WO2022143113A1 - Chromium-rich yeast having high chromium absorption, preparation method therefor and application thereof - Google Patents

Chromium-rich yeast having high chromium absorption, preparation method therefor and application thereof Download PDF

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WO2022143113A1
WO2022143113A1 PCT/CN2021/137249 CN2021137249W WO2022143113A1 WO 2022143113 A1 WO2022143113 A1 WO 2022143113A1 CN 2021137249 W CN2021137249 W CN 2021137249W WO 2022143113 A1 WO2022143113 A1 WO 2022143113A1
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chromium
yeast
saccharomyces cerevisiae
enriched yeast
source
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PCT/CN2021/137249
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French (fr)
Chinese (zh)
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肖明华
翟明
俞灿
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安琪酵母股份有限公司
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Publication of WO2022143113A1 publication Critical patent/WO2022143113A1/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • 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
    • 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
    • C12N1/18—Baker's yeast; Brewer's yeast
    • 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
    • C12N1/36—Adaptation or attenuation of cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/85—Saccharomyces
    • C12R2001/865—Saccharomyces cerevisiae
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

Definitions

  • the invention relates to the technical field of microorganism application, in particular to a chromium-enriched yeast with high chromium absorption rate and a production method and application thereof.
  • Chromium yeast is one of the sources of chromium nutritional supplements designated by the State Food and Drug Administration.
  • Yeast is the earliest and most widely used pure natural nutritional microorganism, and it is also a natural nutritional treasure and an ideal biological carrier.
  • Yeast chromium is to cultivate yeast cells in a medium containing trivalent chromium, and convert inorganic chromium into organic chromium through biotransformation, so as to improve the absorption and utilization rate of chromium in the body, reduce its toxic and side effects, and better play its role. Regulates blood sugar, lipid-lowering and cholesterol-lowering effects.
  • the existing products mainly use Saccharomyces cerevisiae as a strain, add carbon, nitrogen and phosphorus sources, and use inorganic chromium as a chromium source. After fermentation and culture, the chromium-enriched yeast yeast milk is finally obtained, and then the product is obtained by drying.
  • inorganic chromium has the following defects: the absorption rate is low, generally less than 10%; it has no biological activity and needs to be converted into biologically active GTF chromium to regulate metabolism. effect.
  • the body of patients with diabetes and coronary heart disease basically does not have this transformation ability; the harmful effects of inorganic chromium have been found in animal experiments.
  • the problem of the prior art solved by the present invention is as follows: the existing products mainly use Saccharomyces cerevisiae as a strain, add carbon, nitrogen and phosphorus sources, and use inorganic chromium as a chromium source. product.
  • inorganic chromium there are more or less inorganic chromium in the existing products, the absorption rate of inorganic chromium is low, and it needs metabolic regulation to play a role, and some patients cannot even use it.
  • inorganic chromium is easy to produce and trace amount of hexavalent chromium in the process product of chromium source, and trace amount of hexavalent chromium also exists in the chromium-enriched yeast in the present technology. Hexavalent chromium is harmful to human and animal health.
  • the invention adopts organic chromium as the chromium source, eliminates the inorganic chromium source, the main raw material of the present invention (the chromium source is organic chromium) from the source, and uses the organic chromium raw material to produce the production process of chromium-enriched yeast, and there is no inorganic chromium in the finished product. , and the product has higher chromium content and chromium absorption rate.
  • the present invention proposes the following technical solutions.
  • the invention provides a chromium-enriched yeast, wherein the mass content of chromium in the chromium-enriched yeast is greater than 38,000 ppm, the organic chromium ratio in the chromium-enriched yeast is greater than 97% by mass, and the chromium-enriched yeast has a chromium absorption rate by mass. >50%, there is no hexavalent chromium in the chromium-rich yeast.
  • the mass content of chromium in the chromium-enriched yeast is greater than 40,000 ppm
  • the organic chromium ratio in the chromium-enriched yeast is greater than 95% by mass
  • the chromium-enriched yeast is more than 60% by mass in the absorption rate of chromium.
  • the present invention also provides the preparation method of the above-mentioned chromium-enriched yeast, characterized in that it comprises the following steps:
  • Step 1 Amplify Saccharomyces cerevisiae to provide Saccharomyces cerevisiae seeds required for fermentation;
  • Step 2 transfer the Saccharomyces cerevisiae seeds obtained in step 1 into a 50L fermentation tank to continue fermentation, and when the wet weight of Saccharomyces cerevisiae reaches 60-90g/L, add chromium source;
  • Step 3 growing the Saccharomyces cerevisiae obtained in Step 2, when the wet weight of the yeast reaches 120-140g/L, continue fermentation to obtain yeast milk;
  • Step 4 separate the yeast milk obtained in step 3, and then dry to obtain the chromium-enriched yeast;
  • the chromium source is organic chromium
  • the organic chromium is selected from one or more of chromium nicotinate, chromium picolinate and chromium methionine.
  • the fermentation temperature is 25-35° C., preferably, the dissolved oxygen is less than or equal to 30 ⁇ mol/L, and more preferably, the concentration of ethanol is less than or equal to 0.3%.
  • the chromium source is added in flow, preferably, the amount of the chromium source is 2-4mol, the chromium The source is all added within 2-10h.
  • the pH is controlled to be 3.9-4.5. 18h.
  • the pH is raised to 5-8 within 1-4 hours before the end of the fermentation, until the end of the fermentation, to obtain yeast milk.
  • Saccharomyces cerevisiae described in step 1 is selected from Saccharomyces cerevisiae Z1.3 (Saccharomyces cerevisiae HANSEN Z1.3), Saccharomyces cerevisiae Z2.1 (Saccharomyces cerevisiae HANSEN Z2.1) and Saccharomyces cerevisiae FX-2 (Saccharomyces cerevisiae HANSEN Z2.1) One or more of FX-2).
  • the components of the medium used in the culture in steps 1-3 contain one or more of carbon sources, nitrogen sources, phosphorus sources and trace elements.
  • the carbon source is selected from molasses and/or glucose
  • the nitrogen source is selected from one or more of ammonium sulfate, peptone, yeast extract and aqueous ammonia
  • the phosphorus source is selected from dihydrogen phosphate Potassium
  • the trace element is selected from zinc sulfate and/or magnesium sulfate.
  • the concentration of the molasses is 20-40% by mass, and the concentration of the glucose is 20-40% by mass.
  • the purity of the potassium dihydrogen phosphate is greater than 96%, the purity of the ammonium sulfate is greater than 99%, preferably, the purity of the zinc sulfate is greater than 99%, and the purity of the magnesium sulfate is greater than 99% %.
  • the carbon source, the nitrogen source and the phosphorus source are added by means of feed addition.
  • the present invention also provides a chromium-enriched yeast, which is characterized in that it is produced by the above-mentioned method.
  • the present invention also provides the application of the chromium-enriched yeast in the field of animal husbandry, preferably the application in the field of feed or feed additive.
  • the invention adopts organic chromium to replace traditional inorganic chromium as the chromium source, the absorption rate of chromium by yeast is greatly improved, and the absorption rate of the shake flask is increased from 1% to 2% to more than 60%;
  • the total chromium of the chromium-enriched yeast can easily break through the upper limit of the traditional chromium-enriched yeast total chromium of 8000ppm, the minimum content is also above 38000ppm, can reach above 40000ppm, as high as 43096ppm, the mass content of organic chromium is more than 97%, and even can reach 98% The above; there is inorganic chromium that is difficult to wash in traditional chromium-enriched yeast.
  • the new chromium-enriched yeast eliminates inorganic chromium from the source, and what remains is organic chromium, which is more conducive to human absorption and animal absorption; organic chromium sources are used to produce rich Chromium yeast can greatly reduce the chromium content in production wastewater and reduce environmental pressure; there is no hexavalent chromium in the chromium-enriched yeast produced by this process.
  • the strain Saccharomyces cerevisiae Z1.3 (Saccharomyces cerevisiae Hansen Z1.3) used in the present invention has been biologically preserved in the China Center for Type Culture Collection (CCTCC, Wuhan University, zip code 430072) on October 25, 2005, The deposit number is CCTCC M 205125, and this strain has been recorded in the patent publication with the application number CN200610066365.X.
  • Saccharomyces cerevisiae FX-2 Saccharomyces cerevisiae FX-2 used in the present invention was deposited in the China Center for Type Culture Collection (CCTCC) on August 1, 2016, and the deposit number is CCTCC NO: M2016418, and the deposit address is China. Wuhan. Wuhan University, Postal Code: 430072; Tel: (027)-68754052, the strain has been recorded in the patent publication with the application number CN201611141122.8.
  • the strain Saccharomyces cerevisiae Z2.1 (Saccharomyces cerevisiae Hansen Z2.1) used in the present invention was preserved in the China Center for Type Culture Collection on October 25, 2005, the strain preservation number is CCTCC NO: M205127, and the preservation address is: China . Wuhan. Wuhan University, Postcode: 430072, Tel: (027) 68752319, the strain has been recorded in the patent publication with the application number CN201710522840.8.
  • the invention provides a chromium-enriched yeast, wherein the mass content of chromium in the chromium-enriched yeast is greater than 38,000 ppm, the organic chromium ratio in the chromium-enriched yeast is greater than 97% by mass, and the chromium-enriched yeast has a chromium absorption rate by mass. >50%, there is no hexavalent chromium in the chromium-rich yeast.
  • the mass content of chromium in the chromium-enriched yeast is greater than 40,000 ppm
  • the organic chromium ratio in the chromium-enriched yeast is greater than 95% by mass
  • the chromium-enriched yeast is more than 60% by mass in the absorption rate of chromium.
  • the present invention also provides the preparation method of the above-mentioned chromium-enriched yeast, characterized in that it comprises the following steps:
  • Step 1 Amplify Saccharomyces cerevisiae to provide Saccharomyces cerevisiae seeds required for fermentation;
  • Step 2 transfer the Saccharomyces cerevisiae seeds obtained in step 1 into a 50L fermentation tank to continue fermentation, and when the wet weight of Saccharomyces cerevisiae reaches 60-90g/L, add chromium source;
  • Step 3 growing the Saccharomyces cerevisiae obtained in step 2, when the wet weight of the yeast reaches 120-140 g/L, continue fermentation to obtain yeast milk;
  • Step 4 separate the yeast milk obtained in step 3, and then dry to obtain the chromium-enriched yeast;
  • the chromium source is organic chromium
  • the organic chromium is selected from one or more of chromium nicotinate, chromium picolinate and chromium methionine.
  • the present invention also provides a chromium-enriched yeast, which is characterized in that it is produced by the above-mentioned method.
  • the present invention also provides the application of the chromium-enriched yeast in the field of animal husbandry, preferably the application in the field of feed or feed additive.
  • the mass content of chromium in the chromium-enriched yeast is 0-40000 ppm, that is, the amount of chromium absorption is calculated by mass, and the chromium content per gram of yeast is 0-0.04 g;
  • Chromium absorption rate total chromium mass content in yeast (product) / mass content of chromium in the added chromium source;
  • organic chromium ratio intracellular organic chromium mass content/yeast (product) total chromium mass content
  • the chromium content in the chromium-enriched yeast of the present invention refers to the chromium mass content of beneficial trivalent chromium, and at the same time, hexavalent chromium cannot exist.
  • Chromium source chromium nicotinate or chromium methionine or chromium pyridinecarboxylate
  • the yeast milk is dusted by a dusting and drying tower to obtain a chromium-enriched yeast product.
  • Step 1 the Saccharomyces cerevisiae Chromium-enriched Saccharomyces cerevisiae Z1.3 (Saccharomyces cerevisiae HANSEN Z1.3) is picked a ring to carry out shake-flask culture—the step-by-step amplification culture of seed culture, to provide the Saccharomyces cerevisiae seeds required for fermentation;
  • the components of the culture medium in the shake flask are: 100 g of sucrose, 20 g of yeast extract, 1 g of magnesium sulfate, 1 g of potassium dihydrogen phosphate, pH 4.8, cultured at 31°C for 24 hours, 5L shake flask with a liquid volume of 1L, and the shaker rotation speed 180rpm/ min;
  • Step 2 transfer the Saccharomyces cerevisiae seeds of step 1 gained into 50L fermentor, control fermentation temperature to be 35 °C, dissolved oxygen 30 ⁇ mol/L, after 2 hours of fermentation, flow the carbon source (molasses concentration is 30%) of 4000g.
  • Step 3 add 8000g carbon source (molasses concentration is 30%), 200g nitrogen source (ammonium sulfate), 200g phosphorus source (potassium dihydrogen phosphate), control the ethanol concentration to be 0.07% ⁇ 0.03%, for the growth of Saccharomyces cerevisiae , when the wet weight of the yeast reaches 120g/L, continue to maintain the pH at 3.9, control the dissolved oxygen at 30 ⁇ mol/L by ventilation and stirring, continue the fermentation for 9h, start 4h before the end of the fermentation, and gradually increase the pH to 6 with ammonia water , until the end of fermentation, to obtain yeast milk;
  • Step 4 The chromium-enriched yeast product A is obtained by spray-drying the yeast milk obtained in step 3 after separating impurities.
  • the chromium content of the prepared chromium-enriched yeast product A is 40563 ppm; the organic chromium ratio is 98.39%; the chromium absorption rate is 66.45%, and the prepared Chromium-enriched yeast product A has no hexavalent chromium.
  • the steps are the same as those in Example 1, except that the chromium source fed in the step 2 is 3 mol of chromium methionine.
  • the chromium content of the prepared chromium-enriched yeast product B was 38451 ppm; the organic chromium ratio was 97.78%; the chromium absorption rate was 61.03%, and the prepared chromium-enriched yeast product B had no hexavalent chromium.
  • the steps are the same as in Example 1, except that the chromium source streamed in the step 2 is 3mol chromium picolinate.
  • the prepared chromium-enriched yeast product C has a chromium content of 43096 ppm, an organic chromium ratio of 98.78%, and a chromium absorption rate of 68.4%.
  • the prepared chromium-enriched yeast product C has no hexavalent chromium.
  • Example 2 The rest of the experimental steps are the same as in Example 1, except that when the wet weight of Saccharomyces cerevisiae reaches 90 g/L in step 2, after adjusting the pH to 3.9, the chromium source (0.01 mol/L chromium nicotinate for a total of 3 mol , the chromium source is all added within 10h, and then the chromium-enriched yeast product D is obtained.
  • the prepared chromium-enriched yeast product D has a chromium content of 41,025 ppm, an organic chromium ratio of 98.78%, and a chromium absorption rate of 65.18%. There is no hexavalent chromium in the chromium-enriched yeast product D.
  • step 3 when the wet weight of the yeast reaches 140 g/L, the pH is controlled to 3.9, the dissolved oxygen is controlled to 30 ⁇ mol/L by ventilation and stirring, and the fermentation is continued for 18 h.
  • the pH was raised to 6 within 4 hours before the end of the fermentation until the end of the fermentation to obtain yeast milk, and then the chromium-enriched yeast product E was obtained.
  • the prepared chromium-enriched yeast product E has a chromium content of 40127 ppm, an organic chromium ratio of 98.67%, and a chromium absorption rate of 63.76%.
  • the prepared chromium-enriched yeast product E has no hexavalent chromium.
  • step 2 the fermentation temperature of the Saccharomyces cerevisiae seeds obtained in step 1 is controlled to be 25°C; afterward, the chromium-enriched yeast product F is obtained.
  • the prepared chromium-enriched yeast product F has a chromium content of 41357 ppm, an organic chromium ratio of 98.88%, and a chromium absorption rate of 65.71%.
  • the prepared chromium-enriched yeast product F has no hexavalent chromium.
  • step 2 when the wet weight of Saccharomyces cerevisiae reaches 60 g/L, after adjusting the pH to 4.5, maintain the pH at 4.5 (the pH drops and then adds ammonia water to raise the pH to 4.5). ); and then the chromium-enriched yeast product G was obtained.
  • the prepared chromium-enriched yeast product G has a chromium content of 40357 ppm, an organic chromium ratio of 98.38%, and a chromium absorption rate of 64.12%.
  • the prepared chromium-enriched yeast product G has no hexavalent chromium.
  • Example 2 The rest of the experimental conditions are the same as those in Example 1, except that the chromium source fed in in step 2 is chromium chloride; the chromium-enriched yeast product D1 is then obtained.
  • the chromium content of the prepared chromium-enriched yeast product D1 was 2044.78ppm; the organic chromium ratio was 89.71%; the chromium absorption rate was 2.22%; and the hexavalent chromium was 0.933ppm.
  • step 2 chromium nicotinate is not added, and in step 3, when the wet weight reaches 120 g/L, 3 mol of chromium nicotinate is added within 1 hour; then the chromium-enriched yeast product D2 is obtained. .
  • the total chromium content in the chromium-enriched yeast product D2 was 32647ppm, the absorption rate of chromium was 51.87%, and the proportion of organic chromium was 98.52%.
  • Example 2 The rest of the experimental conditions are the same as in Example 1, except that 3 mol of chromium nicotinate was added at the beginning of the fermentation in step 3, and the total amount of chromium nicotinate added was exactly the same as in Example 1; the chromium-enriched yeast product D3 was obtained later.
  • the total chromium content in the chromium-enriched yeast product D3 was 35612ppm, the absorption rate of chromium was 56.58%, and the proportion of organic chromium was 98.41%.
  • Example 2 The rest of the experimental conditions are the same as in Example 1, the difference is only that the fermentation pH is controlled at 5.5 in Steps 2 and 3, and then the chromium-enriched yeast product D4 is obtained.
  • the total chromium content in the chromium-enriched yeast product D4 was 31985ppm, the absorption rate of chromium was 50.82%, and the proportion of organic chromium was 97.89%.
  • Example 2 The rest of the experimental conditions are the same as in Example 1, the only difference is that the fermentation temperature is controlled at 38° C. in Steps 2 and 3; then the chromium-enriched yeast product D5 is obtained.
  • the total chromium content in the chromium-enriched yeast product D5 was 26854ppm, the absorption rate of chromium was 42.67%, and the proportion of organic chromium was 98.14%.
  • step 3 the alcohol of the fermentation broth is controlled at 1.0%; then the chromium-enriched yeast product D6 is obtained.
  • the total chromium content in the chromium-enriched yeast product D6 is 19688ppm, the absorption rate of chromium is 31.28%, and the proportion of organic chromium is 98.36%.
  • step 3 the dissolved oxygen in the fermentation broth is controlled at 60 ⁇ mol/L; then the chromium-enriched yeast product D7 is obtained.
  • the total chromium content in the chromium-enriched yeast product D7 was 22457ppm, the absorption rate of chromium was 35.68%, and the proportion of organic chromium was 98.41%.
  • Example 1 40563 98.39 66.45
  • Example 2 38451 97.78 61.03
  • Example 3 43096 98.78 68.4 Example 4 41025 98.78 65.18
  • Example 5 40127 98.67 63.76
  • Example 6 41357 98.88 65.71
  • Example 7 40357 98.38 64.12 Comparative Example 1 2044.78 89.71 2.22
  • Comparative Example 2 32647 98.52 51.87
  • Comparative Example 3 35612 98.41 56.58 Comparative Example 4 31985 97.89 50.82
  • Comparative Example 6 19688 98.36 31.28 Comparative Example 7 22457 98.41 35.68
  • Comparative Example 4 shows that fermentation pH has a significant impact on the absorption rate of chromium; Comparative Example 5 is compared with Example 1, and it can be seen that fermentation temperature has a significant impact on the absorption rate of chromium; Comparative Example 6 Compared with Example 1, it can be seen that the alcohol content of the fermentation broth has a significant impact on the absorption rate of chromium; in Comparative Example 7, compared with Example 1, it can be seen that the dissolved oxygen in the fermentation broth has a significant impact on the absorption rate of chromium.

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Abstract

Chromium-rich yeast, a preparation method therefor and an application thereof. The chromium-rich yeast has a chromium mass content of >38000 ppm, an organic chromium proportion of >97% by mass, and a chromium absorption rate of >50% by mass, and no hexavalent chromium is present. The preparation method for the chromium-rich yeast uses organic chromium to replace conventional inorganic chromium as a chromium source. The chromium absorption of the yeast is significantly increased, chromium absorption is high, and the chromium content in produced wastewater is greatly reduced, reducing environmental protection stress.

Description

高铬吸收率的富铬酵母及其制备方法和应用Chromium-enriched yeast with high chromium absorption rate and preparation method and application thereof 技术领域technical field
本发明涉及微生物应用技术领域，具体涉及一种高铬吸收率的富铬酵母及其生产方法和应用。The invention relates to the technical field of microorganism application, in particular to a chromium-enriched yeast with high chromium absorption rate and a production method and application thereof.
背景技术Background technique
酵母铬(富铬酵母)是国家食品药品监督管理局指定的铬营养补充来源之一。酵母是人类利用最早、最广泛的纯天然营养型微生物，也是天然的营养宝藏和理想的生物载体。酵母铬是将酵母细胞培养在含三价铬的培养基中，通过生物转化将无机铬转转变成有机铬，从而提高铬在机体内吸收利用率，降低其毒副作用，更好地发挥其调节血糖、降脂及降胆固醇的作用。Chromium yeast (chromium-enriched yeast) is one of the sources of chromium nutritional supplements designated by the State Food and Drug Administration. Yeast is the earliest and most widely used pure natural nutritional microorganism, and it is also a natural nutritional treasure and an ideal biological carrier. Yeast chromium is to cultivate yeast cells in a medium containing trivalent chromium, and convert inorganic chromium into organic chromium through biotransformation, so as to improve the absorption and utilization rate of chromium in the body, reduce its toxic and side effects, and better play its role. Regulates blood sugar, lipid-lowering and cholesterol-lowering effects.
现有产品主要以酿酒酵母为菌株，添加碳氮磷源，以无机铬为铬源，经发酵培养，最终得到富铬酵母酵母乳，再经干燥得到产品。但现有产品中或多或少会存在无机铬，无机铬存在以下缺陷：吸收率低，一般都低于10％；无生物活性，需要转化成有生物活性的GTF铬，才有调节代谢的作用。但糖尿病及冠心病患者的机体基本没有这种转化能力；无机铬的有害作用，在动物实验中发现，被喂无机铬的大鼠，其腹腔网膜上有大量的脂肪堆积，在肾脏的基底细胞有无机铬聚集。而服无机铬的肾衰患者在做透析时，发现其血液中无机铬的浓度远远高于未服无机铬者。The existing products mainly use Saccharomyces cerevisiae as a strain, add carbon, nitrogen and phosphorus sources, and use inorganic chromium as a chromium source. After fermentation and culture, the chromium-enriched yeast yeast milk is finally obtained, and then the product is obtained by drying. However, there are more or less inorganic chromium in the existing products. Inorganic chromium has the following defects: the absorption rate is low, generally less than 10%; it has no biological activity and needs to be converted into biologically active GTF chromium to regulate metabolism. effect. However, the body of patients with diabetes and coronary heart disease basically does not have this transformation ability; the harmful effects of inorganic chromium have been found in animal experiments. In the rats fed with inorganic chromium, there is a large amount of fat accumulation on the peritoneal omentum, which is at the base of the kidney. Cells have inorganic chromium aggregates. The renal failure patients who took inorganic chromium found that the concentration of inorganic chromium in their blood was much higher than that of those who did not take inorganic chromium during dialysis.
发明内容SUMMARY OF THE INVENTION
本发明解决的现有技术的问题是：现有产品主要以酿酒酵母为菌株，添加碳氮磷源，以无机铬为铬源，经发酵培养，最终得到富铬酵母酵母乳，再经干燥得到产品。但现有产品中或多或少会存在无机铬，无机铬吸收率低，且需要代谢调节才能发挥作用，部分病人甚至无法利用。且无机铬为铬源的工艺产品中容易产生及其微量的六价铬，现 在技术中的富铬酵母中也存在微量六价铬。而六价铬对人和动物的健康是有危害的。The problem of the prior art solved by the present invention is as follows: the existing products mainly use Saccharomyces cerevisiae as a strain, add carbon, nitrogen and phosphorus sources, and use inorganic chromium as a chromium source. product. However, there are more or less inorganic chromium in the existing products, the absorption rate of inorganic chromium is low, and it needs metabolic regulation to play a role, and some patients cannot even use it. And inorganic chromium is easy to produce and trace amount of hexavalent chromium in the process product of chromium source, and trace amount of hexavalent chromium also exists in the chromium-enriched yeast in the present technology. Hexavalent chromium is harmful to human and animal health.
降低富铬酵母产品中无机铬比例、提升有机铬占比已成大势所趋。本发明采用有机铬作为铬源，从源头上消除无机铬来源、本发明的主要原料(铬源为有机铬)，以及使用有机铬原料生产富铬酵母的生产工艺，成品中不会存在无机铬，且产品具有更高的铬含量和铬吸收率。It has become a general trend to reduce the proportion of inorganic chromium and increase the proportion of organic chromium in chromium-enriched yeast products. The invention adopts organic chromium as the chromium source, eliminates the inorganic chromium source, the main raw material of the present invention (the chromium source is organic chromium) from the source, and uses the organic chromium raw material to produce the production process of chromium-enriched yeast, and there is no inorganic chromium in the finished product. , and the product has higher chromium content and chromium absorption rate.
具体来说，本发明提出了如下技术方案。Specifically, the present invention proposes the following technical solutions.
本发明提供了一种富铬酵母，所述富铬酵母中铬的质量含量＞38000ppm，以质量计所述富铬酵母中有机铬比例＞97％，所述富铬酵母以质量计铬吸收率＞50％，所述富铬酵母中无六价铬。The invention provides a chromium-enriched yeast, wherein the mass content of chromium in the chromium-enriched yeast is greater than 38,000 ppm, the organic chromium ratio in the chromium-enriched yeast is greater than 97% by mass, and the chromium-enriched yeast has a chromium absorption rate by mass. >50%, there is no hexavalent chromium in the chromium-rich yeast.
优选的，所述富铬酵母中铬的质量含量＞40000ppm，以质量计所述富铬酵母中有机铬比例＞95％，所述富铬酵母以质量计铬吸收率＞60％。Preferably, the mass content of chromium in the chromium-enriched yeast is greater than 40,000 ppm, the organic chromium ratio in the chromium-enriched yeast is greater than 95% by mass, and the chromium-enriched yeast is more than 60% by mass in the absorption rate of chromium.
本发明还提供了上述的富铬酵母的制备方法，其特征在于，包括如下步骤：The present invention also provides the preparation method of the above-mentioned chromium-enriched yeast, characterized in that it comprises the following steps:
步骤1：将酿酒酵母菌进行放大培养，提供发酵所需的酿酒酵母菌种子；Step 1: Amplify Saccharomyces cerevisiae to provide Saccharomyces cerevisiae seeds required for fermentation;
步骤2：将步骤1所得的酿酒酵母菌种子转入50L发酵罐中继续发酵，当酿酒酵母菌的湿重达到60-90g/L时，流加铬源；Step 2: transfer the Saccharomyces cerevisiae seeds obtained in step 1 into a 50L fermentation tank to continue fermentation, and when the wet weight of Saccharomyces cerevisiae reaches 60-90g/L, add chromium source;
步骤3：使步骤2得到的酿酒酵母菌生长，酵母菌的湿重达到120-140g/L时，持续发酵，得到酵母乳；Step 3: growing the Saccharomyces cerevisiae obtained in Step 2, when the wet weight of the yeast reaches 120-140g/L, continue fermentation to obtain yeast milk;
步骤4：将步骤3中得到的酵母乳分离，后干燥即得到所述富铬酵母；Step 4: separate the yeast milk obtained in step 3, and then dry to obtain the chromium-enriched yeast;
其中，所述铬源为有机铬，所述有机铬选自烟酸铬，吡啶甲酸铬和蛋氨酸铬中的一种或两种以上。Wherein, the chromium source is organic chromium, and the organic chromium is selected from one or more of chromium nicotinate, chromium picolinate and chromium methionine.
优选的，所述步骤2中发酵温度为25-35℃，优选的，溶氧≤30μmol/L，更优选的，乙醇浓度为≤0.3％。Preferably, in the step 2, the fermentation temperature is 25-35° C., preferably, the dissolved oxygen is less than or equal to 30 μmol/L, and more preferably, the concentration of ethanol is less than or equal to 0.3%.
优选的，所述步骤2中当酿酒酵母菌的湿重达到60-90g/L时， 将pH调整至3.9-4.5后，流加铬源，优选的，铬源的量为2-4mol，铬源在2-10h内全部加完。Preferably, in the step 2, when the wet weight of Saccharomyces cerevisiae reaches 60-90g/L, after the pH is adjusted to 3.9-4.5, the chromium source is added in flow, preferably, the amount of the chromium source is 2-4mol, the chromium The source is all added within 2-10h.
优选的，所述步骤3中酵母菌的湿重达到120-140g/L时，控制pH为3.9-4.5，优选的，通过通风和搅拌将溶氧控制在0-30μmol/L，持续发酵9-18h。Preferably, in the step 3, when the wet weight of the yeast reaches 120-140g/L, the pH is controlled to be 3.9-4.5. 18h.
优选的，所述步骤3中在发酵结束前1-4h内将ph提升至5-8，直至发酵结束，得到酵母乳。Preferably, in the step 3, the pH is raised to 5-8 within 1-4 hours before the end of the fermentation, until the end of the fermentation, to obtain yeast milk.
优选的，步骤1中所述的酿酒酵母菌选自酿酒酵母Z1.3(Saccharomyces cerevisiae HANSEN Z1.3)，酿酒酵母Z2.1(Saccharomyces cerevisiae HANSEN Z2.1)和酿酒酵母FX-2(Saccharomyces cerevisiae FX-2)中的一种或两种以上。Preferably, the Saccharomyces cerevisiae described in step 1 is selected from Saccharomyces cerevisiae Z1.3 (Saccharomyces cerevisiae HANSEN Z1.3), Saccharomyces cerevisiae Z2.1 (Saccharomyces cerevisiae HANSEN Z2.1) and Saccharomyces cerevisiae FX-2 (Saccharomyces cerevisiae HANSEN Z2.1) One or more of FX-2).
优选的，步骤1-3中培养所使用的培养基的成分含有碳源，氮源，磷源和微量元素中的一种或两种以上。Preferably, the components of the medium used in the culture in steps 1-3 contain one or more of carbon sources, nitrogen sources, phosphorus sources and trace elements.
优选的，所述碳源选自糖蜜和/或葡萄糖，所述氮源选自硫酸铵，蛋白胨，酵母抽提物和氨水中的一种或两种以上，所述磷源选自磷酸二氢钾，所述微量元素选自硫酸锌和/或硫酸镁。Preferably, the carbon source is selected from molasses and/or glucose, the nitrogen source is selected from one or more of ammonium sulfate, peptone, yeast extract and aqueous ammonia, and the phosphorus source is selected from dihydrogen phosphate Potassium, the trace element is selected from zinc sulfate and/or magnesium sulfate.
优选的，所述糖蜜的浓度为20-40％质量浓度，所述葡萄糖的浓度为20％-40％质量浓度。Preferably, the concentration of the molasses is 20-40% by mass, and the concentration of the glucose is 20-40% by mass.
优选的，所述磷酸二氢钾的纯度为大于96％，所述硫酸铵的纯度为大于99％，优选的，所述硫酸锌的纯度为大于99％，所述硫酸镁的纯度为大于99％。Preferably, the purity of the potassium dihydrogen phosphate is greater than 96%, the purity of the ammonium sulfate is greater than 99%, preferably, the purity of the zinc sulfate is greater than 99%, and the purity of the magnesium sulfate is greater than 99% %.
优选的，步骤2和3中加入碳源，氮源和磷源的方式采用流加。Preferably, in steps 2 and 3, the carbon source, the nitrogen source and the phosphorus source are added by means of feed addition.
本发明还提供了一种富铬酵母，其特征在于，由所上述的方法生产得到。The present invention also provides a chromium-enriched yeast, which is characterized in that it is produced by the above-mentioned method.
本发明还提供了所述的富铬酵母在畜牧业领域中的应用，优选为饲料或饲料添加剂领域中的应用。The present invention also provides the application of the chromium-enriched yeast in the field of animal husbandry, preferably the application in the field of feed or feed additive.
本发明所取得的有益效果是：本发明采用有机铬替代传统无机铬作铬源，酵母对铬吸收率有极大提升，摇瓶吸收率从1％-2％提升到60％以上；本发明所述的富铬酵母总铬可轻易突破传统富铬酵母总铬 8000ppm的上限，最低含量也在38000ppm以上，可以达到40000ppm以上，高至43096ppm，有机铬质量含量大于97％，甚至能达到98％以上；传统的富铬酵母中存在着难以洗涤的无机铬，新型富铬酵母从源头上消除了无机铬，保留下来的都是有机铬，更利于人体吸收和动物吸收；采用有机铬源生产富铬酵母能大幅度减少生产废水中铬含量，减少环保压力；该工艺生产的富铬酵母中无六价铬存在。The beneficial effects obtained by the invention are as follows: the invention adopts organic chromium to replace traditional inorganic chromium as the chromium source, the absorption rate of chromium by yeast is greatly improved, and the absorption rate of the shake flask is increased from 1% to 2% to more than 60%; The total chromium of the chromium-enriched yeast can easily break through the upper limit of the traditional chromium-enriched yeast total chromium of 8000ppm, the minimum content is also above 38000ppm, can reach above 40000ppm, as high as 43096ppm, the mass content of organic chromium is more than 97%, and even can reach 98% The above; there is inorganic chromium that is difficult to wash in traditional chromium-enriched yeast. The new chromium-enriched yeast eliminates inorganic chromium from the source, and what remains is organic chromium, which is more conducive to human absorption and animal absorption; organic chromium sources are used to produce rich Chromium yeast can greatly reduce the chromium content in production wastewater and reduce environmental pressure; there is no hexavalent chromium in the chromium-enriched yeast produced by this process.
菌株保藏信息strain preservation information
本发明所用的菌种酿酒酵母Z1.3(Saccharomyces cerevisiae Hansen Z1.3)，其已经于2005年10月25日在中国典型培养物保藏中心(CCTCC，武汉大学，邮编430072)进行了生物保藏，保藏编号为CCTCC M 205125，该菌株在申请号为CN200610066365.X的专利公开文本中已有记载。The strain Saccharomyces cerevisiae Z1.3 (Saccharomyces cerevisiae Hansen Z1.3) used in the present invention has been biologically preserved in the China Center for Type Culture Collection (CCTCC, Wuhan University, zip code 430072) on October 25, 2005, The deposit number is CCTCC M 205125, and this strain has been recorded in the patent publication with the application number CN200610066365.X.
本发明所用的菌种酿酒酵母FX-2(Saccharomyces cerevisiae FX-2)于2016年8月1日保藏在中国典型培养物保藏中心(CCTCC)，保藏编号为CCTCC NO:M2016418，保藏地址：中国.武汉.武汉大学，邮政编码：430072；电话：(027)-68754052，该菌株在申请号为CN201611141122.8的专利公开文本中已有记载。The strain of Saccharomyces cerevisiae FX-2 (Saccharomyces cerevisiae FX-2) used in the present invention was deposited in the China Center for Type Culture Collection (CCTCC) on August 1, 2016, and the deposit number is CCTCC NO: M2016418, and the deposit address is China. Wuhan. Wuhan University, Postal Code: 430072; Tel: (027)-68754052, the strain has been recorded in the patent publication with the application number CN201611141122.8.
本发明所用的菌种酿酒酵母Z2.1(Saccharomyces cerevisiae Hansen Z2.1)，于2005年10月25日保藏于中国典型培养物保藏中心，菌株保藏编号为CCTCC NO:M205127，保藏地址为：中国.武汉.武汉大学，邮编：430072，电话：(027)68752319，该菌株在申请号为CN201710522840.8的专利公开文本中已有记载。The strain Saccharomyces cerevisiae Z2.1 (Saccharomyces cerevisiae Hansen Z2.1) used in the present invention was preserved in the China Center for Type Culture Collection on October 25, 2005, the strain preservation number is CCTCC NO: M205127, and the preservation address is: China . Wuhan. Wuhan University, Postcode: 430072, Tel: (027) 68752319, the strain has been recorded in the patent publication with the application number CN201710522840.8.
具体实施方式Detailed ways
本发明提供了一种富铬酵母，所述富铬酵母中铬的质量含量＞38000ppm，以质量计所述富铬酵母中有机铬比例＞97％，所述富铬酵母以质量计铬吸收率＞50％，所述富铬酵母中无六价铬。The invention provides a chromium-enriched yeast, wherein the mass content of chromium in the chromium-enriched yeast is greater than 38,000 ppm, the organic chromium ratio in the chromium-enriched yeast is greater than 97% by mass, and the chromium-enriched yeast has a chromium absorption rate by mass. >50%, there is no hexavalent chromium in the chromium-rich yeast.
优选的，所述富铬酵母中铬的质量含量＞40000ppm，以质量计 所述富铬酵母中有机铬比例＞95％，所述富铬酵母以质量计铬吸收率＞60％。Preferably, the mass content of chromium in the chromium-enriched yeast is greater than 40,000 ppm, the organic chromium ratio in the chromium-enriched yeast is greater than 95% by mass, and the chromium-enriched yeast is more than 60% by mass in the absorption rate of chromium.
本发明还提供了上述的富铬酵母的制备方法，其特征在于，包括如下步骤：The present invention also provides the preparation method of the above-mentioned chromium-enriched yeast, characterized in that it comprises the following steps:
步骤1：将酿酒酵母菌进行放大培养，提供发酵所需的酿酒酵母菌种子；Step 1: Amplify Saccharomyces cerevisiae to provide Saccharomyces cerevisiae seeds required for fermentation;
步骤2：将步骤1所得的酿酒酵母菌种子转入50L发酵罐中继续发酵，当酿酒酵母菌的湿重达到60-90g/L时，流加铬源；Step 2: transfer the Saccharomyces cerevisiae seeds obtained in step 1 into a 50L fermentation tank to continue fermentation, and when the wet weight of Saccharomyces cerevisiae reaches 60-90g/L, add chromium source;
步骤3：使步骤2得到的酿酒酵母菌生长，酵母菌的湿重达到120-140g/L时，持续发酵，得到酵母乳；Step 3: growing the Saccharomyces cerevisiae obtained in step 2, when the wet weight of the yeast reaches 120-140 g/L, continue fermentation to obtain yeast milk;
步骤4：将步骤3中得到的酵母乳分离，后干燥即得到所述富铬酵母；Step 4: separate the yeast milk obtained in step 3, and then dry to obtain the chromium-enriched yeast;
其中，所述铬源为有机铬，所述有机铬选自烟酸铬，吡啶甲酸铬和蛋氨酸铬中的一种或两种以上。Wherein, the chromium source is organic chromium, and the organic chromium is selected from one or more of chromium nicotinate, chromium picolinate and chromium methionine.
本发明还提供了一种富铬酵母，其特征在于，由所上述的方法生产得到。The present invention also provides a chromium-enriched yeast, which is characterized in that it is produced by the above-mentioned method.
本发明还提供了所述的富铬酵母在畜牧业领域中的应用，优选为饲料或饲料添加剂领域中的应用。The present invention also provides the application of the chromium-enriched yeast in the field of animal husbandry, preferably the application in the field of feed or feed additive.
名词解释：Glossary:
铬在富铬酵母中的质量含量为0-40000ppm，即铬吸收量以质量计，其为每克酵母中铬含量为0-0.04g；The mass content of chromium in the chromium-enriched yeast is 0-40000 ppm, that is, the amount of chromium absorption is calculated by mass, and the chromium content per gram of yeast is 0-0.04 g;
铬吸收率为：铬吸收率＝酵母(产品)中总铬质量含量/投加的铬源中铬的质量含量；Chromium absorption rate: Chromium absorption rate = total chromium mass content in yeast (product) / mass content of chromium in the added chromium source;
有机铬的比例为：有机铬比例＝胞内有机铬质量含量/酵母(产品)总铬质量含量；The ratio of organic chromium is: organic chromium ratio = intracellular organic chromium mass content/yeast (product) total chromium mass content;
本发明的富铬酵母中的铬含量是指有益的三价铬的铬质量含量，且同时不能存在六价铬。The chromium content in the chromium-enriched yeast of the present invention refers to the chromium mass content of beneficial trivalent chromium, and at the same time, hexavalent chromium cannot exist.
实验原料(取自中式平台车间生产原料)：葡萄糖(或糖蜜)、酵母粉、磷酸二氢钾、硫酸铵、硫酸镁、氨水。Experimental raw materials (taken from the production raw materials of the Chinese platform workshop): glucose (or molasses), yeast powder, potassium dihydrogen phosphate, ammonium sulfate, magnesium sulfate, ammonia water.
铬源：烟酸铬或蛋氨酸铬或砒啶甲酸铬Chromium source: chromium nicotinate or chromium methionine or chromium pyridinecarboxylate
发酵控制过程：Fermentation control process:
1.以酿酒酵母为起始菌株；1. Using Saccharomyces cerevisiae as the starting strain;
2.控制发酵温度25-35℃，溶氧30μmol/L以下发酵，湿重达到60—90g/L时候开始流加铬源，维持酒精保持在一个较低水平，在烟酸铬流加前一小时将PH调整至3.9-4.5；2. Control the fermentation temperature at 25-35°C, ferment with dissolved oxygen below 30μmol/L, and start to add chromium source when the wet weight reaches 60-90g/L, and maintain the alcohol at a low level. Adjust the pH to 3.9-4.5 for hours;
4.合理控制补糖量，控制酵母生长速度，使烟酸铬流加结束后湿重达到130g±10g/L，随后减风减糖至少持续发酵9个小时，保证酵母对铬的吸收；最后三小时将PH缓慢上升至6.0。4. Reasonably control the amount of sugar supplementation, and control the growth rate of yeast, so that the wet weight of chromium nicotinate reaches 130g±10g/L after the addition of chromium niacin, and then reduce the wind and sugar for at least 9 hours to ensure the absorption of chromium by yeast; The pH was slowly raised to 6.0 over three hours.
5.酵母乳经喷粉干燥塔喷粉得到富铬酵母产品。5. The yeast milk is dusted by a dusting and drying tower to obtain a chromium-enriched yeast product.
下面对本实施例所用的原料及设备的生产厂家，以及产品分析使用的设备和分析方法进行说明如下，其中所述的化学物质没有标明的均为常规试剂的化学纯级别。其中，实施例和对比例中所用到的原料的信息如下表所示。The manufacturers of the raw materials and equipment used in this example, as well as the equipment and analysis methods used for product analysis are described below. The chemical substances described therein are all chemically pure grades of conventional reagents that are not marked. The information of the raw materials used in the examples and comparative examples is shown in the following table.
表1 本发明所用到的原料和仪器的信息Table 1 Information of raw materials and instruments used in the present invention
试剂reagent 纯度/型号Purity/Type 生产厂商manufacturer
葡萄糖glucose 食品级food grade 济南圣丰工贸有限公司Jinan Shengfeng Industry and Trade Co., Ltd.
酵母抽提物yeast extract 食品级food grade 安琪酵母股份有限公司Angel Yeast Co., Ltd.
磷酸二氢钾Potassium dihydrogen phosphate 食品级food grade 山东东润生物科技有限公司Shandong Dongrun Biotechnology Co., Ltd.
硫酸镁Magnesium sulfate 食品级food grade 连云港冠苏实业有限公司Lianyungang Guansu Industrial Co., Ltd.
蔗糖sucrose 食品级food grade 北京康普汇维科技有限公司Beijing CommScope Huiwei Technology Co., Ltd.
硫酸锌Zinc sulfate 食品级food grade 连云港冠苏实业有限公司Lianyungang Guansu Industrial Co., Ltd.
摇床shaker ZHWY-211DZHWY-211D 上海智诚科技有限公司Shanghai Zhicheng Technology Co., Ltd.
离心机centrifuge TG16-WSTG16-WS 上海诺民生物科技有限公司Shanghai Nuomin Biotechnology Co., Ltd.
抽滤机Suction filter SHB-D(III)ASHB-D(III)A 河南新发现科技有限公司Henan New Discovery Technology Co., Ltd.
酿酒酵母Z1.3Saccharomyces cerevisiae Z1.3    安琪酵母股份有限公司Angel Yeast Co., Ltd.
酿酒酵母FX-2Saccharomyces cerevisiae FX-2    安琪酵母股份有限公司Angel Yeast Co., Ltd.
酿酒酵母Z2.1Saccharomyces cerevisiae Z2.1    安琪酵母股份有限公司Angel Yeast Co., Ltd.
糖蜜molasses    安琪酵母股份有限公司Angel Yeast Co., Ltd.
实施例1Example 1
富铬酵母的制备方法：Preparation method of chromium-enriched yeast:
步骤1：将酿酒酵母菌富铬酿酒酵母Z1.3(Saccharomyces cerevisiae HANSEN Z1.3)挑取一环进行摇瓶培养—种子培养的逐级放大培养，以提供发酵所需的酿酒酵母菌种子；摇瓶培养的培养基的成分为：蔗糖100g，酵母抽提物20g，硫酸镁1g，磷酸二氢钾1g，pH4.8，31℃培养24h，5L摇瓶装液量1L，摇床转速180rpm/min；Step 1: the Saccharomyces cerevisiae Chromium-enriched Saccharomyces cerevisiae Z1.3 (Saccharomyces cerevisiae HANSEN Z1.3) is picked a ring to carry out shake-flask culture—the step-by-step amplification culture of seed culture, to provide the Saccharomyces cerevisiae seeds required for fermentation; The components of the culture medium in the shake flask are: 100 g of sucrose, 20 g of yeast extract, 1 g of magnesium sulfate, 1 g of potassium dihydrogen phosphate, pH 4.8, cultured at 31°C for 24 hours, 5L shake flask with a liquid volume of 1L, and the shaker rotation speed 180rpm/ min;
步骤2：将步骤1所得的酿酒酵母菌种子，转入50L发酵罐中，控制发酵温度为35℃，溶氧30μmol/L，发酵2小时之后，流加4000g的碳源(糖蜜浓度为30％)，100g氮源(硫酸铵)，100g磷源(磷酸二氢钾)，控制乙醇浓度为0.07％±0.03％，供酵母生长，当酿酒酵母菌的湿重达到60g/L时，将pH调整至3.9后，之后维持pH在3.9(pH下降则加入氨水将pH提升至3.9)，并流加铬源(0.01mol/L烟酸铬共3mol)，铬源在10h内全部加完；Step 2: transfer the Saccharomyces cerevisiae seeds of step 1 gained into 50L fermentor, control fermentation temperature to be 35 ℃, dissolved oxygen 30 μmol/L, after 2 hours of fermentation, flow the carbon source (molasses concentration is 30%) of 4000g. ), 100g nitrogen source (ammonium sulfate), 100g phosphorus source (potassium dihydrogen phosphate), control ethanol concentration to be 0.07% ± 0.03%, for yeast growth, when the wet weight of Saccharomyces cerevisiae reaches 60g/L, adjust pH After reaching 3.9, the pH was maintained at 3.9 (after the pH dropped, ammonia water was added to raise the pH to 3.9), and the chromium source (0.01mol/L chromium nicotinate, a total of 3mol) was added simultaneously, and the chromium source was all added within 10h;
步骤3：流加8000g的碳源(糖蜜浓度为30％)，200g氮源(硫酸铵)，200g磷源(磷酸二氢钾)，控制乙醇浓度为0.07％±0.03％，供酿酒酵母菌生长，酵母菌的湿重达到120g/L时，继续维持pH在3.9，通过通风和搅拌将溶氧控制在30μmol/L，持续发酵9h，在发酵结束前4h开始，用氨水逐渐将ph提升至6，直至发酵结束，得到酵母乳；Step 3: add 8000g carbon source (molasses concentration is 30%), 200g nitrogen source (ammonium sulfate), 200g phosphorus source (potassium dihydrogen phosphate), control the ethanol concentration to be 0.07%±0.03%, for the growth of Saccharomyces cerevisiae , when the wet weight of the yeast reaches 120g/L, continue to maintain the pH at 3.9, control the dissolved oxygen at 30μmol/L by ventilation and stirring, continue the fermentation for 9h, start 4h before the end of the fermentation, and gradually increase the pH to 6 with ammonia water , until the end of fermentation, to obtain yeast milk;
步骤4：将步骤3中得到的酵母乳分离杂质后通过喷雾干燥即得到所述富铬酵母产品A。Step 4: The chromium-enriched yeast product A is obtained by spray-drying the yeast milk obtained in step 3 after separating impurities.
使用GB 5009.123-2014食品中铬的测定中的检测方法，经检测，制备得到的富铬酵母产品A铬含量为40563ppm；有机铬比例为98.39％；铬吸收率为66.45％，经检测得到制备得到的富铬酵母产品A中无六价铬。Using the detection method in GB 5009.123-2014 Determination of Chromium in Food, after testing, the chromium content of the prepared chromium-enriched yeast product A is 40563 ppm; the organic chromium ratio is 98.39%; the chromium absorption rate is 66.45%, and the prepared Chromium-enriched yeast product A has no hexavalent chromium.
实施例2Example 2
步骤同实施例1，区别仅仅为步骤2中流加的铬源为3mol蛋氨酸铬。The steps are the same as those in Example 1, except that the chromium source fed in the step 2 is 3 mol of chromium methionine.
经检测，制备得到的富铬酵母产品B铬含量为38451ppm；有机铬比例为97.78％；铬吸收率为61.03％，制备得到的富铬酵母产品B中无六价铬。After testing, the chromium content of the prepared chromium-enriched yeast product B was 38451 ppm; the organic chromium ratio was 97.78%; the chromium absorption rate was 61.03%, and the prepared chromium-enriched yeast product B had no hexavalent chromium.
实施例3Example 3
步骤同实施例1，区别仅仅为步骤2中流加的铬源为3mol吡啶甲酸铬。The steps are the same as in Example 1, except that the chromium source streamed in the step 2 is 3mol chromium picolinate.
经检测，制备得到的富铬酵母产品C铬含量为43096ppm；有机铬比例为98.78％；铬吸收率为68.4％，制备得到的富铬酵母产品C中无六价铬。After testing, the prepared chromium-enriched yeast product C has a chromium content of 43096 ppm, an organic chromium ratio of 98.78%, and a chromium absorption rate of 68.4%. The prepared chromium-enriched yeast product C has no hexavalent chromium.
实施例4Example 4
其余实验步骤与实施例1均相同，区别仅仅在于当步骤2中酿酒酵母菌的湿重达到90g/L时，将pH调整至3.9后，流加铬源(0.01mol/L烟酸铬共3mol，铬源在10h内全部加完，后得到富铬酵母产品D。The rest of the experimental steps are the same as in Example 1, except that when the wet weight of Saccharomyces cerevisiae reaches 90 g/L in step 2, after adjusting the pH to 3.9, the chromium source (0.01 mol/L chromium nicotinate for a total of 3 mol , the chromium source is all added within 10h, and then the chromium-enriched yeast product D is obtained.
经检测，制备得到的富铬酵母产品D铬含量为41025ppm，有机铬比例为98.78％，铬吸收率为65.18％，富铬酵母产品D中无六价铬。After testing, the prepared chromium-enriched yeast product D has a chromium content of 41,025 ppm, an organic chromium ratio of 98.78%, and a chromium absorption rate of 65.18%. There is no hexavalent chromium in the chromium-enriched yeast product D.
实施例5Example 5
其余实验步骤与实施例1均相同，区别仅仅在于步骤3中，酵母菌的湿重达到140g/L时，控制pH为3.9，通过通风和搅拌将溶氧控制在30μmol/L，持续发酵18h，在发酵结束前4h内将ph提升至6，直至发酵结束，得到酵母乳，后得到富铬酵母产品E。The rest of the experimental steps are the same as in Example 1, except that in step 3, when the wet weight of the yeast reaches 140 g/L, the pH is controlled to 3.9, the dissolved oxygen is controlled to 30 μmol/L by ventilation and stirring, and the fermentation is continued for 18 h. The pH was raised to 6 within 4 hours before the end of the fermentation until the end of the fermentation to obtain yeast milk, and then the chromium-enriched yeast product E was obtained.
经检测，制备得到的富铬酵母产品E铬含量为40127ppm，有机铬比例为98.67％，铬吸收率为63.76％，制备得到的富铬酵母产品E 中无六价铬。After testing, the prepared chromium-enriched yeast product E has a chromium content of 40127 ppm, an organic chromium ratio of 98.67%, and a chromium absorption rate of 63.76%. The prepared chromium-enriched yeast product E has no hexavalent chromium.
实施例6Example 6
其余实验步骤与实施例1均相同，区别仅仅在于步骤2中，将步骤1所得的酿酒酵母菌种子，控制发酵温度为25℃；后得到富铬酵母产品F。The rest of the experimental steps are the same as those in Example 1, except that in step 2, the fermentation temperature of the Saccharomyces cerevisiae seeds obtained in step 1 is controlled to be 25°C; afterward, the chromium-enriched yeast product F is obtained.
经检测，制备得到的富铬酵母产品F铬含量为41357ppm，有机铬比例为98.88％，铬吸收率为65.71％，制备得到的富铬酵母产品F中无六价铬。After testing, the prepared chromium-enriched yeast product F has a chromium content of 41357 ppm, an organic chromium ratio of 98.88%, and a chromium absorption rate of 65.71%. The prepared chromium-enriched yeast product F has no hexavalent chromium.
实施例7Example 7
其余步骤与实施例1相同，区别仅仅在于步骤2中当酿酒酵母菌的湿重达到60g/L时，将pH调整至4.5后，之后维持pH在4.5(pH下降则加入氨水将PH提升至4.5)；后得到富铬酵母产品G。The remaining steps are the same as in Example 1, except that in step 2, when the wet weight of Saccharomyces cerevisiae reaches 60 g/L, after adjusting the pH to 4.5, maintain the pH at 4.5 (the pH drops and then adds ammonia water to raise the pH to 4.5). ); and then the chromium-enriched yeast product G was obtained.
经检测，制备得到的富铬酵母产品G铬含量为40357ppm，有机铬比例为98.38％，铬吸收率为64.12％，制备得到的富铬酵母产品G中无六价铬。After testing, the prepared chromium-enriched yeast product G has a chromium content of 40357 ppm, an organic chromium ratio of 98.38%, and a chromium absorption rate of 64.12%. The prepared chromium-enriched yeast product G has no hexavalent chromium.
对比例Comparative ratio
对比例1Comparative Example 1
其余实验条件与实施例1都相同，区别仅在于在步骤2中流加的铬源为氯化铬；后得到富铬酵母产品D1。The rest of the experimental conditions are the same as those in Example 1, except that the chromium source fed in in step 2 is chromium chloride; the chromium-enriched yeast product D1 is then obtained.
经检测，制备得到的富铬酵母产品D1的铬含量为2044.78ppm；有机铬比例为89.71％；铬吸收率为2.22％；其中六价铬为0.933ppm。After testing, the chromium content of the prepared chromium-enriched yeast product D1 was 2044.78ppm; the organic chromium ratio was 89.71%; the chromium absorption rate was 2.22%; and the hexavalent chromium was 0.933ppm.
对比例2Comparative Example 2
其余实验条件与实施例1都相同，区别仅仅在于在步骤2中不流加烟酸铬，在步骤3中湿重达到120g/L时1h内加入3mol烟酸铬；后得到富铬酵母产品D2。The rest of the experimental conditions are the same as in Example 1, except that in step 2, chromium nicotinate is not added, and in step 3, when the wet weight reaches 120 g/L, 3 mol of chromium nicotinate is added within 1 hour; then the chromium-enriched yeast product D2 is obtained. .
经检测，富铬酵母产品D2中总铬含量32647ppm，铬吸收率51.87％；有机铬的比例为98.52％。After testing, the total chromium content in the chromium-enriched yeast product D2 was 32647ppm, the absorption rate of chromium was 51.87%, and the proportion of organic chromium was 98.52%.
对比例3Comparative Example 3
其余实验条件与实施例1都相同，区别仅仅在于在步骤3中在发酵开始时加入3mol烟酸铬，且烟酸铬添加总量与实施例1完全相同；后得到富铬酵母产品D3。The rest of the experimental conditions are the same as in Example 1, except that 3 mol of chromium nicotinate was added at the beginning of the fermentation in step 3, and the total amount of chromium nicotinate added was exactly the same as in Example 1; the chromium-enriched yeast product D3 was obtained later.
经检测，富铬酵母产品D3中总铬含量35612ppm，铬吸收率56.58％，有机铬比例为98.41％。After testing, the total chromium content in the chromium-enriched yeast product D3 was 35612ppm, the absorption rate of chromium was 56.58%, and the proportion of organic chromium was 98.41%.
对比例4Comparative Example 4
其余实验条件与实施例1都相同，区别仅仅在于在步骤2及步骤3中发酵pH控制在5.5；后得到富铬酵母产品D4。The rest of the experimental conditions are the same as in Example 1, the difference is only that the fermentation pH is controlled at 5.5 in Steps 2 and 3, and then the chromium-enriched yeast product D4 is obtained.
经检测，富铬酵母产品D4中总铬含量31985ppm，铬吸收率50.82％；有机铬比例为97.89％。After testing, the total chromium content in the chromium-enriched yeast product D4 was 31985ppm, the absorption rate of chromium was 50.82%, and the proportion of organic chromium was 97.89%.
对比例5Comparative Example 5
其余实验条件与实施例1都相同，区别仅在于在步骤2及步骤3中发酵温度控制在38℃；后得到富铬酵母产品D5。The rest of the experimental conditions are the same as in Example 1, the only difference is that the fermentation temperature is controlled at 38° C. in Steps 2 and 3; then the chromium-enriched yeast product D5 is obtained.
经检测，富铬酵母产品D5中总铬含量26854ppm，铬吸收率42.67％，有机铬比例为98.14％。After testing, the total chromium content in the chromium-enriched yeast product D5 was 26854ppm, the absorption rate of chromium was 42.67%, and the proportion of organic chromium was 98.14%.
对比例6Comparative Example 6
其余实验条件与实施例1都相同，区别仅在于在步骤3中发酵液酒精控制在1.0％；后得到富铬酵母产品D6。The rest of the experimental conditions are the same as those in Example 1, except that in step 3, the alcohol of the fermentation broth is controlled at 1.0%; then the chromium-enriched yeast product D6 is obtained.
经检测，富铬酵母产品D6中总铬含量19688ppm，铬吸收率31.28％，有机铬比例为98.36％。After testing, the total chromium content in the chromium-enriched yeast product D6 is 19688ppm, the absorption rate of chromium is 31.28%, and the proportion of organic chromium is 98.36%.
对比例7Comparative Example 7
其余实验条件与实施例1都相同，区别仅在于在步骤3中发酵液 溶氧控制在60μmol/L；后得到富铬酵母产品D7。The rest of the experimental conditions are the same as in Example 1, except that in step 3, the dissolved oxygen in the fermentation broth is controlled at 60 μmol/L; then the chromium-enriched yeast product D7 is obtained.
经检测，富铬酵母产品D7中总铬含量22457ppm，铬吸收率35.68％，有机铬比例为98.41％。After testing, the total chromium content in the chromium-enriched yeast product D7 was 22457ppm, the absorption rate of chromium was 35.68%, and the proportion of organic chromium was 98.41%.
表2 实施例1-6和对比例1-7中富铬酵母产品的指标参数总结Table 2 Summary of index parameters of chromium-enriched yeast products in Examples 1-6 and Comparative Examples 1-7
富铬酵母产品Chromium-enriched yeast products 总铬含量/ppmTotal chromium content/ppm 有机铬比例/％Organic chromium ratio/% 铬吸收率/％Chromium absorption rate/%
实施例1Example 1 4056340563 98.3998.39 66.4566.45
实施例2Example 2 3845138451 97.7897.78 61.0361.03
实施例3Example 3 4309643096 98.7898.78 68.468.4
实施例4Example 4 4102541025 98.7898.78 65.1865.18
实施例5Example 5 4012740127 98.6798.67 63.7663.76
实施例6Example 6 4135741357 98.8898.88 65.7165.71
实施例7Example 7 4035740357 98.3898.38 64.1264.12
对比例1Comparative Example 1 2044.782044.78 89.7189.71 2.222.22
对比例2Comparative Example 2 3264732647 98.5298.52 51.8751.87
对比例3Comparative Example 3 3561235612 98.4198.41 56.5856.58
对比例4Comparative Example 4 3198531985 97.8997.89 50.8250.82
对比例5Comparative Example 5 2685426854 98.1498.14 42.6742.67
对比例6Comparative Example 6 1968819688 98.3698.36 31.2831.28
对比例7Comparative Example 7 2245722457 98.4198.41 35.6835.68
根据表2的结果可见，对比例1采用氯化铬(无机铬)作铬源，实施例1、2、3中采用有机铬作铬源，对比实施例1、2、3和对比例1数据可见，本发明采用有机铬作铬源时，铬的吸收率较高，在60％以上，高达68.4％；产品中有机铬比例较高，有机铬质量含量大于97％，制备的大多数富铬酵母产品有机铬含量为98％以上；整体而言，本发明制备的富铬酵母中铬的质量含量最低也在38000ppm以上，高达43096ppm；氯化铬(无机铬)作铬源时，铬的吸收率较低， 产品中有机铬比例较低。且有机铬作铬源铬时产品中无六价铬，而氯化铬(无机铬)作铬源时产中检测出六价铬。对比例4与实施例1相比，可以看出发酵pH对铬的吸收率有显著影响；对比例5与实施例1相比，可以看出发酵温度对铬的吸收率有显著影响；对比例6与实施例1相比，可以看出发酵液酒精含量对铬的吸收率有显著影响；对比例7与实施例1相比，可以看出发酵液溶氧对铬的吸收率有显著影响。According to the results in Table 2, it can be seen that in Comparative Example 1, chromium chloride (inorganic chromium) was used as the chromium source, and organic chromium was used as the chromium source in Examples 1, 2, and 3, and the data of Comparative Examples 1, 2, 3 and Comparative Example 1 were It can be seen that when the organic chromium is used as the chromium source in the present invention, the absorption rate of chromium is relatively high, above 60%, up to 68.4%; the proportion of organic chromium in the product is relatively high, the mass content of organic chromium is greater than 97%, and most of the prepared chromium-rich The organic chromium content of the yeast product is more than 98%; on the whole, the mass content of chromium in the chromium-enriched yeast prepared by the present invention is at least 38,000 ppm and as high as 43,096 ppm; when chromium chloride (inorganic chromium) is used as a chromium source, the absorption of chromium The rate is low, and the proportion of organic chromium in the product is low. And when organic chromium is used as chromium source, there is no hexavalent chromium in the product, but when chromium chloride (inorganic chromium) is used as chromium source, hexavalent chromium is detected in the product. Compared with Example 1, Comparative Example 4 shows that fermentation pH has a significant impact on the absorption rate of chromium; Comparative Example 5 is compared with Example 1, and it can be seen that fermentation temperature has a significant impact on the absorption rate of chromium; Comparative Example 6 Compared with Example 1, it can be seen that the alcohol content of the fermentation broth has a significant impact on the absorption rate of chromium; in Comparative Example 7, compared with Example 1, it can be seen that the dissolved oxygen in the fermentation broth has a significant impact on the absorption rate of chromium.
以上所述，仅是本发明实施的较佳实施例，并非对本发明做任何形式上的限制，凡在本发明的精神和原则之内所做的修改、等同替换和改进等，均需要包含在本发明的保护范围之内。The above are only preferred embodiments of the present invention, and are not intended to limit the present invention in any form. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention shall be included in the within the protection scope of the present invention.

Claims (15)

  1. 一种富铬酵母，其特征在于，所述富铬酵母中铬的质量含量＞38000ppm，以质量计所述富铬酵母中有机铬比例＞97％，所述富铬酵母以质量计铬吸收率＞50％，所述富铬酵母中无六价铬。A chromium-enriched yeast, characterized in that the mass content of chromium in the chromium-enriched yeast is greater than 38,000 ppm, the organic chromium ratio in the chromium-enriched yeast is greater than 97% by mass, and the chromium-enriched yeast is chromium-absorbed by mass. >50%, there is no hexavalent chromium in the chromium-rich yeast.
  2. 根据权利要求1所述的富铬酵母，其中，所述富铬酵母中铬的质量含量＞40000ppm，以质量计所述富铬酵母中有机铬比例＞95％，所述富铬酵母以质量计铬吸收率＞60％。The chromium-enriched yeast according to claim 1, wherein the mass content of chromium in the chromium-enriched yeast is greater than 40,000 ppm, the organic chromium ratio in the chromium-enriched yeast is greater than 95% by mass, and the chromium-enriched yeast is based on mass Chromium absorption rate> 60%.
  3. 权利要求1或2所述的富铬酵母的制备方法，其特征在于，包括如下步骤：The preparation method of the described chromium-enriched yeast of claim 1 or 2, is characterized in that, comprises the steps:
    步骤1：将酿酒酵母菌进行放大培养，提供发酵所需的酿酒酵母菌种子；Step 1: Amplify Saccharomyces cerevisiae to provide Saccharomyces cerevisiae seeds required for fermentation;
    步骤2：将步骤1所得的酿酒酵母菌种子继续发酵，当酿酒酵母菌的湿重达到60-90g/L时，流加铬源；Step 2: continue fermentation of the Saccharomyces cerevisiae seeds obtained in step 1, when the wet weight of Saccharomyces cerevisiae reaches 60-90 g/L, add chromium source;
    步骤3：使步骤2得到的酿酒酵母菌生长，酵母菌的湿重达到120-140g/L时，持续发酵，得到酵母乳；Step 3: growing the Saccharomyces cerevisiae obtained in Step 2, when the wet weight of the yeast reaches 120-140g/L, continue fermentation to obtain yeast milk;
    步骤4：将步骤3中得到的酵母乳分离，后干燥即得到所述富铬酵母；Step 4: separate the yeast milk obtained in step 3, and then dry to obtain the chromium-enriched yeast;
    其中，所述铬源为有机铬，所述有机铬选自烟酸铬，吡啶甲酸铬和蛋氨酸铬中的一种或两种以上。Wherein, the chromium source is organic chromium, and the organic chromium is selected from one or more of chromium nicotinate, chromium picolinate and chromium methionine.
  4. 根据权利要求3所述的方法，其中，所述步骤2中发酵温度为25-35℃，优选的，溶氧≤30μmol/L，更优选的，乙醇浓度为≤0.3％。The method according to claim 3, wherein in the step 2, the fermentation temperature is 25-35°C, preferably, the dissolved oxygen is less than or equal to 30 μmol/L, and more preferably, the concentration of ethanol is less than or equal to 0.3%.
  5. 根据权利要求3或4所述方法，其中，所述步骤2中当酿酒酵母菌的湿重达到60-90g/L时，将pH调整至3.9-4.5后，流加铬源，优选的，铬源的量为2-4mol，铬源在2-10h内全部加完。The method according to claim 3 or 4, wherein, in the step 2, when the wet weight of Saccharomyces cerevisiae reaches 60-90g/L, after adjusting the pH to 3.9-4.5, a chromium source, preferably, chromium The amount of the source is 2-4mol, and the chromium source is all added within 2-10h.
  6. 根据权利要求3-5中任一项所述的方法，其中，所述步骤3中酵母菌的湿重达到120-140g/L时，控制pH为3.9-4.5，优选的，通过通风和搅拌将溶氧控制在0-30μmol/L，持续发酵9-18h。The method according to any one of claims 3-5, wherein, in the step 3, when the wet weight of the yeast reaches 120-140g/L, the pH is controlled to be 3.9-4.5, preferably, by ventilation and stirring The dissolved oxygen was controlled at 0-30μmol/L, and the fermentation was continued for 9-18h.
  7. 根据权利要求3-6中任一项所述的方法，其中，所述步骤3中在发酵结束前1-4h内将ph提升至5-8，直至发酵结束，得到酵母乳。The method according to any one of claims 3-6, wherein, in the step 3, the pH is raised to 5-8 within 1-4h before the end of the fermentation, until the end of the fermentation, to obtain yeast milk.
  8. 根据权利要求3-7中任一项所述的方法，其中，步骤1中所述的酿酒酵母菌选自酿酒酵母Z1.3(Saccharomyces cerevisiae HANSEN Z1.3)，酿酒酵母Z2.1(Saccharomyces cerevisiae HANSEN Z2.1)和酿酒酵母FX-2(Saccharomyces cerevisiae FX-2)中的一种或两种以上。The method according to any one of claims 3-7, wherein the Saccharomyces cerevisiae described in step 1 is selected from Saccharomyces cerevisiae Z1.3 (Saccharomyces cerevisiae HANSEN Z1.3), Saccharomyces cerevisiae Z2.1 (Saccharomyces cerevisiae One or more of HANSEN Z2.1) and Saccharomyces cerevisiae FX-2 (Saccharomyces cerevisiae FX-2).
  9. 根据权利要求3-8中任一项所述的方法，其中，步骤1-3中培养所使用的培养基的成分含有碳源，氮源，磷源和微量元素中的一种或两种以上。The method according to any one of claims 3-8, wherein the components of the medium used for culturing in steps 1-3 contain one or more of carbon sources, nitrogen sources, phosphorus sources and trace elements .
  10. 根据权利要求9所述的方法，其中，所述碳源选自糖蜜和/或葡萄糖，所述氮源选自硫酸铵，蛋白胨，酵母抽提物和氨水中的一种或两种以上，所述磷源选自磷酸二氢钾，所述微量元素选自硫酸锌和/或硫酸镁。The method according to claim 9, wherein the carbon source is selected from molasses and/or glucose, the nitrogen source is selected from one or more of ammonium sulfate, peptone, yeast extract and aqueous ammonia, and the The phosphorus source is selected from potassium dihydrogen phosphate, and the trace element is selected from zinc sulfate and/or magnesium sulfate.
  11. 根据权利要求9或10所述的方法，其中，所述糖蜜的浓度为20-40％质量浓度，所述葡萄糖的浓度为20％-40％质量浓度。The method according to claim 9 or 10, wherein the concentration of the molasses is 20-40% by mass, and the concentration of the glucose is 20-40% by mass.
  12. 根据权利要求9-11中任一项所述的方法，其中，所述磷酸二氢钾的纯度为大于96％，所述硫酸铵的纯度为大于99％，优选的，所述硫酸锌的纯度为大于99％，所述硫酸镁的纯度为大于99％。The method according to any one of claims 9-11, wherein the purity of the potassium dihydrogen phosphate is greater than 96%, the purity of the ammonium sulfate is greater than 99%, preferably, the purity of the zinc sulfate is greater than 99%, and the purity of the magnesium sulfate is greater than 99%.
  13. 根据权利要求9-12中任一项所述的方法，其中，步骤2和3中加入碳源，氮源和磷源的方式采用流加。The method according to any one of claims 9-12, wherein, in steps 2 and 3, the carbon source, the nitrogen source and the phosphorus source are added in a fed manner.
  14. 一种富铬酵母，其特征在于，由权利要求3-13中任一项所述的方法制备得到。A chromium-enriched yeast is characterized in that, it is prepared by the method described in any one of claims 3-13.
  15. 权利要求1或2所述富铬酵母或权利要求14所述的富铬酵母在畜牧业领域中的应用，优选为饲料或饲料添加剂领域中的应用。The application of the chromium-enriched yeast of claim 1 or 2 or the chromium-enriched yeast of claim 14 in the field of animal husbandry, preferably the application in the field of feed or feed additives.
PCT/CN2021/137249 2020-12-30 2021-12-10 Chromium-rich yeast having high chromium absorption, preparation method therefor and application thereof WO2022143113A1 (en)

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