CN107581616A - A kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom - Google Patents
A kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom Download PDFInfo
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- CN107581616A CN107581616A CN201710711137.1A CN201710711137A CN107581616A CN 107581616 A CN107581616 A CN 107581616A CN 201710711137 A CN201710711137 A CN 201710711137A CN 107581616 A CN107581616 A CN 107581616A
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Abstract
The invention discloses a kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom, comprise the following steps：S1. mushroom is crushed, extracts degreasing, dried；S2. add water to stir the dry material after degreasing, sequentially add beta glucan enzyme and alkali cellulose enzyme is digested；S3. solution after the enzymolysis of acquisition is subjected to ultrafiltration, obtained respectively through solution and retention solution；S4. solution is passed through by what step S3 was obtained, through resin desalination, heating in vacuum concentration, is spray-dried to obtain mushroom oligosaccharide；S5. retention solution step S3 obtained, pH to 4.0~4.5 is adjusted, precipitated, centrifugation, the crude protein of acquisition is through washing, being dried in vacuo and obtaining protein from lentinus edodes.The inventive method synchronously prepares protein from lentinus edodes and mushroom oligosaccharide, can obtain abundant protein from lentinus edodes and mushroom oligosaccharides simultaneously, and yield is high, fully extracts the nutritional ingredient in mushroom, can extend mushroom industrial chain, increases mushroom added value.
Description
Technical field
The invention belongs to health product technology field, more particularly, to one kind using mushroom synchronously prepare protein from lentinus edodes with
The method of mushroom oligosaccharide.
Background technology
Mushroom (Lentinus edodes), also known as flower mushroom, mushroom, fragrant letter, fragrant bacterium, dried mushroom, are Pleurotaceae plant mushroom
Fructification.Mushroom is second-biggest-in-the-world edible mushroom, and one of China's special product, in the title among the people for being known as " mountain delicacy ".Mushroom taste
Delicious, fragrance make people mentally refreshing is nutritious.The main component of mushroom has lentinan, eritadenine, nucleotides, vitamin, triterpenes
Compound and substantial amounts of protein and amino acid.Dried thin mushroom containing glutamic acid, alanine, histidine, leucine, phenylalanine,
The several amino acids such as valine, there is higher nutritive value and medical value.In China, mushroom is mainly with dry product or fresh goods pin
Sell, deep processing is relatively small, and added value is small, directly affects the income of mushroom grower.
The extraction of lentinan is to study one of more mushroom deep process at present.Lentinan has apparent
Antitumor activity, enhancing host to the resistivity of bacterium, fungi, virus and parasitic infection, enhancing NK and
The various biologicals such as cytotoxic t cell activity and immunologic competence, it is one of important component of mushroom.Therefore, existing perfume (or spice)
Mushroom deep processing is concentrated mainly on lentinan field, and what is utilized is also only lentinan, other abundant nutrition in mushroom
Composition is not utilized well, is a kind of invisible wasting of resources.
Mushroom oligosaccharide is also one of nutritional ingredient of mushroom, while can also be obtained by the hydrolysis of lentinan.It is oligomeric
2~4 monose are formed by connecting small aggressiveness by sugar typically by glycosidic bond.Oligosaccharide can not by human body hydrochloric acid in gastric juice destroy, also without
Method is digested enzyme decomposition, but it can be utilized by the bacterial fermentation in intestines, be converted into short chain fatty acids and lactic acid, suppress enteral
Salmonella and the growth of spoilage organisms, adjust gastrointestinal function.Mushroom oligosaccharide can be used for improving micro-ecological environment in human body, have
Beneficial to Bifidobacterium and the propagation of other beneficial bacteriums；Improve human body blood lipid metabolism, strengthen body immunity, caloric value is very low, very
Fat is converted into less, and work can be played in the low-energy foods such as diet food, diabetic food or hypertensive patient's food
With.But at present, also rarely have the research report on extracting mushroom oligosaccharide.
In addition, fresh mushroom in addition to aqueous 85-90%, contains crude protein 19.9% in solid content, protein content is high.At present for perfume (or spice)
The research of mushroom albumen is still in the junior stage, existing test result indicates that protein from lentinus edodes has to the cancer cell u14 of in vitro culture
Direct CDCC, hence it is evident that suppress the growth of tumour cell, and in vitro can direct killing tumour cell, and can induce
Apoptosis of tumor cells.In addition, also there are some researches show, protein from lentinus edodes can activate the immune system of body, macrophage, thin is improved
The action effect of intracellular cytokine, to realize its antitumor action；Protein from lentinus edodes can also effectively suppress the activity of HIV1-RT
With the propagation of leukaemia.Directly use alkalinity extraction more for the extracting method of protein from lentinus edodes, but the yield of albumen and
Activity is not very good, and a large amount of industrial alcohol is applied to the obstacle applied for protein from lentinus edodes in field of food.
In view of the shortcomings that existing protein from lentinus edodes, mushroom oligosaccharide extraction process are present, also, it is not favourable in traditional handicraft
The method for producing protein from lentinus edodes and mushroom oligosaccharide simultaneously with mushroom.It is high that the present invention proposes a kind of high income, content height, purity
The method for synchronously preparing protein from lentinus edodes and mushroom oligosaccharide.
The content of the invention
For above-mentioned deficiency of the prior art, there is provided one kind synchronously prepares protein from lentinus edodes using mushroom and mushroom is oligomeric
The method of sugar, preparation technology is simple, and high income, Product Activity are high, and safe preparation process is nontoxic.
The above-mentioned purpose of the present invention is achieved by the following technical programs.
A kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom, is comprised the following steps：
S1. mushroom is crushed, extracts degreasing, dried；
S2. add water to stir the dry material after degreasing, sequentially add 1,4 beta-glucanase and alkali cellulose enzyme carries out enzyme
Solution；
S3. solution after the enzymolysis of acquisition is subjected to ultrafiltration, obtained respectively through solution and retention solution；
S4. pass through solution by what step S3 was obtained, through resin desalination, heating in vacuum concentration, be spray-dried mushroom is oligomeric
Sugar；
S5. retention solution step S3 obtained, pH to 4.0~4.5 is adjusted, precipitated, centrifugation, the crude protein of acquisition is through water
Wash, be dried in vacuo to obtain protein from lentinus edodes.
The 1,4 beta-glucanase and alkali cellulose enzyme can use existing product.
Preferably, the addition of 1,4 beta-glucanase described in step S2 is the 0.3~1% of the dry material, enzyme digestion reaction
45~55 DEG C of temperature, pH are 4.5~6.0, persistently digest 0.5~1h.
Preferably, the addition of alkali cellulose enzyme described in step S2 is the 0.9~3% of the dry material, and enzymolysis is anti-
45~55 DEG C of temperature is answered, pH is 10.0~11.0, persistently digests 2~3h.
Preferably, the step S1 includes：By mushroom fruiting body powder, 1 is added:The n-hexane of 5~10 mass ratioes, temperature
30~60 DEG C, 1~3h is extracted, extraction 1~3 time is repeated, filtering, by dry materials after extraction, obtains mushroom fruiting body powder after degreasing.
Preferably, the mushroom fruiting body powder be by mushroom fruiting body in 40~60 DEG C drying after, be crushed to 40~80
Mesh obtains.
Preferably, the ultrafiltration retaining molecular weight of ultrafiltration described in step S3 is 10kD, and ultrafiltration is described through molten to what is obtained
Liquid is 0.4~1 with retention liquor capacity ratio：1.
Preferably, the step S4 includes：The pH through solution is adjusted to 7.0, through anion and cation tree
Ester exchange desalination, heating in vacuum is concentrated into 1/4~1/3 volume, spray-dried to obtain mushroom oligosaccharide.
A kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom, is comprised the following steps：
S1. pre-treatment and degreasing：By mushroom fruiting body after 40~60 DEG C of drying, 40~80 mesh are crushed to, add 1:5~
The food grade n-hexane of 10 mass ratioes, 30~60 DEG C, 1~3h of extraction time of temperature, extraction 1~3 time is repeated, filtering, will be extracted
Material heat drying afterwards, obtain mushroom fruiting body powder after degreasing；
S2. digest：Mushroom fruiting body powder, adds 1 after the degreasing that step S1 is obtained：The water of 10~20 mass ratioes, after stirring
The 1,4 beta-glucanase of degreasing weight of material 0.3~1% is added, for adjustment temperature to 45~55 DEG C, pH is 4.5~6.0, lasting enzymolysis
0.5~1h；Then adjust to 45~55 DEG C, pH is 10.0~11.0, adds 0.9~3% alkali cellulose enzyme, then persistently enzyme
Solve 2~3h；
S3. ultrafiltration：Solution carries out ultrafiltration after the enzymolysis that S2 is obtained, and ultrafiltration retaining molecular weight is 10kD, and ultrafiltration is to saturating
It is 0.4 that solution, which is crossed, with retention liquor capacity ratio:1~1:Untill 1, obtain respectively through solution and retention solution；
S4. solution is passed through by what step S3 was obtained, through anion and cation resin exchange desalination, heating in vacuum is concentrated into
1/4~1/3 volume, it is spray-dried to obtain mushroom oligosaccharide；
S5. retention solution step S3 obtained, pH to 4.0~4.5 is adjusted, after protein precipitation, centrifugation, the thick egg of acquisition
Bai Jing washings, vacuum drying obtain protein from lentinus edodes.
Compared with prior art, beneficial effect of the present invention is：Provide and a kind of synchronously prepare protein from lentinus edodes and mushroom is low
The method of glycan, abundant protein from lentinus edodes and mushroom oligosaccharide can be obtained simultaneously, yield is high, fully extracts the nutrition in mushroom
Composition, mushroom industrial chain can be extended, increase mushroom added value.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.Unless stated otherwise, the reagent of the invention used, method and apparatus is the art conventional reagent, methods
And equipment.
The inventive method is further described by taking specific implementation condition as an example below.
Embodiment 1
A kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom, is comprised the following steps：
S1. pre-treatment and degreasing：By mushroom fruiting body after 55 DEG C of drying, 70 mesh are crushed to, add 1:10 mass ratioes
Food grade n-hexane, 40 DEG C, extraction time 1.5h of temperature, extraction 2 times is repeated, filtering, material heat drying after extraction is obtained de-
Mushroom fruiting body powder after fat；
S2. digest：Mushroom fruiting body powder, adds 1 after the degreasing that step S1 is obtained：The water of 20 mass ratioes, added after stirring
The 1,4 beta-glucanase of degreasing weight of material 1%, adjustment temperature pH 4.5, persistently digest 0.5h to 55 DEG C；Then adjust to 55
DEG C, pH 11.0 adds 3% alkali cellulose enzyme, then persistently digests 2h；
S3. ultrafiltration：Solution carries out ultrafiltration after the enzymolysis that S2 is obtained, and ultrafiltration retaining molecular weight is 10kD, and ultrafiltration is to saturating
It is 0.4 that solution, which is crossed, with retention liquor capacity ratio:Untill 1, obtain respectively through solution and retention solution；
S4. solution is passed through by what step S3 was obtained, through anion and cation resin exchange desalination, heating in vacuum is concentrated into
1/3 volume, it is spray-dried to obtain mushroom oligosaccharide；
S5. retention solution step S3 obtained, pH to 4.0~4.5 is adjusted, after protein precipitation, centrifugation, the thick egg of acquisition
Bai Jing washings, vacuum drying obtain protein from lentinus edodes.
Embodiment 2
A kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom, step S1, S3~S5 and embodiment 1
Identical, step S2 is specially in the present embodiment：
S2. digest：Mushroom fruiting body powder, adds 1 after the degreasing that step S1 is obtained：The water of 10 mass ratioes, added after stirring
The 1,4 beta-glucanase of degreasing weight of material 0.3%, adjustment temperature pH 6.0, persistently digest 1h to 45 DEG C；Then adjust to 45
DEG C, pH 10.0 adds 0.9% alkali cellulose enzyme, then persistently digests 3h；
Embodiment 3
A kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom, is comprised the following steps：
S1. pre-treatment and degreasing：By mushroom fruiting body after 60 DEG C of drying, 80 mesh are crushed to, add 1:The food of 5 mass ratioes
With level n-hexane, 30 DEG C, extraction time 3h of temperature repeats extraction 1 time, filtering, by material heat drying after extraction, after obtaining degreasing
Mushroom fruiting body powder；
S2. digest：Mushroom fruiting body powder, adds 1 after the degreasing that step S1 is obtained：The water of 15 mass ratioes, added after stirring
The 1,4 beta-glucanase of degreasing weight of material 0.8%, adjustment temperature pH 5.5, persistently digest 0.6h to 50 DEG C；Then adjust to
50 DEG C, pH 10.5,2% alkali cellulose enzyme is added, then persistently digests 2.5h；
S3. ultrafiltration：Solution carries out ultrafiltration after the enzymolysis that S2 is obtained, and ultrafiltration retaining molecular weight is 10kD, and ultrafiltration is to saturating
It is 1 that solution, which is crossed, with retention liquor capacity ratio:Untill 1, obtain respectively through solution and retention solution；
S4. solution is passed through by what step S3 was obtained, through anion and cation resin exchange desalination, heating in vacuum is concentrated into
1/4 volume, it is spray-dried to obtain mushroom oligosaccharide；
S5. retention solution step S3 obtained, pH to 4.0~4.5 is adjusted, after protein precipitation, centrifugation, the thick egg of acquisition
Bai Jing washings, vacuum drying obtain protein from lentinus edodes.
Embodiment 4
A kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom, step S2~S5 and the phase of embodiment 3
Together, step S1 is specifically included：
S1. pre-treatment and degreasing：By mushroom fruiting body after 40 DEG C of drying, 40 mesh are crushed to, add 1:The food of 8 mass ratioes
With level n-hexane, temperature 60 C, extraction time 1h repeats extraction 3 times, filtering, by material heat drying after extraction, after obtaining degreasing
Mushroom fruiting body powder.
Embodiment 5
A kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom, is comprised the following steps：
S1. pre-treatment and degreasing：By mushroom fruiting body after 65 DEG C of drying, 80 mesh are crushed to, add 1:15 mass ratioes
Food grade n-hexane, 65 DEG C, extraction time 1h of temperature, extraction 2 times is repeated, filtering, by material heat drying after extraction, obtains degreasing
Mushroom fruiting body powder afterwards；
S2. digest：Mushroom fruiting body powder, adds 1 after the degreasing that step S1 is obtained：The water of 25 mass ratioes, added after stirring
The 1,4 beta-glucanase of degreasing weight of material 1.5%, adjustment temperature pH 6.0, persistently digest 0.5h to 55 DEG C；Then adjust to
55 DEG C, pH 10.0,3.5% alkali cellulose enzyme is added, then persistently digests 2h；
S3. ultrafiltration：Solution carries out ultrafiltration after the enzymolysis that S2 is obtained, and ultrafiltration retaining molecular weight is 10kD, and ultrafiltration is to saturating
It is 1 that solution, which is crossed, with retention liquor capacity ratio:Untill 1, obtain respectively through solution and retention solution；
S4. solution is passed through by what step S3 was obtained, through anion and cation resin exchange desalination, heating in vacuum is concentrated into
1/4 volume, it is spray-dried to obtain mushroom oligosaccharide；
S5. retention solution step S3 obtained, pH to 4.0~4.5 is adjusted, after protein precipitation, centrifugation, the thick egg of acquisition
Bai Jing washings, vacuum drying obtain protein from lentinus edodes.
Embodiment 6
It is a kind of protein from lentinus edodes synchronously to be prepared using mushroom and the method for mushroom oligosaccharide is substantially same as Example 1, it is different
Part is that 1,4 beta-glucanase is different with the order of alkali cellulose enzyme in step S2, and specific steps S2 comprises the following steps：Will
Mushroom fruiting body powder, adds 1 after the degreasing that step S1 is obtained：The water of 20 mass ratioes, the alkali of degreasing weight of material 3% is added after stirring
Property cellulase, adjusts to 55 DEG C, pH 11.0, persistently digests 2h；Then temperature is adjusted to 55 DEG C, pH 4.5, adds 1%
1,4 beta-glucanase, then persistently digest 0.5h.
Using the dried thin mushroom of same batch as raw material, synchronously the mushroom is prepared according to the method for embodiment 1~6 respectively
Albumen and mushroom oligosaccharide, and by Kjeldahl nitrogen determination protein content, contained by high effective liquid chromatography for measuring oligosaccharide
Amount.As a result it is as follows：
| Protein from lentinus edodes recovery rate (mg/g) | Mushroom oligosaccharide recovery rate (mg/g) | |
| Embodiment 1 | 74.65 | 38.79 |
| Embodiment 2 | 72.28 | 35.34 |
| Embodiment 3 | 73.46 | 37.18 |
| Embodiment 4 | 73.13 | 36.70 |
| Embodiment 5 | 66.59 | 30.55 |
| Embodiment 6 | 58.97 | 28.63 |
After testing, the present invention synchronously prepares protein from lentinus edodes and the method for mushroom oligosaccharide can be effectively by the egg in mushroom
In vain, oligosaccharide separates with other compositions, and will be separated between albumen and oligosaccharide, and the albumen and oligosaccharide purity of extraction are high.
This method is digested by two steps and controlled, and can obtain the protein from lentinus edodes of high-purity, and causes the output increased of mushroom oligosaccharide,
Avoid as oligosaccharides is destroyed by further hydrolysis in conventional method.The molecular weight for the protein from lentinus edodes that this method obtains concentrates on 10~
Between 40kD, mainly including acidic protein；The mushroom oligosaccharide main component of acquisition is β -1,3- Portugals oligosaccharides, the albumen of acquisition
Recovery rate and oligosaccharide recovery rate are significantly higher than the albumen of conventional method/oligomeric respectively up to 74.65mg/g and 38.79mg/g
Sugared recovery rate.
Implementation of the invention described in detail above, still, the present invention are not limited to specific thin in above-mentioned embodiment
Section, in the range of the technology design of the present invention, a variety of simple variants can be carried out to technical scheme, these simple changes
Type belongs to protection scope of the present invention.
Claims (7)
- A kind of 1. method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom, it is characterised in that comprise the following steps：S1. mushroom is crushed, extracts degreasing, dried；S2. add water to stir the dry material after degreasing, sequentially add 1,4 beta-glucanase and alkali cellulose enzyme is digested；S3. solution after the enzymolysis of acquisition is subjected to ultrafiltration, obtained respectively through solution and retention solution；S4. solution is passed through by what step S3 was obtained, through resin desalination, heating in vacuum concentration, is spray-dried to obtain mushroom oligosaccharide；S5. retention solution step S3 obtained, adjust pH to 4.0~4.5, precipitate, centrifugation, the crude protein of acquisition through washing, It is dried in vacuo to obtain protein from lentinus edodes.
- 2. a kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom according to claim 1, it is special Sign is that the addition of 1,4 beta-glucanase described in step S2 is the 0.3~1% of the dry material, enzyme digestion reaction temperature 45~ 55 DEG C, pH is 4.5~6.0, persistently digests 0.5~1h.
- 3. a kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom according to claim 2, it is special Sign is that the addition of alkali cellulose enzyme described in step S2 is the 0.9~3% of the dry material, enzyme digestion reaction temperature 45 ~55 DEG C, pH is 10.0~11.0, persistently digests 2~3h.
- 4. a kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom according to claim 1, it is special Sign is that the step S1 includes：By mushroom fruiting body powder, 1 is added:The n-hexane of 5~10 mass ratioes, temperature 30~60 DEG C, 1~3h is extracted, extraction 1~3 time is repeated, filtering, by dry materials after extraction, obtains mushroom fruiting body powder after degreasing.
- 5. a kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom according to claim 4, it is special Sign is, the mushroom fruiting body powder be by mushroom fruiting body after 40~60 DEG C of drying, be crushed to the acquisition of 40~80 mesh.
- 6. a kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom according to claim 1, it is special Sign is that the ultrafiltration retaining molecular weight of ultrafiltration described in step S3 is 10kD, and ultrafiltration is described through solution and retention to what is obtained Liquor capacity ratio is 0.4~1：1.
- 7. a kind of method that protein from lentinus edodes and mushroom oligosaccharide are synchronously prepared using mushroom according to claim 1, it is special Sign is that the step S4 includes：The pH through solution is adjusted to 7.0, taken off through anion and cation resin exchange Salt, heating in vacuum is concentrated into 1/4~1/3 volume, spray-dried to obtain mushroom oligosaccharide.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109010349A (en) * | 2018-10-29 | 2018-12-18 | 湖北中医药大学 | Bletilla striata oligosaccharides is improving the application in intestinal microecology |
| CN113106138A (en) * | 2021-03-16 | 2021-07-13 | 中国农业科学院农产品加工研究所 | Preparation method for extracting and separating anti-tumor active protein from shiitake mushrooms |
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| CN109010349A (en) * | 2018-10-29 | 2018-12-18 | 湖北中医药大学 | Bletilla striata oligosaccharides is improving the application in intestinal microecology |
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| CN113106138A (en) * | 2021-03-16 | 2021-07-13 | 中国农业科学院农产品加工研究所 | Preparation method for extracting and separating anti-tumor active protein from shiitake mushrooms |
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Application publication date: 20180116 |