CN104845893B - One plant of rhodotorula mucilaginosa and its application in the red astaxanthin in fermenting and producing sea - Google Patents
One plant of rhodotorula mucilaginosa and its application in the red astaxanthin in fermenting and producing sea Download PDFInfo
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
Abstract
This application discloses one plant of rhodotorula mucilaginosa (Rhodotorula mucilaginosa), deposit number：CGMCC No.10123, and its application in the red astaxanthin in fermenting and producing sea, are realized by following steps：Rhodotorula mucilaginosa seed liquor is cultivated, 2.5 4L nutrient solution is added into fermentation tank, nutrient solution includes：The 120g/L of one kind 6 in glucose, sucrose, fructose, ethanol or glycerine, the 5g/L of dusty yeast 1, the 0.5g/L of potassium dihydrogen phosphate 0.1, the 0.5g/L of dipotassium hydrogen phosphate 0.1, the 5g/L of urea 2, the 9g/L of sodium chloride 2, medium pH value is between 5.5 7.5, the rhodotorula mucilaginosa seed liquor of access 2 10% into the nutrient solution, fermentation tank rotating speed is 160 300r/min, is fermented 24 days within the temperature range of 28 33 DEG C.Using the rhodotorula mucilaginosa in the present invention, the yield of the red astaxanthin in sea is high.
Description
Technical field
The present invention relates to saccharomycete technical field, is to be related to one plant of rhodotorula mucilaginosa and its in fermenting and producing sea specifically
Application in red astaxanthin.
Background technology
The red astaxanthin in sea is the nutrition pigment and antioxidant of a kind of great exploitation potential, have strong anti-oxidation, anti-aging,
Many important biological functions such as antiviral, anti-body coloring, anticancer, in food, medicine, cosmetics, aquaculture, livestock and poultry
The industries such as cultivation, chemical industry have broad application prospects.The red astaxanthin in sea, a kind of carotenoid with high anti-oxidation ability,
Its mobility approaches with natural astaxanthin ester during thin-layer chromatography, and oxidation resistance is also close with natural left-handed astaxanthin and gains the name,
DPPH methods detect its oxidation resistance and are better than natural left-handed astaxanthin 20~40% or so.
The preparation method of astaxanthin, it is known to chemical synthesis, method, the profit extracted from the extraction of aquatic products waste or algae
Method produced with microorganism etc..Chemical synthesis, production cost is higher, and many countries do not allow to use synthesizing astaxanthin
For in terms of feed addictive；Extracted from aquatic products waste, raw material sources are extensive, cost is low, but production is easily subject to seasonal restrictions, and produce
Condition requires harsh, and production cost is high, and yield is relatively low, and product purity is not high；Extracted from algae, the content astaxanthin in algae
Higher, cost of material is relatively low.But the growth conditions of algae is harsh, and the requirement to water quality, environment and light is very high, mass produce according to
So difficulty be present；The method produced using microorganism, using saccharine material fermenting and producing, given birth to beneficial to large scale and high density
Production, and do not limited by region and weather, production cost relative moderate.
In recent years, it is increasing to the demand of astaxanthin both at home and abroad, there are wide market prospects.At present the U.S., plus
Put on airs, many biotech companies of the state such as European Union are just being directed to Development and Production this kind of product, but far from meeting the market demand,
Therefore price is higher.Therefore step up the development to astaxanthin product, for enhancing our international competitiveness, there is highly important meaning
Justice.Because microbial reproduction speed is fast, growth cycle is short, and zymotechnique is also more ripe, is easy to grasp, and yeast is inherently
A kind of good health material, nutritive value is higher, therefore is currently ideal side using yeast fermenting and producing astaxanthin
Method, it may have broader market prospects.
The content of the invention
In order to solve the above technical problems, the invention provides one plant of rhodotorula mucilaginosa (Rhodotorula
Mucilaginosa), deposit number：CGMCC No.10123.
The invention also discloses application of the rhodotorula mucilaginosa in the red astaxanthin in fermenting and producing sea.
Application of the rhodotorula mucilaginosa in the red astaxanthin in fermenting and producing sea, is realized by following steps：Cultivate rhodotorula mucilaginosa
Seed liquor, 2.5-4L nutrient solution is added into fermentation tank, and nutrient solution includes：In glucose, sucrose, fructose, ethanol or glycerine
A kind of 6-120g/L, dusty yeast 1-5g/L, potassium dihydrogen phosphate 0.1-0.5g/L, dipotassium hydrogen phosphate 0.1-0.5g/L, urea 2-
5g/L, sodium chloride 2-9g/L, medium pH value access 2-10% rhodotorula mucilaginosa into the nutrient solution between 5.5-7.5
Seed liquor, fermentation tank rotating speed are 160-300r/min, are fermented 2-4 days within the temperature range of 28-33 DEG C；
The culture rhodotorula mucilaginosa seed liquor is realized by following steps：Rhodotorula mucilaginosa thalline one is provoked from test tube slant
Ring is inoculated in the triangular flask equipped with 50mL seed culture mediums, the shaking table culture 22- under the conditions of 25-33 DEG C, 150-300r/min
24h obtains primary seed solution, takes seed liquor 5mL to be inoculated in the triangular flask equipped with 100mL seed culture mediums, 25-33 DEG C, 150-
300r/min condition shaking table cultures 22-24h obtains secondary seed solution, and the secondary seed solution is the rhodotorula mucilaginosa seed liquor.
Preferably, the seed culture medium includes：Glucose 6-48g/L, dusty yeast 1-5g/L, urea 2-5g/L, chlorine
Change sodium 2-9g/L, pH value 5.5-7.5.
The Rhodotorula mucilaginose (Rhodotorula mucilaginosa) of the present invention, is preserved in Chinese microorganism strain preservation
Administration committee's common micro-organisms center, preservation address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postcode：100101, protect
Hide the date：On December 01st, 2014, deposit number：CGMCCNo.10123.
The 18S rRNA gene orders of the bacterial strain separated to the present invention are sequenced, by the 18SrRNA gene sequences of acquisition
All sequences in database are compared analysis by row input GenBank by Blastn programs, as a result find the present invention's
The existing type strain of 18S rRNA gene orders and rhodotorula mucilaginosa category has higher similitude, such as table 1, wherein with
Rhodotorula mucilaginosa isolate ZH9 sequence similarity be 100%, and with the sequence of more plants of rhodotorula mucilaginosas
Row similitude is 98%, so that it is determined that the bacterial strain GH101 separated is the new strains system of rhodotorula mucilaginosa category, and it is red to be named as glue
Yeast (Rhodotorula mucilaginosa) GH101.
18S rRNA amplifications are carried out, sequencing sequence is：
CATCCGTATTGTTCAAATTTTAAGCATAACAGTGAAACTGCGAATGGCTCATTAAATCAGTCATAGTTTATTTGATG
GTACCTTACTACATGGATAACTGTGGTAATTCTAGAGCTAATACATGCTGAAAAATCCCGACTTCTGGAAGGGATGT
ATTTATTAGATCCAAAACCAATGGCCTTCGGGTCCCTATGGTGAATCATGATAACTGCTCGAATCGCATGGCCTTGC
GCCGGCGATGCTTCATTCAAATATCTGCCCTATCAACTTTCGATGGTAGGATAGAGGCCTACCATGGTGATGACGGG
TAACGGGGAATAAGGGTTCGATTCCGGAGAGAGGGCCTGAGAAACGGCCCTCAGGTCTAAGGACACGCAGCAGGCGC
GCAAATTATCCCCTGGCAACACTTTGCCGAGATAGTGACAATAAATAACAATGCAGGGCTCTTACGGGTCTTGCAAT
TGGAATGAGTACAATTTAAATCCCTTAACGAGGATCAATTGGAGGGCAAGTCTGGTGCCAGCAGCCGCGGTAATTCC
AGCTCCAATAGCGTATATTAAAATTGTTGCCGTTAAAAAGCTCGTAGTCGAACTTCGGGCTCTGTCAGCCGGTCCGC
CTTCTTGGTGTGTACTTGTTTGACGGAGCCTTACCTCCTGGTGAACGGCGATGTCCTTTACTGGGTGTCGTCGCAAA
CCAGGACTTTTACTTTGAAAAAATTAGAGTGTTCAAAGCAGGCCTTTGCCCGAATACATTAGCATGGAATAATAAAA
TAGGACGCGCGTTCCCATTTTGTTGGTTTCTGAGATCGCCGTAATGATTAATAGGGATAGTTGGGGGCATTTGTATT
CCGTCGTCAGAGGTGAAATTCTTGGATTGCCGGAAGACAAACTACTGCGAAAGCATTTGCCAAGGATGTTTTCATTG
ATCAAGAACGAAGGAAGGGGGATCGAAAACGATTAGATACCGTTGTAGTCTCTTCTGTAAACTATGCCAATTGGGGA
TCGGTACAGGATTTTTAATGACTGTATCGGCACCCGAAGAGAAATCTTTAAATGAGGTTCGGGGGGGAGTATGGTCG
CAAGGCTGAAACTTAAAGGAATTGACGGAAGGGCACCACCAGGTGTGGAGCCTGCGGCTTAATTTGACTCAACACGG
GGAAACTCACCAGGTCCAGACACAATAAGGATTGACAGATTGATAGCTCTTTCTTGATCTTGTGGTTGGTGGTGCAT
GGCCGTTCTTAGTTGGTGGAGTGATTTGTCTGGTTAATTCCGATAACGAACGAG.By Blast sequence alignments
The gene order of table 1 compares
Compared with prior art, application of the rhodotorula mucilaginosa of the present invention in the red astaxanthin in fermenting and producing sea, is utilized
Rhodotorula mucilaginosa in the present invention, the yield of the red astaxanthin in sea is high, and compared to working as fermentation medium in prior art
When middle addition 5g/L dusty yeast and 2g/L urea are as fermentation nitrogen source, the yield highest of the red astaxanthin in sea.
Brief description of the drawings
Accompanying drawing described herein is used for providing a further understanding of the present invention, forms the part of the present invention, this hair
Bright schematic description and description is used to explain the present invention, does not form inappropriate limitation of the present invention.In the accompanying drawings：
Fig. 1 is influence of the different carbon source to the red astaxanthin yield in sea；
Fig. 2 is influence of the different concentration of glucose to the red astaxanthin yield in sea；
Fig. 3 is the flow chart using the red astaxanthin in rhodotorula mucilaginosa fermenting and producing sea of the present invention.
Embodiment
Some vocabulary has such as been used to censure specific components among specification and claim.Those skilled in the art should
It is understood that hardware manufacturer may call same component with different nouns.This specification and claims are not with name
The difference of title is used as the mode for distinguishing component, but is used as the criterion of differentiation with the difference of component functionally.Such as logical
The "comprising" of piece specification and claim mentioned in is an open language, therefore should be construed to " include but do not limit
In "." substantially " refer in receivable error range, those skilled in the art can be described within a certain error range solution
Technical problem, basically reach the technique effect.In addition, " coupling " one word is herein comprising any direct and indirect electric property coupling
Means.Therefore, if the first device of described in the text one is coupled to a second device, representing the first device can directly electrical coupling
The second device is connected to, or the second device is electrically coupled to indirectly by other devices or coupling means.Specification
Subsequent descriptions for implement the present invention better embodiment, so it is described description be by illustrate the present invention rule for the purpose of,
It is not limited to the scope of the present invention.Protection scope of the present invention is worked as to be defined depending on appended claims institute defender.
The present invention is described in further detail below in conjunction with accompanying drawing, but it is not as a limitation of the invention.
Embodiment 1：
Cultivate rhodotorula mucilaginosa seed liquor：The ring of rhodotorula mucilaginosa thalline one is provoked from test tube slant to be inoculated in equipped with 50mL seeds
In the 250mL triangular flasks of culture medium, 25 DEG C, 150r/min condition shaking table cultures 22h obtain primary seed solution, with this seed liquor 5mL
Be inoculated in the 500mL triangular flasks equipped with 100mL seed culture mediums, 25 DEG C, 150r/min condition shaking table cultures 22h obtain two level kind
Sub- liquid, the secondary seed solution are the rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 6g/L, dusty yeast 1g/L, sodium chloride 2g/L, pH value 5.5.
2.5L nutrient solution is added into 5L fermentation tanks, nutrient solution includes：Glucose 6g/L, dusty yeast 1g/L, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.1g/L, dipotassium hydrogen phosphate 0.1g/L, urea 2g/L, sodium chloride 2g/L, medium pH value 5.5, into the nutrient solution
The rhodotorula mucilaginosa seed liquor of access 2%, fermentation tank rotating speed are 160r/min, and 28 DEG C ferment 2 days.
Embodiment 2：
Cultivate rhodotorula mucilaginosa seed liquor：The ring of rhodotorula mucilaginosa thalline one is provoked from test tube slant to be inoculated in equipped with 50mL seeds
In the 250mL triangular flasks of culture medium, 33 DEG C, 300r/min condition shaking table cultures 24h obtain primary seed solution, with this seed liquor 5mL
Be inoculated in the 500mL triangular flasks equipped with 100mL seed culture mediums, 33 DEG C, 300r/min condition shaking table cultures 24h obtain two level kind
Sub- liquid, the secondary seed solution are the rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 48g/L, dusty yeast 8g/L, sodium chloride 9g/L, pH value 7.5.
4L nutrient solution is added into 5L fermentation tanks, nutrient solution includes：Glucose 120g/L, dusty yeast 5g/L, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, urea 5g/L, sodium chloride 9g/L, medium pH value 7.5, into the nutrient solution
The rhodotorula mucilaginosa seed liquor of access 10%, fermentation tank rotating speed are 300r/min, and 33 DEG C ferment 4 days.
Embodiment 3：
Cultivate rhodotorula mucilaginosa seed liquor：The ring of rhodotorula mucilaginosa thalline one is provoked from test tube slant to be inoculated in equipped with 50mL seeds
In the 250mL triangular flasks of culture medium, 30 DEG C, 180r/min condition shaking table cultures 24h obtain primary seed solution, with this seed liquor 5mL
Be inoculated in the 500mL triangular flasks equipped with 100mL seed culture mediums, 30 DEG C, 180r/min condition shaking table cultures 24h obtain two level kind
Sub- liquid, the secondary seed solution are the rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 27g/L, dusty yeast 4g/L, sodium chloride 5g/L, pH value 6.5.
4L nutrient solution is added into 5L fermentation tanks, nutrient solution includes：Glucose 73g/L, dusty yeast 3g/L, biphosphate
Potassium 0.3g/L, dipotassium hydrogen phosphate 0.3g/L, urea 3.5g/L, sodium chloride 5g/L, medium pH value 6.5, into the nutrient solution
The rhodotorula mucilaginosa seed liquor of access 6%, fermentation tank rotating speed are 230r/min, and 33 DEG C ferment 3 days.
Embodiment 4：
Cultivate rhodotorula mucilaginosa seed liquor：The ring of rhodotorula mucilaginosa thalline one is provoked from test tube slant to be inoculated in equipped with 50mL seeds
In the 250mL triangular flasks of culture medium, 28 DEG C, 240r/min condition shaking table cultures 24h obtain primary seed solution, with this seed liquor 5mL
Be inoculated in the 500mL triangular flasks equipped with 100mL seed culture mediums, 28 DEG C, 240r/min condition shaking table cultures 24h obtain two level kind
Sub- liquid, the secondary seed solution are the rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 27g/L, dusty yeast 4g/L, sodium chloride 5g/L, pH value 6.5.
3.5L nutrient solution is added into 5L fermentation tanks, nutrient solution includes：Glucose 70g/L, dusty yeast 5g/L, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, urea 2g/L, sodium chloride 2g/L, medium pH value 6.5, into the nutrient solution
The rhodotorula mucilaginosa seed liquor of access 3%, fermentation tank rotating speed are 250r/min, and 30 DEG C ferment 3 days.
Dry cell weight 18g/L, the red astaxanthin yield 216mg/L in sea are obtained in the present embodiment.
Embodiment 5：
Cultivate rhodotorula mucilaginosa seed liquor：The ring of rhodotorula mucilaginosa thalline one is provoked from test tube slant to be inoculated in equipped with 50mL seeds
In the 250mL triangular flasks of culture medium, 28 DEG C, 240r/min condition shaking table cultures 24h obtain primary seed solution, with this seed liquor 5mL
Be inoculated in the 500mL triangular flasks equipped with 100mL seed culture mediums, 28 DEG C, 240r/min condition shaking table cultures 24h obtain two level kind
Sub- liquid, the secondary seed solution are the rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, dusty yeast 3.7g/L, sodium chloride 4.5g/L, pH value 6.2.
3.5L nutrient solution is added into 5L fermentation tanks, nutrient solution includes：Glucose 42g/L, dusty yeast 4g/L, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium chloride 4g/L, medium pH value 6, connects into the nutrient solution
Enter 3% rhodotorula mucilaginosa seed liquor, fermentation tank rotating speed is 220r/min, and 30 DEG C ferment 3 days.Using saccharine material fermentation Hai Hong
During astaxanthin, the presence of saccharic carbon source is the necessary condition that growing microorganism and product effectively accumulate in culture medium.
Embodiment 6：
Cultivate rhodotorula mucilaginosa seed liquor：The ring of rhodotorula mucilaginosa thalline one is provoked from test tube slant to be inoculated in equipped with 50mL seeds
In the 250mL triangular flasks of culture medium, 28 DEG C, 180r/min condition shaking table cultures 24h obtain primary seed solution, with this seed liquor 5mL
Be inoculated in the 500mL triangular flasks equipped with 100mL seed culture mediums, 28 DEG C, 180r/min condition shaking table cultures 24h obtain two level kind
Sub- liquid, the secondary seed solution are the rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, dusty yeast 3.7g/L, sodium chloride 4.5g/L, pH value 6.2.
3.5L nutrient solution is added into 5L fermentation tanks, nutrient solution includes：Glucose 50g/L, dusty yeast 4g/L, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium chloride 4g/L, medium pH value 6, connects into the nutrient solution
Enter 3% rhodotorula mucilaginosa seed liquor, fermentation tank rotating speed is 220r/min, and 30 DEG C ferment 3 days.
Embodiment 7
Cultivate rhodotorula mucilaginosa seed liquor：The ring of rhodotorula mucilaginosa thalline one is provoked from test tube slant to be inoculated in equipped with 50mL seeds
In the 250mL triangular flasks of culture medium, 28 DEG C, 180r/min condition shaking table cultures 24h obtain primary seed solution, with this seed liquor 5mL
Be inoculated in the 500mL triangular flasks equipped with 100mL seed culture mediums, 28 DEG C, 180r/min condition shaking table cultures 24h obtain two level kind
Sub- liquid, the secondary seed solution are the rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, dusty yeast 3.7g/L, sodium chloride 4.5g/L, pH value 6.2.
3.5L nutrient solution is added into 5L fermentation tanks, nutrient solution includes：Sucrose 50g/L, dusty yeast 4g/L, biphosphate
Potassium 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium chloride 4g/L, medium pH value 6, accessed into the nutrient solution
3% rhodotorula mucilaginosa seed liquor, fermentation tank rotating speed are 220r/min, and 30 DEG C ferment 3 days.
Embodiment 8
Cultivate rhodotorula mucilaginosa seed liquor：The ring of rhodotorula mucilaginosa thalline one is provoked from test tube slant to be inoculated in equipped with 50mL seeds
In the 250mL triangular flasks of culture medium, 28 DEG C, 180r/min condition shaking table cultures 24h obtain primary seed solution, with this seed liquor 5mL
Be inoculated in the 500mL triangular flasks equipped with 100mL seed culture mediums, 28 DEG C, 180r/min condition shaking table cultures 24h obtain two level kind
Sub- liquid, the secondary seed solution are the rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, dusty yeast 3.7g/L, sodium chloride 4.5g/L, pH value 6.2.
3.5L nutrient solution is added into 5L fermentation tanks, nutrient solution includes：Fructose 50g/L, dusty yeast 4g/L, biphosphate
Potassium 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium chloride 4g/L, medium pH value 6, accessed into the nutrient solution
3% rhodotorula mucilaginosa seed liquor, fermentation tank rotating speed are 220r/min, and 30 DEG C ferment 3 days.
Embodiment 9
Cultivate rhodotorula mucilaginosa seed liquor：The ring of rhodotorula mucilaginosa thalline one is provoked from test tube slant to be inoculated in equipped with 50mL seeds
In the 250mL triangular flasks of culture medium, 28 DEG C, 180r/min condition shaking table cultures 24h obtain primary seed solution, with this seed liquor 5mL
Be inoculated in the 500mL triangular flasks equipped with 100mL seed culture mediums, 28 DEG C, 180r/min condition shaking table cultures 24h obtain two level kind
Sub- liquid, the secondary seed solution are the rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, dusty yeast 3.7g/L, sodium chloride 4.5g/L, pH value 6.2.
3.5L nutrient solution is added into 5L fermentation tanks, nutrient solution includes：Ethanol 50g/L, dusty yeast 4g/L, biphosphate
Potassium 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium chloride 4g/L, medium pH value 6, accessed into the nutrient solution
3% rhodotorula mucilaginosa seed liquor, fermentation tank rotating speed are 220r/min, and 30 DEG C ferment 3 days.
Embodiment 10
Cultivate rhodotorula mucilaginosa seed liquor：The ring of rhodotorula mucilaginosa thalline one is provoked from test tube slant to be inoculated in equipped with 50mL seeds
In the 250mL triangular flasks of culture medium, 28 DEG C, 180r/min condition shaking table cultures 24h obtain primary seed solution, with this seed liquor 5mL
Be inoculated in the 500mL triangular flasks equipped with 100mL seed culture mediums, 28 DEG C, 180r/min condition shaking table cultures 24h obtain two level kind
Sub- liquid, the secondary seed solution are the rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, dusty yeast 3.7g/L, sodium chloride 4.5g/L, pH value 6.2.
3.5L nutrient solution is added into 5L fermentation tanks, nutrient solution includes：Glycerine 50g/L, dusty yeast 4g/L, biphosphate
Potassium 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium chloride 4g/L, medium pH value 6, accessed into the nutrient solution
3% rhodotorula mucilaginosa seed liquor, fermentation tank rotating speed are 220r/min, and 30 DEG C ferment 3 days.
As can be seen from Figure 1, in different carbon source, sucrose and the advantageous synthesis in astaxanthin of glucose, wherein glucose
It is most favourable, the red astaxanthin yield highest in sea.
Embodiment 11
On the basis of embodiment 6, the implementation case illustrates influence of the different concentration of glucose to the red astaxanthin yield in sea,
Add the grape that concentration is 0g/L, 40g/L, 50g/L, 60g/L, 70g/L, 80g/L and 90g/L respectively in the fermentation medium
Sugar, fermentation temperature are 30 DEG C, rotating speed 180r/min, and ferment 3d.The accumulation of the red astaxanthin in sea is with the increasing of concentration of glucose
Add and be proportionate, but the increase of later stage addition, the yield increase of astaxanthin are little.
Embodiment 12
On the basis of embodiment 6, fermented, the fermentation condition all same in addition to nitrogen source, the implementation case is said
Bright different nitrogen sources produce the influence of the red astaxanthin yield in sea to rhodotorula mucilaginosa, and nitrogen source is that protein and nucleic acid etc. contain in synthesis thalline
The main source of nitrogen material, therefore its importance is only second to carbon source in microbial cultivation process.Fermented in rhodotorula mucilaginosa and produce sea
During red astaxanthin, dusty yeast and urea are to be beneficial to the organic nitrogen source of the red astaxanthin accumulation in sea and inorganic nitrogen-sourced respectively, in order to
Ensure in whole fermentation process, the offer nitrogen source that can continue, this experiment have chosen the inorganic nitrogen-sourced and organic nitrogen of various concentrations
The mixture in source have studied its influence to the red astaxanthin yield in sea as fermentation nitrogen source.
As a result as shown in Table 1, when addition 5g/L dusty yeast in fermentation medium and 2g/L urea are as fermentation nitrogen source
When, the yield highest of the red astaxanthin in sea.
Table 1：Different organic nitrogen sources are with inorganic nitrogen-sourced with the influence for comparing the red astaxanthin yield in sea
| Nitrogen source species and concentration/(g/L) | The red astaxanthin yield (%) in sea | Thalline yield (%) |
| Dusty yeast 7 | 0.66 | 11.3 |
| Dusty yeast 6+ urea 1 | 1.06 | 14.8 |
| Dusty yeast 5+ urea 2 | 1.14 | 18.25 |
| Dusty yeast 4+ urea 3 | 1.09 | 16.7 |
| Dusty yeast 3+ urea 4 | 0.92 | 14.1 |
| Dusty yeast 2+ urea 5 | 0.75 | 10.9 |
| Urea 7 | 0.38 | 9.6 |
Some preferred embodiments of the present invention have shown and described in described above, but as previously described, it should be understood that the present invention
Be not limited to form disclosed herein, be not to be taken as the exclusion to other embodiment, and available for various other combinations,
Modification and environment, and above-mentioned teaching or the technology or knowledge of association area can be passed through in the scope of the invention is set forth herein
It is modified., then all should be in this hair and the change and change that those skilled in the art are carried out do not depart from the spirit and scope of the present invention
In the protection domain of bright appended claims.
Claims (2)
1. one plant of rhodotorula mucilaginosa (Rhodotorula mucilaginosa) application in the red astaxanthin in fermenting and producing sea, it is special
Sign is, the rhodotorula mucilaginosa deposit number：CGMCC No.10123, are realized by following steps：Rhodotorula mucilaginosa seed liquor is cultivated,
2.5-4L nutrient solution is added into fermentation tank, nutrient solution includes：Glucose 70g/L, dusty yeast 5g/L, potassium dihydrogen phosphate 0.1-
0.5g/L, dipotassium hydrogen phosphate 0.1-0.5g/L, urea 2g/L, sodium chloride 2-9g/L, medium pH value between 5.5-7.5, to
2-10% rhodotorula mucilaginosa seed liquor is accessed in the nutrient solution, fermentation tank rotating speed is 160-300r/min, in 28-33 DEG C of temperature
Fermented 2-4 days in the range of degree；
The culture rhodotorula mucilaginosa seed liquor is realized by following steps：The ring of rhodotorula mucilaginosa thalline one is provoked from test tube slant to connect
Kind is in the triangular flask equipped with 50mL seed culture mediums, and shaking table culture 22-24h is obtained under the conditions of 25-33 DEG C, 150-300r/min
Primary seed solution, seed liquor 5mL is taken to be inoculated in the triangular flask equipped with 100mL seed culture mediums, 25-33 DEG C, 150-300r/
Min condition shaking table cultures 22-24h obtains secondary seed solution, and the secondary seed solution is the rhodotorula mucilaginosa seed liquor.
2. application of the rhodotorula mucilaginosa according to claim 1 in the red astaxanthin in fermenting and producing sea, it is characterised in that described
Seed culture medium includes：Glucose 6-48g/L, dusty yeast 1-5g/L, urea 2-5g/L, sodium chloride 2-9g/L, pH value 5.5-
7.5。
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