CN101575617B - Chromium-rich yeast culture and fermentation process thereof - Google Patents
Chromium-rich yeast culture and fermentation process thereof Download PDFInfo
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- CN101575617B CN101575617B CN2009100376131A CN200910037613A CN101575617B CN 101575617 B CN101575617 B CN 101575617B CN 2009100376131 A CN2009100376131 A CN 2009100376131A CN 200910037613 A CN200910037613 A CN 200910037613A CN 101575617 B CN101575617 B CN 101575617B
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- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 title claims abstract description 78
- 239000011651 chromium Substances 0.000 title claims abstract description 73
- 229910052804 chromium Inorganic materials 0.000 title claims abstract description 73
- 238000000855 fermentation Methods 0.000 title claims abstract description 38
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 37
- 230000004151 fermentation Effects 0.000 title claims abstract description 33
- 239000007787 solid Substances 0.000 claims abstract description 46
- 239000007788 liquid Substances 0.000 claims abstract description 19
- 238000010564 aerobic fermentation Methods 0.000 claims abstract description 10
- 238000004519 manufacturing process Methods 0.000 claims abstract description 8
- 230000009466 transformation Effects 0.000 claims abstract description 7
- 238000001035 drying Methods 0.000 claims abstract description 6
- 241000222178 Candida tropicalis Species 0.000 claims abstract description 5
- 230000001580 bacterial effect Effects 0.000 claims description 15
- 240000007594 Oryza sativa Species 0.000 claims description 12
- 235000007164 Oryza sativa Nutrition 0.000 claims description 12
- 235000009566 rice Nutrition 0.000 claims description 12
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 9
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 9
- 240000008042 Zea mays Species 0.000 claims description 8
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 8
- 235000015097 nutrients Nutrition 0.000 claims description 7
- 241001465754 Metazoa Species 0.000 claims description 6
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 6
- 235000005822 corn Nutrition 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 229920000742 Cotton Polymers 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 4
- 235000015099 wheat brans Nutrition 0.000 claims description 4
- 235000017060 Arachis glabrata Nutrition 0.000 claims description 3
- 244000105624 Arachis hypogaea Species 0.000 claims description 3
- 235000010777 Arachis hypogaea Nutrition 0.000 claims description 3
- 235000018262 Arachis monticola Nutrition 0.000 claims description 3
- 235000010469 Glycine max Nutrition 0.000 claims description 3
- 240000006240 Linum usitatissimum Species 0.000 claims description 3
- 235000004431 Linum usitatissimum Nutrition 0.000 claims description 3
- 230000009286 beneficial effect Effects 0.000 claims description 3
- 235000012054 meals Nutrition 0.000 claims description 3
- 238000010899 nucleation Methods 0.000 claims description 3
- 235000020232 peanut Nutrition 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 235000016068 Berberis vulgaris Nutrition 0.000 claims description 2
- 241000335053 Beta vulgaris Species 0.000 claims description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 2
- 244000068988 Glycine max Species 0.000 claims description 2
- 240000003183 Manihot esculenta Species 0.000 claims description 2
- 235000016735 Manihot esculenta subsp esculenta Nutrition 0.000 claims description 2
- 229920002472 Starch Polymers 0.000 claims description 2
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims description 2
- 230000009977 dual effect Effects 0.000 claims description 2
- 235000009973 maize Nutrition 0.000 claims description 2
- 238000011160 research Methods 0.000 claims description 2
- 235000019698 starch Nutrition 0.000 claims description 2
- 239000008107 starch Substances 0.000 claims description 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 abstract description 30
- 238000000034 method Methods 0.000 abstract description 13
- 230000008569 process Effects 0.000 abstract description 8
- 244000005700 microbiome Species 0.000 abstract description 6
- 239000001963 growth medium Substances 0.000 abstract description 3
- 230000000813 microbial effect Effects 0.000 abstract description 3
- 244000144972 livestock Species 0.000 abstract description 2
- 238000012216 screening Methods 0.000 abstract description 2
- JOPOVCBBYLSVDA-UHFFFAOYSA-N chromium(6+) Chemical compound [Cr+6] JOPOVCBBYLSVDA-UHFFFAOYSA-N 0.000 abstract 1
- 244000144977 poultry Species 0.000 abstract 1
- 239000006041 probiotic Substances 0.000 abstract 1
- 230000000529 probiotic effect Effects 0.000 abstract 1
- 235000018291 probiotics Nutrition 0.000 abstract 1
- 230000001131 transforming effect Effects 0.000 abstract 1
- 239000000047 product Substances 0.000 description 17
- 238000005516 engineering process Methods 0.000 description 9
- 239000002609 medium Substances 0.000 description 9
- 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 description 7
- 239000008103 glucose Substances 0.000 description 7
- 230000001954 sterilising effect Effects 0.000 description 7
- 239000000243 solution Substances 0.000 description 6
- 238000004659 sterilization and disinfection Methods 0.000 description 6
- 239000001888 Peptone Substances 0.000 description 5
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- 230000008901 benefit Effects 0.000 description 5
- 229940041514 candida albicans extract Drugs 0.000 description 5
- 235000019319 peptone Nutrition 0.000 description 5
- 108090000765 processed proteins & peptides Proteins 0.000 description 5
- 239000012138 yeast extract Substances 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 210000005253 yeast cell Anatomy 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
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- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 229910021556 Chromium(III) chloride Inorganic materials 0.000 description 2
- 235000019750 Crude protein Nutrition 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 102000004877 Insulin Human genes 0.000 description 2
- 108090001061 Insulin Proteins 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 230000002180 anti-stress Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 229960000359 chromic chloride Drugs 0.000 description 2
- QSWDMMVNRMROPK-UHFFFAOYSA-K chromium(3+) trichloride Chemical compound [Cl-].[Cl-].[Cl-].[Cr+3] QSWDMMVNRMROPK-UHFFFAOYSA-K 0.000 description 2
- 239000013530 defoamer Substances 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 229910017604 nitric acid Inorganic materials 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 235000015277 pork Nutrition 0.000 description 2
- 229910052700 potassium Inorganic materials 0.000 description 2
- 239000011591 potassium Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- 229930195730 Aflatoxin Natural products 0.000 description 1
- XWIYFDMXXLINPU-UHFFFAOYSA-N Aflatoxin G Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1C1C=COC1O2 XWIYFDMXXLINPU-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000272525 Anas platyrhynchos Species 0.000 description 1
- 208000035404 Autolysis Diseases 0.000 description 1
- 206010057248 Cell death Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000005409 aflatoxin Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- HSFWRNGVRCDJHI-UHFFFAOYSA-N alpha-acetylene Natural products C#C HSFWRNGVRCDJHI-UHFFFAOYSA-N 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 230000002358 autolytic effect Effects 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 230000036983 biotransformation Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229940046374 chromium picolinate Drugs 0.000 description 1
- QOWZHEWZFLTYQP-UHFFFAOYSA-K chromium(3+);triformate Chemical compound [Cr+3].[O-]C=O.[O-]C=O.[O-]C=O QOWZHEWZFLTYQP-UHFFFAOYSA-K 0.000 description 1
- OZKRURPNXOGYGD-UHFFFAOYSA-N chromium;pyridine-2-carboxylic acid Chemical group [Cr].OC(=O)C1=CC=CC=N1 OZKRURPNXOGYGD-UHFFFAOYSA-N 0.000 description 1
- GJYSUGXFENSLOO-UHFFFAOYSA-N chromium;pyridine-2-carboxylic acid Chemical compound [Cr].OC(=O)C1=CC=CC=N1.OC(=O)C1=CC=CC=N1.OC(=O)C1=CC=CC=N1 GJYSUGXFENSLOO-UHFFFAOYSA-N 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
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- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 229910003460 diamond Inorganic materials 0.000 description 1
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- 238000007865 diluting Methods 0.000 description 1
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- 239000012153 distilled water Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 125000002534 ethynyl group Chemical group [H]C#C* 0.000 description 1
- 238000012262 fermentative production Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000008642 heat stress Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- TVBSSDNEJWXWFP-UHFFFAOYSA-N nitric acid perchloric acid Chemical compound O[N+]([O-])=O.OCl(=O)(=O)=O TVBSSDNEJWXWFP-UHFFFAOYSA-N 0.000 description 1
- 235000021590 normal diet Nutrition 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a microorganism and a microbial fermentation process thereof, in particular to a chromium-rich yeast culture and a fermentation process thereof. The process is characterized by selecting and screening Candida tropicalis and saccharomyces cerevisiae tolerant of high chromium and carrying out production through a liquid-solid two-phase fermentation process. The process flow comprises that: high-chromium-tolerant (trivalent) strains are screened; the strains are progressively expanded and cultured; liquid fermentation is performed to transform inorganic chromium, wherein the content of the inorganic chromium in a culture medium is 0.05 to 0.4 percent, preferably 0.1 to 0.2 percent, and the time of fermentation and transformation is 12 to 48 hours, preferably 18 to 26hours; after the liquid fermentation is over, culture solution and a solid culture medium are mixed according to the weight ratio of 1:3-1.5:1, preferably 1:1, and are subjected to solid aerobic fermentation for 16 to 36 hours, preferably 16 to 24 hours; after the solid aerobic fermentation is over, solid anaerobic fermentation is performed to break the wall of yeast at a temperature kept between40 and 60 DEG C for 6 to 24 hours, preferably at 50 DEG C for 6 to 10 hours; and a finished product after drying contains organic chromium accounting for 0.05 to 0.3 percent. The proportion of transforming the inorganic chromium into the organic chromium by utilizing a technique of the invention is more than 85 percent, and no hexavalent chromium is detected. The chromium-rich yeast culture produced in the invention has the characteristics of simple process, low production cost, good product quality and the like, has the double function of organic chromium and probiotic feed, and can be used for feed of livestock, poultry and aquatic products.
Description
Technical field
The present invention relates to microorganism and microorganism fermentation process thereof, specifically a kind of chromium-rich yeast culture and zymotechnique thereof.
Background technology
Chromium is glucose tolerance factor (Glucose To1erance Factor, active integral part GTF), main effect performance biological effectiveness by enhancing Regular Insulin.Chromium (trivalent) is the essential trace element of animal.Chromium content is low in the normal diet raw material; The extra inorganic chromium utilization ratio of adding is lower than 1%, and kidney can not heavily absorb it, runs off rapidly, consumes bigger especially under stress situation.Therefore, advocate in feed and use organic chromium, the organic chromium absorption rate is between 10～40%.At present the organic chromium of using mainly contains chromium picolinate, chromic formate, amino acid chromium and yeast chromium, and it is best wherein pork pig to be raised the effect of yeast chromium, and the recommendation addition in the daily ration is 0.2～0.4ppm (Lindemann et al., 2008).Yeast chromium is that yeast cell is cultivated in containing chromic substratum, by bio-transformation inorganic chromium is transformed into organic chromium, wherein 80% above chromium is incorporated into protoplasma, main combining form is chromium-pyridine carboxylic acid-gsh (Cr-dinicotinic acid-glutathione), low molecular peptide chromium polymkeric substance (Chromodulin) is also arranged, and peptide contains glycine, halfcystine, L-glutamic acid, aspartic acid.
Yeast chromium has mitigation (Guan et al., 2000) with the collaborative lowering blood glucose of Regular Insulin to diabetes; But the yeast chromium anti-stress strengthens the animal immune function, improves vaccinated tiring (Moonise-Shageer and Mowat, 1993; Kegley and Spears, 1995; Gatta et al., 2001); Yeast chromium can improve pork pig (Grela et al., 1997), broiler chicken (Guo Yu Ming etc. 1998), sow (Leethongdee et al., 2000), meat duck (Li Lili etc. 2001), milk cow breeding performonce fo animals such as (Al-Saiadyet al.2004).
Yeast chromium product technology in the world at present, how by liquid fermenting, solid absorption technology is produced.Such technology has the following disadvantages: 1. the benefit generation of yeast fermentation generation divides limited; 2. it is big to invest amount; 3. production cost height; 4. there is waste liquid to discharge, is necessary further to be improved.
The relevant yeast chromium patent of internal feed can search out an example, and name is called: cow heat stress alleviating agent and its preparation method and application (CN1618316), be characterised in that how to use yeast chromium, and do not relate to zymotechnique.The present domestic patented technology that also yeast culture and chromium conversion is combined.
Yeast culture (Yeast culture) technology is different from common viable cell yeast, to after reaching some amount, thalline carry out broken wall, it does not contain many viable yeast bacterium, be complicated leavened prod, comprise yeast-leavened meta-bolites, sex change substratum, yeast cells wall and yeast content.People know that for a long time the autolysis of utilizing yeast cell disintegrates cell as the effective means (Nolf that obtains component in the cell, 1991), aqtocytolysis is owing to triggered (Chiu etc. 1997) due to the Decomposition of autolytic enzyme that cell can digest self structure under certain condition.Under proper temperature, the breaking yeast cellule membrane of fermentation certain hour can self-dissolving patent 200610150964X such as () Pan Jun.What have only that U.S. Diamond V (Da Nongwei) company claims that its yeast culture product benefit health " XP " takes in production technique at present is liquid-solid two-phase zymotechnique.
The technology that the present invention takes is on the basis of liquid fermenting, introduces solid fermentation, adopts liquid-solid two phase fermentation methods to produce yeast chromium.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of liquid-solid two-phase fermentative production yeast culture technology that can realize abundant rich chromium.Technological process is as follows:
(1) bacterial screening: with candida tropicalis or yeast saccharomyces cerevisiae, determine bacterial strain uses therefor through the performance of the growth in high density trivalent inorganic chromium (0.1%～0.5%) substratum;
(2) bacterial classification enlarged culturing step by step: through the inclined-plane, shake bottle and seeding tank enlarged culturing;
(3) liquid fermenting transforms: trivalent inorganic chromium content is 0.05%～0.4% in the liquid nutrient medium, preferred 0.1%～0.2%; The liquid fermenting transformation time is 12～48 hours, preferred 18～26 hours;
(4) after liquid fermentation liquid and the solid medium mixing liquid fermentation ends, nutrient solution gives solid medium by 1: 3～1.5: 1, and part by weight mixed in preferred 1: 1, entered the solid fermentation stage.Solid medium described here refers to contain one or more mixtures in dregs of beans (cake), cotton dregs (cake), the dish dregs of rice (cake), peanut meal (cake), the flax dregs of rice (cake), beet pulp (cake), wheat bran, rice chaff, inferior powder, Tapioca Starch, soybean hulls, maize alcohol lees, corn, the rice, one or more mixtures in preferred dregs of beans (cake), cotton dregs (cake), the dish dregs of rice (cake), corn, the wheat bran;
Solid fermentation described here refers to solid medium directly or after the sterilising treatment, the fermentation production process that utilizes stacking, container, bag or solid fermentation reactor to carry out is divided into two stages of aerobic and anaerobically fermenting again.
(5) solid aerobic fermentation: aerobic fermentation 16～36 hours, preferred 16～24 hours;
(6) solid anaerobic fermentation: the solid aerobic fermentation carries out broken wall to yeast after finishing, and condition is to keep 40～60 ℃, solid anaerobic fermentation 6～24 hours, preferred 50 ℃ of fermentations 6～10 hours;
(7) finished product: the solid fermentation product is carried out drying obtain finished product; Drying described here refers to fluidized-bed, air-flow, roller drying.Moisture content of finished products is lower than 15%, preferably is lower than 13%, organic chromium 0.05%～0.3%, fragrant odour.
By solid fermentation, microorganism continues propagation in solid medium, enlarges benefits such as product sex change substratum, meta-bolites, yeast cells wall, content and generates the branch ratio, and chromium is able to further evenly disperse along with the yeast fragmentation, promotes the product biological value.
Embodiment
By the following examples the present invention is further described, but the present invention is not limited to these embodiment.
Embodiment 1:
1, bacterial classification: select the candida tropicalis AS2.637 that buys from the Microbiology Research Inst., Guangzhou City for use
2, substratum: glucose 2.0%, peptone 1.0%, yeast extract paste 0.5%, pH=6.8, inorganic chromium (deriving from chromium trichloride) 0.2%, cultivate 24h, confirm that bacterial classification can keep well-grown in above-mentioned nutrient solution.
Embodiment 2:
1, bacterial classification: with embodiment 1.
2, slant medium and culture condition:
Substratum is glucose 2.0%, peptone 1.0%, yeast extract paste 0.5%, agar 2.0%.PH=6.8,115 ℃ the sterilization 18 minutes.Test tube strains is expanded numerous to culture dish, put into 30 ℃ of constant incubators and cultivate 24h.
Embodiment 3:
1, bacterial classification is with embodiment 1.
2, slant culture is with embodiment 2.
3, shake-flask seed substratum and culture condition:
Substratum is glucose 2.0%, peptone 1.0%, yeast extract paste 0.5%.Initial pH=6.8,115 ℃ the sterilization 18 minutes.Bottle is shaken in the access of culture dish bacterial classification, put into 30 ℃ of constant temperature shaking tables, 200r/min shaking culture 24h.
Embodiment 4:
1, bacterial classification is with embodiment 1.
2, slant culture is with embodiment 2.
3, shake-flask seed is cultivated with embodiment 3.
4, first class seed pot expands breeding culture medium and culture condition:
Substratum is dregs of beans 1.0%, corn 0.5%, sucrose 2.0%, peptone 1.0%, yeast extract paste 0.5%, potassium primary phosphate 0.1%, ammonium sulfate 0.2%, sodium-chlor 0.05%, defoamer 0.05%.Initial pH=6.8,121 ℃ the sterilization 30 minutes.Shake bottle and go into jar to the seeding tank inoculum size by 5% inoculation, the seed tank culture time is 24h.
Embodiment 5:
1, bacterial classification is with embodiment 1.
2, slant culture is with embodiment 2.
3, shake-flask seed is cultivated with embodiment 3.
4, first class seed pot expands numerous cultivation with embodiment 4.
5, secondary seed jar chromatize is cultivated:
Substratum is dregs of beans 1.0%, corn 0.5%, sucrose 2.0%, peptone 1.0%, yeast extract paste 0.5%, potassium primary phosphate 0.1%, ammonium sulfate 0.2%, sodium-chlor 0.05%, defoamer 0.05%, inorganic chromium (deriving from chromium trichloride) 0.1%.Initial pH=5.0,121 ℃ the sterilization 30 minutes.Insert first class seed pot by 5% inoculum size again, cultivate 35h.
Embodiment 6:
1, bacterial classification is with embodiment 1.
2, slant culture is with embodiment 2.
3, shake-flask seed is cultivated with embodiment 3.
4, first class seed pot expands numerous cultivation with embodiment 4.
5, secondary seed jar chromatize is cultivated with embodiment 5.
6, solid fermentation:
Dregs of beans is heated to 85～100 ℃, keeps sterilization in 5 minutes, be cooled to 40 ℃; Secondary seed jar chromatize nutrient solution is mixed with the sterilization dregs of beans, and adjustment solid water content reaches 45% and begins fermentation.The fermentation materials temperature is monitored, made temperature not surpass 45 ℃; Enter anaerobically fermenting behind the aerobic fermentation 24h, temperature reaches more than 50 ℃, 6～10 hours broken walls that ferment, and microscopy broken wall situation determines when to finish to carry out drying.
Embodiment 7:
1, bacterial classification is with embodiment 1.
2, slant culture is with embodiment 2.
3, shake-flask seed is cultivated with embodiment 3.
4, first class seed pot expands numerous cultivation with embodiment 4.
5, secondary seed jar chromatize is cultivated with embodiment 5.
6, the solid fermentation broken wall is with embodiment 6.
7, product detects and effect
Sample is cleared up under the high-temperature and high-pressure conditions in perchloric acid-nitric acid mix acid liquor, and digestion solution is after suitably diluting, with aas determination chromium content.
(1) measure total chromium:
Get 0.2～0.5g product of the present invention in the digestion bottle, add 8ml HC1O4-HNO3 (4: 1) mixed solution and on electric furnace, digest.When becoming colorless, solution can stop digestion.(150 watts of frequencies 20KHZ) are handled 30min, will change liquid afterwards and transfer in the volumetric flask of 10ml, with 5.0% concentration HNO3 solution constant volume with ultrasonator in the cooling back.Adopt atomic absorption method to measure, test condition is: lamp current I=12mA, passband AA=1.6nm, wavelength X 357.8nm, burner height=7.5mm, air flow quantity=9.4min, acetylene gas flow=2.5L/min.Can draw the total chrome content for the treatment of in the test sample according to typical curve.Bag organic chromium and inorganic chromium in the total chrome content of surveying.
(2) separation determination of product organic chromium and inorganic chromium
0.2～0.3g product adding of the present invention is filled in the centrifuge tube of 9ml distilled water, stir, leave standstill 12h, the centrifugal 20min of 3500r/min rotating speed draws supernatant liquor and measures inorganic chromium content with atom suction method.Lower sediment is shifted out centrifuge tube, and the digestion constant volume adopts aas determination to go out organic chromium content.
(3) the transformation efficiency organic chromium content=total chrome content-inorganic chromium content of organic chromium content and chromium, the transformation efficiency of chromium=organic chromium content/total chrome content.
(4) the main nutritive value index of product of the present invention
Measure organic chromium content＞0.1% as stated above; Inorganic chromium changes into ratio＞85% of organic chromium, and sexavalent chrome does not detect; Crude protein content 10%～50% is according to the peptide transformation efficiency (the medium and small peptide content of crude protein)＞8% of national soy peptide powder industry standard (QB/T2653-2004) mensuration; Aflatoxin＜30 μ g/kg.
Product of the present invention and technological advantage
(1) the present invention draws the advantage of related microorganism preparation production method, adopting known candida tropicalis bacterium and S. cervisiae is bacterial classification, be raw material with glucose, dregs of beans (cake), cotton dregs (cake), the dish dregs of rice (cake), peanut meal (cake), the flax dregs of rice (cake) etc., wide material sources, technological process is simple, and product price is cheap.
(2) the present invention adopts microbial liquid-solid two-phase new process for fermenting, by the heavy dose inoculation, can solve in the solid fermentation process, and fermentation period is long, problems such as easy pollution, the constant product quality of production.
(3) microorganism continues propagation at solid medium, and chromium further evenly disperses.
(4) the present invention adopts microbial liquid-solid two-phase new process for fermenting, has not only finished inorganic chromium and has been converted into the organic chromium process, also by technology for broken wall, produces a large amount of yeast culture compositions, has the characteristics of beneficial uncooked feed.
(5) chromium-rich yeast culture produced of the present invention has the dual function of organic chromium and beneficial uncooked feed concurrently, is used for aquatic feed for domestic animal, can improve the animal anti-stress ability, improves the female livestock breed performance, improves meat matter.
(6) the present invention all contain chromium bacterium liquid and all sneak into the solid material, continue fermentation and make finished product, do not have liquid and waste slag produced discharging, fully compliance with environmental protection requirements.
Claims (3)
1. the zymotechnique of a chromium-rich yeast culture, carry out according to following steps:
(1) bacterial classification enlarged culturing step by step: through the inclined-plane, shake-flask seed and seeding tank enlarged culturing;
(2) liquid fermenting: inorganic chromium content is 0.05%～0.4% in the liquid nutrient medium, and the liquid fermenting transformation time is 12～48 hours, and inorganic chromium described here is trivalent chromium;
(3) solid fermentation: after liquid fermenting finished, nutrient solution mixed by 1: 3～1.5: 1 part by weight with solid medium, enters the solid fermentation stage; Solid medium described here is characterized in that containing one or more mixtures in dregs of beans, cotton dregs, the dish dregs of rice, peanut meal, the flax dregs of rice, beet pulp, wheat bran, rice chaff, inferior powder, Tapioca Starch, soybean hulls, maize alcohol lees, corn, the rice; Solid fermentation described here refers to the production process of utilizing stacking, container, bag or solid fermentation reactor to carry out be divided into two stages of aerobic and anaerobically fermenting again;
(4) the aerobic fermentation stage in the solid: aerobic fermentation 16～36 hours;
(5) the anaerobically fermenting stage in the solid: the solid aerobic fermentation carries out broken wall to yeast after finishing, and condition is to keep 40～60 ℃, solid anaerobic fermentation 6～24 hours;
(6) drying: the solid fermentation product behind the broken wall is dried to moisture below 15%, obtains finished product; Wherein, the candida tropicalis AS2.637 of described bacterial classification for being bought by the Microbiology Research Inst., Guangzhou City.
2. the zymotechnique of chromium-rich yeast culture according to claim 1, it is characterized in that: inorganic chromium content is 0.1% in the substratum; The liquid fermenting transformation time is 18 hours; Nutrient solution mixes with solid medium by 1: 1 part by weight; Solid medium is characterized in that containing one or more mixtures in dregs of beans, cotton dregs, the dish dregs of rice, corn, the wheat bran; Solid aerobic fermentation 24 hours; 50 ℃ of solid anaerobic fermentations, 10 hours; Finished product is dried to moisture below 13%.
3. the described chromium-rich yeast culture of claim 1 has the dual function of organic chromium and beneficial uncooked feed concurrently, can be used for aquatic feed for domestic animal.
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| CN101792720B (en) * | 2009-05-12 | 2014-06-04 | 广州市博善生物饲料有限公司 | Production method of selenium-enriched yeast culture |
| CN101914460B (en) * | 2010-07-13 | 2013-09-11 | 广州市博善生物饲料有限公司 | Method for fermenting phodotorula benthica culture |
| CN102643864A (en) * | 2012-04-27 | 2012-08-22 | 广州市富泉生物科技有限公司 | Process for preparing yeast cultures |
| CN103396955B (en) * | 2013-07-15 | 2015-03-04 | 浙江深友生物技术有限公司 | Yeast and method for producing chromium-enriched yeast by utilizing yeast via high-density fermentation |
| CN104489347A (en) * | 2014-12-12 | 2015-04-08 | 韦文峰 | Chromium-rich microbial feed additive for dairy cow and preparation method of chromium-rich microbial feed additive |
| CN105062905B (en) * | 2015-07-22 | 2018-11-09 | 山东宝来利来生物工程股份有限公司 | One plant of saccharomyces cerevisiae and its application for solid fermentation |
| CN106987530A (en) * | 2017-03-21 | 2017-07-28 | 高唐华农生物工程有限公司 | A kind of preparation method of yeast culture |
| CN111041050A (en) * | 2019-12-03 | 2020-04-21 | 同济大学 | Method for converting hexavalent chromium in tea into trivalent chromium by utilizing food-derived microbial fermentation |
| CN114540214B (en) * | 2020-11-27 | 2023-07-04 | 北京岳泰华生物科技有限公司 | Microorganism for high-yield organic chromium and application thereof |
| CN114686383B (en) * | 2020-12-30 | 2023-12-19 | 安琪酵母股份有限公司 | Chromium-enriched yeast with high chromium absorptivity, and preparation method and application thereof |
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