CN102864111B - Schizochytrium limacinum strain for producing docosahexaenoic acid - Google Patents
Schizochytrium limacinum strain for producing docosahexaenoic acid Download PDFInfo
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- CN102864111B CN102864111B CN 201210380900 CN201210380900A CN102864111B CN 102864111 B CN102864111 B CN 102864111B CN 201210380900 CN201210380900 CN 201210380900 CN 201210380900 A CN201210380900 A CN 201210380900A CN 102864111 B CN102864111 B CN 102864111B
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Abstract
The invention relates to a schizochytrium limacinum strain for producing docosahexaenoic acid and belongs to the technical field of bioengineering. The strain is named as slug type schizochytrium limacinum JN168 and preserved at China Center for Type Culture Collection, and the preserving number is CCTCCNO: M2012074. By performing separation screening on water samples of the Yellow Sea water area of Qingdao, Shandong Province, the schizochytrium limacinum strain which is used for producing docosahexaenoic acid (DHA) and is stable in characteristics is obtained, and high yield of docosahexaenoic acid can be achieved by liquid state or solid state fermentation at 25-30 DEG C.
Description
Technical field
The schizochytrium limacinum bacterial strain of docosahexenoic acid is produced in one strain, belongs to technical field of bioengineering.The present invention relates to a kind of schizochytrium limacinum bacterial strain of high yield docosahexenoic acid, this strain growth speed is fast, output is high, for the suitability for industrialized production of docosahexenoic acid significance is arranged.
Background technology
The English DHA that is called for short of docosahexenoic acid is the important a member in the n-3 series polyunsaturated fatty acid, and system is called all-cis formula △-4,7,10,13,16,19-docosahexenoic acid.Because contain the special construction of five unsaturated link(age)s, DHA has special effect and significant effects for the health of human body.DHA mainly is present in the pallium and retina of human body.Studies show that: aspects such as DHA is synthetic at prevention cerebral thrombosis, promotion baby's brain, infant's intelligence growth, anti-inflammatory, enhancing body immunity, anti-arrhythmia have vital role, existing scholar is devoted to the anticancer research of DHA at present in addition, report that DHA can cancer cell specific induction of apoptosis, in the process of cancer therapy drug research and development, have broad application prospects.Based on these vital role of docosahexenoic acid, it has been widely used in aspects such as food, medicine, feed.DHA mainly is distributed in the marine organisms, especially lives in the fish in the low marine site of water temperature.Human body can not be by self synthetic DHA, and can only derive by dietary supplementation or by alpha-linolenic acid gets, but because the human body carbochain prolongs the shortage of enzyme and desaturase, this conversion process is very slow and efficient is very low.Present industrialized DHA refines gained from fish oil, but the influence of from fish oil, extracting DHA and having many drawbacks for example fishy smell is heavy, cholesterol level is high, being subjected to weather and change in location, the content instability of DHA in the fish oil.DHA often coexists with EPA in the fish oil in addition, the sepn process complexity, and extraction cost is higher.These shortcomings have seriously hindered the industrialization development of DHA.Form and simply, do not contain fishy smell, extract advantages such as easy owing to utilize fermentative Production DHA to have content height, quality product and stable yield, lipid acid, enjoy Chinese scholars to pay close attention to.DHA extensively is present in bacterium, fungi and the little algae.Because to produce the DHA bacterium all be from the polar region or the waters, deep-sea is separated and obtained, it has a liking for high pressure, low temperature, and bacterium DHA yields poorly, and extracts difficulty, studies less.What research was more at present is fungi and algae, the Saprolegniales of fungi, Entomophthorales, mucorales, and the diatoms of little algae, chrysophyceae class, green algae, dinoflagellate class, Crypthecodinium cohnii class, blue algae all can fermentative production DHA.Schizochytrium limacinum
(Schizochytrium)Be to be under the jurisdiction of Mycophyta
(Eumycota), Oomycete
(Oomycetes), Saprolegniales
(Saprolegniales), thraustochytriale section
(Thraustochytriaceae)A class thalassiomycetes since its have growth rapidly, DHA content height, advantage such as to be easy to cultivate be to study the optimum strain of DHA fermentation at present.Use schizochytrium limacinum abroad
(Schizochytrium)The research of carrying out DHA has early just been used schizochytrium limacinum as far back as the omega biotech company of the U.S. in 1991
(Schizochytrium)Carry out the suitability for industrialized production of DHA.Japan Nagase Biochemicals (" the Optimization of docosahexaenoic acid productionby of company limited
SchizochytriumLimacinum SR21 ", Applied Microbiology and Biotechnology, 1998,49,72-76) by optimizing bacterial strain
Schizochytrium limacinumThe culture condition of SR21, final biomass reaches 59.2g/L, and DHA output reaches 15.5g/L.Domestic in recent years research institution and colleges and universities have also begun schizochytrium limacinum
(Schizochytrium)The research of fermentative production DHA, but yield and quality is compared with developed country big gap is still arranged.The Zhou.L(of Xiamen University " Enhanced production of docosahexaenoic acid using
SchizochytriumSp by optimization of medium components ", Journal of chemical engineering of Japan, 2007,40 (12), 1093-1100) pass through bacterial strain
SchizochytriumSp carries out the substratum compositional optimization, and the DHA ultimate capacity can reach 13.8g/L, (" schizochytrium limacinums such as the Zhang Juanmei of Fujian Normal University
SchizochytriumThe research of sp.FJU-512 cell grease ", 2007,23 (2), 75-80) utilize the Pollen Pini technology of fishing successfully to isolate a strain DHA high yield bacterium
SchizochytriumSp.FJU-512 adopts high-density culture in the 1000L fermentor tank, the DHA production peak can reach 5.42g/L.
Docosahexenoic acid (DHA) structural formula
Patent US6607900 discloses a kind of method that adopts two-step fermentation to produce DHA, adopts batchwise to cultivate
SchizochytriumSp, and in culturing process, keep constant sugared concentration by adding maize treacle.Make oxygen concn maintain high level in the cell enlargement stage, in the lipid accumulation stage oxygen concn is maintained low-levelly, final biomass can reach 200g/L, and DHA output is up to 40-45g/L.
Patent CN101892160A discloses that screening obtains a strain DHA superior strain schizochytrium limacinum LX0809 in the rotten soil of gathering from beach, Zhao Quan He Xiang Red sea, Panjin City Dawa County, Liaoning Province, by fed-batch fermentation after 100 hours, schizochytrium limacinum LX0809 dry cell weight can reach 72.5g/L, and the output of DHA reaches 15.3g/L.
Patent CN1916156A discloses a kind of schizochytrium limacinum WZU4771 and the application in preparation DHA powder and DHA grease thereof.Cultivated 5 days by optimizing the back, the thalline biomass reaches 42.5g/L, and DHA output is 14.1g/L.
But the present invention screens from Qingdao of Shandong province Huanghai Sea waters and to have obtained that proterties is stable, fermentation period is short, at the schizochytrium limacinum of 25-30 ℃ of high yield docosahexenoic acid
Schizochytrium limacinumJN168.Has good industrial application value.
Summary of the invention
The purpose of this invention is to provide a strain and produce the schizochytrium limacinum bacterial strain of docosahexenoic acid, it is stable to have obtained proterties, produces the schizochytrium limacinum bacterial strain of docosahexenoic acid, can produce docosahexenoic acid by solid-state or liquid state fermentation.
Technical scheme of the present invention: the schizochytrium limacinum bacterial strain of docosahexenoic acid is produced in a strain, classification called after slug type schizochytrium limacinum
Schizochytrium limacinumJN168 has been preserved in Chinese typical culture collection center, and preserving number is CCTCC NO:M2012074.
This bacterial strain main metabolites is docosahexenoic acid (DHA).Its 18S rDNA sequence is:
SEQ ID NO：1。
The available carbon source of this bacterial strain is one or more in glucose, fructose, maltose, sucrose, lactose, glycerine, the Zulkovsky starch.
The available nitrogenous source of this bacterial strain is one or more in inorganic ammonium salt, inorganic nitrate, yeast powder, yeast extract paste, peptone, the corn steep liquor.
Gather seawater sample in Qingdao of Shandong province Huanghai Sea waters, with the mode of filtration sterilization except degerming, to place the culture dish of the bacterium of having gone out in advance except the seawater sample after degerming, to wherein adding Pollen Pini through ultraviolet sterilization as bait, cultivate 3d for 25 ℃, with transfering loop dip in get enrichment halobiontic pollen carry out plate streaking, be placed on and cultivate in 25 ℃ the biochemical incubator until there being bacterium colony to produce, picking list bacterium colony carries out plate streaking repeatedly, until the pure growth that obtains color, form unanimity.Separate and with substratum be: glucose 20g, peptone 10g, yeast extract paste 5g, agar 20g, sea crystal 20g, distilled water 1000mL, penicillin 20mg, pH7.0.The pure growth that obtains is carried out shake-flask culture.The mycelium suction filtration of results shake-flask culture, lyophilize is to constant weight, grinds mycelium, extracts grease with cable-styled extraction method, to carry grease carry out the MS-GS coupling and analyze the lipid acid composition.Adopt above method repeatedly, through repeatedly screening the schizochytrium limacinum that obtains the docosahexenoic acid high yield
Schizochytrium limacinumJN168.The bacterial classification that the present invention relates to has following feature:
1. schizochytrium limacinum
Schizochytrium limacinumThe JN168 culture condition
This culture of strains normal condition has been that air culture is supported.The saccharine material that is used for the cultivation bacterial classification can be glucose, fructose, maltose, sucrose, lactose, glycerine, Zulkovsky starch.The nitrogenous source of cultivating usefulness can be inorganic ammonium salt, inorganic nitrate, yeast powder, yeast extract paste, peptone, corn steep liquor.The growth temperature range of this bacterial strain is 10 ℃-35 ℃, and the best accumulation of docosahexenoic acid temperature is 25 ℃, and bacterial strain can be grown in pH is the 2-9 scope and docosahexenoic acid is produced in metabolism, and the best accumulation of docosahexenoic acid pH is 6.0.
2. schizochytrium limacinum
Schizochytrium limacinumThe form of JN168 is described
The bacterial strain microscopic morphology feature of utilizing scanning electron microscope and perspective electron microscopic observation to arrive is as follows: vegetative cell ellipse, monokaryon, diameter 8-15cm, cell aggregation cluster on the liquid medium within, bacterium colony intermediate cell circle, the irregular cell of visible a little elongation in edge.Cell walls is made of scale closely.Form radial expolasm net from expolasm net form adult.To schizochytrium limacinum
Schizochytrium limacinumJN168 carries out continuous observed and recorded as can be known: be inoculated into the continuous bipartition of schizochytrium limacinum vegetative cell evolution on the fresh culture, and then nucleus division of cytokinesis forms tetraploid cell.Form the sporocyst of zoospore in telophase, the spore cyst wall begins to fluff then, discharges zoospore, and the spore oval has two laterally inserted flagellums.Former minister's flagellum has the tubulose hair, and every hair all has base, tubular shafts, terminal filament.Tubular shafts does not smoothly have lateral filament.
3. schizochytrium limacinum
Schizochytrium limacinumThe 18S rDNA molecular biological characteristics of JN168
See sequence table SEQ ID NO:1.
4. can adopt conventional liquid state fermentation method, utilize above-mentioned bacterial classification production.Substratum can be with containing ordinary culture mediums such as different carbon sources, nitrogenous source, mineral ion.
Also can add an amount of nutrition to the special needs of employed microorganism growth in the substratum, generalized case, these nutrition are present in the above-mentioned natural nutrition source.
This bacterial strain begins to produce docosahexenoic acid after 24 hours in the liquid state fermentation process, docosahexenoic acid output reaches maximum value after 72 hours.
Beneficial effect of the present invention: schizochytrium limacinum
Schizochytrium limacinumThe JN168 main metabolites is docosahexenoic acid, and docosahexenoic acid content reaches more than 50% less side products in the mycelia.
Also can add an amount of nutrition to the special needs of employed microorganism growth in the substratum, generalized case, these nutrition are present in the above-mentioned natural nutrition source.
The optimum carbon source that docosahexenoic acid is produced in this bacterial strain liquid state fermentation is glucose, and optimum nitrogen source is peptone.Can reach 23g/L with this substratum in the output of 25-30 ℃ of following docosahexenoic acid.
In this bacterial strain liquid state fermentation process, begin to produce docosahexenoic acid after 24 hours, docosahexenoic acid output reaches maximum after 72 hours.
The biological material specimens preservation: the schizochytrium limacinum bacterial strain of docosahexenoic acid is produced in a strain, classification called after slug type schizochytrium limacinum
Schizochytrium limacinumJN168, preservation and Chinese typical culture collection center, be called for short CCTCC, the address: Chinese Wuhan Wuhan University, preserving number is CCTCC NO:M2012074, preservation date is on 03 10th, 2012.
Embodiment
Embodiment 1
The schizochytrium limacinum fatty acid compositional analysis
Fermentation strain is schizochytrium limacinum
Schizochytrium limacinumJN168 adopts liquid state fermentation, and substratum consists of: glucose 20g, peptone 10g, yeast extract paste 5g, sea crystal 20g, distilled water 1000mL, pH7.0.25 ℃ of leavening temperatures, secondary fermentation in 5 days finishes.Suction filtration is collected mycelium, and lyophilize grinds mycelium with mortar to constant weight.Get the mycelium of drying about 0.1g, join in the test tube of lid.To the potassium hydroxide that wherein adds 0.4mol/L-methanol solution 5mL, in 50 ℃ of water-baths, keep 1h, again to wherein adding 5mL 14% boron trifluoride-methanol solution, in 50 ℃ of water-baths, keep 1h, add the extracting of 5mL normal hexane, add the washing of 2mL saturated nacl aqueous solution at last, pipette upper solution and adopt the GC-MS coupling to detect.
Chromatographic condition: the Gas Chromatography-mass Spectrometer (GCMS) Finnigan Trace MS(U.S.).Adopt DB-MAX capillary column (30m * 0.25mm * 0.25 μ m).180 ℃ of initial column temperatures keep 1min, are elevated to 230 ℃ with 5 ℃/min, keep 8min.260 ℃ of injector temperatures, carrier gas are helium, and flow velocity is 0.8ml/min, and splitting ratio is 10 ︰ 1.
The mass spectrometric detection condition: EI+(70Ev), transmitter current 200 μ A, 350 ℃ of ion source temperatures, 260 ℃ of interface temperature.
Table 1 lipid acid is formed
| Title | C16:0 | C18:0 | C18:1 | C18:2 | C20:4 | C20:5 | C22:5 | C22:6 | Other |
| Account for lipid acid proportion of composing (%) | 35.3 | 1.36 | 0.64 | 0.17 | 0.29 | 0.70 | 9.4 | 50.1 | 2.04 |
Embodiment 2
Determining of optimum carbon source
The bacterial classification that is kept at-70 ℃ is transferred on the flat board that contains an amount of seawater, adds the Pollen Pini of an amount of ultraviolet sterilization, cultivated 4 days for 25 ℃.Get the bacterium liquid after the 3mL activation, be linked into the 250mL that the 50mL seed culture medium is housed and shake in the bottle, 25 ℃, 200r/min shaking table were cultivated 2 days.With 4% inoculum size seed liquor is inoculated in the fermention medium, 25 ℃, 200r/min shaking table were cultivated 72 hours.Use single factor to investigate the optimum carbon source of docosahexenoic acid accumulation, the glucose that uses fructose, maltose, sucrose, lactose, glycerine, Zulkovsky starch to substitute in the basic fermention medium respectively carries out liquid state fermentation, measures dry weight and docosahexenoic acid output.Basic fermention medium is formed (w/v): glucose 90, yeast powder 10, sea crystal 20, pH7.0,115 ℃ of sterilization 20min.The optimum carbon source of strain fermentation product docosahexenoic acid is glucose as can be seen from Table 2.
The docosahexenoic acid condition of production under the different carbon sources of table 2
| Carbon source kind | Glucose | Fructose | Maltose | Sucrose | Lactose | Glycerine | Zulkovsky starch |
| Dry cell weight (g/l) | 59.4 | 50.0 | 48.7 | 36.7 | 35.5 | 29.8 | 37.3 |
| Output (g/l) | 13.50 | 11.23 | 11.08 | 8.11 | 8.56 | 7.84 | 9.30 |
Embodiment 3
Determining of optimum nitrogen source
The bacterial classification that is kept at-70 ℃ is transferred on the flat board that contains an amount of seawater, adds the Pollen Pini of an amount of ultraviolet sterilization, cultivated 4 days for 25 ℃.Get the bacterium liquid after 3mL activates, be linked into the 250mL that the 50mL seed culture medium is housed and shake in the bottle, 25 ℃, the 200r/min shaking table was cultivated 2 days.With 4% inoculum size seed liquor is inoculated in the fermention medium, 25 ℃, the 200r/min shaking table was cultivated 72 hours.Use single factor to investigate the optimum nitrogen source of docosahexenoic acid accumulation, the yeast powder that uses ammonium sulfate, ammonium nitrate, yeast extract paste, peptone, corn steep liquor to substitute in the basic fermention medium respectively carries out liquid state fermentation, measures dry weight and docosahexenoic acid output.Basic fermention medium is formed (w/v): glucose 90, yeast powder 10, sea crystal 20, pH7.0,115 ℃ of sterilization 20min.The optimum nitrogen source of strain fermentation product docosahexenoic acid is peptone as can be seen from Table 3.
The colored docosahexenoic acid condition of production under table 3 different nitrogen sources
| The nitrogenous source kind | Ammonium sulfate | Ammonium nitrate | Yeast powder | Yeast extract paste | Peptone | Corn steep liquor |
| Dry cell weight (g/l) | 38.3 | 24.5 | 58.5 | 59.0 | 67.7 | 49.0 |
| Output (g/l) | 8.75 | 5.69 | 13.42 | 12.73 | 15.71 | 10.63 |
Embodiment 4
Best pH determines
The bacterial classification that is kept at-70 ℃ is transferred on the flat board that contains an amount of seawater, adds the Pollen Pini of an amount of ultraviolet sterilization, cultivated 4 days for 25 ℃.Get the bacterium liquid after 3mL activates, be linked into the 250ml that the 50mL seed culture medium is housed and shake in the bottle, 25 ℃, the 200r/min shaking table was cultivated 2 days.With 4% inoculum size seed liquor is inoculated in the fermention medium, 25 ℃, the 200r/min shaking table was cultivated 72 hours.Use single factor to investigate the best pH of docosahexenoic acid accumulation, the pH that will ferment adjusts to 4.0,5.0,6.0,7.0,8.0,9.0 and measures dry weight and docosahexenoic acid output.Basic fermention medium is formed (w/v): glucose 90, peptone 10,20,115 ℃ of sterilizations of sea crystal 20min.The best pH of strain fermentation product docosahexenoic acid is 6.0 as can be seen from Table 4.
The docosahexenoic acid condition of production under the different pH values of table 4
| pH | 4.0 | 5.0 | 6.0 | 7.0 | 8.0 | 9.0 |
| Dry cell weight (g/l) | 48.9 | 59.5 | 70.4 | 68.4 | 53.4 | 38.2 |
| Output (g/l) | 9.77 | 13.15 | 16.54 | 15.63 | 12.55 | 8.60 |
Embodiment 5
Temperature is to the influence of docosahexenoic acid
The output of the docosahexenoic acid under table 4 differing temps
| Temperature ℃ | 15 | 20 | 25 | 30 | 35 |
| Dry cell weight g/L | 35.2 | 51.3 | 71.3 | 72.2 | 63.1 |
| Output (g/L) | 10.22 | 15.33 | 16.21 | 16.74 | 9.31 |
The bacterial classification that is kept at-70 ℃ is transferred on the flat board that contains an amount of seawater, adds the Pollen Pini of an amount of ultraviolet sterilization, cultivated 4 days for 30 ℃.Get the bacterium liquid after 3mL activates, be linked into the 250ml that the 50mL seed culture medium is housed and shake in the bottle, pH=7,30 ℃, the 200r/min shaking table was cultivated 2 days.With 4% inoculum size seed liquor is inoculated in the fermention medium, pH=7, the 200r/min shaking table was cultivated 72 hours.Use single factor to investigate the optimum temps of docosahexenoic acid accumulation, leavening temperature is adjusted to 15,20, measure dry weight and docosahexenoic acid output for 25,30,35 ℃.Basic fermention medium is formed (w/v): glucose 90, peptone 10,20,115 ℃ of sterilizations of sea crystal 20min.Bacterial strain is best in the situation of 25-30 ℃ of fermentation product docosahexenoic acid as can be seen from Table 5.
Embodiment 6
The fermentor tank amplification test
With the 5L fermentor tank schizochytrium limacinum is carried out amplification test.Get the preservation inclined-plane of schizochytrium limacinum, add the 5mL sterilized water, scrape gently and get lawn, draw 2.5mL bacterium liquid and join the 250mL that the 50ml seed culture medium is housed and shake in the bottle, 25 ℃, 200r/min are cultivated 8h, the bacterium liquid of getting after 10mL activates joins the 500mL shake-flask culture 32h that 100mL fresh seeds substratum is housed, seed liquor and fermention medium are formed (w/v): glucose 90, peptone 10, sea crystal 20, pH6.0,115 ℃ of sterilization 20min.With 1% inoculum size seed liquor is inoculated in the 5L fermentor tank.Tank pressure: 0.55kg/m
3, air flow: 4L/min.With speed adjustment dissolved oxygen saturation ratio 40%, control dissolved oxygen saturation ratio is 25% after the fermentation to 48 hour, use 2mol/L HCL or 2mol/L NaOH to regulate the pH value stabilization 6.0, when glucose concn in the fermented liquid drops to about 15g/L, begin stream and add the glucose solution of 90g/L, glucose concn is maintained about 15g/L, and fermenting stopped after 72 hours.Biomass reaches 100g/L, and docosahexenoic acid output is up to 22.9g/L.
Embodiment 7
Adopt solid state fermentation, substratum is formed (weight %): 30 purpose Semen Maydis powder, 35,30 purpose wheat-flours 35, wheat bran 8.7, rice bran 20, glucose 1, KH
2PO
40.25, MgSO
47H
2O 0.05; It is wetting to add the water spice, in the wide-necked bottle of packing into, and the 0.1MPa 45-60min that sterilizes, the cooling back is inserted liquid seeds and is mixed all.30 ℃ of leavening temperatures, pH7, after 7 days fermentation ends, the content of docosahexenoic acid is 7.3% in the fermention medium.
＜110〉Southern Yangtze University
＜120〉the schizochytrium limacinum bacterial strain of docosahexenoic acid is produced in a strain
<160>1
<210>1
<211>1683
<212>DNA
＜213〉schizochytrium limacinum (
Schizochytriumlimacinum) JN168
<400>1
ccaacctggt tgatcctgcc ccagctccag tgctcgtctc aaagattaag ccatgcatgt 60
gtaagtataa gcgattgtac tgtgagactg cgaccggctt attatatcag taataatttc 120
ttcggtagtt tcttttatat ggatacctgc agtaattctg gaaataatac atgctgtaag 180
agccctgtat ggggctgcac ggtcgatgaa gaagccgatt ttattggtga atcatgataa 240
ttgagcagat tgactatttt tcgtttgagt ttctgcccca tcagttgtcg ttattagatt 300
acggtagtgt attggactac ggtgactata acgggtgacg gagagttagg gctcgactcc 360
ggagagggag aggtcgtatg ggctaccata tccaaggata gcagcaggcg cgtaaattac 420
ccactgtgga ctccacgagg tagtgacgag aaatttcggt gcgaagcgtg tatgcgtttt 480
gctatcggaa tgagagcaat gtaaaaccct catcgaggat caactggagg gcaagtctgg 540
tgccagcagc cgcggtaatt aagcatatgc taaagttgtt gcagttaaaa agtagtcata 600
agctcgtagt tgaatttctg gcatgggcga ccggtgcttt ccctgaatgg ggattgattg 660
tctgtgttgc cttggccatc tttctcatgc tgttattggt atgagatctt tcactgtaat 720
caaagcagag tgttccaagc accggtattt ttattatggg atgataagat cctgagagac 780
aggacttggg tgctattttg ttggtttgcc cgcctgagta atggttaata ggaacagttg 840
ggggtattcg tatttaggag ctagaggtga aattcttgga tttccgaaag acgaactaga 900
gcgaaggcat ttaccaagca tgttttcatt aatcaagaac gaaagtctgg ggatcgaaga 960
tgattagata ccatcgtagt ctagaccgta aacgatgccg acttgcgatt gttgggtgct 1020
tttttatggg cctcagcagc agcacatgag aaatcaaagt ctttgggttc cggggggagt 1080
atggtcgcaa ggctgaaact taaaggaatt gacggaaggg caccaccagg agtggagcct 1140
gcggcttaat ctgtgatgcc accgaaaaac ttaccaggtc cagacatagg taggattgac 1200
aaattgagag ctctttcatg attctatggg tggtggtgca tggccgttct tagttggtgg 1260
agtgatttgt ctggttaatt ccgttaacga acgggacctc ggcctactaa atagtgcgtg 1320
gtatggcaac atagtacgtt tttaacttct tagagggaca tgtccggttt acgggcagga 1380
agttcgaggc tataacaggt cttagatgtt ctgggccgca cgcgcgctac ttgactcaac 1440
actgatgggt tcatcgggtt ttatttctgt ttttatggaa ttgagtgctt ggtcggaagg 1500
cctggctaat ccttggaacg ctcatcgtgc tggggctaga tttttgcaat tattaatctc 1560
caacgaggaa ttcctagtaa acgcaagtca tcagcttgca ttgaatacgt ccctgccctt 1620
tgtacacacc gcccgtcgca cctaccgatt gaacggtccg atgaaaccat gggatgtttc 1680
tgt 1683
Claims (2)
1. the schizochytrium limacinum bacterial strain of docosahexenoic acid DHA is produced in a strain, its classification called after slug type schizochytrium limacinum (
Schizochytrium limacinum) JN168, being preserved in Chinese typical culture collection center, preserving number is CCTCC NO:M2012074.
2. the application of the described schizochytrium limacinum bacterial strain of claim 1, it is characterized in that described bacterial strain is used for fermentation and produces docosahexenoic acid, fermentation is in glucose, fructose, maltose, sucrose, lactose, glycerine, the Zulkovsky starch one or more with carbon source, and fermenting with nitrogenous source is in inorganic ammonium salt, inorganic nitrate, yeast powder, yeast extract paste, peptone, the corn steep liquor one or more.
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| CN103602591B (en) * | 2013-10-17 | 2015-08-26 | 华中科技大学 | A kind of schizochytrium limacinum and the method for the production of docosahexaenoic acid grease |
| CN103911292B (en) * | 2013-12-05 | 2016-04-06 | 中国科学院天津工业生物技术研究所 | The schizochytrium limacinum of one strain tolerance less salt and application thereof |
| CN104031843B (en) * | 2014-05-14 | 2016-03-30 | 中国科学院青岛生物能源与过程研究所 | A kind of schizochytrium limacinum and application thereof |
| CN103981106B (en) * | 2014-06-03 | 2016-08-24 | 上海来益生物药物研究开发中心有限责任公司 | DHA superior strain and application thereof |
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