CN104694392A - Wall breaking and nano preparation method for se-enriched yeast - Google Patents
Wall breaking and nano preparation method for se-enriched yeast Download PDFInfo
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- CN104694392A CN104694392A CN201510153242.9A CN201510153242A CN104694392A CN 104694392 A CN104694392 A CN 104694392A CN 201510153242 A CN201510153242 A CN 201510153242A CN 104694392 A CN104694392 A CN 104694392A
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- selenium
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- enriched yeast
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/06—Lysis of microorganisms
Abstract
The invention discloses a wall breaking and nano preparation method for se-enriched yeast. The method comprises the technical steps that A, se-enriched yeast is dissolved in water to obtain suspension liquid with the concentration being 10%-30% (w/w); B, the se-enriched yeast suspension liquid with the concentration being well adjusted is placed in a ball mill; C, agate balls are used as impacting balls and placed in the se-enriched yeast suspension liquid in the ball mill, the ball mill is rotated, and then the agate balls impact and crush se-enriched yeast in the se-enriched yeast suspension liquid, so that the se-enriched yeast is crushed in a wall-breaking mode; D, the suspension liquid obtained in the step C is added into an ultrasonic breaking instrument which is used for further refining and homogenizing the suspension liquid; E, the suspension liquid obtained in the step D is subjected to freeze drying to form powdered finished products. By means of the preparation method, the walls of the se-enriched yeast can be broken by 100%, nutrient substance in the se-enriched yeast is completely released, and the se-enriched yeast can be further homogenized and nano-crystallized so that human bodies can have a better effect of absorbing bioselenium.
Description
Technical field
The present invention relates to biological technical field, relate in particular to the preparation method of a kind of yeast rich in selenium broken wall and nanometer.
Background technology
Yeast is a kind of single celled eukaryotic microorganism, its cell wall thickness is about 0.1 ~ 0.3 μm, structure is tough and tensile, and main component has dextran, twenty dew glycan, protein, chitin, lipid etc., and wherein the content of dextran and mannosans accounts for about 30% of cell walls dry weight respectively; Yeast cell matter contains abundant nutritive substance, as protein, Yeast Nucleic Acid, and vitamin B group and abundant amino acid etc.β in yeast cells wall-(1,3)-dextran can strengthen mammiferous immune vigor, anticancer, antibacterium, antiviral, antimycotic, parasiticide, reduction cholesterol and the ability such as blood fat, promotion wound healing, is a kind of good biological response modifiers.Therefore, after breaking yeast cellule membrane cracking, both can discharge nutritive substance in born of the same parents, can cell wall polysaccharides be obtained again.
Because yeast cells wall complex structure is tough and tensile, broken wall is comparatively difficult.Conventional wall-breaking method has the chemical process such as the physical methods such as high pressure homogenization, supersonic method, freeze-thaw method and organic solvent method, acid-base method, enzyme process.Wherein the large speed of high pressure homogenization process yeast cell amount is fast, but needs to repeat fragmentation, and in treating processes, the rising of temperature easily causes the inactivation of sensitive active materials.Also there is heat radiation difficulty in supersonic method, produces the problem of localized hyperthermia, be only suitable for laboratory scale application.Acid-base method broken wall needs to use a large amount of strong acid and strong bases, easily to environment.Enzymatic cleavage broken wall has a lot of outstanding advantage compared with other physico-chemical process, as low in energy consumption, low to nutritive ingredient destructiveness, environmental protection etc.But existing enzymatic shell-broken method also exists a lot of defect, as large in enzyme dosage, enzyme cost is high, enzyme easy in inactivation and need poisonous sulfhydryl compound pre-treatment yeast cell etc.
Yeast rich in selenium is saccharomycetic one, selenium element is added exactly in the process of culturing yeast, selenium has been absorbed during yeast growth, the protein of selenium in yeast body and polysaccharide organic are combined and are converted into biological selenium, thus eliminate chemical selenium (as Sodium Selenite) toxic side effects of human body and stomach are stimulated, selenium more efficiently, is more safely absorbed by the body utilization.Yeast rich in selenium be also up to now domestic the most efficiently, the selenium-replenishing preparation that the safest, nutrition is the most balanced.
The same broken wall of yeast rich in selenium is comparatively difficult, and because of the non-broken wall of yeast rich in selenium, yeast is arrived also to be needed through this procedure of hydrochloric acid in gastric juice broken wall in stomach, and the activity of yeast can bring a little untoward reaction simultaneously; Soda acid in the chemical method of existing wall breaking technology and enzymatic shell-broken, easy black yeast content, unfriendly to environment.High pressure homogenization in the Physical of existing wall breaking technology, supersonic method are broken, sand mill is broken, and supersonic method is very little to the effect of yeast broken wall, sporoderm-broken rate about 25%; High-pressure homogeneous fragmentation is 100% (with reference to from CN101195893A) for fresh activity yeast sporoderm-broken rate, and only has about 50% to non-living dried yeast sporoderm-broken rate; Sand mill Breaking Yeast sporoderm-broken rate 60% ~ 94% (with reference to from CN102174407A), absolutely sporoderm-broken rate can not be reached, yeast cells wall is just given fragmentation by above-mentioned wall breaking technology simultaneously, Nano grade can not be reached by super-refinement, yeast rich in selenium suspension particle particle diameter is larger, and size heterogeneity, make small intestine insufficient to its absorption, cause effective constituent to waste.
Therefore, a kind of method of industrial urgent need exploitation, this method both can, to yeast rich in selenium very broken wall, make wherein nutritive substance thoroughly discharge, and can carry out homogenization, nanometer again, make human body obtain better effect to biological selenium absorption to it.
Summary of the invention
The object of the invention is to the defect for above-mentioned prior art, the preparation method of a kind of yeast rich in selenium broken wall and nanometer is provided, can to yeast rich in selenium very broken wall, wherein nutritive substance is thoroughly discharged, also can carry out homogenization, nanometer to it, make human body obtain better effect to biological selenium absorption.
For achieving the above object, the technical solution used in the present invention is:
A preparation method for yeast rich in selenium broken wall and nanometer, the processing step of the method is:
A, yeast rich in selenium is soluble in water, make the suspension of 10% ~ 30% (w/w) concentration;
B, the yeast rich in selenium suspension mixing up concentration is placed in ball mill;
C, choose agate bead and make impact bead, be positioned in the yeast rich in selenium suspension of ball mill, rotating ball mill, described agate bead carries out shock to the yeast rich in selenium in yeast rich in selenium suspension and pulverizes, thus the broken wall realizing yeast rich in selenium is pulverized;
D, then the suspension that step C obtains is joined in Ultrasonic Cell Disruptor, with the further refinement of Ultrasonic Cell Disruptor and homogenization;
E, the powdered finished product of suspension lyophilize that step D is obtained.
As the improvement to technique scheme, described agate bead is two kinds, and the diameter of two kinds of described agate beads is respectively 0.5 ~ 1cm and 2 ~ 3cm.
As the improvement to technique scheme, the rotating speed of described ball mill is 300 ~ 500r/min, and the time that described shock is pulverized is 0.5 ~ 2h.
As the improvement to technique scheme, the power of described Ultrasonic Cell Disruptor is 200 ~ 400w, and the refinement of described Ultrasonic Cell Disruptor and homogenization time are 10 ~ 20min.
Under different ball milling conditions, ultrasound condition, the sporoderm-broken rate of yeast rich in selenium and the Data Representation of water power size are as shown in Table 1.
Table one
The yeast rich in selenium of non-broken wall carries out the result of SEM energy spectrum analysis as shown in Table 2,
Table two
| Element | Weight percent | Atomic percent |
| C | 57.68 | 65.05 |
| N | 12.92 | 12.50 |
| O | 23.88 | 20.22 |
| Na | 0.40 | 0.24 |
| Mg | 0.26 | 0.14 |
| Si | 0.15 | 0.07 |
| P | 1.64 | 0.72 |
| S | 0.43 | 0.18 |
| Cl | 0.23 | 0.09 |
| K | 2.15 | 0.74 |
| Se | 0.25 | 0.04 |
Yeast rich in selenium is after method broken wall of the present invention, and the result of SEM energy spectrum analysis as shown in Table 3.
Table three
| Element | Weight percent | Atomic percent |
| C | 61.97 | 69.80 |
| O | 33.64 | 28.44 |
| Na | 0.34 | 0.20 |
| Mg | 0.23 | 0.13 |
| P | 1.34 | 0.58 |
| S | 0.38 | 0.16 |
| Cl | 0.21 | 0.08 |
| K | 1.65 | 0.57 |
| Se | 0.25 | 0.04 |
Learnt with comparing of table three by table two, after method broken wall of the present invention, yeast rich in selenium obtains selenium element
Content is 0.25% (mass ratio), 0.04% (atomic ratio), identical with the content of selenium element in yeast rich in selenium before the yeast rich in selenium of non-broken wall, nanometer.Prove that the method can not destroy yeast rich in selenium intracellular organic matter, also can not bring objectionable impurities into.Biological selenium amount remains intact, and will greatly improve human body absorbing yeast rich in selenium.
Compared with prior art, the advantage that has of the present invention and positively effect are:
The preparation method of yeast rich in selenium broken wall of the present invention and nanometer, by the determination to Ball-milling Time, rotating speed, suspension concentration and ultrasonic power, time and suspension concentration, significantly improves yeast rich in selenium broken wall nanometer degree.The broken great efforts of ball mill, broken wall thoroughly, not only makes yeast rich in selenium content all discharge but also its impact force is pulverized further to the yeast rich in selenium after broken wall, such that second step is ultrasonic to carry out smoothly, is conducive to nanometer and pulverizes.Coordinate extensive ball mill and ultrasonication machine, easily can realize the extensive nanometer continuously of yeast rich in selenium, meet the demand of industrialized production; And this process is pure physical method, does not have chemical reaction, yeast rich in selenium intracellular organic matter can not be destroyed, also can not bring objectionable impurities into, clean environmental protection; In addition, crushing medium also can Reusability, and nanometer cost is low.The product sporoderm-broken rate obtained under Optimizing Technical reaches 100%, and size is less than 400 nanometers, and biological selenium amount remains intact, and will greatly improve human body absorbing yeast rich in selenium.
Accompanying drawing explanation
Fig. 1 is the SEM shape appearance figure of yeast rich in selenium;
Fig. 2 be yeast rich in selenium ultrasonic after water power dimensioned drawing;
Fig. 3 is the SEM shape appearance figure of yeast rich in selenium after dry ball milling;
Fig. 4 be yeast rich in selenium through dry ball milling and ultrasonic after water power dimensioned drawing;
Fig. 5 is the SEM shape appearance figure of yeast rich in selenium after wet ball grinding;
Fig. 6 be yeast rich in selenium through wet ball grinding and ultrasonic after water power dimensioned drawing.
Embodiment
Below in conjunction with specific embodiments, be clearly and completely described technical scheme of the present invention, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art are not making the every other embodiment obtained under creative work prerequisite, and any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Embodiment one
Take yeast rich in selenium dry powder 4g, be dissolved in certain volume water, make the suspension of 10% (w/w) concentration, the yeast rich in selenium suspension mixing up concentration is placed in ball mill; Choose particle diameter between 0.5 ~ 1.0cm and 2 ~ 3cm, two kinds of agate beads, 300r/min wet ball grinding 1h.Then the suspension Ultrasonic Cell Disruptor Fine by Ultrasonic dispersion will obtained, 200w power ultrasonic 15min, makes it nanometer.Observe pattern by scanning electronic microscope (SEM), percentage of damage is 75%.Measure water power size with Dynamic laser scattering instrument (DLS), size is about 700nm.
Embodiment two
Take yeast rich in selenium dry powder 5g, be dissolved in certain volume water, make the suspension of 12.5% (w/w) concentration, the yeast rich in selenium suspension mixing up concentration is placed in ball mill; Choose particle diameter between 0.5 ~ 1.0cm and 2 ~ 3cm, two kinds of agate beads, 400r/min wet ball grinding 2h.Then with the dispersion of Ultrasonic Cell Disruptor Fine by Ultrasonic, 400w power ultrasonic 15min, makes it nanometer.Observe pattern with SEM, percentage of damage is 85%.Measure water power size with DLS, size is about 550nm.
Embodiment three
Take yeast rich in selenium dry powder 4g, 8g and 12g, be dissolved in respectively in certain volume water, make the suspension of 10%, 20% and 30% (w/w) concentration, the yeast rich in selenium suspension mixing up concentration is placed in ball mill; Choose particle diameter between 0.5 ~ 1.0cm and 2 ~ 3cm, two kinds of agate beads, 300r/min ball milling 0.5h.Then the suspension Ultrasonic Cell Disruptor Fine by Ultrasonic dispersion will obtained, 200w power ultrasonic 10min, makes it nanometer.Observe pattern with SEM, three kinds of concentration yeast rich in selenium suspension sporoderm-broken rates are about 70%.Adopt DLS to measure water power size, size is followed successively by about 765nm, 770nm and 760nm.
Embodiment four
Take yeast rich in selenium dry powder 4g, 8g and 12g, be dissolved in respectively in certain volume water, make the suspension of 10%, 20% and 30% (w/w) concentration, the yeast rich in selenium suspension mixing up concentration is placed in ball mill; Choose particle diameter between 0.5 ~ 1.0cm and 2 ~ 3cm, two kinds of agate beads, 400r/min ball milling 1h.Then the suspension Ultrasonic Cell Disruptor Fine by Ultrasonic dispersion will obtained, 300w power ultrasonic 15min, makes it nanometer.Observe pattern with SEM, three kinds of concentration yeast rich in selenium suspension sporoderm-broken rates are about 80%.Adopt DLS to measure water power size, size is followed successively by about 650nm, 645nm and 665nm.
Embodiment five
Take yeast rich in selenium dry powder 4g, 8g and 12g, be dissolved in respectively in certain volume water, make the suspension of 10%, 20% and 30% (w/w) concentration, the yeast rich in selenium suspension mixing up concentration is placed in ball mill; Choose particle diameter between 0.5 ~ 1.0cm and 2 ~ 3cm, two kinds of agate beads, 500r/min ball milling 2h.Then the suspension Ultrasonic Cell Disruptor Fine by Ultrasonic dispersion will obtained, 400w power ultrasonic 20min, makes it nanometer.Observe pattern with SEM, three kinds of concentration yeast rich in selenium suspension sporoderm-broken rates are almost 100%.Adopt DLS to measure water power size, size is followed successively by about 355nm, 365nm and 350nm.SEM does power spectrum ultimate analysis, and obtaining selenium constituent content is 0.25% (w/w), almost constant compared with selenium constituent content before broken wall.To redissolving dispersion after its freeze-drying, SEM observes pattern, DLS observes water power size, considerable change does not occur compared with non-freeze-drying sample.
The yeast rich in selenium profile of non-broken wall is intact, and become elliposoidal, size is between 2 ~ 5 microns.It is soluble in water, and ultrasonic disperse, water power size is at 4.5 microns.Its basic condition as depicted in figs. 1 and 2.
The yeast rich in selenium gathering that dry ball milling obtains is sticked serious, and not easily subsequent disposal, sporoderm-broken rate also reduces greatly, and its basic condition as shown in Figure 3 and Figure 4.Under dry ball milling state, its percentage of damage is about 50%, DLS measurement water power size is about 800nm.If do not adopt ball milling, only adopt ultrasonic disperse, then almost can not broken wall.
Yeast rich in selenium is after wet ball grinding, and yeast rich in selenium percentage of damage reaches as high as 100%, and content all comes out.350 ran are of a size of by recording its water power after its ultrasonic disperse nanometer.Its basic condition as shown in Figure 5 and Figure 6.
The preparation method of yeast rich in selenium broken wall of the present invention and nanometer, by the determination to Ball-milling Time, rotating speed, suspension concentration and ultrasonic power, time and suspension concentration, significantly improves yeast rich in selenium broken wall nanometer degree.The broken great efforts of ball mill, broken wall thoroughly, not only makes yeast rich in selenium content all discharge but also its impact force is pulverized further to the yeast rich in selenium after broken wall, such that second step is ultrasonic to carry out smoothly, is conducive to nanometer and pulverizes.Coordinate extensive ball mill and ultrasonication machine, easily can realize the extensive nanometer continuously of yeast rich in selenium, meet the demand of industrialized production; And this process is pure physical method, does not have chemical reaction, yeast rich in selenium intracellular organic matter can not be destroyed, also can not bring objectionable impurities into, clean environmental protection; In addition, crushing medium also can Reusability, and nanometer cost is low.The product sporoderm-broken rate obtained under Optimizing Technical reaches 100%, and size is less than 400 nanometers, and biological selenium amount remains intact, and will greatly improve human body absorbing yeast rich in selenium.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.
Claims (4)
1. a preparation method for yeast rich in selenium broken wall and nanometer, is characterized in that: the processing step of the method is:
A, yeast rich in selenium is soluble in water, make the suspension of 10% ~ 30% (w/w) concentration;
B, the yeast rich in selenium suspension mixing up concentration is placed in ball mill;
C, choose agate bead and make impact bead, be positioned in the yeast rich in selenium suspension of ball mill, rotating ball mill, described agate bead carries out shock to the yeast rich in selenium in yeast rich in selenium suspension and pulverizes, thus the broken wall realizing yeast rich in selenium is pulverized;
D, then the suspension that step C obtains is joined in Ultrasonic Cell Disruptor, with the further refinement of Ultrasonic Cell Disruptor and homogenization;
E, the powdered finished product of suspension lyophilize that step D is obtained.
2. the preparation method of yeast rich in selenium broken wall according to claim 1 and nanometer, is characterized in that: described agate bead is two kinds, and the diameter of two kinds of described agate beads is respectively 0.5 ~ 1cm and 2 ~ 3cm.
3. the preparation method of yeast rich in selenium broken wall according to claim 1 and 2 and nanometer, is characterized in that: the rotating speed of described ball mill is 300 ~ 500r/min, and the time that described shock is pulverized is 0.5 ~ 2h.
4. the preparation method of yeast rich in selenium broken wall according to claim 3 and nanometer, is characterized in that: the power of described Ultrasonic Cell Disruptor is 200 ~ 400w, and the refinement of described Ultrasonic Cell Disruptor and homogenization time are 10 ~ 20min.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113519855A (en) * | 2021-05-26 | 2021-10-22 | 威海特伦斯生物工程有限公司 | Nano organic selenium tablet and preparation method thereof |
| CN114507601A (en) * | 2021-11-05 | 2022-05-17 | 南京郢天健康管理有限公司 | Wall breaking method for preparing nanoscale selenium-enriched yeast |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101906399A (en) * | 2010-07-19 | 2010-12-08 | 江苏丘陵地区镇江农业科学研究所 | Method for breaking walls of cells of bran by wet grinding |
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2015
- 2015-04-01 CN CN201510153242.9A patent/CN104694392A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101906399A (en) * | 2010-07-19 | 2010-12-08 | 江苏丘陵地区镇江农业科学研究所 | Method for breaking walls of cells of bran by wet grinding |
Non-Patent Citations (2)
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| 孔林等: "富锌酵母的选育及其培养工艺", 《食品与生物技术学报》 * |
| 李威等: "脆壁克鲁维酵母乳糖酶的提取与发酵优化", 《酿酒》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113519855A (en) * | 2021-05-26 | 2021-10-22 | 威海特伦斯生物工程有限公司 | Nano organic selenium tablet and preparation method thereof |
| CN114507601A (en) * | 2021-11-05 | 2022-05-17 | 南京郢天健康管理有限公司 | Wall breaking method for preparing nanoscale selenium-enriched yeast |
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