CN1322109C - Drying production technique of fluidized bed by starch adsorption of dry oceanic rhodotorula - Google Patents
Drying production technique of fluidized bed by starch adsorption of dry oceanic rhodotorula Download PDFInfo
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- CN1322109C CN1322109C CNB021391785A CN02139178A CN1322109C CN 1322109 C CN1322109 C CN 1322109C CN B021391785 A CNB021391785 A CN B021391785A CN 02139178 A CN02139178 A CN 02139178A CN 1322109 C CN1322109 C CN 1322109C
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- 241000223252 Rhodotorula Species 0.000 title claims abstract description 56
- 238000001035 drying Methods 0.000 title claims abstract description 20
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 17
- 229920002472 Starch Polymers 0.000 title claims abstract description 14
- 235000019698 starch Nutrition 0.000 title claims abstract description 14
- 239000008107 starch Substances 0.000 title claims abstract description 14
- 238000001179 sorption measurement Methods 0.000 title abstract 3
- 238000000034 method Methods 0.000 title description 10
- 239000007787 solid Substances 0.000 claims abstract description 16
- 230000001580 bacterial effect Effects 0.000 claims description 18
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 18
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 15
- 239000004202 carbamide Substances 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 14
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 11
- 239000008103 glucose Substances 0.000 claims description 11
- 206010001497 Agitation Diseases 0.000 claims description 10
- 238000013019 agitation Methods 0.000 claims description 10
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 9
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 9
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 9
- 235000010333 potassium nitrate Nutrition 0.000 claims description 9
- 239000004323 potassium nitrate Substances 0.000 claims description 9
- 229910019142 PO4 Inorganic materials 0.000 claims description 8
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 8
- 229940041514 candida albicans extract Drugs 0.000 claims description 8
- 239000001963 growth medium Substances 0.000 claims description 8
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 8
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 8
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 8
- 239000010452 phosphate Substances 0.000 claims description 8
- 239000011591 potassium Substances 0.000 claims description 8
- 229910052700 potassium Inorganic materials 0.000 claims description 8
- 239000012138 yeast extract Substances 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 7
- 241000223253 Rhodotorula glutinis Species 0.000 claims description 7
- 229930006000 Sucrose Natural products 0.000 claims description 7
- 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 claims description 7
- 239000012141 concentrate Substances 0.000 claims description 7
- 239000005720 sucrose Substances 0.000 claims description 7
- 238000005406 washing Methods 0.000 claims description 7
- 229920001817 Agar Polymers 0.000 claims description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 6
- 239000008272 agar Substances 0.000 claims description 6
- 210000000481 breast Anatomy 0.000 claims description 6
- 229910052799 carbon Inorganic materials 0.000 claims description 6
- 238000000855 fermentation Methods 0.000 claims description 6
- 230000004151 fermentation Effects 0.000 claims description 6
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 5
- 238000005138 cryopreservation Methods 0.000 claims description 5
- 239000012531 culture fluid Substances 0.000 claims description 5
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 5
- 235000015097 nutrients Nutrition 0.000 claims description 5
- 239000002245 particle Substances 0.000 claims description 5
- 238000012807 shake-flask culturing Methods 0.000 claims description 5
- 238000003860 storage Methods 0.000 claims description 5
- 208000011580 syndromic disease Diseases 0.000 claims description 5
- 239000008399 tap water Substances 0.000 claims description 5
- 235000020679 tap water Nutrition 0.000 claims description 5
- 230000008719 thickening Effects 0.000 claims description 5
- 238000011534 incubation Methods 0.000 claims description 2
- 239000002609 medium Substances 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 238000009423 ventilation Methods 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims 1
- 241000238557 Decapoda Species 0.000 abstract description 16
- 210000004027 cell Anatomy 0.000 abstract description 8
- 230000000694 effects Effects 0.000 abstract description 7
- 238000005516 engineering process Methods 0.000 abstract description 3
- 238000004321 preservation Methods 0.000 abstract description 3
- 230000036541 health Effects 0.000 abstract description 2
- 238000005273 aeration Methods 0.000 abstract 1
- 238000009395 breeding Methods 0.000 abstract 1
- 230000001488 breeding effect Effects 0.000 abstract 1
- 230000002265 prevention Effects 0.000 abstract 1
- 210000005253 yeast cell Anatomy 0.000 abstract 1
- 239000000047 product Substances 0.000 description 19
- 238000012258 culturing Methods 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 238000010564 aerobic fermentation Methods 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000008676 import Effects 0.000 description 3
- 239000012263 liquid product Substances 0.000 description 3
- 241000238553 Litopenaeus vannamei Species 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
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- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- JEBFVOLFMLUKLF-IFPLVEIFSA-N Astaxanthin Natural products CC(=C/C=C/C(=C/C=C/C1=C(C)C(=O)C(O)CC1(C)C)/C)C=CC=C(/C)C=CC=C(/C)C=CC2=C(C)C(=O)C(O)CC2(C)C JEBFVOLFMLUKLF-IFPLVEIFSA-N 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- RMMPZDDLWLALLJ-UHFFFAOYSA-N Thermophillin Chemical compound COC1=CC(=O)C(OC)=CC1=O RMMPZDDLWLALLJ-UHFFFAOYSA-N 0.000 description 1
- GZCGUPFRVQAUEE-SLPGGIOYSA-N aldehydo-D-glucose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-SLPGGIOYSA-N 0.000 description 1
- MQZIGYBFDRPAKN-ZWAPEEGVSA-N astaxanthin Chemical compound C([C@H](O)C(=O)C=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)C(=O)[C@@H](O)CC1(C)C MQZIGYBFDRPAKN-ZWAPEEGVSA-N 0.000 description 1
- 229940022405 astaxanthin Drugs 0.000 description 1
- 235000013793 astaxanthin Nutrition 0.000 description 1
- 239000001168 astaxanthin Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical class 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000002354 daily effect Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000009313 farming Methods 0.000 description 1
- 235000021050 feed intake Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 229940057059 monascus purpureus Drugs 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 235000015170 shellfish Nutrition 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 210000003934 vacuole Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/16—Particles; Beads; Granular material; Encapsulation
- C12M25/20—Fluidized bed
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention discloses a drying production technology of a starch adsorption fluidized bed of solid oceanic rhodotorula, which adopts a drying method of continuous stream plus deep aeration culture and a starch adsorption fluidized bed, and dry solid oceanic rhodotorula products of transport prevention and easy preservation can be obtained. The yeast cell concentration can finally reach 50 to 100 hundred million /gram by the design of fed batch culture technology, and the yield can be effectively enhanced. Solid oceanic rhodotorula products of more or less 30% of living cells can be obtained by the drying method. Preferable effect can be obtained by applying the products in breeding and adult cultivation of aquatic products, such as prawns, sea crabs, etc. The present invention can effectively enhance the health state of aquatic products, reduce mortality and enhance cultivation efficiency.
Description
Technical field
The present invention relates to a kind of production method of ocean rhodotorula, particularly a kind of starch absorption fluidised bed drying production method of solid ocean rhodotorula.
Background technology
Ocean rhodotorula is widely used at present as a kind of biological feed in growing seedlings of fishery products such as shrimps, crab class, shellfish and the adult breed.Chinese patent discloses a kind of denomination of invention and has been " live single-cell sea red-yeast ecological bait and production method thereof ", and publication number is CN1146290A, and open day is on April 24th, 1997.This invention discloses the production method of ocean rhodotorula yeast culture and liquid product in specification sheets, this method relates to a kind of ocean rhodotorula separates, makes after once feed intake liquid fermenting, the post-processed liquid product from occurring in nature method.This method exists the speed of growth of ocean rhodotorula slow, and the production cycle is long, and the cell concn of the ocean rhodotorula that of fermenting is low, and the final liquid product quality guaranteed period is short, needs refrigeration to wait the shortcoming that is unfavorable for scale operation, prolonged preservation and long-distance transport.
Summary of the invention
The purpose of this utility model is exactly the starch absorption fluidised bed drying production method that a kind of solid ocean rhodotorula will be provided, its adopts the ventilate method of continuous feeding culture ocean rhodotorula of deep liquid, improved the output of ocean rhodotorula, made it can carry out industrialized production.And on post-processed, adopt starch absorption fluid-bed drying, can obtain having certain active ocean rhodotorula solid phase prod, thereby the quality guaranteed period of ocean rhodotorula product is prolonged, reach the purpose of convenient transportation.
The purpose of this utility model is achieved in that
The microorganism of using
The microbial strains that is used for producing ocean rhodotorula solid phase prod of the present invention is a rhodotorula glutinis, and the Latin formal name used at school is Rhodotorula glutinis.The bacterial classification purposes is food and fodder yeast.Bacterial classification is available from China Committee for Culture Collection of Microorganisms common micro-organisms center, and deposit number is CGMCC No.2.703.
The rhodotorula glutinis bacterial strain has following character
1, morphological specificity
Circle or oval, individual less, be about 3-4.2
*4-5.4um.Microscopically can be seen typical yeast vacuole.
2, the feature on substratum
30 ℃ cultivate inoculation after, observed in 24 hours, in wort liquid is cultivated,, and do not break away from after the polygon budding of cell if it is very slow to leave standstill then growth, a circle Pu shape thing is arranged in the place near liquid level, shake bottle or air agitation fermentation microscopy and survey and mostly be individual cells.
30 ℃ cultivate inoculation after, observed in 48 hours, the chromatogram (nineteen fifty-seven version) of publishing with Science Press is as the standard of color description.Be the circular bacterium colony of scarlet on the wort agar substratum, the surface is glossy, and neat in edge is opaque, and is long to the few middle bump that has of later stage bacterium.
3, physiological and biochemical property
(1) fermenting carbohydrate: glucose-maltose-semi-lactosi-sucrose-lactose-Mi disaccharides-melizitose-synanthrin-Zulkovsky starch-
(2) assimilation carbon source: glucose+lactose-ethanol-maltose+sucrose+Zulkovsky starch-semi-lactosi-Mi disaccharides-L-arabinose-
(3) assimilation nitrogenous source: saltpetre+ammonium sulfate+urea+Sodium Nitrite-L-Methionin+
(4) assimilation inositol :-
(5) produce the kind of starch compound :-
(6) decomposing urea :+
(7) produce the ester reaction :-
(8) anti-height oozes reaction: 50% glucose+60% glucose-
(9) anti-ethanol: 3%
(10) fatal temperature: 56 ℃
4, utilization of carbon source
Glucose maltose sucrose
The production method of solid ocean rhodotorula of the present invention may further comprise the steps:
(1) going down to posterity of bacterial classification: rhodotorula glutinis (Rhodotorula glutinis) CGMCC No.2.703 adopts the inclined-plane to go down to posterity, substratum is the wort agar substratum of 8-12Be, culture temperature is 26 ℃-31 ℃, and incubation time is 26-72 hour, to growing pink circular bacterium colony; Colony diameter is 1-2mm;
(2) strain expanded culture: the bacterial classification of (1) step is connected in the triangular flask of the wort nutrient solution that the sterilized 8-12Be of 150ml is housed, at 26 ℃-31 ℃, carried out under the 100-250rpm condition shake-flask culture 36-48 hour, insert then in the Ka Shi jar of two wort aseptic culture fluids that 20 liters of 8-12Be respectively are housed, 26 ℃ of-31 ℃ of magnetic agitation were cultivated 36-48 hour, again two Ka Shi jar bacterial classifications are inserted in the pure culture fermentor tank that 3-5 ton substratum is housed of having sterilized, 26 ℃ of-31 ℃ of aeration-agitations were cultivated 36-48 hour, and air flow is controlled at 90-130m
3/ hr;
(3) the continuous feeding culture of fermentation deep ventilation: under aseptic condition, the seed liquor of turning out is inserted fermentor tank, 26 ℃-31 ℃ of controlled temperature, aeration-agitation was cultivated 30-48 hour, and air flow is controlled at 90-130m
3/ hr, continuing to flow simultaneously adds carbon source and nitrogenous source.Carbon source is a sucrose, and concentration is 2%-5% (weight ratio), and nitrogenous source is a urea, and concentration is 0.1%-0.3% (weight ratio);
(4) separating, washing concentrates: under the separating machine of 3000-5000rpm rotating speed fermented liquid is concentrated, yeast-lactic after concentrating concentrates after dissolving with tap water once more, to with ocean rhodotorula thalline washes clean, the storage tank cryopreservation of the yeast-lactic behind the thickening and washing;
(5) drying: the ocean rhodotorula breast with W-Gum or yam starch and after concentrating is by 2: 1-3: the physical condition homogeneous is mixed, is stirred to 2 (weight ratios); making granularity by nodulizer is 10-60 purpose small-particle; by fluidized-bed in 70 ℃-90 ℃ wind-warm syndrome rapid drying 5-20 minute, get product then.
The culture medium prescription of pure culture fermentor tank and big fermentor tank is: (weight ratio)
Glucose 2-5%
Magnesium sulfate heptahydrate 0.05-0.25%
Yeast extract 0.1-0.3%
Potassium primary phosphate 0.5-1%
Urea 0.1-0.3%
Ammonium sulfate 0.05-0.2%
Saltpetre 0.05-0.2%
PH value 4.0-6.0
The temperature of medium sterilization is 121 ℃, and the time is 30 minutes.
The solid ocean rhodotorula that adopts the present invention to produce has following feature:
1, shape: be by W-Gum or yam starch and the red granules shape that mixes of the ocean rhodotorula breast after concentrating
Material.
2, size: granularity is the 10-60 order.
3, form: W-Gum or yam starch content are 85%-95% (weight ratio) in the product, and ocean rhodotorula content is 5%-15% (weight ratio).
4, biological activity: the ocean rhodotorula cell concn is 50-100 hundred million/gram in the product, and cytoactive is 10%-30%.
The present invention is owing to adopt the ventilate production method of continuous feeding culture ocean rhodotorula of deep liquid, so that the output of ocean rhodotorula increases substantially, bring up to 20-30 hundred million/ml by original 5-10 hundred million/ml; Owing to adopt starch absorption fluid-bed drying, the liquid ocean rhodotorula after concentrating is dried to the solid ocean rhodotorula again, is keeping having improved the preservation period of product on the certain active basis of ocean rhodotorula like this, and be convenient to transportation.The product that the present invention produces by the result of use in prawn culturing as can be seen, it has improved the state of health of prawn, reduces the mortality ratio of prawn, has improved the quality of prawn, for the shrimp farming has brought economic benefit.
Below with regard to manufacturing of the present invention, the test example illustrate.
Production Example 1
At substratum is on the wort agar substratum test tube slant of 8Be, will stick the rhodotorula bacterial classification and be inoculated on aseptic condition in this substratum, and the control culture temperature is 30 ℃, cultivates 48 hours, and to grow pink circular bacterium colony, colony diameter is 1.5mm.Then above-mentioned bacterial classification is connected under aseptic condition in two 500ml triangular flasks, (dress 150ml wort nutrient solution in the triangular flask) carries out enlarged culturing, 30 ℃ of control culture temperature, and shake-flask culture is 48 hours under 200rpm.Above-mentioned triangular flask bacterial classification is inserted respectively in the Ka Shi jar of two wort aseptic culture fluids that 20 liters of 8Be respectively are housed, 30 ℃ of magnetic agitation were cultivated 48 hours again.At last above-mentioned cultured Ka Shi jar seed is inserted the aerobic fermentation that once feeds intake in the pure culture jar that 5 tons of substratum are housed of having sterilized and cultivate, controlled temperature is 27 ℃, and air flow is 100m
3/ hr, cultivated 48 hours, culture medium prescription is: glucose 2%, magnesium sulfate heptahydrate 0.25%, yeast extract 0.2%, potassium primary phosphate 0.5%, urea 0.3%, ammonium sulfate 0.05%, saltpetre 0.05%, PH are 4.0-6.0, and substratum was sterilized 30 minutes down at 121 ℃.
With 5 tons in above-mentioned seed, all insert in 100 tons the fermentor tank and carry out continuous feeding culture, 27 ℃ of control culture temperature, air flow is 100m
3/ hr, Continuous Flow adds 2% sucrose and 0.3% urea in the cultivation, cultivates 48 hours.Culture medium prescription is: glucose 2%, magnesium sulfate heptahydrate 0.25%, yeast extract 0.2%, potassium primary phosphate 0.5%, urea 0.3%, ammonium sulfate 0.05%, saltpetre 0.05%, PH are 4.0-6.0, and substratum was sterilized 30 minutes down at 121 ℃.After the fermentation ends, fermented liquid is pumped into the high speed butterfly chip separating machine (model is FEUX510, ALFA-LAVAL produce) of import, under the rotating speed of 4000rpm, fermented liquid is concentrated, yeast-lactic after concentrating concentrates after dissolving with tap water once more, can be with ocean rhodotorula thalline washes clean.Yeast-lactic behind the thickening and washing is delivered to the storage tank cryopreservation with transferpump.Ocean rhodotorula breast after 2 tons of W-Gums and 1 ton concentrated mixes; be stirred to the physical condition homogeneous; (model is FLG120 by one-step-granulating method again; luxuriant source pharmaceutical machine factory produces) to make granularity be 30 purpose small-particles; (model is HF20 by fluidized-bed at last; changzhou city medical matters drying plant factory produces); rapid drying is 20 minutes in 70 ℃ wind-warm syndrome; can obtain 2.2 tons of solid ocean rhodotorulas; wherein W-Gum contains 91% (weight ratio); ocean rhodotorula contains 9% (weight ratio); the activity of ocean rhodotorula is 30% in this product, and cell concn reaches 10,000,000,000/gram.
The said products is bundled into 500 gram or 10 kilograms products, is put in shady and cool dry place, can effectively preserve 2 years.
Production Example 2
At substratum is on the wort agar substratum test tube slant of 10Be, will stick the rhodotorula bacterial classification and be inoculated on aseptic condition in this substratum, and the control culture temperature is 26 ℃, cultivates 26 hours, and to grow pink circular bacterium colony, colony diameter is 1mm.Then above-mentioned bacterial classification is connected under aseptic condition in two 500ml triangular flasks, (dress 150ml wort nutrient solution in the triangular flask) carries out enlarged culturing, 26 ℃ of control culture temperature, and shake-flask culture is 36 hours under 100rpm.Above-mentioned triangular flask bacterial classification is inserted respectively in the Ka Shi jar of two wort aseptic culture fluids that 20 liters of 8Be respectively are housed, 26 ℃ of magnetic agitation were cultivated 36 hours again.At last above-mentioned cultured Ka Shi jar seed is inserted the aerobic fermentation that once feeds intake in the pure culture jar that 3 tons of substratum are housed of having sterilized and cultivate, controlled temperature is 26 ℃, and air flow is 90m
3/ hr, cultivated 36 hours, culture medium prescription is: glucose 3.5%, magnesium sulfate heptahydrate 0.15%, yeast extract 0.1%, potassium primary phosphate 0.75%, urea 0.2%, ammonium sulfate 0.1%, saltpetre 0.1%, PH are 4.0-6.0, and substratum was sterilized 30 minutes down at 121 ℃.
With 3 tons in above-mentioned seed, all insert in 100 tons the fermentor tank and carry out continuous feeding culture, 26 ℃ of control culture temperature, air flow is 90m
3/ hr, Continuous Flow adds 3% sucrose and 0.2% urea in the cultivation, cultivates 30 hours.Culture medium prescription is: glucose 3.5%, magnesium sulfate heptahydrate 0.15%, yeast extract 0.1%, potassium primary phosphate 0.75%, urea 0.2%, ammonium sulfate 0.1%, saltpetre 0.1%, PH are 4.0-6.0, and substratum was sterilized 30 minutes down at 121 ℃.After the fermentation ends, fermented liquid is pumped into the high speed butterfly chip separating machine (model is FEUX510, ALFA-LAVAL produce) of import, under the rotating speed of 3000rpm, fermented liquid is concentrated, yeast-lactic after concentrating concentrates after dissolving with tap water once more, can be with ocean rhodotorula thalline washes clean.Yeast-lactic behind the thickening and washing is delivered to the storage tank cryopreservation with transferpump.Ocean rhodotorula breast after 3 tons of W-Gums and 2 tons are concentrated mixes; be stirred to the physical condition homogeneous; (model is FLG120 by one-step-granulating method again; luxuriant source pharmaceutical machine factory produces) to make granularity be 10 purpose small-particles; (model is HF20 by fluidized-bed at last; changzhou city medical matters drying plant factory produces); rapid drying is 10 minutes in 80 ℃ wind-warm syndrome; can obtain 3.4 tons of solid ocean rhodotorulas; wherein W-Gum contains 85% (weight ratio); ocean rhodotorula contains 15% (weight ratio); the activity of ocean rhodotorula is 20% in this product, and cell concn reaches 9,000,000,000/gram.
The said products is bundled into 500 gram or 10 kilograms products, is put in shady and cool dry place, can effectively preserve 2 years.
Production Example 3
At substratum is on the wort agar substratum test tube slant of 12Be, will stick the rhodotorula bacterial classification and be inoculated on aseptic condition in this substratum, and the control culture temperature is 31 ℃, cultivates 72 hours, and to grow pink circular bacterium colony, colony diameter is 2mm.Then above-mentioned bacterial classification is connected under aseptic condition in two 500ml triangular flasks, (dress 150ml wort nutrient solution in the triangular flask) carries out enlarged culturing, 31 ℃ of control culture temperature, and shake-flask culture is 45 hours under 250rpm.Above-mentioned triangular flask bacterial classification is inserted respectively in the Ka Shi jar of two wort aseptic culture fluids that 20 liters of 8Be respectively are housed, 31 ℃ of magnetic agitation were cultivated 45 hours again.At last above-mentioned cultured Ka Shi jar seed is inserted the aerobic fermentation that once feeds intake in the pure culture jar that 4 tons of substratum are housed of having sterilized and cultivate, controlled temperature is 31 ℃, and air flow is 130m
3/ hr, cultivated 45 hours, culture medium prescription is: glucose 5%, magnesium sulfate heptahydrate 0.05%, yeast extract 0.3%, potassium primary phosphate 1.0%, urea 0.1%, ammonium sulfate 0.2%, saltpetre 0.2%, PH are 4.0-6.0, and substratum was sterilized 30 minutes down at 121 ℃.
With 4 tons in above-mentioned seed, all insert in 100 tons the fermentor tank and carry out continuous feeding culture, 31 ℃ of control culture temperature, air flow is 130m
3/ hr, Continuous Flow adds 5% sucrose and 0.1% urea in the cultivation, cultivates 45 hours.Culture medium prescription is: glucose 5%, magnesium sulfate heptahydrate 0.05%, yeast extract 0.3%, potassium primary phosphate 1.0%, urea 0.1%, ammonium sulfate 0.2%, saltpetre 0.2%, PH are 4.0-6.0, and substratum was sterilized 30 minutes down at 121 ℃.After the fermentation ends, fermented liquid is pumped into the high speed butterfly chip separating machine (model is FEUX510, ALFA-LAVAL produce) of import, under the rotating speed of 4000rpm, fermented liquid is concentrated, yeast-lactic after concentrating concentrates after dissolving with tap water once more, can be with ocean rhodotorula thalline washes clean.Yeast-lactic behind the thickening and washing is delivered to the storage tank cryopreservation with transferpump.Ocean rhodotorula breast after 2.5 tons of W-Gums and 1.5 tons are concentrated mixes; be stirred to the physical condition homogeneous; (model is FLG120 by one-step-granulating method again; luxuriant source pharmaceutical machine factory produces) to make granularity be 60 purpose small-particles; (model is HF20 by fluidized-bed at last; changzhou city medical matters drying plant factory produces); rapid drying is 5 minutes in 90 ℃ wind-warm syndrome; can obtain 2.8 tons of solid ocean rhodotorulas; wherein W-Gum contains 95% (weight ratio); ocean rhodotorula contains 5% (weight ratio); the activity of ocean rhodotorula is 10% in this product, and cell concn reaches 5,000,000,000/gram.
The said products is bundled into 500 gram or 10 kilograms products, is put in shady and cool dry place, can effectively preserve 2 years.
Test the effect test of routine product solid ocean rhodotorula of the present invention in Penaeus vannamei is cultured.
Ocean rhodotorula is except that rich in proteins, also have astaxanthin (carotenoid a kind of) and a large amount of unsaturated fatty acidss, be used for aquaculture, can obviously improve the survival rate of output, fishes and shrimps and improve body colour, and can avoid residual in human body of the edible chemical pigment of tradition in culturing, microbiotic and the harm that brings.In order to understand the effect of product solid ocean rhodotorula of the present invention in prawn culturing, we test on the shrimp pool that North Sea group woods biotechnology company limited provides, and situation is as follows:
One, materials and methods
1, experiment pool and contrast pond: select 2 of test tanks, the pond number is 15
#, 16
#, area is respectively 9 mu, 10 mu.2 in pond of contrast, the pond number is 17
#, 18
#, being respectively 9 mu, 11 mu, shrimp pond auxiliary facility and cultivating condition are basic identical.
2, breed variety and putting in a suitable place to breed: breed variety is a Penaeus vannamei, and the shrimp seedling is from local seedling field, puts the date in a suitable place to breed and density sees Table 1
Table 1 test tank is put situation in a suitable place to breed with the contrast pond
| Pond number | Area | Put the date (month, day) in a suitable place to breed | Put quantity (ten thousand tails) in a suitable place to breed | Mu is put (ten thousand tails) | |
| The contrast pond | 17 # | 9 | August 4 calendar year 2001 | 22.5 | 2.5 |
| 18 # | 11 | August 4 calendar year 2001 | 27.5 | 2.5 | |
| Test tank | 15 # | 9 | August 4 calendar year 2001 | 22.5 | 2.5 |
| 16 # | 10 | August 4 calendar year 2001 | 25 | 2.5 | |
3, cultural method: press the 2% interpolation product solid ocean rhodotorula of the present invention of feed grain in the daily ration of test tank every day, control group does not add, and other aquaculture management measure is identical.
Two, interpretation of result
A) results situation (seeing Table 2):
Each pond results situation of table 2
| Pond number | The breed fate (my god) | Gross output (kilogram) | Per mu yield (kilogram) | Survival rate (%) | Specification (tail/kilogram) | |
| The contrast pond | 17 # | 101 | 1350 | 150 | 50 | 66 |
| 18 # | 102 | 1672 | 152 | 52 | 67 | |
| Test tank | 15 # | 95 | 1440 | 160 | 56 | 60 |
| 16 # | 96 | 1630 | 163 | 58 | 59 | |
B) Economic and Efficiency Analysis: the test tank comparison is according to 10 kilograms of the average mu volume increase in pond, about 6.7%.
C) surviving rate situation analysis: as can be seen from Table 2, volume increase is mainly reflected in the surviving rate raising, illustrates that rhodotorula is strengthening the prawn vigor, has very strong effect on the surviving rate of raising prawn.
D) mature stage analysis: can find out obviously that from table 2 becoming the shrimp specification preferably under the situation, about 6 days of mature stage are shortened in test tank average specific contrast pond.
Claims (1)
1. the starch of a solid ocean rhodotorula adsorbs the fluidised bed drying production method, and it is characterized in that: it may further comprise the steps:
(1) bacterial classification goes down to posterity: rhodotorula glutinis (Rhodotorula glutinis) CGMCC No.2.703 adopts the inclined-plane to go down to posterity, substratum is the wort agar substratum of 8-12Be, culture temperature is 26 ℃-31 ℃, incubation time is 26-72 hour, to growing pink circular bacterium colony, colony diameter is 1-2mm;
(2) strain expanded culture: the bacterial classification of step (1) is connected in the triangular flask of the wort nutrient solution that the sterilized 8-12Be of 150ml is housed, at 26 ℃-31 ℃, carried out under the 100-250rpm condition shake-flask culture 36-48 hour, insert then in the Ka Shi jar of two wort aseptic culture fluids that 20 liters of 8-12Be respectively are housed, 26 ℃ of-31 ℃ of magnetic agitation were cultivated 36-48 hour, again two Ka Shi jar bacterial classifications are inserted in the pure culture fermentor tank that 3-5 ton substratum is housed of having sterilized, 26 ℃ of-31 ℃ of aeration-agitations were cultivated 36-48 hour, and air flow is controlled at 90-130m
3/ hr;
(3) the continuous feeding culture of fermentation deep ventilation: under aseptic condition, the seed liquor of turning out is inserted fermentor tank, 26 ℃-31 ℃ of controlled temperature, aeration-agitation was cultivated 30-48 hour, and air flow is controlled at 9-130m
3/ hr, continuing to flow simultaneously adds carbon source and nitrogenous source; Carbon source is a sucrose, and by weight, concentration is 2%-5%; Nitrogenous source is a urea, and by weight, concentration is 0.1%-0.3%;
(4) separating, washing concentrates: under the separating machine of 3000-5000rpm rotating speed fermented liquid is concentrated, yeast-lactic after concentrating concentrates after dissolving with tap water once more, to with ocean rhodotorula thalline washes clean, the storage tank cryopreservation of the yeast-lactic behind the thickening and washing;
(5) drying: the ocean rhodotorula breast with W-Gum or yam starch and after concentrating is by weight 2: 1-3: 2 mix, are stirred to the physical condition homogeneous, making granularity by nodulizer is 10-60 purpose small-particle, by fluidized-bed in 70 ℃-90 ℃ wind-warm syndrome rapid drying 5-20 minute, get product then;
The culture medium prescription of fermentor tank in pure culture fermentor tank and the step (3) in the step (2) wherein, count by weight:
Glucose 2-5%
Magnesium sulfate heptahydrate 0.05-0.25%
Yeast extract 0.1-0.3%
Potassium primary phosphate 0.5-1%
Urea 0.1-0.3%
Ammonium sulfate 0.05-0.2%
Saltpetre 0.05-0.2%
PH value 4.0-6.0
The temperature of medium sterilization is 121 ℃, and the time is 30 minutes.
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| CN102919597A (en) * | 2012-11-08 | 2013-02-13 | 黑龙江省轻工科学研究院 | Production method of red-enhancing feed for ornamental red parrot fish |
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| CN103374530B (en) * | 2012-04-23 | 2016-03-30 | 安琪酵母股份有限公司 | A kind of Semi-dry yeast and preparation method thereof |
| CN102676408B (en) * | 2012-06-13 | 2016-03-23 | 北京大北农科技集团股份有限公司 | The method of ocean rhodotorula is produced in a kind of high density liquid submerged fermentation |
| CN103614307B (en) * | 2013-11-13 | 2016-01-20 | 中国水产科学研究院南海水产研究所 | A kind of solid ocean rhodotorula preparation and its preparation method and application |
| KR20160095049A (en) * | 2013-12-06 | 2016-08-10 | 디에스엠 아이피 어셋츠 비.브이. | Biomass formulation |
| CN105502679B (en) * | 2015-01-28 | 2019-02-05 | 大连玉兔岛海洋生物科技有限公司 | The preparation method of high-activity ocean yeast dry powder |
| CN104774900A (en) * | 2015-05-05 | 2015-07-15 | 昌邑市明兴饲料有限责任公司 | Technology of using ocean phaffiarodozyma for producing feed additive astaxanthin in fermentation mode |
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| CN1050975A (en) * | 1990-11-13 | 1991-05-01 | 沈阳市沈雪饲料加工厂 | Refined granular yeast fodder and preparation method |
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| CN102919597A (en) * | 2012-11-08 | 2013-02-13 | 黑龙江省轻工科学研究院 | Production method of red-enhancing feed for ornamental red parrot fish |
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