CN102978186A - Method for preparing lysozyme, sensory basic flavoring and egg oil by using salted egg - Google Patents
Method for preparing lysozyme, sensory basic flavoring and egg oil by using salted egg Download PDFInfo
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- CN102978186A CN102978186A CN2012105011658A CN201210501165A CN102978186A CN 102978186 A CN102978186 A CN 102978186A CN 2012105011658 A CN2012105011658 A CN 2012105011658A CN 201210501165 A CN201210501165 A CN 201210501165A CN 102978186 A CN102978186 A CN 102978186A
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Abstract
The invention discloses a method for preparing lysozyme, sensory basic flavoring and egg oil by using a salted egg. According to the method, by using salted egg white or salted duck egg white as a raw material, egg white precipitation is subjected to acid and thermal treatment, egg white enzymatic hydrolysis sensitivity is improved, and liquor containing lysozyme and protein precipitation are obtained through centrifugal filtration. The liquor is subjected to ultrafiltration and freeze-drying so as to obtain the lysozyme. The protein precipitation is hydrolyzed by using protease to obtain the sensory basic flavoring and egg oil with high hydrolysis degree and high protein recovering rate, and finally the purpose of comprehensively utilizing the salted egg white at a higher degree is achieved.
Description
Technical field
The present invention relates to the technical field of the deep processing of salted egg albumen, be specifically related to utilize salted egg albumen to produce the preparation method of N,O-Diacetylmuramidase, base of flavour development and Oil of egg yolk.
Background technology
Salted egg albumen is the by product of preserved egg yellow Production of Moon Cake, and nearly ten thousand tons of annual production generally are light yellow, mainly is owing to unavoidably cause a part of preserved egg yellow to be sneaked in the egg white in salted egg albumen and the preserved egg yellow sepn process.Because the production time of moon cake relatively concentrates, quantity is large, brings great difficulty for the processing of salted egg albumen.In the salted egg albumen protein content (N,O-Diacetylmuramidase that comprises 0.1-0.2%), salt content about 9-11% about 10%, Oil of egg yolk content is at 1-4%, is the low value resource of production N,O-Diacetylmuramidase, protein and the Oil of egg yolk of high-quality.Yet meat product is made weighting agent to salted egg albumen or as the feed, most abandoneds cause serious pollution to surrounding environment, water source except part is used for all the time.Utilize at present egg white or salted egg albumen to produce the existing many pieces of bibliographical informations of N,O-Diacetylmuramidase, disclose a kind of employing deamination acylation chitosan gel affinity adsorption such as patent " from salted egg albumen, extracting N,O-Diacetylmuramidase method (number of patent application 01107411.6) " and from salted egg albumen, extracted the N,O-Diacetylmuramidase method." extraction of lysozyme from egg white and the comprehensive utilization of albumen (number of patent application: 200710187139.1) " discloses a kind of novel method of extracting N,O-Diacetylmuramidase from birds, beasts and eggs (egg, duck's egg or goose egg), unites by series of process such as the separation of hydrochlorate (hydrochloric acid or lactic acid, Repone K) temperature control combined method enzyme extraction, cooling, centrifugal, adjust pH, the concentrated purification of hollow fiber ultrafiltration membrane method enzyme, spray-drying processes and produces egg white lysozyme.
Take egg white as raw material, preparing anti-oxidation peptide, Antihypertensive Peptides or egg white hydrolysate by protease hydrolysis also has many pieces of bibliographical informations both at home and abroad.But because egg white contains multiple to the inhibited material of proteolytic enzyme, such as the solid albumen of ovomucoid, ovum protein enzyme inhibitor, L-Cysteine HCL Anhydrous inhibition and ovum, cause above-mentioned research all to have the problems such as protein degree is low, the zymin consumption is large, protein recovery is low, production cost is high.
Extracting at present the Oil of egg yolk main method both at home and abroad has: organic solvent method, adopted by industrial, and its Oil of egg yolk quality product is better, and extraction yield is higher, but has certain dissolvent residual, and especially some solvent also has toxicity, in addition its complex process, time-consuming; Dry distillation is China's folk tradition method, and its preparation is easy, but oil yield is very low, second-rate, and a large amount of productions easily make environmental pollution; Supercritical carbon dioxide extraction method is a modern high technology, can be from the yolk powder raw material effective separation go out not contain Oil of egg yolk, egg phospholipids and the yolk protein of phosphatide, its good product quality, but the method is very high to yolk powder raw materials quality and the requirement of extraction plant pressure withstanding degree, and production cost is higher.In recent years, the existing novel process research that utilizes modern biotechnology Enzymatic Extraction Oil of egg yolk of Chinese scholars, adopt protease A and Cathepsin B from dried hen egg yolk, to extract Oil of egg yolk (Wuxi Light Industry Univ.'s journal, 2003,22 (1): 102-105) such as people such as Wang Hui.Have not yet to see the report that utilizes proteolytic enzyme from salted egg albumen, to reclaim Oil of egg yolk.
Summary of the invention
The object of the invention provides a kind of method of utilizing salted egg albumen to prepare N,O-Diacetylmuramidase, base of flavour development and Oil of egg yolk.
Utilize salted egg albumen to prepare the method for N,O-Diacetylmuramidase, base of flavour development and Oil of egg yolk, comprise the steps:
(1) salted egg albumen is mixed with the water of 1-5 times of weight after gauze filters, and adjust pH is to 2-4, and 60 ℃ of-80 ℃ of thermal treatment 10min-30min are centrifugal, filter to get supernatant liquor and albumen precipitation;
(2) described supernatant liquor is crossed the ultra-filtration membrane that molecular weight is 10000Da, gets the liquid that dams, and lyophilize gets N,O-Diacetylmuramidase; Add aspartic protease in the described albumen precipitation, 45 ℃-60 ℃ are hydrolyzed 24-72 hour, centrifugal, get subnatant and Oil of egg yolk;
(3) the subnatant adjust pH is to 5.5-7.5, and the enzyme that goes out filters, and gets base of flavour development.
Further optimize, the described pH value of step (1) adopts hydrochloric acid, citric acid or phosphoric acid solution to regulate.
Further optimize, the described fungi aspartic protease of step (2) addition is the 1%-3% of the albumen weight of substrate protein.
Further optimize, the described centrifugal condition of step (2) is the centrifugal 15min-30min of 5000g-10000g.
Further optimize, the described enzyme condition of going out of step (3) is 70 ℃-90 ℃ and processes 15min-30min.
Further optimize, the described pH value of step (3) adopts sodium hydroxide or potassium hydroxide solution to regulate.
Further optimize, step (2) also to described albumen precipitation squeeze to albumen precipitation moisture weight content be 70% ~ 90%.
The present invention has adopted above technical scheme, compared with prior art has following advantage and effect:
(1) the invention provides the method that a kind of more easy full use salted egg albumen prepares N,O-Diacetylmuramidase, base of flavour development and Oil of egg yolk, the method technique is simple, easy to operate, is suitable for suitability for industrialized production.
(2) contriver finds under acidic conditions salted egg albumen to be heat-treated, not only can make ovomacroglobulin (proteinase inhibitor) inactivation that contains in the salted egg albumen, and can make the fine and close protein structure of egg white script become loose, peptide bond exposes, and is conducive to the effect of proteolytic enzyme.This technique can significantly improve the albuminised degree of hydrolysis of salted egg and protein recovery, reduces the zymin consumption, and provides the basis for the centrifugation of base of flavour development and Oil of egg yolk.
(3) the present invention heat-treats salted egg albumen under acidic conditions, and gained precipitation albumen pH value is added the fungi aspartic protease and is hydrolyzed for a long time substantially risk without microbial contamination between 3-4.
Embodiment
Below in conjunction with example enforcement of the present invention is described further, but the invention process and protection domain are not limited to this.
The measuring method of N,O-Diacetylmuramidase enzyme activity: turbidimetry
(1) preparation of substrate: with 0.2mol/L pH6.2 phosphate buffered saline buffer the micrococcus lysodeikticus thalline on the solid slant culture base is washed out, make suspension, be diluted to OD450 and be 0.8 ~ 0.9, now with the current.
(2) sample tests: get an amount of sample dissolution in 0.2mol/L pH6.2 phosphate buffered saline buffer, then place sample solution and micrococcus lysodeikticus suspension the 25C water-bath to be incubated, reaction system sees Table 1, reading A1, A2 when writing down reaction 15s and 75s, descending by per minute OD450 value 0.001 is the definition of 1 unit of activity (U), and every milligram of bacteriolyze enzyme activity calculates with following formula in the sample solution:
Wherein, Δ E
450Poor for 450nm wavelength place absorbance A 1, A2, C is the content (mg/mL) of N,O-Diacetylmuramidase in the sample solution.
Table 1 egg white solution assaying reaction system mL
Oil of egg yolk yield=Oil of egg yolk weight/salted egg albumen weight * 100%
Degree of hydrolysis=hydrolyzed solution ammonia-nitrogen content/hydrolyzed solution total nitrogen content * 100%
Protein recovery=hydrolyzed solution total nitrogen content/salted egg albumen total nitrogen content * 100%
Embodiment 1
A kind of method of utilizing salted egg albumen to prepare N,O-Diacetylmuramidase, base of flavour development and Oil of egg yolk:
(1) salted egg albumen is purchased in the market of farm produce, Guangzhou.Get the salted egg albumen 100g after the filtered through gauze, add 400g water and its mixing, with 1mol/L hydrochloric acid soln accent pH to 4.0,60 ℃ of thermal treatment 30min, centrifugal, filter to get supernatant liquor and albumen precipitation;
(2) supernatant liquor is crossed the ultra-filtration membrane that molecular weight is 10000Da, gets the liquid that dams, and lyophilize gets N,O-Diacetylmuramidase, and recording antalzyme activity with turbidimetry is 23000U/mg;
(3) to albumen precipitation squeeze to the albumen precipitation moisture content be 90%, add the fungi aspartic protease, addition is 3% of substrate protein weight, 60 ℃ of lower hydrolysis 36 hours, the centrifugal 20min of 8000g gets subnatant and Oil of egg yolk, and wherein the degree of hydrolysis of clear liquid is 38.3%, protein recovery is 88.7%, and the Oil of egg yolk yield is 3.3%;
(4) the subnatant sodium hydroxide solution adjust pH to 6.0 of 1mol/L, 70 ℃ of enzyme 30min that go out filter, and get base of flavour development, find that by subjective appreciation its delicate flavour is stronger.The salt content of base of flavour development only is 1.6%.
Embodiment 2
A kind of method of utilizing salted egg albumen to prepare N,O-Diacetylmuramidase, base of flavour development and Oil of egg yolk:
(1) get salted egg albumen 200g after the filtered through gauze, add 600g water and its mixing, with 1mol/L citric acid solution accent pH to 3.5,70 ℃ of thermal treatment 25min, centrifugal, filter to get supernatant liquor and albumen precipitation;
(2) supernatant liquor is crossed the ultra-filtration membrane that molecular weight is 10000Da, gets the liquid that dams, and lyophilize gets N,O-Diacetylmuramidase, and recording antalzyme activity with turbidimetry is 21000U/mg;
(3) to albumen precipitation squeeze to the albumen precipitation moisture content be 80%, add the fungi aspartic protease in the albumen precipitation, addition is 2% of substrate protein, 55 ℃ of hydrolysis 72 hours, centrifugal, get subnatant and Oil of egg yolk, wherein degree of hydrolysis is 47.9%, protein recovery is 73.5%, and the Oil of egg yolk yield is 3.8%;
(4) the subnatant potassium hydroxide solution adjust pH to 5.5 of 1mol/L, 80 ℃ of enzyme 20min that go out filter, and get base of flavour development, find that by subjective appreciation its delicate flavour is stronger.The salt content of base of flavour development only is 2.1%.
Embodiment 3
A kind of method of utilizing salted egg albumen to prepare N,O-Diacetylmuramidase, base of flavour development and Oil of egg yolk:
(1) get salted egg albumen 1kg after the filtered through gauze, add 2kg water and its mixing, with 1mol/L phosphoric acid solution accent pH to 3.0,75 ℃ of thermal treatment 20min, centrifugal, filter to get supernatant liquor and albumen precipitation;
(2) supernatant liquor is crossed the ultra-filtration membrane that molecular weight is 10000Da, gets the liquid that dams, and lyophilize gets N,O-Diacetylmuramidase, and recording antalzyme activity with turbidimetry is 20500U/mg;
(3) to albumen precipitation squeeze to the albumen precipitation moisture content be 75%, add the fungi aspartic protease in the albumen precipitation, addition is 2.5% of substrate protein, 50 ℃ of hydrolysis 60 hours, centrifugal, get subnatant and Oil of egg yolk, wherein degree of hydrolysis is 41.1%, protein recovery is 85.6%, and the Oil of egg yolk yield is 3.9%;
(4) the subnatant sodium hydroxide solution adjust pH to 7.0 of 1mol/L, 80 ℃ of enzyme 30min that go out filter, and get base of flavour development, find that by subjective appreciation its delicate flavour is stronger.The salt content of base of flavour development only is 2.3%.
Embodiment 4
A kind of method of utilizing salted egg albumen to prepare N,O-Diacetylmuramidase, base of flavour development and Oil of egg yolk:
(1) get salted egg albumen 2kg after the filtered through gauze, add 4kg water and its mixing, with 1mol/L hydrochloric acid soln accent pH to 3.0,70 ℃ of thermal treatment 30min, centrifugal, filter to get supernatant liquor and albumen precipitation;
(2) supernatant liquor is crossed the ultra-filtration membrane that molecular weight is 10000Da, gets the liquid that dams, and lyophilize gets N,O-Diacetylmuramidase, and recording antalzyme activity with turbidimetry is 19500U/mg;
(3) to albumen precipitation squeeze to the albumen precipitation moisture content be 80%, add the fungi aspartic protease in the albumen precipitation, addition is 3% of substrate protein, 55 ℃ of hydrolysis 48 hours, centrifugal, get subnatant and Oil of egg yolk, wherein degree of hydrolysis is 39.6%, protein recovery is 82.7.6%, and the Oil of egg yolk yield is 3.6%;
(4) the subnatant potassium hydroxide solution adjust pH to 6.0 of 1mol/L, 90 ℃ of enzyme 10min that go out filter, and get base of flavour development, find that by subjective appreciation its delicate flavour is stronger.The salt content of base of flavour development only is 2.4%.
Claims (7)
1. utilize salted egg albumen to prepare the method for N,O-Diacetylmuramidase, base of flavour development and Oil of egg yolk, it is characterized in that comprising the steps:
(1) salted egg albumen is mixed with the water of 1-5 times of weight after gauze filters, and adjust pH is to 2-4, and 60 ℃-80 ℃ thermal treatment 10 min-30 min are centrifugal, filter to get supernatant liquor and albumen precipitation;
(2) described supernatant liquor is crossed the ultra-filtration membrane that molecular weight is 10000Da, gets the liquid that dams, and lyophilize gets N,O-Diacetylmuramidase; Add the fungi aspartic protease in the described albumen precipitation, 45 ℃-60 ℃ are hydrolyzed 24-72 hour, centrifugal, get subnatant and Oil of egg yolk;
(3) the subnatant adjust pH is to 5.5-7.5, and the enzyme that goes out filters, and gets base of flavour development.
2. method according to claim 1 is characterized in that the described pH value of step (1) adopts hydrochloric acid, citric acid or phosphoric acid solution to regulate.
3. method according to claim 1 is characterized in that the described fungi aspartic protease of step (2) addition is the 1%-3% of the albumen weight of substrate protein.
4. method according to claim 1 is characterized in that the described centrifugal condition of step (2) is the centrifugal 15min-30min of 5000g-10000g.
5. method according to claim 1 is characterized in that the described enzyme condition of going out of step (3) is 70 ℃-90 ℃ and processes 15min-30min.
6. method according to claim 1 is characterized in that the described pH value of step (3) adopts sodium hydroxide or potassium hydroxide solution to regulate.
7. method according to claim 1, it is characterized in that step (2) also to described albumen precipitation squeeze to albumen precipitation moisture weight content be 70% ~ 90%.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104164298A (en) * | 2014-07-22 | 2014-11-26 | 山西中医学院 | Preparation method of yolk oil containing Harman alkaloid |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104164298A (en) * | 2014-07-22 | 2014-11-26 | 山西中医学院 | Preparation method of yolk oil containing Harman alkaloid |
| CN104164298B (en) * | 2014-07-22 | 2016-04-27 | 山西中医学院 | A kind of Oil of egg yolk preparation method containing Harman alkaloid |
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Application publication date: 20130320 Assignee: Tai Hui bio tech ltd, Guangzhou Assignor: South China University of Technology Contract record no.: 2015440000625 Denomination of invention: Method for preparing lysozyme, sensory basic flavoring and egg oil by using salted egg Granted publication date: 20150311 License type: Exclusive License Record date: 20150916 |
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Effective date of registration: 20170508 Address after: 511458 A1 building, building 4, Nansha science and technology innovation center, Guangzhou, Guangdong, Nansha District, China Patentee after: Tai Hui bio tech ltd, Guangzhou Address before: 510640 Tianhe District, Guangdong, No. five road, No. 381, Patentee before: South China University of Technology |