CN107557417A - Yeast polypeptides and its preparation method and application - Google Patents
Yeast polypeptides and its preparation method and application Download PDFInfo
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- CN107557417A CN107557417A CN201610510577.6A CN201610510577A CN107557417A CN 107557417 A CN107557417 A CN 107557417A CN 201610510577 A CN201610510577 A CN 201610510577A CN 107557417 A CN107557417 A CN 107557417A
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Abstract
The present invention relates to yeast polypeptides field, and in particular to a kind of yeast polypeptides and its preparation method and application.The preparation method of the yeast polypeptides, comprises the following steps：(1) Yeast protein in raw material is extracted；(2) alkali protease enzymolysis is added, then adds compound protease enzymolysis, supernatant is collected and obtains enzymolysis product；(3) separate, decolourize, obtain yeast polypeptides.The yeast polypeptides obtained using the present invention are colourless, tasteless, content of peptides more than 80%, and more than 85% polypeptide molecular weight below 2000, it is practically free of free amino acid, and the bitter taste having without peptide products such as general Soybean Peptides, available for food and health products, flavour of food products can be improved, help postexercise recovery muscle power etc..And if the yeast polypeptides that the present invention obtains dust without drying, are a excellent cosmetics Essence, have and nourish antiwrinkle active, may apply in cosmetics, color, perfume (or spice) without influenceing cosmetics etc., and there is senile-resistant efficacy.
Description
Technical field
The present invention relates to polypeptide field, and in particular to a kind of yeast polypeptides and preparation method and application.
Background technology
The molecular weight of peptide is typically between 180-5000Da.Peptide of its middle-molecular-weihydroxyethyl between 180-1000Da is referred to as small peptide
Or oligopeptides or oligopeptide, also referred to as micromolecule active polypeptide.Polypeptide has some physiological activities, including immunological regulation, antibacterial,
Antitumor and raising immunity of organisms etc..It is considered that protein be only digested breaks down into amino acids into human body could be by people
Body is absorbed, and research recently is found really not so.Human body takes in protein after digesting enzyme effect, not only with free amino acid
Form absorb, be more in the form of small peptide absorb.And small peptide specific ionization amino acid digestion is faster, more, this table is absorbed
The biological value and nutritive value of bright peptide are higher than free amino acid.
Research to polypeptide at present is mainly based upon the polypeptide of plant origin and animal origin, for example, Soybean Peptide, corn peptide,
Small beef liver peptide etc..It was verified that addition polypeptide can change the quality of food, flavor, it can also increase feature.Such as Soybean Peptide
Cholesterol generation can be suppressed, increase lipid-metabolism；Pea peptide is good milk allergy resistance agent.Except these plant sources
Outside polypeptide and animal sources polypeptide, also antimicrobial polypeptide, such as yeast polypeptides.Compared with the former two, the albumen of yeast sources or more
Peptide also has more advantages：In the absence of pollution by pesticides, do not influenceed by weather and seasonal variations, be not in may have to human body
Hormone, antibiotic and artificial color of influence etc., in the absence of transgenosis problem, also in the absence of avian influenza virus, crazy heifer disease virus,
The pollution channels such as hepatitis B.
Presently commercially available yeast polypeptides powder is to decompose its cell membrane broken wall using yeast bio self-dissolving, then through digesting, spraying
Mist is dried and formed, in faint yellow powdered to yellow.In addition to polypeptide, also a large amount of undecomposed protein, zymosan,
The materials such as amino acid, nucleotides and vitamin.The content of polypeptide wherein is at most 50% or so, and this polypeptide powder is mainly used in
Animal feed.The plant polypeptide of high-purity be widely used in food and health products and yeast polypeptides be used for human food substantially not
See.
Yeast polypeptides cannot be only used for feed, food, also have the effect of fine applied to cosmetic field.Small molecule is more
Peptide can directly be absorbed by skin, supplement nutrition needed for skin, improve Skin Cell immunity, reduce wound of the external environment to skin
Evil.In addition, hair product is applied also for, addition polypeptide can nourish reparation damaged hair in shampoo or hair conditioner,
Reduce withered bifurcated.
Document and patent on the yeast polypeptides this respect of preparation high-purity are delivered seldom, patent CN102550802A
A kind of method that polypeptide and amino acid are extracted from beer waste yeast is disclosed, the scheme digested using high-pressure homogeneous rear extracting,
Finally obtain polypeptide and amino acid finished product.But how much do not disclose in final products polypeptide and each accounting of amino acid, and exist and take out
Carry the time it is longer the problem of.Wu Xinying etc. uses autolysis method enzymolysis and extraction yeast polypeptides product from brewer's yeast, has obtained excellent
The enzymolysis process of change, but the purity and molecular weight distribution of polypeptide are not disclosed.
The content of the invention
Problem of the prior art solved by the invention is：The technique of the existing polypeptide that higher degree is prepared from yeast
For：Yeast enzymolysis, extracting, centrifugation, purifying obtain polypeptide, are related to the techniques such as enzymolysis, centrifugation, polishing purification, decolouring.Extraction obtains
Polypeptide products purity it is relatively low, the impurity such as nucleic acid, polysaccharide and salinity is more, and the molecular weight ranges of polypeptide are indefinite, color, gas
The sensory properties such as taste and mouthfeel is poor.
For these problems, it is an object of the invention to improve the yield and purity of yeast polypeptides as far as possible.
Specifically, the invention provides a kind of preparation method of yeast polypeptides, comprise the following steps：
(1) Yeast protein in raw material is extracted；
(2) alkali protease enzymolysis is added, then adds compound protease enzymolysis, supernatant is collected and obtains enzymolysis product；
(3) separate, decolourize, obtain yeast polypeptides.
Preferably, Yeast protein is extracted in the basic conditions to step (1) raw material, wherein alkaline ph values are 9-13, effect
Temperature is 30-70 DEG C.
It is furthermore preferred that Yeast protein is extracted in the basic conditions to step (1) raw material, wherein, operative temperature 50-70
℃。
Preferably, supernatant is taken to the Yeast protein separation obtained under alkalescence condition, it is 3.0-7.0 then to adjust pH value, point
Precipitated from paste is obtained.
Preferably, before extracting the Yeast protein in raw material, the raw material of step (1) is configured into mass fraction first is
2%-20% solution.
It is furthermore preferred that before extracting the Yeast protein in raw material, the raw material of step (1) is configured into mass fraction first is
5%-15% solution.
Preferably, in step (2), the action condition of alkali protease is pH7-10,50-65 DEG C of temperature；Alkali protease
The mass fraction for accounting for dry matter is 0.1%-2%.
It is furthermore preferred that in step (2), the action condition of alkali protease is pH8-9,55-60 DEG C of temperature；Alkali protease
The mass fraction for accounting for dry matter is 0.5%-1%.
Preferably, in step (2), the action condition of compound protease is pH7-10, and temperature is 40-60 DEG C；Compound protein
The mass fraction that enzyme accounts for dry matter is 0.5%-3%.
It is furthermore preferred that in step (2), the action condition of compound protease is pH8-9, and temperature is 45-55 DEG C, compound protein
The mass fraction that enzyme accounts for dry matter is 1%-2%.
Preferably, the compound protease is selected from neutral proteinase, Protamex, Thermoase C, Protin SD
One or more in NY, papain or bromelain.
Preferably, after step (2) compound protease enzymolysis, it is 4.2-4.7 to add acid for adjusting pH, is warming up to 70-80 DEG C of guarantor
Hold 20-30min.
Preferably, it is configured to the solution that mass fraction is 2%-10% before step (2) enzymolysis.
It is furthermore preferred that the solution that mass fraction is 5%-8% is configured to before step (2) enzymolysis.
Preferably, enzymolysis liquid is separated using chlorination aluminum flocculation, film centrifugation or diatomite filtering in step (3).
It is furthermore preferred that enzymolysis liquid is separated using film centrifugation in step (3).
It is further preferred that molecular cut off is used to be centrifuged for 2000-20000D film in step (3).
Still more preferably, enzymolysis liquid is carried out using the film using molecular cut off 2000-5000D in step (3)
Separation.
Preferably, the pH value of decolorization is 2.0-6.0 in step (3), and the temperature of decolorization is 10-70 DEG C.
It is furthermore preferred that the pH value of decolorization is 3.0-4.0 in step (3), the temperature of decolorization is 20-40 DEG C.
Preferably, the preparation method also includes the yeast polypeptides that step (3) obtains processing is dried, and obtains powdery
Yeast polypeptides.
It is furthermore preferred that the drying process is spray drying or freeze-drying.
Preferably, the raw material in step (1) is the raw material containing yeast.
It is furthermore preferred that the raw material containing yeast is in brewer's yeast, saccharomyces cerevisiae, Saccharomyces cerevisiae or yeast extract
It is a kind of.
Invention also provides the yeast polypeptides obtained using above-described preparation method.
To prepare cosmetics in addition, the invention provides described yeast polypeptides or prepare commodity or prepare daily skin
Application in skin repair medicine field and field of food.
The beneficial effects of the invention are as follows：(1) present invention using first extraction Yeast protein and then is digested to obtain to albumen
Colourless, tasteless, the yeast polypeptides of content of peptides more than 80% are prepared in the technique of yeast polypeptides, and are obtained using the present invention
The polypeptide molecular weight of yeast polypeptides more than 85% is practically free of free amino acid below 2000, and without general Soybean Peptide etc.
The bitter taste that peptide product has.Improve flavour of food products, help postexercise recovery muscle power etc. available for that in food and health products, can play
Effect；(2) if the yeast polypeptides liquid that the present invention obtains dusts without drying, it is a excellent cosmetics Essence, has
Antiwrinkle active is nourished, may apply in cosmetics, color, perfume (or spice) without influenceing cosmetics etc., and there is senile-resistant efficacy.May be used also
It is applied in the commodity such as shampoo, hair conditioner, maintenance can be played and be damaged healthy hair, improves the effect of withered bifurcated.
Embodiment
As described above, it is an object of the invention to：A kind of method for producing high-purity yeast polypeptides product is provided and its obtained
The yeast polypeptides arrived and application.
Wherein, in a kind of embodiment of the present invention, there is provided a kind of preparation method of yeast polypeptides, including such as
Lower step：
(1) Yeast protein in raw material is extracted；
(2) alkali protease enzymolysis is added, then adds compound protease enzymolysis, supernatant is collected and obtains enzymolysis product；
(3) separate, decolourize, obtain yeast polypeptides.
Wherein, the action time for Yeast protein being extracted under the conditions of step (1) neutral and alkali is 2-6h.
Wherein, step (1) takes supernatant to the Yeast protein separation obtained under alkalescence condition, regulation pH value is 3.0-7.0
Afterwards, 10-30 DEG C stands 1-5h, then isolated paste precipitation.
Wherein, the enzymolysis time of step (2) neutral and alkali protease is 1-5h, preferably 2-4h.
The enzymolysis time of compound protease is 1-20h, preferably 4-14h in step (2).
Wherein, after step (2) compound protease enzymolysis, acid adding regulation PH is 4.2-4.7, is warming up to 70-80 DEG C and protects
Hold 20-30min.
In another embodiment of the present invention, there is provided the ferment that a kind of preparation method using the present invention obtains
Female polypeptide.
The present invention raw material be the raw material containing yeast, including but not limited to saccharomyces cerevisiae, brewer's yeast, Saccharomyces cerevisiae,
Yeast extract etc., the soda acid for adjusting pH are food-grade, and species is unlimited.
With reference to embodiment, the present invention is further detailed explanation.
Reagent and device information used are as shown in table 1 in the embodiment of the present invention：
The reagent of table 1 and production firm
Ultra high temperature short time sterilization machine, model：RSCG01-1, producer：The grand light industry and machinery Co., Ltd of Wenzhou mayor；
High performance liquid chromatograph, model Prominence LC-20A：Producer：Japanese Shimadzu；
Moisture teller, model：ZS-201, producer：Su Sai Electronic Science and Technology Co., Ltd.s of Shenzhen.
Embodiment one
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 500kg beer yeast powder 10kg, add alkali to adjust pH12.5, temperature 60 C, carry
Take 2h, 10000rpm centrifuging and taking supernatants, pH to 4.7 is adjusted with watery hydrochloric acid, refrigerate to standing 1h, 10000rpm centrifugations after 20 DEG C.
(2) egg white icing after centrifuging adds 100kg purifying water washings and centrifuged again, and obtaining egg white icing is about
25kg, moisture teller is used to determine crude protein content as the 12% of cream weight, crude protein is to remove the content after moisture.
(3) it is 60L crude protein to be dissolved in water to volume, and heating stirring is uniform, 70 DEG C of holding 30min, is cooled to 55 DEG C
Afterwards, pH8.0 is adjusted, adds alkali protease 15g, 50 DEG C is cooled to after digesting 2h, adds neutral proteinase 15g, papain
30g digests 10h.
PH4.5 is adjusted, is heated to 80 DEG C of holding 20min, centrifugation point takes supernatant to precipitate.
(4) supernatant adjusts pH4.0 after diatomite filters, and 40 DEG C of temperature, adds 0.5% activated carbon decolorizing 30min, mistake
Filter, repeats this step once, obtains destainer 48kg.
(5) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
Wherein, in the present invention in yeast polypeptides determining content of peptides with reference to People's Republic of China (PRC) light industry standard QB/T
Measure in 2653-2004 soy peptide powders on Soybean Peptide.I.e. hmw protein is easily deposited in acid condition, relatively low point
The protein hydrolysate of son amount is the molten protein of acid, dissolves in acid solution (wherein comprising peptide and free amino acid).Sample passes through
After acidifying, the sour molten protein content in filtrate subtracts the content that free aminoacid content is peptide.Wherein, it is solvable in supernatant
The assay of protein is measured according to the assay method of GB/T 22492-2008 Appendix B peptide contents, and free amino acid contains
It is fixed with reference to GB/T 5009.124-2003 standard tests to measure.Content of peptides is calculated using equation below (I).
The content (I) of content-free amino acid of the molten protein of content of peptides=acid
Purity is further calculated using equation below (II)：
Purity (%)=(content of content-free amino acid of the molten protein of acid) ÷ dry matter content × 100%
(Ⅱ)
Wherein, dry matter content is the amount that butt is scaled after destainer is dried in formula (II).
Polypeptide yield is further calculated using equation below (III)：
Polypeptide yield (%)=(content of content-free amino acid of the molten protein of acid) original yeast powder contents of ÷ ×
100% (III)
Wherein, the assay method of peptide molecular weight distribution uses People's Republic of China (PRC) light industry standard QB/T2653-
High performance gel filtration chromatography in 2004 appendix As, i.e., using porous filler as stationary phase, according to sample component molecular volume size
Difference separated, detected under the conditions of the UV absorption wavelength 220nm of peptide bond, use gel chromatography measure molecular weight distribution
Exclusive data processing it is soft (i.e. GPC softwares), chromatogram and its data are handled, the average molecular of polypeptide is calculated
Measure size and distribution.And count percentage of the content of peptides below molecular weight 2000D in total content of peptides.
After measured, in the yeast polypeptides liquid that embodiment one is prepared, content of peptides is 1.28kg (meter of giving money as a gift), and polypeptide is pure
Degree 82%, molecular weight are 90% in below 2000D content of peptides.Polypeptide yield is 12.5%.
Embodiment two
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 50kg yeast extract 10kg, adjust pH11,55 DEG C of temperature, after extracting 6h
5000rpm centrifuging and taking supernatants.Supernatant acid adding adjusts pH to 4.5 protein precipitations.5h, 5000rpm centrifugations are stood after being cooled to 30 DEG C.
(2) egg white icing after centrifuging adds 50kg purifying water washing centrifugations.Egg white icing 30.3kg is obtained, crude protein content is about
9.3% is weighed for cream.
(3) crude protein is dissolved in water to 60kg, is heated to 80 DEG C of insulation 25min, is cooled to 55 DEG C or so and adds alkali to adjust
PH7.8, after adding 3g alkali proteases enzymolysis 3h, add papain 35g enzymolysis 16h.
PH value of solution is adjusted to 4.7, is then heated to 75 DEG C of holding 20min, centrifuging and taking supernatant.
Supernatant uses molecular cut off to carry out UF membrane for 2000D ceramic membrane, and collection passes through supernatant, obtains supernatant
50kg。
(4) supernatant uses salt acid for adjusting pH as 4.0, and temperature adjustment is 40 DEG C, 1% activated carbon decolorizing is added, after filtering
Collect destainer, common 49kg.
(5) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
After measured, content of peptides is 1.372kg (meter of giving money as a gift) in the yeast polypeptides product being prepared, and Purity is
90%, polypeptide yield 13.7%, molecular weight is 85% in below 2000D content of peptides.
Embodiment three
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 90kg Saccharomyces cerevisiae powder 10kg, add alkali to adjust pH13, temperature 70 C, extraction
4h, centrifuging and taking supernatant, pH to 4.3 is adjusted with watery hydrochloric acid, 3h is stood after being cooled to room temperature, is centrifuged.
(2) egg white icing after centrifuging adds 100kg purifying water washings and centrifuged.It is about 28kg to obtain egg white icing, crude protein
Content is that cream weighs 12%.
(3) it is 40L crude protein to be dissolved in water to volume, and heating stirring is uniform, is heated to 60 DEG C, adjusts pH9.0, adds alkali
Property protease 3 0g, be cooled to 50 DEG C after digesting 2h, add neutral proteinase 30g, papain 60g enzymolysis 6h.
It is 4.5 to adjust solution ph, is heated to 80 DEG C of holding 20min, and centrifugation point takes supernatant to precipitate.
(4) supernatant adjusts pH4.0 after diatomite filters, and temperature 50 C, adds 0.5% activated carbon decolorizing 30min, mistake
Filter, repeats this step once.Obtain destainer 36kg.
(5) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
After measured, in the yeast polypeptides liquid being prepared, content of peptides is 1.476kg (meter of giving money as a gift), and Purity is
80.8%, molecular weight is 81% in below 2000D content of peptides, polypeptide yield 14.7%.
Example IV
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 90kg beer yeast powder 10kg, add alkali to adjust pH13,65 DEG C of temperature, extraction
4h, centrifuging and taking supernatant, pH to 4.3 is adjusted with watery hydrochloric acid, after being cooled to room temperature, centrifuged after standing 4h.
(2) egg white icing after centrifuging adds 90kg purifying water washings and centrifuged.It is about 27kg to obtain egg white icing, and crude protein contains
Measure and weigh 12% for cream.
(3) it is 35L crude protein to be dissolved in water to volume, and heating stirring is uniform, is heated to 60 DEG C, adjusts pH9.0, adds alkali
Property protease 3 0g, be cooled to 50 degree after digesting 2h, add neutral proteinase 30g, papain 60g enzymolysis 4h.
It is 4.5 to adjust pH value, is heated to 80 DEG C of holding 20min, and centrifugation point takes supernatant to precipitate.
(4) supernatant decolourizes by macroreticular resin after diatomite filtering is cooled to room temperature, obtains eluent 33L.
(5) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
After measured, in the yeast polypeptides liquid being prepared, polypeptide total content is 1.28kg (meter of giving money as a gift), and Purity is
81%, molecular weight is 83% in below 2000D content of peptides, polypeptide yield 12.8%.
Embodiment five
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 200kg yeast extract 20kg, add alkali to adjust pH12.8,55 DEG C of temperature, carry
4h is taken, centrifuging and taking supernatant, pH to 4.3 is adjusted with watery hydrochloric acid, is centrifuged after 10 DEG C of refrigeration 5h.
(2) egg white icing after centrifuging adds 90kg purifying water washings and centrifuged.It is about 47 kilograms to obtain egg white icing, crude protein
Content is that cream weighs 13%.
(3) it is 75L crude protein to be dissolved in water to volume, and heating stirring is uniform, is heated to 60 DEG C, adjusts pH9.0, adds alkali
Property protease 3 0g, be cooled to 50 DEG C after digesting 4h, add neutral proteinase 30g, papain 60g enzymolysis 4h.
It is 4.5 to adjust pH value, is heated to 80 DEG C of holding 20min, and centrifugation point takes supernatant to precipitate.
(4) supernatant decolourizes by macroreticular resin after diatomite filtering is cooled to room temperature, obtains eluent 70L.
(5) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
After measured, in the yeast polypeptides liquid being prepared, polypeptide total content is 2.59kg (meter of giving money as a gift), and Purity is
81.3%, molecular weight is 81% in below 2000D content of peptides, polypeptide yield 12.9%.
Embodiment six
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 200kg Saccharomyces cerevisiae powder 16kg, add alkali to adjust pH12.8, temperature 60 C, carry
4h is taken, centrifuging and taking supernatant, pH to 4.5 is adjusted with watery hydrochloric acid, is centrifuged after 10 DEG C of refrigeration 5h.
(2) egg white icing after centrifuging adds 100kg purifying water washings and centrifuged.It is about 40kg to obtain egg white icing, crude protein
Content is that cream weighs 12%.
(3) it is 75L crude protein to be dissolved in water to volume, and heating stirring is uniform, is heated to 55 DEG C, adjusts pH8.5, adds alkali
Property protease 24g, be cooled to 45 DEG C after digesting 2h, add neutral proteinase 48g, papain 72g enzymolysis 4h.
It is 4.5 to adjust pH value, is heated to 80 DEG C of holding 20min, and centrifugation point takes supernatant to precipitate.
(4) after supernatant is cooled to room temperature, aggregated chlorination aluminum flocculation centrifugation, supernatant adjusts pH3.0, adds amount of liquid
1% powder activity carbon decoloring 1h, filters after decolouring, obtains filtrate 72L.
(5) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
After measured, in the yeast polypeptides liquid being prepared, polypeptide total content is 2.88kg (meter of giving money as a gift), and Purity is
80.6%, molecular weight is 80% in below 2000D content of peptides, polypeptide yield 14.4%.
Embodiment seven
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 100kg Saccharomyces cerevisiae powder 10kg, add alkali to adjust pH9.0,30 DEG C of temperature, carry
5h is taken, centrifuging and taking supernatant, adjusts pH to 7.0, room temperature to be centrifuged after placing 2h with watery hydrochloric acid.
(2) egg white icing after centrifuging adds 20kg purifying water washings and centrifuged.It is about 11.5kg to obtain egg white icing, crude protein
Content is that cream weighs 13%.
(3) it is 14.95L crude protein to be dissolved in water to volume, and heating stirring is uniform, is heated to 55 DEG C, adjusts pH7.0, adds
Enter alkali protease 27g, be cooled to 40 DEG C after digesting 1h, add bromelain 8g, digest 20h.
It is 4.3 to adjust pH value, is heated to 70 DEG C of holding 30min, and centrifugation point takes supernatant to precipitate.
(4) after supernatant is cooled to room temperature, aggregated chlorination aluminum flocculation centrifugation, supernatant adjusts pH3.0, adds amount of liquid
1% powder activity carbon decoloring 1h, filters after decolouring, obtains filtrate 13.5L.
(5) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
After measured, in the yeast polypeptides liquid being prepared, polypeptide total content is 1.215kg (meter of giving money as a gift), and Purity is
81.1%, molecular weight is 85% in below 2000D content of peptides, polypeptide yield 12%.
Embodiment eight
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 100kg Saccharomyces cerevisiae powder 10kg, add alkali to adjust pH10.0,40 DEG C of temperature, carry
3h is taken, centrifuging and taking supernatant, adjusts pH to 6.0, room temperature to be centrifuged after placing 4h with watery hydrochloric acid.
(2) egg white icing after centrifuging adds 30kg purifying water washings and centrifuged.It is about 15.4kg to obtain egg white icing, crude protein
Content is that cream weighs 13%.
(3) it is 100L crude protein to be dissolved in water to volume, and heating stirring is uniform, is heated to 50 DEG C, adjusts pH10.0, is added
Alkali protease 18g, 60 DEG C are warming up to after digesting 5h, add Thermosase C 100 10g, the 10g of Protin NY 100,
Digest 12h.
It is 4.3 to adjust pH value, is heated to 70 DEG C of holding 30min, and centrifugation point takes supernatant to precipitate.
(4) after supernatant is cooled to room temperature, aggregated chlorination aluminum flocculation centrifugation, supernatant adjusts pH3.0, adds amount of liquid
1% powder activity carbon decoloring 1h, filters after decolouring, obtains filtrate 78L.
(5) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
After measured, in the yeast polypeptides liquid being prepared, polypeptide total content is 1.40kg (meter of giving money as a gift), and Purity is
81.1%, molecular weight is 83.5% in below 2000D content of peptides, polypeptide yield 14%.
Comparative example one
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 100kg Saccharomyces cerevisiae powder 11kg, add alkali to adjust pH13.0, temperature 70 C, carry
4h is taken, centrifuging and taking supernatant, pH to 4.3 is adjusted with watery hydrochloric acid, is centrifuged after 4 DEG C of placement 6h.
(2) egg white icing after centrifuging adds 40kg purifying water washings and centrifuged.It is about 26.4kg to obtain egg white icing, crude protein
Content is that cream weighs 12.5%.
(3) it is 60L crude protein to be dissolved in water to volume, and heating stirring is uniform, is heated to 50 DEG C, pH7.0 is adjusted, in addition
Property protease 3 0g, papain 60g, digest 15h.
It is 4.5 to adjust pH value, is heated to 80 DEG C of holding 20min, and centrifugation point takes supernatant to precipitate.
(4) after supernatant is cooled to room temperature, aggregated chlorination aluminum flocculation centrifugation, supernatant adjusts pH4.0, adds amount of liquid
0.5% powder activity carbon decoloring 30min, filters after decolouring, obtains filtrate 50L.
(5) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
After measured, in the yeast polypeptides liquid being prepared, polypeptide total content is 1kg (meter of giving money as a gift), and Purity is
70.4%, molecular weight is 67% in below 2000D content of peptides, polypeptide yield 10%.
Comparative example two
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 90kg Saccharomyces cerevisiae powder 10kg, add alkali to adjust pH13,55 DEG C of temperature, extraction
8h, centrifuging and taking supernatant, pH to 4.3 is adjusted with watery hydrochloric acid, is centrifuged after 4 DEG C of refrigeration 30min.
(2) egg white icing after centrifuging adds 100kg purifying water washings and centrifuged.It is about 28kg to obtain egg white icing, crude protein
Content is that cream weighs 12%.
(3) it is 40L crude protein to be dissolved in water to volume, and heating stirring is uniform, is heated to 60 DEG C, adjusts pH9.0, adds alkali
Property proteinase 9 0g, be cooled to 50 DEG C after digesting 2h, add papain 10g enzymolysis 10h.
It is 4.5 to adjust solution ph, is heated to 80 DEG C of holding 20min, and centrifugation point takes supernatant to precipitate.
(4) supernatant adjusts pH4.0 after diatomite filters, and temperature 50 C, adds 0.5% activated carbon decolorizing 30min, mistake
Filter, repeats this step once.Obtain destainer 32kg.
(5) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
After measured, in the yeast polypeptides liquid being prepared, polypeptide total content is 1.024kg (meter of giving money as a gift), and Purity is
69.1%, molecular weight is 58% in below 2000D content of peptides, polypeptide yield 10.2%.
Comparative example three
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 90kg Saccharomyces cerevisiae powder 10kg, add alkali to adjust pH8,80 DEG C of temperature, extraction
4h, centrifuging and taking supernatant, pH to 4.3 is adjusted with watery hydrochloric acid, is centrifuged after 4 DEG C of refrigeration 8h.
(2) egg white icing after centrifuging adds 10kg purifying water washings and centrifuged.It is about 10kg to obtain egg white icing, and crude protein contains
Measure and weigh 12% for cream.
(3) it is 20L crude protein to be dissolved in water to volume, and heating stirring is uniform, is heated to 50 DEG C, pH7.0 is adjusted, in addition
Property protease 3 0g, papain 60g enzymolysis 10h.After adjust pH8.0, add alkali protease 30g, digest 2h.
It is 4.5 to adjust solution ph, is heated to 80 DEG C of holding 20min, and centrifugation point takes supernatant to precipitate.
(4) supernatant adjusts pH4.0 after diatomite filters, and temperature 50 C, adds 1% activated carbon decolorizing 30min, filters,
Repeat this step once.Obtain destainer 15kg.
(5) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
After measured, in the yeast polypeptides liquid being prepared, polypeptide total content is 0.57kg (meter of giving money as a gift), and Purity is
65.3%, molecular weight is 79% in below 2000D content of peptides, polypeptide yield 5.7%.
Comparative example four
The method for preparing yeast polypeptides, its step are as follows：
(1) add pure water to be configured to solution 200kg yeast extract 10kg, add alkali to adjust pH8, be heated to 55 DEG C, add
Enter alkali protease 50g, digest 2h, rear 50 DEG C of temperature regulating, add neutral proteinase 50g, papain 100g, digest 16h.
It is 4.5 to adjust solution ph, is heated to 80 DEG C of holding 20min, and centrifugation point takes supernatant to precipitate.
(2) supernatant adjusts pH4.0 after diatomite filters, and temperature 50 C, adds 1% activated carbon decolorizing 30min, filters,
Repeat this step once.Obtain destainer 120kg.
(3) sterilized using ultra high temperature short time sterilization machine, 121 DEG C, heat 3-5s, carried out after sterilizing filling.
After measured, in the yeast polypeptides liquid being prepared, polypeptide total content is 2.16kg (meter of giving money as a gift), and Purity is
55.3%, molecular weight is 65% in below 2000D content of peptides, polypeptide yield 21.6%.
After measured, the yeast polypeptides being prepared according to the preparation method of the embodiment of the present invention one to embodiment eight, its is pure
Degree is more than 80%, polypeptide yield is higher, and molecular weight accounts for more than 80% in below 2000D content of peptides, even
It is more than 85%.Although for comparative example four without the extraction of Yeast protein before enzymolysis, obtained polypeptide yield is higher, more
Peptide purity and below 2000D content of peptides are relatively low, and value is not high.The yeast obtained using preparation method of the present invention
Polypeptide, colorless and odorless, available in food and health products, improving flavour of food products, helping postexercise recovery muscle power and other effects, separately
Outside, if yeast polypeptides liquid dusts without drying, it may be used on preparing in cosmetics and skin care item, there is high value.
Present pre-ferred embodiments are the foregoing is only, are not used to the limitation present invention, it is all in the spiritual and former of the present invention
Modifications, equivalent substitutions and improvements done within then etc., it is required within the protection domain of invention.
Claims (15)
1. a kind of preparation method of yeast polypeptides, it is characterised in that comprise the following steps：
(1) Yeast protein in raw material is extracted；
(2) alkali protease enzymolysis is added, then adds compound protease enzymolysis, supernatant is collected and obtains enzymolysis product；
(3) separate, decolourize, obtain yeast polypeptides.
2. preparation method according to claim 1, it is characterised in that ferment is extracted in the basic conditions to step (1) raw material
Female albumen, wherein alkaline ph values are 9-13, and operative temperature is 30-70 DEG C, preferably 50-70 DEG C.
3. preparation method according to claim 2, it is characterised in that taken to the Yeast protein separation obtained under alkalescence condition
Supernatant, it is 3.0-7.0 then to adjust pH value, isolated paste precipitation.
4. according to the preparation method any one of claim 1-3, it is characterised in that extraction raw material in Yeast protein it
Before, the raw material of step (1) is configured to the solution that mass fraction is 2%-20% first, preferred mass fraction is 5%-15%.
5. according to the preparation method any one of claim 1-4, it is characterised in that in step (2), alkali protease
Action condition is pH7-10, preferably pH8-9,50-65 DEG C of temperature, preferably 55-60 DEG C；Alkali protease accounts for the quality of dry matter
Fraction is 0.1%-2%, is preferably 0.5%-1%.
6. according to the preparation method any one of claim 1-5, it is characterised in that in step (2), compound protease
Action condition is pH7-10, preferably pH8-9, and temperature is 40-60 DEG C, is preferably 45-55 DEG C；Compound protease accounts for dry matter
Mass fraction is 0.5%-3%, is preferably 1%-2%.
7. according to the preparation method any one of claim 1-6, it is characterised in that the compound protease is selected from neutrality
Protease, Protamex, Thermoase C, Protin SD NY, papain or one kind or several in bromelain
Kind.
8. according to the preparation method any one of claim 1-7, it is characterised in that step (2) compound protease digests
Afterwards, it is 4.2-4.7 to add acid for adjusting pH, is warming up to 70-80 DEG C.
9. according to the preparation method any one of claim 1-8, it is characterised in that be configured to before step (2) enzymolysis
Mass fraction is 2%-10% solution, and preferred mass fraction is 5%-8%.
10. according to the preparation method any one of claim 1-9, it is characterised in that wadded a quilt with cotton in step (3) using aluminium chloride
Solidifying, film centrifugation or diatomite filtering separate to enzymolysis liquid, and preferably film centrifuges, more preferably using molecular cut off
2000-20000D film is separated, and further preferred 2000-5000D film separates to enzymolysis liquid.
11. according to the preparation method any one of claim 1-10, it is characterised in that decolorization in step (3)
PH value is 2.0-6.0, and preferable ph 3.0-4.0, the temperature of decolorization is 10-70 DEG C, and preferable temperature is 20-40 DEG C.
12. according to the preparation method any one of claim 1-11, it is characterised in that the preparation method also includes will
Processing is dried in the yeast polypeptides that step (3) obtains, and obtains dusty yeast polypeptide, and drying process is preferably spray drying or cold
It is lyophilized dry.
13. according to the preparation method any one of claim 1-12, it is characterised in that raw material in step (1) be containing
There is the raw material of yeast, the one kind being preferably selected from brewer's yeast, saccharomyces cerevisiae, Saccharomyces cerevisiae or yeast extract.
14. the yeast polypeptides obtained using the preparation method described in claim any one of 1-13.
15. yeast polypeptides described in claim 14 are preparing cosmetics or are preparing commodity or prepare daily skin repair medicine
Application in thing field and field of food.
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Effective date of registration: 20210316 Address after: 443003 No.168, Chengdong Avenue, Wujiagang District, Yichang City, Hubei Province Applicant after: Angel Nutt Co.,Ltd. Address before: 443003 No. 168, Chengdong Avenue, Yichang, Hubei Applicant before: Angel Yeast Co.,Ltd. |
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Application publication date: 20180109 |