CN102020705B - Method for preparing spirulina protein isolate - Google Patents
Method for preparing spirulina protein isolate Download PDFInfo
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- CN102020705B CN102020705B CN2009100920991A CN200910092099A CN102020705B CN 102020705 B CN102020705 B CN 102020705B CN 2009100920991 A CN2009100920991 A CN 2009100920991A CN 200910092099 A CN200910092099 A CN 200910092099A CN 102020705 B CN102020705 B CN 102020705B
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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 tenaculat Habenaria protein isolates, belong to the Food Science and Engineering technical field.
Background technology:
Tenaculat Habenaria (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.Tenaculat Habenaria originates in Africa and NA, 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 tenaculat Habenaria 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 tenaculat Habenaria is because of kind difference difference to some extent, contains extremely that rich in protein is one of prominent feature of nutritive component of spirulina, and the protein of spirulina powder is about 50%～70%%; And the amino acid of hydrolysis of protein spirulina composition A wide selection of colours and designs, 8 kinds of indispensable amino acid balanced proportion of human body need pattern very for similar with 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(Phe) 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 tenaculat Habenaria is a kind of high-quality full price nutrient protein source, the contained Mierocrystalline cellulose of its cell walls is less, and its proteinic digestibility can reach 75.0%; Bioavailability can reach 68.0%, still, no matter is directly edible; Still use as protein supplements, all exist some limitation: the first, the kind of tenaculat Habenaria product is comparatively single; Form with sheet or pill occurs mostly, is prone to cause the sensation of taking medicine to the human consumer; The second, tenaculat Habenaria 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 contnt, digestibility and bioavailability.These have brought considerable restraint all for tenaculat Habenaria processing products and comprehensive utilization.In view of this; The present invention carries out cytoclasis, decolouring, degreasing with tenaculat Habenaria and takes off raw meat and handle, and preparation becomes protein isolates, 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 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 deficiency of prior art; The object of the present invention is to provide a kind of preparation method of tenaculat Habenaria protein isolates; Use that the tenaculat Habenaria protein isolates that this method produces has lighter color, fishy smell is light, protein contnt is high, the good and digestible advantage of solvability, can be used as a kind of high-quality complete protein supplementary or additive and use.
The present invention realizes through following technical scheme:
A kind of preparation method of tenaculat Habenaria protein isolates may further comprise the steps:
(1) preparation of the broken decoloration deodorization degreasing of spirulina cells material
Adopting the ultra-high speed colloidal mill to carry out micronizing spirulina raw material (algae mud) handles; Make cell walls broken, 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; 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, spinning then makes spirulina protein weak brine extracting solution.To extract centrifugal 2 times by above-mentioned condition again through the tenaculat Habenaria material slag that extracts after centrifugal, obtain spirulina protein weak brine extracting solution respectively, and with the spirulina protein weak brine extracting solution that obtains for 3 times merge, subsequent use.
(3) pH gradient segmentation acid adjustment protein precipitation is neutral with adjusting
Use HCl solution adjusting pH to be acidity the spirulina protein weak brine extracting solution of gained; Adopt pH gradient segmentation acid heavy, leave standstill, make albumen precipitation complete; Spinning goes out tenaculat Habenaria Acid precipitation albumen; Acid precipitation albumen is scattered in it in water after washing again, and the pH that uses NaOH solution to regulate this tenaculat Habenaria Acid precipitation albumen dispersion liquid is 7.0;
(4) Ultra High Temperature Short Time and spraying drying
With the pH of gained is that 7.0 tenaculat Habenaria Acid precipitation albumen dispersion liquid carries out Ultra High Temperature Short Time, and spraying drying is carried out in cooling then, promptly gets product tenaculat Habenaria protein isolates.
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 spirulina cells broken, and the rotating speed of ultra-high speed colloidal mill is 6000～8500rpm; 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.
The concentration of the said 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 from height in gradient, divides 3 times precipitating proteins, and precipitate time of repose is 10～30min at every turn; The spinning rotating speed is 4000～6000rpm; 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 said Ultra High Temperature Short Time of step (4) is: 135～143 ℃ of sterilising temps, and the time is 2～15s; Said 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 that spirulina cells is broken; 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 tenaculat Habenaria material that cytoclasis and organic solvent extraction are prepared into, the stripping of tenaculat Habenaria protein isolates when helping strengthening weak brine and extracting; Through weak brine extraction, pH gradient segmentation acid adjustment deposition, Ultra High Temperature Short Time and spray drying treatment, make protein that sex change to a certain degree take place, improved the digestibility and the solvability of protein isolates.
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, higher than water, diluted acid water, diluted alkaline water and extraction using alcohol albumen solubility rate, sex change is littler.
4, in the iso-electric point scope, adopt the pH gradient segmentation acid adjustment precipitator method to make the protein multiple spot separate out, fully precipitating proteins further improves recovery of protein rate in the extracting solution.
Embodiment:
The present invention is further described the present invention through following examples:
Embodiment 1:
Use the high speed colloidal mill under the condition of rotating speed, to handle tenaculat Habenaria (algae mud) feed liquid as 6500rpm; Carry out cytoclasis, in cytoclastic tenaculat Habenaria 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, removes 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, is under 60 ℃ the condition in temperature; Extract 2 hours postcooling to room temperature; Spinning then, centrifuge speed are 3000rpm, obtain spirulina protein weak brine extracting solution.To extract centrifugal 2 times by above-mentioned condition again through the tenaculat Habenaria material slag that extracts after centrifugal, obtain spirulina protein weak brine extracting solution respectively, and with the spirulina protein weak brine extracting solution that obtains for 3 times merge, subsequent use.
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 tenaculat Habenaria Acid precipitation albumen with the rotating speed of 4000rpm, the heavy albumen of 3 hypo acids is merged, use with the water washing of volume 2 times with whizzer; Acid precipitation albumen is modulated into aqueous dispersions, and using this Acid precipitation albumen dispersion liquid of NaOH solution adjusting pH of 0.5mol/L again is 7.0;
With the pH of gained is that 7.0 tenaculat Habenaria Acid precipitation albumen dispersion liquid is in ultrahigh-temperature (UHT) instantaneous sterilizing device; The 5s that under 135 ℃ temperature, sterilizes is cooled to 55 ℃ rapidly, carries 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 tenaculat Habenaria separated protein powder.
Embodiment 2:
Use the high speed colloidal mill under the condition of rotating speed, to handle tenaculat Habenaria (algae mud) feed liquid as 7000rpm; Carry out cytoclasis, in cytoclastic tenaculat Habenaria 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, removes 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; Spinning then, centrifuge speed 3500rpm obtains spirulina protein weak brine extracting solution.To extract centrifugal 2 times by above-mentioned condition again through the tenaculat Habenaria material slag that extracts after centrifugal, obtain spirulina protein weak brine extracting solution respectively, and with the spirulina protein weak brine extracting solution that obtains for 3 times merge, subsequent use.
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 tenaculat Habenaria Acid precipitation albumen with the rotating speed of 5000rpm, the heavy albumen of 3 hypo acids is merged, use with the water washing of volume 2 times with whizzer; Acid precipitation albumen is modulated into aqueous dispersions, and using this Acid precipitation albumen dispersion liquid of NaOH solution adjusting pH of 0.5mol/L again is 7.0;
With the pH of gained is that 7.0 tenaculat Habenaria Acid precipitation albumen dispersion liquid is in ultrahigh-temperature (UHT) instantaneous sterilizing device; The 4s that under 136 ℃ temperature, sterilizes is cooled to 55 ℃ rapidly, carries 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 tenaculat Habenaria separated protein powder.
Embodiment 3:
Use the high speed colloidal mill under the condition of rotating speed, to handle tenaculat Habenaria (algae mud) feed liquid, carry out cytoclasis, in cytoclastic tenaculat Habenaria algae pug liquid, add gasoline then as 7500rpm; 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 gets the broken feed liquid of decoloration deodorization spirulina cells after the degassing, subsequent use;
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; Spinning then, centrifuge speed 4000rpm obtains spirulina protein weak brine extracting solution.To extract centrifugal 1 time by above-mentioned condition again through the tenaculat Habenaria material slag that extracts after centrifugal, obtain spirulina protein weak brine extracting solution respectively, and with the spirulina protein weak brine extracting solution that obtains for 2 times merge, subsequent use.
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 tenaculat Habenaria Acid precipitation albumen with the rotating speed of 6000rpm, the heavy albumen of 3 hypo acids is merged, use with the water washing of volume 2 times with whizzer; Acid precipitation albumen is modulated into aqueous dispersions, and using this Acid precipitation albumen dispersion liquid of NaOH solution adjusting pH of 0.5mol/L again is 7.0;
With the pH of gained is that 7.0 tenaculat Habenaria Acid precipitation albumen dispersion liquid is in ultrahigh-temperature (UHT) instantaneous sterilizing device; The 3s that under 137 ℃ temperature, sterilizes is cooled to 55 ℃ rapidly, carries 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 tenaculat Habenaria separated protein powder.
Claims (5)
1. the preparation method of a tenaculat Habenaria protein isolates is characterized in that: may further comprise the steps:
(1) preparation of the broken decoloration deodorization degreasing of spirulina cells material
Adopting the ultra-high speed colloidal mill to carry out micronizing spirulina raw material handles; Make cell walls broken, 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; 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, spinning then makes spirulina protein weak brine extracting solution; To extract centrifugal 2 times by above-mentioned condition again through the tenaculat Habenaria material slag that extracts after centrifugal, obtain spirulina protein weak brine extracting solution respectively, and with the spirulina protein weak brine extracting solution that obtains for 3 times merge, subsequent use;
(3) pH gradient segmentation acid adjustment protein precipitation is neutral with adjusting
Use HCl solution adjusting pH to be acidity the spirulina protein weak brine extracting solution of gained; Adopt pH gradient segmentation acid heavy, leave standstill, make albumen precipitation complete; Spinning goes out tenaculat Habenaria Acid precipitation albumen; Acid precipitation albumen is scattered in it in water after washing again, and the pH that uses NaOH solution to regulate this tenaculat Habenaria Acid precipitation albumen dispersion liquid is 7.0;
(4) Ultra High Temperature Short Time and spraying drying
With the pH of gained is that 7.0 tenaculat Habenaria Acid precipitation albumen dispersion liquid carries out Ultra High Temperature Short Time, and spraying drying is carried out in cooling then, promptly gets product tenaculat Habenaria protein isolates.
2. the preparation method of a kind of tenaculat Habenaria protein isolates 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 spirulina cells broken, the rotating speed of ultra-high speed colloidal mill is 6000～8500rpm; 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 tenaculat Habenaria protein isolates 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.
4. the preparation method of a kind of tenaculat Habenaria protein isolates 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 from height in gradient, divide 3 times precipitating proteins, precipitate time of repose is 10～30min at every turn; The spinning rotating speed is 4000～6000rpm; 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 tenaculat Habenaria protein isolates 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; Said 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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