CN102965421A - Preparation method and application of bioactive peptides for promoting yeast growth and improving ethanol tolerance performance of yeast - Google Patents
Preparation method and application of bioactive peptides for promoting yeast growth and improving ethanol tolerance performance of yeast Download PDFInfo
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- CN102965421A CN102965421A CN2012104307024A CN201210430702A CN102965421A CN 102965421 A CN102965421 A CN 102965421A CN 2012104307024 A CN2012104307024 A CN 2012104307024A CN 201210430702 A CN201210430702 A CN 201210430702A CN 102965421 A CN102965421 A CN 102965421A
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Abstract
The invention discloses a preparation method and application of bioactive peptides for promoting yeast growth and improving ethanol tolerance performance of yeast. The preparation method is as below: soybean isolated protein is used as a raw material, and hydrolyzed by papaya protease and alkaline protease; reaction products are centrifuged to collect soybean peptides; and the soybean peptides pass through ultrafiltration membrane with different molecular weight cutoffs, so as to obtain soybean peptides with different molecular weights. According to the invention, soybean peptides with molecular weight less than 1 ku and addition of 0.1-0.3% can significantly promote the growth of yeast; and soybean peptides with molecular weight of 3-5 ku and addition of 0.1-0.3% can significantly improve the ethanol tolerance performance of yeast.
Description
Technical field
The present invention relates to functional living being bioactive peptide technical field, the preparation method and the application that are specifically related to short yeast growth and improve the biologically active peptides of the anti-alcohol performance of yeast.
Background technology
In recent years along with people to the understanding in depth of peptide, find that some peptide has important regulating effect to different kind organism functional, this part peptide is called as bioactive peptide or functional peptide.At present, increasing research begins to pay close attention to bioactive peptide to the physiological regulating control effect of microorganism.It is reported that a class native peptides has anti-microbial effect, has significant fungistatic effect to gram-positive microorganism.Find that Antibacterial Peptide can effectively kill Gram-negative bacteria and part gram-positive microorganism, but to some gram-positive microorganisms such as streptococcus aureus, genus bacillus unrestraint effect.In addition, bioactive peptide also has report to the promoter action of microbial growth and metabolism more.The enzyme system that Yang Lanli produces with aspergillus oryzae in the dregs of beans Daqu culturing process to dregs of beans carry out Controlled-enzymatic Hydrolysis obtain molecular weight less than 3ku inspire the ferment peptide, can regulate and control the metabolism of aspergillus oryzae, become the generation of bent proteolytic enzyme to have to soy sauce and significantly induce and the raising effect.Zhang Qingli finds that casein papain hydrolysis peptide (molecular weight is less than 3ku) can effectively promote Yoghourt fermentation and improve the quality of sour milk, has obtained the casein peptide that three (6 peptides or 7 peptides) have significantly short lactobacter growth activity through separation and purification step by step.
Soybean protein becomes small molecules through microbial fermentation or protease hydrolysis with macromolecules degradation, forms low molecular peptide.The soybean Toplink promotes the metabolism and growth of microorganism cells.Soybean polypeptide can promote the propagation of milk-acid bacteria, yeast, bifidus bacillus and mould, and can promote the aerogenesis of bread yeast.Oligopeptides can promote the breeding of the resident flora of cud.The report soy bean protein hydrolysates such as Zhang Rongzhen have promoter action to the propagation of milk-acid bacteria, think that the soy bean protein hydrolysate of interpolation 1% can make the rate of propagation of lactobacillus bulgaricus improve one times nearly.The research proteolysis soybean proteins such as Zhao Dongmei obtain the following soybean peptides of 1.4 ku molecular weight sections to bacterium and saccharomycetic culture experiment, find that the growth of yeast cell is had significant promoter action.Therefore use soybean peptides that the yeast saccharomyces cerevisiae physiologically active is regulated and control to have important using value.
Summary of the invention
Preparation method and application thereof that the object of the invention provides a kind of short yeast growth and improves the biologically active peptides of the anti-alcohol performance of yeast.
The preparation method of the biologically active peptides of short yeast growth and the anti-alcohol performance of raising yeast comprises the steps:
(1) soybean protein isolate is pressed 1:15(w/v, g/mL) water-soluble, 80-90 ℃ of thermal treatment 10-30 min;
(2) add an amount of papoid and Sumizyme MP behind 50-55 ℃ of constant temperature enzymolysis 6 ~ 10h, boil the 10-30 min enzyme that goes out, the centrifugal 5-10 min of zymolyte 6000g-10000g;
(3) cross the filter membrane that different molecular weight is held back, collect the soybean peptides of different molecular weight section, lyophilize; The soybean peptides of different molecular weight section comprises: molecular weight is less than the soybean peptides (SP1) of 1ku, molecular weight is the soybean peptides (SP2) of 1 ~ 3ku, molecular weight is the soybean peptides (SP3) of 3 ~ 5ku, molecular weight is the soybean peptides (SP4) of 5 ~ 10ku, more than each section soybean peptides total content account for the soybean protein isolates amount more than 70%.
Further, step (3) finally obtains molecular weight less than the soybean peptides of 1ku and the soybean peptides of molecular weight 3 ~ 5 ku, i.e. described biologically active peptides.
Further, the addition of the described papoid of step (2) and Sumizyme MP is respectively 0.2-0.4% and the 0.4-0.6%(w/w of substrate protein);
The application of the biologically active peptides that above-mentioned preparation method makes is about to biologically active peptides and is added in the fermented liquid.
Further, in the described application, molecular weight can significantly promote yeast growth when being fermented liquid quality 0.1 ~ 0.3% less than the soybean peptides addition of 1ku; The soybean peptides addition of molecular weight 3 ~ 5 ku can significantly improve the anti-alcohol performance of yeast when being fermented liquid quality 0.1 ~ 0.3%.Yeast in the described fermented liquid be yeast saccharomyces cerevisiae (
Saccharomyces pastorianus).
Compared with prior art, the present invention has adopted above technical scheme, has following advantage and effect:
(1) under the synergy of papoid and Sumizyme MP, the gained soybean protein hydrolyate has short yeast growth and improves the anti-ethanol function of yeast at soybean protein in the present invention.
(2) molecular weight can significantly promote yeast growth when being 0.1 ~ 0.3% less than 1 kDa soybean peptides addition; Molecule can significantly improve the anti-alcohol performance of yeast when 3 ~ 5 kDa soybean peptides additions are 0.1 ~ 0.3%.
Description of drawings
Fig. 1 is fermentation yeast saccharomyces cerevisiae FBY0095 upgrowth situation comparison diagram of (interpolation different concentration ethanol) in the YNB substratum under 30 ℃ and anaerobic condition during 24h.
Fig. 2 is fermentation yeast saccharomyces cerevisiae FBY0095 survival rate comparison diagram of (interpolation different concentration ethanol) in the YNB substratum under 30 ° of C and anaerobic condition during 24h.
Embodiment
Below in conjunction with example and accompanying drawing enforcement of the present invention is described further, but enforcement of the present invention and protection domain are not limited to this.
Embodiment 1
The preparation method:
(1) soybean protein isolate is pressed 1:15(w/v, g/mL) water-soluble, 90 ℃ of thermal treatment 10min;
(2) add respectively papoid and the Sumizyme MP of soybean protein isolate weight 0.2% and 0.4%, behind 55 ℃ of constant temperature enzymolysis 6h, boil the 10 min enzyme that goes out, the centrifugal 5min of zymolyte 10000g;
(3) cross the filter membrane that different molecular weight is held back, collect molecular weight less than the soybean peptides (SP1) of 1ku, molecular weight is 1 ~ 3ku(SP2) soybean peptides, molecular weight is 3 ~ 5ku(SP3) soybean peptides, molecular weight is 5 ~ 10ku(SP4) soybean peptides, more than each section soybean peptides total content account for soybean protein isolates amount 72%, lyophilize;
(yeast is yeast saccharomyces cerevisiae to adopt the YNB substratum
Saccharomyces pastorianus): glucose 0.5%(w/v, g/mL), yeast nitrogen substratum (YNB) 0.17%(w/v, g/mL).Fermentation condition: inoculum size is the 4g(yeast slurry)/L, 12 ℃ of anaerobically fermenting 24 h.The fermentation before with each molecular weight soybean peptides with 0.1%(w/v, g/mL) be added in the substratum.Adopt the biomass in the blood red tally method calculating fermenting process, with investigation different molecular weight soybean peptides under the coercing of different concentration ethanol Fig. 1 is seen in the impact of yeast cell growth, wherein, *: the significance of blank when relatively not adding ethanol (
P＜0.05); +: relatively add 4%(w/v, g/mL) significance of blank during ethanol (
P＜0.05); △: relatively add 8%(w/v, g/mL) significance of blank during ethanol (
P＜0.05); : relatively add 12%(w/v, g/mL) significance of blank during ethanol (
P＜0.05).
As shown in Figure 1, the soybean peptides of small molecular weight has promoter action to yeast cell, particularly molecular weight less than the soybean peptides of 1ku have significantly (
P＜0.05) effect.Do not adding under the condition of ethanol, the cell concentration that adds SP1 has improved 22.49% than blank, than adding the SPH(soybean protease hydrolysate) cell concentration improved 9.57%.Along with the alcohol concn that adds in the substratum raises, the growth promoting function of SP1 reduces, and is adding 12%(w/v, g/mL) under the ethanol condition, the cell concentration that adds SP1 has improved 12.07% than blank.
Embodiment 2
The preparation method:
(1) soybean protein isolate is pressed 1:15(w/v, g/mL) water-soluble, 80 ℃ of thermal treatment 30 min;
(2) add an amount of papoid and Sumizyme MP behind 50 ℃ of constant temperature enzymolysis 10h, boil the 30 min enzyme that goes out, centrifugal 10 min of zymolyte 10000g;
(3) cross the filter membrane that different molecular weight is held back, collect molecular weight less than the soybean peptides (SP1) of 1ku, molecular weight is 1 ~ 3ku(SP2) soybean peptides, molecular weight is 3 ~ 5ku(SP3) soybean peptides, molecular weight is 5 ~ 10ku(SP4) soybean peptides, more than each section soybean peptides total content account for soybean protein isolates amount 73%, lyophilize;
(yeast is yeast saccharomyces cerevisiae to adopt the YNB substratum
Saccharomyces pastorianus): glucose 0.5%(w/v, g/mL), yeast nitrogen substratum (YNB) 0.17%(w/v, g/mL).Fermentation condition: inoculum size is the 4g(yeast slurry)/L, 12 ℃ of anaerobically fermenting 24 h.The fermentation before with each molecular weight soybean peptides with 0.3%(w/v, g/mL) be added in the substratum.Adopt the biological activity of yeast in the blue staining monitoring of the methyne fermenting process, to investigate under the coercing of different concentration ethanol the different molecular weight soybean peptides to the impact of yeast cell survival rate, see Fig. 2, wherein *: relatively add 8%(w/v, g/mL) significance of blank during ethanol (
P＜0.05); +: relatively add 12%(w/v, g/mL) significance of blank during ethanol (
P＜0.05).
As shown in Figure 2, at 8%(w/v, g/mL) under the alcohol concn condition, in substratum, add SP2, SP3 and SP4 and make the survival rate of yeast improve 12.56%, 14.60% and 13.03% than blank respectively, and SPH and SP1 to be the survival rate of yeast reduced by 5.96% and 6.59% than blank (Control) respectively.Be 12%(w/v, g/mL when adding alcohol concn in the substratum) time, SP1 make the yeast cell survival rate significantly reduce (
P＜0.05), SP3 make the yeast cell survival rate significantly raise (
P＜0.05).The result shows, coerce at high concentration ethanol lower, small molecular weight titanium (Mw＜1ku) can not make that yeast cell is active to be improved, and macromolecule peptide, particularly molecular weight to be the soybean peptides of 3 ~ 5ku can make significantly that yeast cell is active to be improved.
Only be the part example below, technical scheme provided by the present invention all can realize.
Claims (6)
1. the preparation method of the biologically active peptides of short yeast growth and the anti-alcohol performance of raising yeast is characterized in that comprising the steps:
(1) soybean protein isolate is pressed 1:15(w/v, g/mL) water-soluble, 80-90 ℃ of thermal treatment 10-30 min;
(2) add an amount of papoid and Sumizyme MP behind 50-55 ℃ of constant temperature enzymolysis 6 ~ 10h, boil the 10-30 min enzyme that goes out, the centrifugal 5-10 min of zymolyte 6000g-10000g;
(3) cross the filter membrane that different molecular weight is held back, collect the soybean peptides of different molecular weight section, lyophilize; The soybean peptides of different molecular weight section comprises: molecular weight is less than the soybean peptides (SP1) of 1ku, molecular weight is the soybean peptides (SP2) of 1 ~ 3ku, molecular weight is the soybean peptides (SP3) of 3 ~ 5ku, molecular weight is the soybean peptides (SP4) of 5 ~ 10ku, more than each section soybean peptides total content account for the soybean protein isolates amount more than 70%.
2. preparation method according to claim 1 is characterized in that step (3) finally obtains molecular weight less than the soybean peptides of 1ku and the soybean peptides of molecular weight 3 ~ 5 ku, i.e. described biologically active peptides.
3. preparation method according to claim 1 is characterized in that the addition of the described papoid of step (2) and Sumizyme MP is respectively 0.2-0.4% and the 0.4-0.6%(w/w of substrate protein).
4. the application of the biologically active peptides that makes of each described preparation method of claim 1 ~ 3 is characterized in that biologically active peptides is added in the fermented liquid.
5. application according to claim 4 is characterized in that molecular weight is fermented liquid quality 0.1 ~ 0.3% less than the soybean peptides addition of 1ku; The soybean peptides addition of molecular weight 3 ~ 5 ku is fermented liquid quality 0.1 ~ 0.3%.
6. application according to claim 4, it is characterized in that yeast in the described fermented liquid be yeast saccharomyces cerevisiae (
Saccharomyces pastorianus).
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106243187A (en) * | 2016-09-05 | 2016-12-21 | 华南理工大学 | A kind of method of sharp separation oyster peptide from Concha Ostreae enzymolysis solution |
| CN108165596A (en) * | 2018-01-31 | 2018-06-15 | 华南理工大学 | A kind of preparation method and application for the wheat gluten protein peptide that can improve brewing yeast cell osmotic pressure tolerance |
| CN110229852A (en) * | 2019-06-20 | 2019-09-13 | 东莞理工学院 | A method of utilizing soybean protein hydrolysate fermenting and producing gamma-polyglutamic acid |
| CN114395602A (en) * | 2022-01-21 | 2022-04-26 | 广州大学 | Preparation method and application of peptide-zinc complex for promoting growth and fermentation of lactic acid bacteria |
-
2012
- 2012-11-02 CN CN2012104307024A patent/CN102965421A/en active Pending
Non-Patent Citations (2)
| Title |
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| 万春艳 等: "大豆蛋白水解物对酿酒酵母生长和发酵性能的影响", 《食品与发酵工业》 * |
| 陈星 等: "不同蛋白酶酶解产物活性大豆肽分子量分布状态的研究", 《西南大学学报(自然科学版)》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106243187A (en) * | 2016-09-05 | 2016-12-21 | 华南理工大学 | A kind of method of sharp separation oyster peptide from Concha Ostreae enzymolysis solution |
| CN106243187B (en) * | 2016-09-05 | 2019-08-20 | 华南理工大学 | A method of the quick separating oyster peptide from oyster enzymolysis liquid |
| CN108165596A (en) * | 2018-01-31 | 2018-06-15 | 华南理工大学 | A kind of preparation method and application for the wheat gluten protein peptide that can improve brewing yeast cell osmotic pressure tolerance |
| CN108165596B (en) * | 2018-01-31 | 2021-10-26 | 华南理工大学 | Preparation method and application of wheat gluten protein peptide capable of improving osmotic pressure tolerance of saccharomyces cerevisiae cells |
| CN110229852A (en) * | 2019-06-20 | 2019-09-13 | 东莞理工学院 | A method of utilizing soybean protein hydrolysate fermenting and producing gamma-polyglutamic acid |
| CN110229852B (en) * | 2019-06-20 | 2021-02-23 | 东莞理工学院 | Method for producing gamma-polyglutamic acid by fermenting soybean protein zymolyte |
| CN114395602A (en) * | 2022-01-21 | 2022-04-26 | 广州大学 | Preparation method and application of peptide-zinc complex for promoting growth and fermentation of lactic acid bacteria |
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Application publication date: 20130313 |