CN110656080A - Directional culture method of yeast cells - Google Patents
Directional culture method of yeast cells Download PDFInfo
- Publication number
- CN110656080A CN110656080A CN201911159471.6A CN201911159471A CN110656080A CN 110656080 A CN110656080 A CN 110656080A CN 201911159471 A CN201911159471 A CN 201911159471A CN 110656080 A CN110656080 A CN 110656080A
- Authority
- CN
- China
- Prior art keywords
- parts
- yeast cells
- culture
- temperature
- culture medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 210000005253 yeast cell Anatomy 0.000 title claims abstract description 49
- 238000012136 culture method Methods 0.000 title claims abstract description 10
- 238000003756 stirring Methods 0.000 claims abstract description 32
- 239000001963 growth medium Substances 0.000 claims abstract description 27
- 239000007788 liquid Substances 0.000 claims abstract description 23
- 235000013575 mashed potatoes Nutrition 0.000 claims abstract description 21
- 229910052761 rare earth metal Inorganic materials 0.000 claims abstract description 19
- 150000002910 rare earth metals Chemical class 0.000 claims abstract description 19
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000002360 preparation method Methods 0.000 claims abstract description 17
- 239000008367 deionised water Substances 0.000 claims abstract description 16
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 16
- 230000006698 induction Effects 0.000 claims abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 12
- 239000008103 glucose Substances 0.000 claims abstract description 12
- 229930091371 Fructose Natural products 0.000 claims abstract description 6
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims abstract description 6
- 239000005715 Fructose Substances 0.000 claims abstract description 6
- 238000006243 chemical reaction Methods 0.000 claims abstract description 6
- 230000005855 radiation Effects 0.000 claims description 26
- 238000002156 mixing Methods 0.000 claims description 20
- 239000000463 material Substances 0.000 claims description 17
- 239000011159 matrix material Substances 0.000 claims description 16
- 238000012258 culturing Methods 0.000 claims description 15
- 239000000843 powder Substances 0.000 claims description 14
- 229940070527 tourmaline Drugs 0.000 claims description 14
- 229910052613 tourmaline Inorganic materials 0.000 claims description 14
- 239000011032 tourmaline Substances 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 230000003213 activating effect Effects 0.000 claims description 10
- 238000001914 filtration Methods 0.000 claims description 10
- 239000011573 trace mineral Substances 0.000 claims description 10
- 235000013619 trace mineral Nutrition 0.000 claims description 10
- 238000005406 washing Methods 0.000 claims description 10
- 235000019774 Rice Bran oil Nutrition 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 6
- 239000008165 rice bran oil Substances 0.000 claims description 6
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 5
- CXRFDZFCGOPDTD-UHFFFAOYSA-M Cetrimide Chemical compound [Br-].CCCCCCCCCCCCCC[N+](C)(C)C CXRFDZFCGOPDTD-UHFFFAOYSA-M 0.000 claims description 5
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 5
- 229910017569 La2(CO3)3 Inorganic materials 0.000 claims description 5
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 5
- 244000061456 Solanum tuberosum Species 0.000 claims description 5
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 5
- NZPIUJUFIFZSPW-UHFFFAOYSA-H lanthanum carbonate Chemical compound [La+3].[La+3].[O-]C([O-])=O.[O-]C([O-])=O.[O-]C([O-])=O NZPIUJUFIFZSPW-UHFFFAOYSA-H 0.000 claims description 5
- 229960001633 lanthanum carbonate Drugs 0.000 claims description 5
- ICAKDTKJOYSXGC-UHFFFAOYSA-K lanthanum(iii) chloride Chemical compound Cl[La](Cl)Cl ICAKDTKJOYSXGC-UHFFFAOYSA-K 0.000 claims description 5
- 238000005360 mashing Methods 0.000 claims description 5
- 230000004048 modification Effects 0.000 claims description 5
- 238000012986 modification Methods 0.000 claims description 5
- 235000012015 potatoes Nutrition 0.000 claims description 5
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 claims description 3
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims description 3
- 229910052749 magnesium Inorganic materials 0.000 claims description 3
- 239000011777 magnesium Substances 0.000 claims description 3
- 239000011701 zinc Substances 0.000 claims description 3
- 229910052725 zinc Inorganic materials 0.000 claims description 3
- 229910052742 iron Inorganic materials 0.000 claims description 2
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 claims 1
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 abstract description 22
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 abstract description 22
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 abstract description 22
- 240000004808 Saccharomyces cerevisiae Species 0.000 abstract description 11
- 239000002609 medium Substances 0.000 abstract description 4
- 238000010521 absorption reaction Methods 0.000 abstract description 2
- 235000015097 nutrients Nutrition 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract description 2
- 230000000052 comparative effect Effects 0.000 description 11
- 239000012530 fluid Substances 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- YYGNTYWPHWGJRM-UHFFFAOYSA-N (6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC=C(C)CCC=C(C)CCC=C(C)C YYGNTYWPHWGJRM-UHFFFAOYSA-N 0.000 description 1
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 description 1
- BHEOSNUKNHRBNM-UHFFFAOYSA-N Tetramethylsqualene Natural products CC(=C)C(C)CCC(=C)C(C)CCC(C)=CCCC=C(C)CCC(C)C(=C)CCC(C)C(C)=C BHEOSNUKNHRBNM-UHFFFAOYSA-N 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N dodecahydrosqualene Natural products CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000011177 media preparation Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 150000002840 non-reducing disaccharides Chemical class 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 229940031439 squalene Drugs 0.000 description 1
- TUHBEKDERLKLEC-UHFFFAOYSA-N squalene Natural products CC(=CCCC(=CCCC(=CCCC=C(/C)CCC=C(/C)CC=C(C)C)C)C)C TUHBEKDERLKLEC-UHFFFAOYSA-N 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
Classifications
-
- 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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
-
- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Mycology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a directional culture method of yeast cells, which comprises the following steps: step one, preparation of a culture medium: adding 10-20 parts of glucose, 2-5 parts of citric acid and 2-6 parts of fructose into a reaction kettle, stirring at the rotation speed of 100-200r/min for 20-30min, then adding 15-25 parts of deionized water, and continuing stirring for 10 min. The culture medium is changed, and the induction medium is added before the directional culture, so that the yield of trehalose is fundamentally improved, the absorption of yeast on nutrient substances can be enhanced by adding the activated rare earth liquid into the culture medium, the trehalose generation of the yeast can be enhanced, and simultaneously the added mashed potatoes can play a bridging role in the culture medium after being modified, so that the mashed potatoes are fully mixed into the medium.
Description
Technical Field
The invention relates to the technical field of yeast cell culture, in particular to a directional culture method of yeast cells.
Background
Trehalose is a non-reducing disaccharide, which is formed by connecting glucose with 1, 1-glycosidic bond, widely exists in various organisms, especially in organisms resistant to dehydration, such as some microorganisms (especially yeast), drought-resistant plants and even low-temperature dormant vertebrates, and has the name of "life sugar" because trehalose can improve the resistance of the organisms to adverse environments, such as drought, low temperature, high temperature, pH, high permeability and the like.
Although the existing yeast cell culture method can produce trehalose, the culture effect of the adopted culture medium is not surprising, and the trehalose content needs to be improved by combining the culture medium with the culture method.
The prior Chinese patent document CN108841897A discloses a yeast culture method for producing trehalose, which comprises the following steps: step one, strain preparation, step two, strain screening, step three, culture medium preparation, step four, yeast culture, step five, trehalose content determination; the culture medium adopted by the culture method is simple, so that further improvement treatment is still needed.
Disclosure of Invention
The present invention aims to provide a method for the directed culture of yeast cells to solve the problems set forth in the background art.
In order to achieve the purpose, the invention provides the following technical scheme:
a method for the directed culture of yeast cells, comprising the steps of:
step one, preparation of a culture medium: adding 10-20 parts of glucose, 2-5 parts of citric acid and 2-6 parts of fructose into a reaction kettle, stirring at the rotation speed of 100-200r/min for 20-30min, then adding 15-25 parts of deionized water, continuing to stir for 10min, then adding 1-3 parts of activated rare earth liquid and 1-2 parts of trace elements, continuing to stir for 5-15min, then continuing to add 5-10 parts of modified mashed potato, continuing to stir at the rotation speed of 300-400r/min for 1-2h, and ending stirring to obtain a culture medium;
step two, activating and inoculating yeast cells: activating yeast cells in a glucose solution at the temperature of 22-30 ℃, and then adding the activated yeast cells into the culture medium in the first step, wherein the culture temperature is 38-42 ℃;
step three, preparing an induction matrix: adding 10-20 parts of maltose into 30-40 parts of rice bran oil, and stirring at the rotating speed of 200-500r/min for 20-30min to obtain an induction matrix;
step four, directional culture of yeast cells: and (3) inoculating the activated and inoculated yeast cells in the step two into the induction matrix in the step three to continue the directional culture, firstly culturing for 20h at the temperature of 42-45 ℃, and then culturing for 40h at the temperature of 30-40 ℃ to obtain the yeast cell.
Preferably, the preparation method of the activated rare earth liquid comprises the steps of adding lanthanum carbonate into hydrochloric acid to prepare a lanthanum chloride solution with the mass fraction of 30-40%, and then adopting60CorAt the radiation sourceAnd treating to obtain activated rare earth liquid.
Preferably, the60CorThe total dose of radiation source treatment is 2.0-2.5kGy, the radiation metering rate is 15.0-25.0Gy/min, and the radiation time is 1-2 min.
Preferably, the60CorThe total dose of the radiation source treatment is 2.25kGy, the radiation metering rate is 20.0Gy/min, and the radiation time is 1.5 min.
Preferably, the trace element is a composition of one or more of zinc element, magnesium element, manganese element and iron element.
Preferably, the preparation method of the modified mashed potato comprises the following steps:
(1) mashing potatoes into paste, adding deionized water, washing for 2-3 times, and filtering to obtain a primary material;
(2) then mixing the primary material and the irradiated tourmaline powder according to the weight ratio of 3:1, wherein the mixing speed is 20-100r/min, the mixing time is 10-20min, and after the mixing, sending the mixture into the modification treatment liquid for standing for 20-30min, and the standing temperature is 45-55 ℃;
(3) and washing and filtering the standing mixed material, and drying the obtained filter residue in an oven at the temperature of 50-60 ℃ for 20-30min to obtain the modified mashed potato.
Preferably, the modified treatment fluid comprises the following raw materials in parts by weight: 20-30 parts of tetradecyl trimethyl ammonium bromide and 60-70 parts of deionized water.
Preferably, the irradiated tourmaline powder is irradiated for 10-20min by adopting beta rays, and the irradiation dose is 1.8-2.2 Gy/min.
Preferably, the irradiated tourmaline powder is irradiated for 15min by adopting beta rays, and the irradiation dose is 2.0 Gy/min.
Compared with the prior art, the invention has the following beneficial effects:
the invention improves the yield of the trehalose fundamentally by changing the culture medium and adding the inducing medium before the directional culture, the addition of the activated rare earth liquid in the culture medium can enhance the absorption of the yeast to nutrient substances and enhance the production of the trehalose by the yeast, meanwhile, the added mashed potatoes can play a bridging role in the culture medium after being modified, the mashed potatoes are fully mixed into the medium, the tourmaline powder in the mashed potato can be further matched with activated rare earth liquid, the tourmaline powder and the activated rare earth liquid can play a synergistic and combined role to enhance the generation of trehalose, and rice bran oil in an induced matrix contains various fatty acids, r-oryzanol, plant squalene and other bioactive components, thereby further inducing the yeast to produce trehalose, the trehalose yield of the invention example 3 was 36.3%, while the trehalose yield in comparative example 2 was 21.1%, it was found that the trehalose yield in example 3 of the present invention was improved by 15.2% as compared to comparative example 2.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below with reference to specific embodiments, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1:
the method for directionally culturing the yeast cells comprises the following steps:
step one, preparation of a culture medium: adding 10 parts of glucose, 2 parts of citric acid and 2 parts of fructose into a reaction kettle, stirring at the rotating speed of 100r/min for 20min, then adding 15 parts of deionized water, continuing to stir for 10min, then adding 1 part of activated rare earth liquid and 1 part of trace elements, continuing to stir for 5min, then continuing to add 5 parts of modified mashed potato, continuing to stir at the rotating speed of 300r/min for 1h, and obtaining a culture medium after stirring;
step two, activating and inoculating yeast cells: activating yeast cells in a glucose solution at the temperature of 22 ℃, and then adding the activated yeast cells into the culture medium obtained in the first step, wherein the culture temperature is 38 ℃;
step three, preparing an induction matrix: adding 10 parts of maltose into 30 parts of rice bran oil, and stirring at the rotating speed of 200r/min for 20min to obtain an induction matrix;
step four, directional culture of yeast cells: and (3) inoculating the activated and inoculated yeast cells in the step two into the induction matrix in the step three to continue the directional culture, firstly culturing for 20h at 42 ℃, and then culturing for 40h at 30 ℃ to obtain the yeast cell.
The preparation method of the activated rare earth liquid in this example is to add lanthanum carbonate into hydrochloric acid to prepare a lanthanum chloride solution with a mass fraction of 30%, and then adopt60CorTreating with radiation source to obtain activated rare earth liquid.
Of the present embodiment60CorThe total dose of the radiation source treatment is 2.0kGy, the radiation metering rate is 15.0Gy/min, and the radiation time is 1 min.
The trace element in this example is zinc.
The preparation method of the modified mashed potato of the embodiment comprises the following steps:
(1) mashing potatoes into paste, adding deionized water, washing for 2 times, and filtering to obtain a primary material;
(2) then mixing the primary material and the irradiated tourmaline powder according to the weight ratio of 3:1, wherein the mixing speed is 20r/min, the mixing time is 10min, and after the mixing is finished, sending the mixture into a modification treatment liquid to stand for 20min, and the standing temperature is 45 ℃;
(3) and washing and filtering the standing mixed material, and drying the obtained filter residue in an oven at the temperature of 50 ℃ for 20min to obtain the modified mashed potato.
The modified treatment fluid comprises the following raw materials in parts by weight: 20 parts of tetradecyl trimethyl ammonium bromide and 60 parts of deionized water.
The irradiated tourmaline powder of the embodiment is irradiated for 10min by beta rays, and the irradiation dose is 1.8 Gy/min.
Example 2:
the method for directionally culturing the yeast cells comprises the following steps:
step one, preparation of a culture medium: adding 20 parts of glucose, 5 parts of citric acid and 6 parts of fructose into a reaction kettle, stirring at the rotating speed of 200r/min for 30min, then adding 25 parts of deionized water, continuing to stir for 10min, then adding 3 parts of activated rare earth liquid and 2 parts of trace elements, continuing to stir for 15min, then continuing to add 10 parts of modified mashed potato, continuing to stir at the rotating speed of 400r/min for 2h, and obtaining a culture medium after stirring;
step two, activating and inoculating yeast cells: activating yeast cells in a glucose solution at 30 ℃, and then adding the activated yeast cells into the culture medium obtained in the first step, wherein the culture temperature is 42 ℃;
step three, preparing an induction matrix: adding 20 parts of maltose into 40 parts of rice bran oil, and stirring at the rotating speed of 500r/min for 30min to obtain an induction matrix;
step four, directional culture of yeast cells: and (3) inoculating the activated and inoculated yeast cells in the step two into the induction matrix in the step three to continue directional culture, firstly culturing for 20h at 45 ℃, and then culturing for 40h at 40 ℃ to obtain the yeast cell.
The preparation method of the activated rare earth liquid in this example is to add lanthanum carbonate into hydrochloric acid to prepare a lanthanum chloride solution with a mass fraction of 40%, and then adopt60CorTreating with radiation source to obtain activated rare earth liquid.
Of the present embodiment60CorThe total dose of the radiation source treatment is 2.5kGy, the radiation metering rate is 25.0Gy/min, and the radiation time is 2 min.
The trace element in this example is magnesium.
The preparation method of the modified mashed potato of the embodiment comprises the following steps:
(1) mashing potatoes into paste, adding deionized water, washing for 3 times, and filtering to obtain a primary material;
(2) then mixing the primary material and the irradiated tourmaline powder according to the weight ratio of 3:1, wherein the mixing speed is 100r/min, the mixing time is 20min, and after the mixing is finished, sending the mixture into a modification treatment liquid to stand for 30min, and the standing temperature is 55 ℃;
(3) and washing and filtering the standing mixed material, and drying the obtained filter residue in an oven at the temperature of 60 ℃ for 30min to obtain the modified mashed potato.
The modified treatment fluid comprises the following raw materials in parts by weight: 30 parts of tetradecyl trimethyl ammonium bromide and 70 parts of deionized water.
The irradiated tourmaline powder of the embodiment is irradiated for 20min by adopting beta rays, and the irradiation dose is 2.2 Gy/min.
Example 3:
the method for directionally culturing the yeast cells comprises the following steps:
step one, preparation of a culture medium: adding 15 parts of glucose, 3.5 parts of citric acid and 4 parts of fructose into a reaction kettle, stirring at the rotating speed of 150r/min for 25min, then adding 20 parts of deionized water, continuing to stir for 10min, then adding 2 parts of activated rare earth liquid and 1.5 parts of trace elements, continuing to stir for 10min, then continuing to add 7.5 parts of modified mashed potato, continuing to stir at the rotating speed of 350r/min for 1.5h, and obtaining a culture medium after stirring;
step two, activating and inoculating yeast cells: activating yeast cells in a glucose solution at 26 ℃, and then adding the activated yeast cells into the culture medium obtained in the first step, wherein the culture temperature is 40 ℃;
step three, preparing an induction matrix: adding 15 parts of maltose into 35 parts of rice bran oil, and stirring at the rotating speed of 350r/min for 25min to obtain an induction matrix;
step four, directional culture of yeast cells: and (3) inoculating the activated and inoculated yeast cells in the step two into the induction matrix in the step three to continue the directional culture, firstly culturing for 20h at 43.5 ℃, and then culturing for 40h at 35 ℃ to obtain the yeast cell.
The preparation method of the activated rare earth liquid in this example is to add lanthanum carbonate into hydrochloric acid to prepare a lanthanum chloride solution with a mass fraction of 35%, and then adopt60CorTreating with radiation source to obtain activated rare earth liquid.
Of the present embodiment60CorThe total dose of the radiation source treatment is 2.25kGy, the radiation metering rate is 20.0Gy/min, and the radiation time is 1.5 min.
The trace element in this example is iron
The preparation method of the modified mashed potato of the embodiment comprises the following steps:
(1) mashing potatoes into paste, adding deionized water, washing for 3 times, and filtering to obtain a primary material;
(2) then mixing the primary material and the irradiated tourmaline powder according to the weight ratio of 3:1, wherein the mixing speed is 60r/min, the mixing time is 15min, and after the mixing is finished, sending the mixture into a modification treatment liquid to stand for 25min, and the standing temperature is 50 ℃;
(3) and washing and filtering the standing mixed material, and drying the obtained filter residue in an oven at the temperature of 55 ℃ for 25min to obtain the modified mashed potato.
The modified treatment fluid comprises the following raw materials in parts by weight: 25 parts of tetradecyl trimethyl ammonium bromide and 65 parts of deionized water.
The irradiated tourmaline powder of the embodiment is irradiated for 15min by beta rays, and the irradiation dose is 2.0 Gy/min.
Comparative example 1.
The materials and preparation process were substantially the same as those of example 3, except that the culture medium was not supplemented with the modified mashed potato.
Comparative example 2.
Basically the same materials and preparation process as in example 3, except that Chinese patent document CN108841897A discloses a yeast culture method for producing trehalose, which comprises the following steps of example 2 raw materials and method.
The trehalose yield was obtained by comparing the amount of trehalose produced by the yeasts of examples 1 to 3 and comparative examples 1 to 2 with that produced by the existing yeast.
The results of the performance tests of examples 1 to 3 and comparative examples 1 to 2 are as follows
| Trehalose yield (%) | |
| Example 1 | 35.8 |
| Example 2 | 35.2 |
| Example 3 | 36.3 |
| Comparative example 1 | 28.1 |
| Comparative example 2 | 21.1 |
As shown in examples 1-3 and comparative examples 1-2, the trehalose yield of example 3 according to the present invention was 36.3%, while the trehalose yield of comparative example 2 was 21.1%, indicating that the trehalose yield of example 3 according to the present invention was improved by 15.2% as compared with comparative example 2.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (9)
1. A method for the directed culture of yeast cells, comprising the steps of:
step one, preparation of a culture medium: adding 10-20 parts of glucose, 2-5 parts of citric acid and 2-6 parts of fructose into a reaction kettle, stirring at the rotation speed of 100-200r/min for 20-30min, then adding 15-25 parts of deionized water, continuing to stir for 10min, then adding 1-3 parts of activated rare earth liquid and 1-2 parts of trace elements, continuing to stir for 5-15min, then continuing to add 5-10 parts of modified mashed potato, continuing to stir at the rotation speed of 300-400r/min for 1-2h, and ending stirring to obtain a culture medium;
step two, activating and inoculating yeast cells: activating yeast cells in a glucose solution at the temperature of 22-30 ℃, and then adding the activated yeast cells into the culture medium in the first step, wherein the culture temperature is 38-42 ℃;
step three, preparing an induction matrix: adding 10-20 parts of maltose into 30-40 parts of rice bran oil, and stirring at the rotating speed of 200-500r/min for 20-30min to obtain an induction matrix;
step four, directional culture of yeast cells: and (3) inoculating the activated and inoculated yeast cells in the step two into the induction matrix in the step three to continue the directional culture, firstly culturing for 20h at the temperature of 42-45 ℃, and then culturing for 40h at the temperature of 30-40 ℃ to obtain the yeast cell.
2. The method for directionally culturing yeast cells as claimed in claim 1, wherein the activated rare earth solution is prepared by adding lanthanum carbonate into hydrochloric acid to prepare a lanthanum chloride solution with a mass fraction of 30-40%, and then adopting60CorTreating with radiation source to obtain activated rare earth liquid.
3. The method of claim 2, wherein the yeast cells are cultured in a targeted manner60CorThe total dose of radiation source treatment is 2.0-2.5kGy, the radiation metering rate is 15.0-25.0Gy/min, and the radiation time is 1-2 min.
4. The method of claim 3, wherein the yeast cells are cultured in a targeted manner60CorThe total dose of the radiation source treatment is 2.25kGy, the radiation metering rate is 20.0Gy/min, and the radiation time is 1.5 min.
5. The method of claim 1, wherein the trace element is a combination of one or more of zinc, magnesium, manganese, and iron.
6. The method for directionally culturing yeast cells as claimed in claim 1, wherein the modified mashed potato is prepared by the following steps:
(1) mashing potatoes into paste, adding deionized water, washing for 2-3 times, and filtering to obtain a primary material;
(2) then mixing the primary material and the irradiated tourmaline powder according to the weight ratio of 3:1, wherein the mixing speed is 20-100r/min, the mixing time is 10-20min, and after the mixing, sending the mixture into the modification treatment liquid for standing for 20-30min, and the standing temperature is 45-55 ℃;
(3) and washing and filtering the standing mixed material, and drying the obtained filter residue in an oven at the temperature of 50-60 ℃ for 20-30min to obtain the modified mashed potato.
7. The method for directional culture of yeast cells as claimed in claim 6, wherein the modified treatment solution comprises the following raw materials in parts by weight: 20-30 parts of tetradecyl trimethyl ammonium bromide and 60-70 parts of deionized water.
8. The directional culture method of yeast cells according to claim 1, wherein the irradiated tourmaline powder is irradiated with beta rays for 10-20min, and the irradiation dose is 1.8-2.2 Gy/min.
9. The directional culture method of yeast cells according to claim 8, wherein the irradiated tourmaline powder is irradiated with beta rays for 15min, and the irradiation dose is 2.0 Gy/min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911159471.6A CN110656080A (en) | 2019-11-22 | 2019-11-22 | Directional culture method of yeast cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911159471.6A CN110656080A (en) | 2019-11-22 | 2019-11-22 | Directional culture method of yeast cells |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110656080A true CN110656080A (en) | 2020-01-07 |
Family
ID=69043778
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911159471.6A Pending CN110656080A (en) | 2019-11-22 | 2019-11-22 | Directional culture method of yeast cells |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110656080A (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101818183A (en) * | 2010-04-15 | 2010-09-01 | 山东轻工业学院 | Method for improving trehalose content of yeast cell |
| CN105219663A (en) * | 2015-09-18 | 2016-01-06 | 上海交通大学 | The special strain therefore of trehalose synthesis and the method for the synthesis of trehalose thereof |
| CN105462888A (en) * | 2015-12-23 | 2016-04-06 | 青岛海科生物技术有限公司 | Efficient culture medium as well as preparation method and application thereof |
| CN105602861A (en) * | 2015-10-20 | 2016-05-25 | 珲春市龙裕农业发展集团有限公司 | Germanium-rich yeast additive adopting rice bran as medium, and preparation method thereof |
| CN105695349A (en) * | 2016-04-15 | 2016-06-22 | 湖北工业大学 | Method using phosphorus starvation culture to increase intracellular trehalose of yeast cells |
| CN107502631A (en) * | 2017-09-28 | 2017-12-22 | 上海应用技术大学 | A kind of production method of candida utili β D glucans |
| CN110055293A (en) * | 2019-04-29 | 2019-07-26 | 长沙而道新能源科技有限公司 | A kind of synthetic method of trehalose |
-
2019
- 2019-11-22 CN CN201911159471.6A patent/CN110656080A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101818183A (en) * | 2010-04-15 | 2010-09-01 | 山东轻工业学院 | Method for improving trehalose content of yeast cell |
| CN105219663A (en) * | 2015-09-18 | 2016-01-06 | 上海交通大学 | The special strain therefore of trehalose synthesis and the method for the synthesis of trehalose thereof |
| CN105602861A (en) * | 2015-10-20 | 2016-05-25 | 珲春市龙裕农业发展集团有限公司 | Germanium-rich yeast additive adopting rice bran as medium, and preparation method thereof |
| CN105462888A (en) * | 2015-12-23 | 2016-04-06 | 青岛海科生物技术有限公司 | Efficient culture medium as well as preparation method and application thereof |
| CN105695349A (en) * | 2016-04-15 | 2016-06-22 | 湖北工业大学 | Method using phosphorus starvation culture to increase intracellular trehalose of yeast cells |
| CN107502631A (en) * | 2017-09-28 | 2017-12-22 | 上海应用技术大学 | A kind of production method of candida utili β D glucans |
| CN110055293A (en) * | 2019-04-29 | 2019-07-26 | 长沙而道新能源科技有限公司 | A kind of synthetic method of trehalose |
Non-Patent Citations (1)
| Title |
|---|
| 董永胜: "《谷胱甘肽生产技术》", 31 August 2015, 中国轻工业出版社 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102433255B (en) | Method for producing table vinegar by adopting two-step acetic acid fermentation method | |
| CN101215517B (en) | Technique for producing germinating brown rice vinegar and products thereof | |
| CN101665384B (en) | Se-enriched bio-organic fertilizer and producing method thereof | |
| CN101649286B (en) | Method for preparing hawthorn fruit vinegar | |
| CN103865701A (en) | Beer compound enzyme containing neutral protease | |
| CN106867865A (en) | A kind of preparation method of bletilla vinegar | |
| CN101289680A (en) | Process for producing 2,3-butanediol using american artichoke as raw material by fermentation | |
| CN106495855A (en) | A kind of edible fungus fermented slag selenium-enriched high-calcium ecological organic fertilier | |
| CN111454799A (en) | Flavored malt beer and preparation method thereof | |
| CN101671699A (en) | Method for preparing ethanol by fermenting tobacco leftovers by mixing composite carrier and immobilized yeast | |
| CN103305399B (en) | Aromatic millet vinegar and preparation method thereof | |
| CN106478264A (en) | A kind of fruit fermentation residue selenium-enriched high-calcium environmental protection fertilizer | |
| CN104223296A (en) | Sports functional drink prepared from secondarily fermented pickle liquid | |
| CN105018262A (en) | Cordyceps militaris draft beer and brewing process thereof | |
| CN106495856A (en) | A kind of cassava fermentation residue selenium-enriched high-calcium fertilizer | |
| CN102276305B (en) | Biological organic fertilizer special for pollution-free medlar and preparation method thereof | |
| CN107455723B (en) | Method for producing cooking wine with high amino acid nitrogen content by using yellow wine lees | |
| CN110656080A (en) | Directional culture method of yeast cells | |
| CN101724531A (en) | Method for preparing rice puree spirit | |
| CN106007874A (en) | High-temperature-resistant foliage fertilizer | |
| CN116355618A (en) | Soil conditioner, preparation method thereof and application thereof in improving acid soil | |
| CN104909942A (en) | High-efficiency termitomyces albuminosus culture medium prepared from needle mushroom dregs and preparation method of culture medium | |
| CN102936587A (en) | Method for producing low-temperature pectinase and low-temperature clear apple juice from shaddock peel powder | |
| CN107674813A (en) | A kind of preparation method of low sugar aromatic vinegar | |
| CN112772807A (en) | Preparation method of eucommia ulmoides fermented beverage |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information |
Inventor after: Gu Ting Inventor after: Wen Chunlan Inventor before: Gu Ting |
|
| CB03 | Change of inventor or designer information | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20220316 Address after: Room 409 and 412, building C, Caohu science and Technology Park, Xijiao University, No. 1, Guantang Road, Caohu street, Xiangcheng District, Suzhou, Jiangsu 215100 Applicant after: Suzhou herneng Biotechnology Co.,Ltd. Address before: 730000 403, No. 73, Zhongshan forest, Chengguan District, Lanzhou City, Gansu Province Applicant before: Gu Ting |
|
| TA01 | Transfer of patent application right | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200107 |
|
| RJ01 | Rejection of invention patent application after publication |