirulina is formed A wide selection of colours and designs, 8 kinds of indispensable amino acid balanced proportion of human body, need pattern very for similar to the amino acid that FAO and WHO are recommended, be respectively different bright ammonia 4.1%～4.5%, leucine 5.8%～6.8%, Methionin 4.4%～4.6%, phenylalanine 2.9%～4.4%, methionine(Met) 1.8%～2.4%, Threonine 2.7%～4.1%, tryptophane 1.1%～1.6%, Xie Ansuan 4.8%～6.5%.
Though spirulina is a kind of high-quality full price nutrient protein source, the contained Mierocrystalline cellulose of its cell walls is less, its proteinic digestibility can reach 75.0%, and bioavailability can reach 68.0%, still, no matter be directly edible, still use as protein supplements, all exist some limitation: the first, the kind of spirulina product is comparatively single, form with sheet or pill occurs mostly, easily causes the sensation of taking medicine to the human consumer; The second, spirulina fishy smell is very heavy, has influence on the acceptability of its product to a great extent, can influence the local flavor of product when being applied to other food; The 3rd, as the proteinaceous nutrient additive, to compare with soybean protein isolate, also there are bigger gap in its protein content, digestibility and bioavailability.These have brought considerable restraint all for the processing of spirulina goods and comprehensive utilization.In view of this, the present invention carries out cytoclasis, decolouring, degreasing with spirulina and takes off raw meat and handle, preparation becomes protein isolate, to further raising hydrolysis of protein spirulina content, digestibility and bioavailability, expand its adjustable property and range of application, particularly make it as a kind of high-quality complete protein nutritious supplementary or additive, be applied to other various food (especially being applied to dispensed food for baby), have important practical value.
Summary of the invention:
In order to overcome the deficiencies in the prior art, the object of the present invention is to provide a kind of preparation method of spirulina protein isolate, use that the spirulina protein isolate that this method produces has lighter color, fishy smell is light, protein content is high, the good and digestible advantage of solvability, can be used as a kind of high-quality complete protein nutritious supplementary or additive and use.
The present invention is achieved through the following technical solutions:
A kind of preparation method of spirulina protein isolate may further comprise the steps:
(1) preparation of the broken decoloration deodorization degreasing of spirulina cells material
Spirulina raw material (algae mud) is carried out micronizing to be handled, make the cell walls fragmentation, add organic solvent then, stir or vibration, extraction pigment, lipid and fishy smell material, remove organic solvent layer, the cytoclasis material is carried out vacuum outgas, be prepared into decoloration deodorization degreasing spirulina cells broken material liquid;
(2) rare salts solution extracts albumen and centrifugal treating
The decoloration deodorization degreasing spirulina cells broken material liquid of gained with salt, is adjusted to rare salt dispersion liquid and makes the albumen stripping; With salt is that salt is adjusted to weak brine, and making spirulina cells broken material and weak brine ratio is 1: 10～20; Be under 40～60 ℃ the condition in temperature, rare salt dispersion liquid of gained is extracted 1～2 hour postcooling to room temperature, centrifugation then makes spirulina protein weak brine extracting solution.Spirulina material slag through extracting after centrifugal is extracted centrifugal 2 times by above-mentioned condition again, obtains spirulina protein weak brine extracting solution respectively, and with the spirulina protein weak brine extracting solution that obtains for 3 times merge, standby.
(3) pH gradient segmentation acid adjustment protein precipitation is neutral with adjusting
It is acid that the spirulina protein weak brine extracting solution of gained is regulated pH with HCl solution, adopt pH gradient segmentation acid heavy, leave standstill, make albumen precipitation complete, centrifugation goes out spirulina Acid precipitation albumen, Acid precipitation albumen is scattered in it in water after washing again, and the pH that regulates this spirulina Acid precipitation albumen dispersion liquid with NaOH solution is 7.0;
(4) Ultra High Temperature Short Time and spraying drying
With the pH of gained is that 7.0 spirulina Acid precipitation albumen dispersion liquid carries out Ultra High Temperature Short Time, and spraying drying is carried out in cooling then, promptly gets product spirulina protein isolate.
Micronizing that spirulina raw material (algae mud) is carried out described in the step (1) is handled, and making the cell walls fragmentation is to adopt the ultra-high speed milling treatment of colloid to make the spirulina cells fragmentation, and the rotating speed of ultra-high speed colloidal mill is 6000～8500rpm/min; Described organic solvent is a kind of in sherwood oil, normal hexane and the gasoline or their mixture; When extraction pigment and fishy smell material, the mass volume ratio of organic solvent and spirulina cells broken material is 1: 1～10, and extraction times is 2～3 times; The vacuum tightness of described vacuum outgas is 0.07～0.09MPa.
Described being meant with salt of step (2) adds NaCl, KCl, MgCl
2, Na
2SO
4, NH
4SO
4, Na
2CO
3, Na
2HPO
4In a kind of or their mixture; The concentration of salt is 0.02～0.6mol/L in described rare salt dispersion liquid; Described centrifugal rotational speed is 3000～4500rpm/min.
The concentration of the described HCl solution of step (3) is 2.0mol/L, and regulating the pH value is 3.5～4.2, and pH is changed to low in gradient from height, divides 3 times precipitating proteins, and precipitate time of repose is 10～30min at every turn; The centrifugation rotating speed is 4000～6000rpm/min; The water washing number of times is 2～3 times, and the NaOH strength of solution is 0.5～2.0mol/L.
The processing parameter of the described Ultra High Temperature Short Time of step (4) is: 135～143 ℃ of sterilising temps, and the time is 2～15s; Described spray-dired processing parameter is: feeding temperature is 60～80 ℃, and the hot air inlet temperature is 160～200 ℃, and temperature out is 80～90 ℃.
The present invention utilizes the ultra-high speed colloidal mill with the spirulina cells fragmentation, impel pigment, lipid, fishy smell material and protein fully to discharge, improve the effect that organic solvent extraction removes pigment, lipid and fishy smell composition so greatly, used vacuum outgas then can further remove fishy smell; Through the decoloration deodorization degreasing spirulina material that cytoclasis and organic solvent extraction are prepared into, the stripping of spirulina protein isolate when helping strengthening weak brine and extracting; By weak brine extraction, pH gradient segmentation acid adjustment precipitation, Ultra High Temperature Short Time and spray drying treatment, make protein generation sex change to a certain degree, improved the digestibility and the solvability of protein isolate.
The present invention has following beneficial effect compared with prior art:
1, adopts ultra-high speed colloidal mill replacement high-pressure homogenization or clarifixator to handle broken spirulina cells, simplified technological operation, can significantly enhance productivity, reduce production costs.
2, the method that adopts organic solvent extraction and vacuum outgas to combine is taken off raw meat, and fishy-removing-methods such as specific activity carbon adsorption, cyclodextrine entrapping method, enzymolysis process, heating method and fermentation method are simple, and deodorization effect is better.
3, adopt weak brine to extract, good to protein stability, solubleness is big, littler than water, diluted acid water, diluted alkaline water and extraction using alcohol albumen solubility rate height, sex change.
4, adopt the pH gradient segmentation acid adjustment precipitator method to make the protein multiple spot separate out in the iso-electric point scope, fully precipitating proteins further improves recovery of protein rate in the extracting solution.
Embodiment:
The present invention is further illustrated by following examples in the present invention:
Embodiment 1:
Be to handle under the condition of 6500rpm/min at rotating speed with spirulina (algae mud) feed liquid with the high speed colloidal mill, carry out cytoclasis, in cytoclastic spirulina algae pug liquid, add sherwood oil then, sherwood oil and feed liquid add in 1: 5 ratio, stir or vibration, extraction pigment and fishy smell material, extraction times is 3 times, remove the petroleum ether solvent layer that contains pigment and fishy smell material, is that 0.09MPa outgas with vacuum tightness with lower floor's spirulina cells broken material in vacuum degassing machine, promptly gets the broken feed liquid of decoloration deodorization spirulina cells after the degassing;
The decoloration deodorization spirulina cells broken material liquid of gained is added NaCl, be adjusted to the dispersion liquid that NaCl concentration is 0.15mol/L, making spirulina cells broken material and weak brine ratio is 1: 10, be under 60 ℃ the condition in temperature, extract 2 hours postcooling to room temperature, centrifugation then, centrifuge speed are 3000rpm/min, obtain spirulina protein weak brine extracting solution.Spirulina material slag through extracting after centrifugal is extracted centrifugal 2 times by above-mentioned condition again, obtains spirulina protein weak brine extracting solution respectively, and with the spirulina protein weak brine extracting solution that obtains for 3 times merge, standby.
The spirulina protein weak brine extracting solution of gained is regulated pH4.0, pH 3.8 and pH 3.6 successively with the HCl solution of 2.0mol/L, it is heavy to carry out 3 segmentations acid, leave standstill 15min at every turn, isolate spirulina Acid precipitation albumen with the rotating speed of 4000rpm/min with whizzer, the heavy albumen of 3 hypo acids is merged, with with the water washing of volume 2 times, Acid precipitation albumen is modulated into aqueous dispersions, and regulating this Acid precipitation albumen dispersion liquid pH with the NaOH solution of 0.5mol/L again is 7.0;
With the pH of gained is that 7.0 spirulina Acid precipitation albumen dispersion liquid is in ultrahigh-temperature (UHT) instantaneous sterilizing device, 5s sterilizes under 135 ℃ temperature, be cooled to 55 ℃ rapidly, carry out spraying drying then, spray-dired processing parameter is: feeding temperature is 80 ℃, the hot air inlet temperature is 160 ℃, and temperature out is 80 ℃, so far promptly obtains product spirulina separated protein powder.
Embodiment 2:
Be to handle under the condition of 7000rpm/min at rotating speed with spirulina (algae mud) feed liquid with the high speed colloidal mill, carry out cytoclasis, in cytoclastic spirulina algae pug liquid, add normal hexane then, normal hexane and feed liquid add in 1: 5 ratio, stir or vibration, extraction pigment and fishy smell material, extraction times is 3 times, remove the normal hexane solvent layer that contains pigment and fishy smell material, is that 0.08MPa outgas with vacuum tightness with lower floor's spirulina cells broken material in vacuum degassing machine, promptly gets the broken feed liquid of decoloration deodorization spirulina cells after the degassing;
The decoloration deodorization spirulina cells broken material liquid of gained is added Na
2SO
4, be adjusted to Na
2SO
4Concentration is the dispersion liquid of 0.07mol/L, and making spirulina cells broken material and weak brine ratio is 1: 15, is under 55 ℃ the condition in temperature, extract 1.5 hours postcooling to room temperature, centrifugation then, centrifuge speed 3500rpm/min obtains spirulina protein weak brine extracting solution.Spirulina material slag through extracting after centrifugal is extracted centrifugal 2 times by above-mentioned condition again, obtains spirulina protein weak brine extracting solution respectively, and with the spirulina protein weak brine extracting solution that obtains for 3 times merge, standby.
The spirulina protein weak brine extracting solution of gained is regulated pH4.1, pH 3.9 and pH 3.7 successively with the HCl solution of 2.0mol/L, it is heavy to carry out 3 segmentations acid, leave standstill 20min at every turn, isolate spirulina Acid precipitation albumen with the rotating speed of 5000rpm/min with whizzer, the heavy albumen of 3 hypo acids is merged, with with the water washing of volume 2 times, Acid precipitation albumen is modulated into aqueous dispersions, and regulating this Acid precipitation albumen dispersion liquid pH with the NaOH solution of 0.5mol/L again is 7.0;
With the pH of gained is that 7.0 spirulina Acid precipitation albumen dispersion liquid is in ultrahigh-temperature (UHT) instantaneous sterilizing device, 4s sterilizes under 136 ℃ temperature, be cooled to 55 ℃ rapidly, carry out spraying drying then, spray-dired processing parameter is: feeding temperature is 70 ℃, the hot air inlet temperature is 170 ℃, and temperature out is 80 ℃, so far promptly obtains product spirulina separated protein powder.
Embodiment 3:
Be to handle under the condition of 7500rpm/min at rotating speed with spirulina (algae mud) feed liquid with the high speed colloidal mill, carry out cytoclasis, in cytoclastic spirulina algae pug liquid, add gasoline then, gasoline and feed liquid add in 1: 5 ratio, stir or vibration, extraction pigment and fishy smell material, extraction times is 3 times, remove the petrolic solvent layer that contains pigment and fishy smell material, is that 0.07MPa outgas with vacuum tightness with lower floor's spirulina cells broken material in vacuum degassing machine, promptly get the broken feed liquid of decoloration deodorization spirulina cells after the degassing, standby;
The decoloration deodorization spirulina cells broken material liquid of gained is added Na
2HPO
4, be adjusted to Na
2HPO
4Concentration is the dispersion liquid of 0.20mol/L, and making spirulina cells broken material and weak brine ratio is 1: 20, is under 50 ℃ the condition in temperature, extract 1 hour postcooling to room temperature, centrifugation then, centrifuge speed 4000rpm/min obtains spirulina protein weak brine extracting solution.Spirulina material slag through extracting after centrifugal is extracted centrifugal 1 time by above-mentioned condition again, obtains spirulina protein weak brine extracting solution respectively, and with the spirulina protein weak brine extracting solution that obtains for 2 times merge, standby.
The spirulina protein weak brine extracting solution of gained is regulated pH4.2, pH 4.0 and pH 3.5 successively with the HCl solution of 2.0mol/L, it is heavy to carry out 3 segmentations acid, leave standstill 25min at every turn, isolate spirulina Acid precipitation albumen with the rotating speed of 6000rpm/min with whizzer, the heavy albumen of 3 hypo acids is merged, with with the water washing of volume 2 times, Acid precipitation albumen is modulated into aqueous dispersions, and regulating this Acid precipitation albumen dispersion liquid pH with the NaOH solution of 0.5mol/L again is 7.0;
With the pH of gained is that 7.0 spirulina Acid precipitation albumen dispersion liquid is in ultrahigh-temperature (UHT) instantaneous sterilizing device, 3s sterilizes under 137 ℃ temperature, be cooled to 55 ℃ rapidly, carry out spraying drying then, spray-dired processing parameter is: feeding temperature is 60 ℃, the hot air inlet temperature is 180 ℃, and temperature out is 80 ℃, so far promptly obtains product spirulina separated protein powder.
Claims (5)
1. the preparation method of a spirulina protein isolate is characterized in that: may further comprise the steps:
(1) preparation of the broken decoloration deodorization degreasing of spirulina cells material
Spirulina raw material is carried out micronizing handle, make the cell walls fragmentation, add organic solvent then, stir or vibration, extraction pigment, lipid and fishy smell material are removed organic solvent layer, the cytoclasis material is carried out vacuum outgas, be prepared into decoloration deodorization degreasing spirulina cells broken material liquid;
(2) rare salts solution extracts albumen and centrifugal treating
The decoloration deodorization degreasing spirulina cells broken material liquid of gained with salt, is adjusted to rare salt dispersion liquid; With salt is that salt is adjusted to weak brine, and making spirulina cells broken material and weak brine ratio is 1: 10～20; Be under 40～60 ℃ the condition in temperature, rare salt dispersion liquid of gained is extracted 1～2 hour postcooling to room temperature, centrifugation then makes spirulina protein weak brine extracting solution; Spirulina material slag through extracting after centrifugal is extracted centrifugal 2 times by above-mentioned condition again, obtains spirulina protein weak brine extracting solution respectively, and with the spirulina protein weak brine extracting solution that obtains for 3 times merge, standby;
(3) pH gradient segmentation acid adjustment protein precipitation is neutral with adjusting
It is acid that the spirulina protein weak brine extracting solution of gained is regulated pH with HCl solution, adopt pH gradient segmentation acid heavy, leave standstill, make albumen precipitation complete, centrifugation goes out spirulina Acid precipitation albumen, Acid precipitation albumen is scattered in it in water after washing again, and the pH that regulates this spirulina Acid precipitation albumen dispersion liquid with NaOH solution is 7.0;
(4) Ultra High Temperature Short Time and spraying drying
With the pH of gained is that 7.0 spirulina Acid precipitation albumen dispersion liquid carries out Ultra High Temperature Short Time, and spraying drying is carried out in cooling then, promptly gets product spirulina protein isolate.
2. the preparation method of a kind of spirulina protein isolate according to claim 1, it is characterized in that the micronizing that spirulina raw material is carried out described in the step (1) handles, be to adopt the ultra-high speed milling treatment of colloid to make the spirulina cells fragmentation, the rotating speed of ultra-high speed colloidal mill is 6000～8500rpm/min; Described organic solvent is a kind of in sherwood oil, normal hexane and the gasoline or their mixture; When extraction pigment and fishy smell material, the mass volume ratio of organic solvent and spirulina cells broken material is 1: 1～10, and extraction times is 2～3 times; The vacuum tightness of described vacuum outgas is 0.07～0.09MPa.
3. the preparation method of a kind of spirulina protein isolate according to claim 2 is characterized in that being meant with salt described in the step (2) adds NaCl, KCl, MgCl
2, Na
2SO
4, NH
4SO
4, Na
2CO
3, Na
2HPO
4In a kind of or their mixture; The concentration of salt is 0.02～0.6mol/L in described rare salt dispersion liquid; Described centrifugal rotational speed is 3000～4500rpm/min.
4. the preparation method of a kind of spirulina protein isolate according to claim 3, the concentration that it is characterized in that HCl solution described in the step (3) is 2.0mol/L, regulating the pH value is 3.5～4.2, pH is changed to low in gradient from height, divide 3 precipitating proteins, precipitate time of repose is 10～30min at every turn; The centrifugation rotating speed is 4000～6000rpm/min; The water washing number of times is 2～3 times, and the NaOH strength of solution is 0.5～2.0mol/L.
5. the preparation method of a kind of spirulina protein isolate according to claim 4 is characterized in that the processing parameter of Ultra High Temperature Short Time described in the step (4) is: 135～143 ℃ of sterilising temps, and the time is 2～15s; Described spray-dired processing parameter is: feeding temperature is 60～80 ℃, and the hot air inlet temperature is 160～200 ℃, and temperature out is 80～90 ℃.
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| CN102645357A (en) * | 2012-03-27 | 2012-08-22 | 浙江大学 | Method for preparing spirulina dielectrophoresis analysis protein |
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