CN102174512A - Process method for producing yeast polysaccharide and yeast nucleotide from yeast - Google Patents
Process method for producing yeast polysaccharide and yeast nucleotide from yeast Download PDFInfo
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- CN102174512A CN102174512A CN2011100438243A CN201110043824A CN102174512A CN 102174512 A CN102174512 A CN 102174512A CN 2011100438243 A CN2011100438243 A CN 2011100438243A CN 201110043824 A CN201110043824 A CN 201110043824A CN 102174512 A CN102174512 A CN 102174512A
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Abstract
The invention relates to a process method for producing yeast polysaccharide and yeast nucleotide from yeast, which solves the problems of low yield, long period, high equipment investment and high production cost existing in the conventional process for extracting the yeast polysaccharide and the yeast nucleotide. The yeast polysaccharide and the yeast nucleotide are respectively extracted by taking waste beer yeast as a raw material through the steps of high-pressure homogenization and performing of an enzyme method. The process method has the characteristics of high yield and purity of extracted yeast polysaccharide, high yeast nucleotide content, short extraction time, low enzyme consumption, low equipment investment and production cost and suitability for large-scale industrial production.
Description
Technical field
The present invention relates to a primary yeast treatment process, a primary yeast is produced the processing method of zymosan and Nucleotide specifically.
Background technology
Waste yeast in China's brewing industry production is used as the cheap processing of roughage raw material more, causes the ample resources waste.If waste yeast is fully developed, effective constituent is wherein rationally reclaimed, economic benefit will be very considerable.Processing for waste yeast mainly is to be that raw material adopts acid-base method or enzyme alkaline process to extract zymosan with the waste yeast at present, and acid-base method is abandoned gradually to the seriously polluted of environment; Though and the enzyme alkaline process can obtain the higher zymosan of purity, but required enzyme concn is higher, and enzymolysis and alkaline purification time are grown (enzymolysis time is 12h at least, and the alkaline purification time is 4h at least), thereby improved production cost greatly, influenced production efficiency and industrial scale.Ma Sen etc. disclose a kind of " ultrasonic-enzyme-alkaline process extracts beer waste yeast β-1; research of 3 dextran " in that [brewing science and technology] the 2009th the 2nd is interim, adopt ultrasonic-enzyme-alkaline process from beer waste yeast, to extract β-1, the 3-dextran, in ultrasonic pretreatment and enzymolysis top condition, utilize Response Surface Method to research and analyse NaOH concentration, temperature, consumption and time, the influence of 3-dextran yield, purity and protein content to β-1.Test-results shows that sporoderm-broken rate is 94.22% after the ultrasonication; The protein removal rate is 62.82% behind the enzymolysis; As the NaOH 30.50mL that adds 2.05%, handle 5.7h for 74 ℃, β-1, the yield of 3-dextran is 10.21%, and purity is 88.14%, and protein content is 1.19%.Ultrasonic-enzyme-alkaline process treatment process has β-1,3-dextran yield, purity height, the low short characteristics of extraction time that reach of protein content.This method is to utilize hyperacoustic cavitation effect to reach shell-broken effect, because Ultrasonic Cell Disruptor complicated operation, treatment capacity little (per hour treatment capacity number rise to about tens liters), (microwave treatment high temperature of local moment can reach 5000 ℃ easily to produce high temperature, handling the back product temperature also reaches about 70 ℃, high temperature can make protein denaturation, the activeconstituents loss of activity), processing condition require height, general laboratory and the less place of other treatment capacities of only being used for.This method only terminates in the experimental phase at present, can't realize the heavy industrialization application.And because yeast cells wall is than the ordinary cells wall thickness, can reach 0.1-0.3 μ m, and general bacterial cell wall thickness is only about tens nanometer, for guaranteeing enzymolysis efficiency, usually the addition with hydrolysising protease is controlled at the 200-210U/g dry-matter, and the purchase cost height of hydrolysising protease if the usage quantity of proteolytic enzyme can't be reduced, then can't be realized the purpose of production cost control in fact.
Summary of the invention
The objective of the invention is in order to solve the problems of the technologies described above, providing a kind of is raw material with the yeast, produce the processing method of zymosan and Nucleotide, zymosan yield that this processing method is extracted and purity height, nucleotide content height, but have extraction time short, enzyme dosage is few, the characteristics of the low large-scale industrial production of production cost.
Processing method of the present invention is:
One. yeast is dissolved in water is the yeast suspension liquid of 10-12%;
Two. the yeast suspension liquid is carried out under 50-60Mpa high-pressure homogeneous, the centrifugation of high-pressure homogeneous back, collecting precipitation thing;
Three. add hydrolysising protease in throw out, add-on is the 110-130U/g dry-matter, is constant temperature enzymolysis 5-7h under 5-6, the 50-60 ℃ condition at pH then, carries out centrifugation again and obtains enzymolysis supernatant liquor and enzymolysis throw out;
Four. the enzymolysis supernatant liquor is carried out carrying out spraying drying again behind the enzyme-deactivating obtain yeast Nucleotide;
Five. the enzymolysis throw out is carried out carrying out centrifugation again after the alkaline purification, collect the alkaline purification throw out and repeatedly be washed to neutrality, spraying drying makes zymosan.
The yeast suspension liquid is under 60Mpa high-pressure homogeneous 3 times in the described step 2.
Hydrolysising protease is at least a in papoid and the neutral protease in the described step 3.
Described hydrolysising protease is papoid and neutral protease, and both mass ratioes are 1: 1.
In the step 3, adding hydrolysising protease 120U/g dry-matter in described throw out, is constant temperature enzymolysis 6h under 5,55 ℃ of conditions at pH then.
In the step 5, described alkali treatment method is: add 3%NaOH solution in the enzymolysis throw out, add-on is the 2ml/g dry-matter, handles 2 hours under 90 ℃ of conditions then.
Described dry-matter is: the centrifuged deposit thing is removed the surplus materials behind the moisture, and the amount of indication dry-matter is according to the throw out moisture content conversion gained after centrifugal in the corresponding steps herein.
The present invention has utilized high pressure homogenizer, shear by the intensive that high pressure produces, bump, hole and turbulent eddy effect are carried out high-pressure homogeneous to the yeast suspension liquid, the cell walls physical structure is changed, reach the purpose of cell walls fragmentation, research and experimental results show that being specially adapted to the thicker yeast of pair cell wall carries out broken wall treatment, have and adopt other wall-breaking method excellent effect that is beyond one's reach, compare and have simple to operate than ultrasonic method, no protein denaturation (can not produce moment high temperature, handling the back product temperature maintains about 30-40 ℃), processing condition require low, characteristics of high efficiency, high-pressure homogeneous processing by early stage, can eliminate sterically hindered between enzyme and the substrate, the action site of proteolytic enzyme on yeast cells wall increased, hydrolysis efficiency is improved, and then enzymolysis time can further reduce, time that also can corresponding minimizing alkaline purification; And high pressure homogenizer is simple in structure, purchase far below Ultrasonic Cell Disruptor, but the material treatment capacity can realize large-scale industrial production much larger than Ultrasonic Cell Disruptor (treatment capacity can reach per hour more than 5000 liters).On the other hand, remarkable just because of shell-broken effect, the proteolytic enzyme action site increases, the contriver finds, the addition that significantly reduces hydrolysising protease this moment also can guarantee maximum enzymolysis efficiency, thereby makes the explained hereafter cost obtain real effectively control, has produced beyond thought technique effect.
On the other hand, after centrifugation,, the enzymolysis throw out obtains the zymosan except that being handled, the enzymolysis supernatant liquor further handled to obtain Nucleotide, a technology is extracted simultaneously and is obtained two kinds of products, the application of high pressure homogenization technique makes the technology later stage contain the enzymolysis throw out of zymosan and to contain the enzymolysis supernatant liquor resolution of Nucleotide good, and the content of two products, yield and purity all can be guaranteed.
Further, prove that through experimental study for many years the yeast suspension liquid is high-pressure homogeneous 3 best results under 60Mpa; Sedimentary enzymolysis top condition after the high-pressure homogeneous processing of described process is to add hydrolysising protease 120U/g dry-matter in described throw out, be constant temperature enzymolysis 6h under 5,55 ℃ of conditions at pH then, the addition of hydrolysising protease and enzymolysis time significantly reduce and shell-broken effect obvious, and the follow-up alkaline purification time also foreshorten to 2h; Described hydrolysising protease is choose reasonable as required, preferred papoid and neutral protease add simultaneously, both mass ratioes are 1: 1, its reason is: both optimal ph and the temperature of effect are approaching, and papoid is different with the neutral protease action site, both are used in combination and can obtain very good effect, cooperate high-pressure homogeneous method can further reduce the addition of hydrolysising protease.The waste yeast that described yeast produces in can the beer manufacturing is a raw material, and effective constituent is wherein rationally reclaimed.
The present invention is raw material with the waste yeast, fully extract effective ingredient wherein, a logical technology is extracted two kinds of products, enzymolysis time and alkaline purification time have shortened over half at least than existing enzyme alkaline process in this processing method, enzyme dosage also reduced near half, make whole significantly cripetura of explained hereafter cycle, equipment throw time and production cost declines to a great extent, can realize large-scale industrial production, the zymosan yield and the purity height (approaching) that extract with " ultrasonic-enzyme-alkaline process ", the nucleotide content height has vast market prospect.
Description of drawings
Fig. 1 is a process flow sheet of the present invention.
Embodiment
Embodiment 1
Get yeast powder (beer waste yeast) and add water to the yeast suspension liquid that concentration is 10% (m/m), with yeast suspension liquid high-pressure homogeneous 3 times at 60Mpa, the centrifugal 5min of 3000r/min then, the collecting precipitation thing, add papoid and neutral protease (both mass ratioes are 1: 1) by the 120U/g dry-matter, pH is 5,55 ℃ of constant temperature enzymolysis 6h, the centrifugal 5min of 3000r/min obtains enzymolysis throw out and enzymolysis supernatant liquor then, in the enzymolysis throw out, add 3%NaOH (m/m) solution with the 2ml/g dry-matter, handled 2 hours for 90 ℃, the centrifugal 5min of 3000r/min collects the alkaline purification throw out and repeatedly is washed to neutrality, and spraying drying makes zymosan and (adopts the phenol sulfuric acid process to detect, the product purity of polysaccharide reaches 85.4%, and yield is 10.5%); Enzymolysis supernatant liquor spraying drying behind moment high temperature enzyme-deactivating obtains yeast Nucleotide (crude protein content 47.3%, rna content 15.4%).
Embodiment 2
Getting yeast powder (beer waste yeast), to add water to concentration be 12% yeast suspension liquid, with yeast suspension liquid high-pressure homogeneous 3 times at 55Mpa, the centrifugal 5min of 3500r/min then, the collecting precipitation thing, add papoid and neutral protease (both mass ratioes are 1: 1) by the 130U/g dry-matter, 60 ℃ of constant temperature enzymolysis 7h, the centrifugal 5min of 3000r/min obtains enzymolysis throw out and enzymolysis supernatant liquor then, in the enzymolysis throw out, add 3%NaOH solution with the 2ml/g dry-matter, handled 2 hours for 95 ℃, the centrifugal 5min of 3000r/min collects the alkaline purification throw out and repeatedly is washed to neutrality, and spraying drying makes zymosan and (adopts the phenol sulfuric acid process to detect, the product purity of polysaccharide reaches 80.2%, and yield is 8.3%); Enzymolysis supernatant liquor spraying drying behind moment high temperature enzyme-deactivating obtains yeast Nucleotide (crude protein content 46.2%, rna content 13.4%).
Embodiment 3
Getting yeast powder (beer waste yeast), to add water to concentration be 11% yeast suspension liquid, with yeast suspension liquid high-pressure homogeneous 2 times at 50Mpa, the centrifugal 5min of 3000r/min then, the collecting precipitation thing, press the 110U/g dry-matter and add papoid, 50 ℃ of constant temperature enzymolysis 5h, the centrifugal 5min of 3000r/min obtains enzymolysis throw out and enzymolysis supernatant liquor then,, in the enzymolysis throw out, add 3%NaOH solution with the 2ml/g dry-matter, handled 2 hours for 85 ℃, the centrifugal 5min of 3000r/min collects the alkaline purification throw out and repeatedly is washed to neutrality, and spraying drying makes zymosan and (adopts the phenol sulfuric acid process to detect, the product purity of polysaccharide reaches 77.3%, and yield is 8.0%); Enzymolysis supernatant liquor spraying drying behind moment high temperature enzyme-deactivating obtains yeast Nucleotide (crude protein content 43.2%, rna content 14.5%).
Claims (6)
1. a primary yeast is produced the processing method of zymosan and Nucleotide, it is characterized in that method is:
One. yeast is dissolved in water is the yeast suspension liquid of 10-12%;
Two. the yeast suspension liquid is carried out under 50-60Mpa high-pressure homogeneous, the centrifugation of high-pressure homogeneous back, collecting precipitation thing;
Three. add hydrolysising protease in throw out, add-on is the 110-130U/g dry-matter, is constant temperature enzymolysis 5-7h under 5-6, the 50-60 ℃ condition at pH then, carries out centrifugation again and obtains enzymolysis supernatant liquor and enzymolysis throw out;
Four. the enzymolysis supernatant liquor is carried out carrying out spraying drying again behind the enzyme-deactivating obtain yeast Nucleotide;
Five. the enzymolysis throw out is carried out carrying out centrifugation again after the alkaline purification, collect the alkaline purification throw out and repeatedly be washed to neutrality, spraying drying makes zymosan.
2. yeast as claimed in claim 1 is produced the processing method of zymosan and Nucleotide, it is characterized in that, the yeast suspension liquid is under 60Mpa high-pressure homogeneous 3 times in the described step 2.
3. yeast as claimed in claim 1 is produced the processing method of zymosan and Nucleotide, it is characterized in that, hydrolysising protease is at least a in papoid and the neutral protease in the described step 3.
4. yeast as claimed in claim 2 is produced the processing method of zymosan and Nucleotide, it is characterized in that described hydrolysising protease is papoid and neutral protease, and both mass ratioes are 1: 1.
5. the processing method of producing zymosan and Nucleotide as each described yeast among the claim 1-4, it is characterized in that, in the step 3, adding hydrolysising protease 120U/g throw out in described throw out, is constant temperature enzymolysis 6h under 5,55 ℃ of conditions at pH then.
6. yeast as claimed in claim 5 is produced the processing method of zymosan and Nucleotide, it is characterized in that in the step 5, described alkali treatment method is: in the enzymolysis throw out, add 3%Na0H solution, add-on is the 2ml/g dry-matter, handles 1.5-2.5 hour under 85-95 ℃ of condition then.
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Cited By (4)
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| CN102732509A (en) * | 2012-07-19 | 2012-10-17 | 中粮生物化学（安徽）股份有限公司 | Method for extracting ribonucleic acid from fermentation thalli |
| CN108220280A (en) * | 2016-12-09 | 2018-06-29 | 安琪酵母股份有限公司 | Method for extracting high-purity nucleic acid using yeast and products thereof and application |
| CN109355284A (en) * | 2018-11-28 | 2019-02-19 | 罗锋利 | A kind of extracting method of yeast nucleic acid and its application in production selenium-rich nucleic acid |
| CN110256588A (en) * | 2019-05-28 | 2019-09-20 | 浦江县美泽生物科技有限公司 | Freshwater mussel antioxidant activity polysaccharide and preparation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102732509A (en) * | 2012-07-19 | 2012-10-17 | 中粮生物化学（安徽）股份有限公司 | Method for extracting ribonucleic acid from fermentation thalli |
| CN108220280A (en) * | 2016-12-09 | 2018-06-29 | 安琪酵母股份有限公司 | Method for extracting high-purity nucleic acid using yeast and products thereof and application |
| CN108220280B (en) * | 2016-12-09 | 2021-08-06 | 安琪酵母股份有限公司 | Method for extracting high-purity nucleic acid by using yeast, product and application thereof |
| CN109355284A (en) * | 2018-11-28 | 2019-02-19 | 罗锋利 | A kind of extracting method of yeast nucleic acid and its application in production selenium-rich nucleic acid |
| CN110256588A (en) * | 2019-05-28 | 2019-09-20 | 浦江县美泽生物科技有限公司 | Freshwater mussel antioxidant activity polysaccharide and preparation method thereof |
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Application publication date: 20110907 |