CN104845893A - Rhodotorula mucilaginosa and application thereof to fermentation production of malus micromalus astaxanthin - Google Patents
Rhodotorula mucilaginosa and application thereof to fermentation production of malus micromalus astaxanthin Download PDFInfo
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Abstract
The invention discloses a strain of rhodotorula mucilaginosa, of which the preservation number is CGMCC No. 10123, and application thereof to the fermentation production of malus micromalus astaxanthin. The application is realized through the following steps of culturing a rhodotorula mucilaginosa seed solution, adding 2.5L to 4L of culture solution into a fermentation tank, wherein the culture solution comprises 6g/L to 120g/L of one of glucose, cane sugar, fructose, ethyl alcohol or glycerinum, 1g/L to 5g/L of yeast powder, 0.1g/L to 0.5g/L of monopotassium phosphate, 0.1g/L to 0.5g/L of dipotassium phosphate, 2g/L to 5g/L of urea, and 2g/L to 9g/L of sodium chloride, and the pH value of the culture solution is between 5.5 and 7.5; introducing 2% to 10% of the rhodotorula mucilaginosa seed solution into the culture solution; fermenting for 2 to 4 days at a rotational speed, which is 160r/min to 300r/min, of the fermentation tank and in the temperature range of 28 DEG C to 33 DEG C. By utilizing the rhodotorula mucilaginosa which is provided by the invention, the yield of the malus micromalus astaxanthin is high.
Description
Technical field
The present invention relates to yeast technical field, specifically, relate to a strain rhodotorula mucilaginosa and the application in the red astaxanthin in fermentative production sea thereof.
Background technology
The red astaxanthin in sea is a kind of nutrition pigment and antioxidant of great exploitation potential for its, there is many important biological functions such as strong anti-oxidation, anti-ageing, antiviral, anti-body be painted, anticancer, have broad application prospects in industries such as food, medicine, makeup, aquaculture, livestock and poultry cultivation, chemical industry.The red astaxanthin in sea, a kind of carotenoid with high anti-oxidation ability, during thin-layer chromatography its mobility and natural astaxanthin ester close, resistance of oxidation also with natural left-handed astaxanthin close to and gain the name, DPPH method detects its resistance of oxidation and is better than natural left-handed astaxanthin about 20 ~ 40%.
The preparation method of astaxanthin, there will be a known chemical synthesis, the method extracted from the extraction of aquatic products refuse or algae, utilizes microorganism to carry out the method etc. of producing.Chemical synthesis, production cost is higher, and many countries do not allow to use synthesizing astaxanthin for fodder additives aspect; Extract from aquatic products refuse, raw material sources are extensive, cost is low, but production is easily subject to seasonal restrictions, and working condition requires harsh, and production cost is high, and output is lower, and product purity is not high; Extract from algae, the content astaxanthin in algae is higher, and raw materials cost is lower.But the growth conditions of algae is harsh, very high to the requirement of water quality, environment and light, still there is difficulty in scale operation; Utilize microorganism to carry out the method for producing, adopt saccharine material fermentative production, be beneficial to large scale and high density and produce, and be not subject to the restriction of region and weather, production cost relative moderate.
In recent years, increasing to the demand of astaxanthin both at home and abroad, there are wide market outlook.Many biotech companies of the states such as the current U.S., Canada, European Union are just being devoted to this series products of Development and Production, but far can not meet the need of market, therefore price is higher.Therefore stepping up the development to astaxanthin product, for enhancing our international competitiveness, being of great significance.Because microbial reproduction speed is fast, growth cycle is short, and zymotechnique is also comparatively ripe, be easy to grasp, and yeast inherently a kind of good health material, nutritive value is higher, therefore utilizing yeast fermentation to produce astaxanthin is ideal method at present, also has more wide market outlook.
Summary of the invention
For solving the problems of the technologies described above, the invention provides a strain rhodotorula mucilaginosa (Rhodotorulamucilaginosa), deposit number: CGMCC No.10123.
The invention also discloses the application of this rhodotorula mucilaginosa in the red astaxanthin in fermentative production sea.
The application of this rhodotorula mucilaginosa in the red astaxanthin in fermentative production sea, realized by following steps: cultivate rhodotorula mucilaginosa seed liquor, the nutrient solution of 2.5-4L is added in fermentor tank, nutrient solution comprises: glucose, sucrose, fructose, a kind of 6-120g/L in ethanol or glycerine, yeast powder 1-5g/L, potassium primary phosphate 0.1-0.5g/L, dipotassium hydrogen phosphate 0.1-0.5g/L, urea 2-5g/L, sodium-chlor 2-9g/L, medium pH value is between 5.5-7.5, the rhodotorula mucilaginosa seed liquor of 2-10% is accessed in described nutrient solution, fermentor tank rotating speed is 160-300r/min, ferment 2-4 days in the temperature range of 28-33 DEG C,
Described cultivation rhodotorula mucilaginosa seed liquor is realized by following steps: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in the triangular flask that 50mL seed culture medium is housed, 25-33 DEG C, shaking table is cultivated 22-24h and is obtained primary seed solution under 150-300r/min condition, getting this seed liquor 5mL is inoculated in the triangular flask that 100mL seed culture medium is housed, 25-33 DEG C, 150-300r/min condition shaking table cultivate 22-24h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
Preferably, described seed culture medium comprises: glucose 6-48g/L, yeast powder 1-5g/L, urea 2-5g/L, sodium-chlor 2-9g/L, pH value 5.5-7.5.
Rhodotorula mucilaginose of the present invention (Rhodotorula mucilaginosa), be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, postcode: 100101, preservation date: on December 01st, 2014, deposit number: CGMCCNo.10123.
The 18S rRNA gene order of the bacterial strain that the present invention is separated is checked order, by the 18SrRNA gene order input GenBank obtained, by Blastn program, analysis is compared to all sequences in database, found that the existing type strain that 18S rRNA gene order of the present invention and rhodotorula mucilaginosa belong to has higher similarity, as table 1, be wherein 100% with the sequence similarity of Rhodotorula mucilaginosa isolate ZH9, and with the sequence similarity of many strains rhodotorula mucilaginosa 98%, thus determine that the bacterial strain GH101 separated is the new strains system that rhodotorula mucilaginosa belongs to, and called after rhodotorula mucilaginosa (Rhodotorulamucilaginosa) GH101.
Carry out 18S rRNA amplification, sequencing sequence is:
CATCCGTATTGTTCAAATTTTAAGCATAACAGTGAAACTGCGAATGGCTCATTAAATCAGTCATAGTTTATTTGATGGTACCTTACTACATGGATAACTGTGGTAATTCTAGAGCTAATACATGCTGAAAAATCCCGACTTCTGGAAGGGATGTATTTATTAGATCCAAAACCAATGGCCTTCGGGTCCCTATGGTGAATCATGATAACTGCTCGAATCGCATGGCCTTGCGCCGGCGATGCTTCATTCAAATATCTGCCCTATCAACTTTCGATGGTAGGATAGAGGCCTACCATGGTGATGACGGGTAACGGGGAATAAGGGTTCGATTCCGGAGAGAGGGCCTGAGAAACGGCCCTCAGGTCTAAGGACACGCAGCAGGCGCGCAAATTATCCCCTGGCAACACTTTGCCGAGATAGTGACAATAAATAACAATGCAGGGCTCTTACGGGTCTTGCAATTGGAATGAGTACAATTTAAATCCCTTAACGAGGATCAATTGGAGGGCAAGTCTGGTGCCAGCAGCCGCGGTAATTCCAGCTCCAATAGCGTATATTAAAATTGTTGCCGTTAAAAAGCTCGTAGTCGAACTTCGGGCTCTGTCAGCCGGTCCGCCTTCTTGGTGTGTACTTGTTTGACGGAGCCTTACCTCCTGGTGAACGGCGATGTCCTTTACTGGGTGTCGTCGCAAACCAGGACTTTTACTTTGAAAAAATTAGAGTGTTCAAAGCAGGCCTTTGCCCGAATACATTAGCATGGAATAATAAAATAGGACGCGCGTTCCCATTTTGTTGGTTTCTGAGATCGCCGTAATGATTAATAGGGATAGTTGGGGGCATTTGTATTCCGTCGTCAGAGGTGAAATTCTTGGATTGCCGGAAGACAAACTACTGCGAAAGCATTTGCCAAGGATGTTTTCATTGATCAAGAACGAAGGAAGGGGGATCGAAAACGATTAGATACCGTTGTAGTCTCTTCTGTAAACTATGCCAATTGGGGATCGGTACAGGATTTTTAATGACTGTATCGGCACCCGAAGAGAAATCTTTAAATGAGGTTCGGGGGGGAGTATGGTCGCAAGGCTGAAACTTAAAGGAATTGACGGAAGGGCACCACCAGGTGTGGAGCCTGCGGCTTAATTTGACTCAACACGGGGAAACTCACCAGGTCCAGACACAATAAGGATTGACAGATTGATAGCTCTTTCTTGATCTTGTGGTTGGTGGTGCATGGCCGTTCTTAGTTGGTGGAGTGATTTGTCTGGTTAATTCCGATAACGAACGAG。Through Blast sequence alignment
The comparison of table 1 gene order
Compared with prior art, the application of rhodotorula mucilaginosa of the present invention in the red astaxanthin in fermentative production sea, utilize the rhodotorula mucilaginosa in the present invention, the output of the red astaxanthin in sea is high, and in prior art when the urea of the yeast powder and 2g/L that add 5g/L in fermention medium is as fermentation nitrogen source, the output of extra large red astaxanthin is the highest.
Accompanying drawing explanation
Accompanying drawing described herein is used to provide a further understanding of the present invention, forms a part of the present invention, and schematic description and description of the present invention, for explaining the present invention, does not form inappropriate limitation of the present invention.In the accompanying drawings:
Fig. 1 is the impact of different carbon source on the red astaxanthin yield in sea;
Fig. 2 is the impact of different glucose concn on the red astaxanthin yield in sea;
Fig. 3 is the schema utilizing the red astaxanthin in rhodotorula mucilaginosa fermentative production of the present invention sea.
Embodiment
As employed some vocabulary to censure specific components in the middle of specification sheets and claim.Those skilled in the art should understand, and hardware manufacturer may call same assembly with different noun.This specification and claims are not used as with the difference of title the mode distinguishing assembly, but are used as the criterion of differentiation with assembly difference functionally." comprising " as mentioned in the middle of specification sheets and claim is in the whole text an open language, therefore should be construed to " comprise but be not limited to "." roughly " refer to that in receivable limit of error, those skilled in the art can solve the technical problem within the scope of certain error, reach described technique effect substantially.In addition, " couple " word and comprise directly any and indirectly electric property coupling means at this.Therefore, if describe a first device in literary composition to be coupled to one second device, then represent described first device and directly can be electrically coupled to described second device, or be indirectly electrically coupled to described second device by other devices or the means that couple.Specification sheets subsequent descriptions is for implementing better embodiment of the present invention, and right described description is to illustrate for the purpose of rule of the present invention, and is not used to limit scope of the present invention.Protection scope of the present invention is when being as the criterion depending on the claims person of defining.
Below in conjunction with accompanying drawing, the present invention is described in further detail, but not as a limitation of the invention.
Embodiment 1:
Cultivate rhodotorula mucilaginosa seed liquor: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in and be equipped with in the 250mL triangular flask of 50mL seed culture medium, 25 DEG C, 150r/min condition shaking table cultivate 22h obtain primary seed solution, being inoculated in this seed liquor 5mL is equipped with in the 500mL triangular flask of 100mL seed culture medium, 25 DEG C, 150r/min condition shaking table cultivate 22h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 6g/L, yeast powder 1g/L, sodium-chlor 2g/L, pH value 5.5.
The nutrient solution of 2.5L is added in 5L fermentor tank, nutrient solution comprises: glucose 6g/L, yeast powder 1g/L, potassium primary phosphate 0.1g/L, dipotassium hydrogen phosphate 0.1g/L, urea 2g/L, sodium-chlor 2g/L, medium pH value 5.5, accesses the rhodotorula mucilaginosa seed liquor of 2% in described nutrient solution, fermentor tank rotating speed is 160r/min, and 28 DEG C ferment 2 days.
Embodiment 2:
Cultivate rhodotorula mucilaginosa seed liquor: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in and be equipped with in the 250mL triangular flask of 50mL seed culture medium, 33 DEG C, 300r/min condition shaking table cultivate 24h obtain primary seed solution, being inoculated in this seed liquor 5mL is equipped with in the 500mL triangular flask of 100mL seed culture medium, 33 DEG C, 300r/min condition shaking table cultivate 24h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 48g/L, yeast powder 8g/L, sodium-chlor 9g/L, pH value 7.5.
The nutrient solution of 4L is added in 5L fermentor tank, nutrient solution comprises: glucose 120g/L, yeast powder 5g/L, potassium primary phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, urea 5g/L, sodium-chlor 9g/L, medium pH value 7.5, accesses the rhodotorula mucilaginosa seed liquor of 10% in described nutrient solution, fermentor tank rotating speed is 300r/min, and 33 DEG C ferment 4 days.
Embodiment 3:
Cultivate rhodotorula mucilaginosa seed liquor: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in and be equipped with in the 250mL triangular flask of 50mL seed culture medium, 30 DEG C, 180r/min condition shaking table cultivate 24h obtain primary seed solution, being inoculated in this seed liquor 5mL is equipped with in the 500mL triangular flask of 100mL seed culture medium, 30 DEG C, 180r/min condition shaking table cultivate 24h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 27g/L, yeast powder 4g/L, sodium-chlor 5g/L, pH value 6.5.
The nutrient solution of 4L is added in 5L fermentor tank, nutrient solution comprises: glucose 73g/L, yeast powder 3g/L, potassium primary phosphate 0.3g/L, dipotassium hydrogen phosphate 0.3g/L, urea 3.5g/L, sodium-chlor 5g/L, medium pH value 6.5, accesses the rhodotorula mucilaginosa seed liquor of 6% in described nutrient solution, fermentor tank rotating speed is 230r/min, and 33 DEG C ferment 3 days.
Embodiment 4:
Cultivate rhodotorula mucilaginosa seed liquor: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in and be equipped with in the 250mL triangular flask of 50mL seed culture medium, 28 DEG C, 240r/min condition shaking table cultivate 24h obtain primary seed solution, being inoculated in this seed liquor 5mL is equipped with in the 500mL triangular flask of 100mL seed culture medium, 28 DEG C, 240r/min condition shaking table cultivate 24h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 27g/L, yeast powder 4g/L, sodium-chlor 5g/L, pH value 6.5.
The nutrient solution of 3.5L is added in 5L fermentor tank, nutrient solution comprises: glucose 70g/L, yeast powder 5g/L, potassium primary phosphate 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, urea 2g/L, sodium-chlor 2g/L, medium pH value 6.5, accesses the rhodotorula mucilaginosa seed liquor of 3% in described nutrient solution, fermentor tank rotating speed is 250r/min, and 30 DEG C ferment 3 days.
Dry cell weight 18g/L, extra large red astaxanthin yield 216mg/L is obtained in the present embodiment.
Embodiment 5:
Cultivate rhodotorula mucilaginosa seed liquor: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in and be equipped with in the 250mL triangular flask of 50mL seed culture medium, 28 DEG C, 240r/min condition shaking table cultivate 24h obtain primary seed solution, being inoculated in this seed liquor 5mL is equipped with in the 500mL triangular flask of 100mL seed culture medium, 28 DEG C, 240r/min condition shaking table cultivate 24h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, yeast powder 3.7g/L, sodium-chlor 4.5g/L, pH value 6.2.
The nutrient solution of 3.5L is added in 5L fermentor tank, nutrient solution comprises: glucose 42g/L, yeast powder 4g/L, potassium primary phosphate 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium-chlor 4g/L, medium pH value 6, accesses the rhodotorula mucilaginosa seed liquor of 3% in described nutrient solution, fermentor tank rotating speed is 220r/min, and 30 DEG C ferment 3 days.When using the extra large red astaxanthin of saccharine material fermentation, in substratum, the existence of saccharic carbon source is the prerequisite that growing microorganism and product effectively accumulate.
Embodiment 6:
Cultivate rhodotorula mucilaginosa seed liquor: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in and be equipped with in the 250mL triangular flask of 50mL seed culture medium, 28 DEG C, 180r/min condition shaking table cultivate 24h obtain primary seed solution, being inoculated in this seed liquor 5mL is equipped with in the 500mL triangular flask of 100mL seed culture medium, 28 DEG C, 180r/min condition shaking table cultivate 24h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, yeast powder 3.7g/L, sodium-chlor 4.5g/L, pH value 6.2.
The nutrient solution of 3.5L is added in 5L fermentor tank, nutrient solution comprises: glucose 50g/L, yeast powder 4g/L, potassium primary phosphate 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium-chlor 4g/L, medium pH value 6, accesses the rhodotorula mucilaginosa seed liquor of 3% in described nutrient solution, fermentor tank rotating speed is 220r/min, and 30 DEG C ferment 3 days.
Embodiment 7
Cultivate rhodotorula mucilaginosa seed liquor: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in and be equipped with in the 250mL triangular flask of 50mL seed culture medium, 28 DEG C, 180r/min condition shaking table cultivate 24h obtain primary seed solution, being inoculated in this seed liquor 5mL is equipped with in the 500mL triangular flask of 100mL seed culture medium, 28 DEG C, 180r/min condition shaking table cultivate 24h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, yeast powder 3.7g/L, sodium-chlor 4.5g/L, pH value 6.2.
The nutrient solution of 3.5L is added in 5L fermentor tank, nutrient solution comprises: sucrose 50g/L, yeast powder 4g/L, potassium primary phosphate 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium-chlor 4g/L, medium pH value 6, accesses the rhodotorula mucilaginosa seed liquor of 3% in described nutrient solution, fermentor tank rotating speed is 220r/min, and 30 DEG C ferment 3 days.
Embodiment 8
Cultivate rhodotorula mucilaginosa seed liquor: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in and be equipped with in the 250mL triangular flask of 50mL seed culture medium, 28 DEG C, 180r/min condition shaking table cultivate 24h obtain primary seed solution, being inoculated in this seed liquor 5mL is equipped with in the 500mL triangular flask of 100mL seed culture medium, 28 DEG C, 180r/min condition shaking table cultivate 24h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, yeast powder 3.7g/L, sodium-chlor 4.5g/L, pH value 6.2.
The nutrient solution of 3.5L is added in 5L fermentor tank, nutrient solution comprises: fructose 50g/L, yeast powder 4g/L, potassium primary phosphate 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium-chlor 4g/L, medium pH value 6, accesses the rhodotorula mucilaginosa seed liquor of 3% in described nutrient solution, fermentor tank rotating speed is 220r/min, and 30 DEG C ferment 3 days.
Embodiment 9
Cultivate rhodotorula mucilaginosa seed liquor: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in and be equipped with in the 250mL triangular flask of 50mL seed culture medium, 28 DEG C, 180r/min condition shaking table cultivate 24h obtain primary seed solution, being inoculated in this seed liquor 5mL is equipped with in the 500mL triangular flask of 100mL seed culture medium, 28 DEG C, 180r/min condition shaking table cultivate 24h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, yeast powder 3.7g/L, sodium-chlor 4.5g/L, pH value 6.2.
The nutrient solution of 3.5L is added in 5L fermentor tank, nutrient solution comprises: ethanol 50g/L, yeast powder 4g/L, potassium primary phosphate 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium-chlor 4g/L, medium pH value 6, accesses the rhodotorula mucilaginosa seed liquor of 3% in described nutrient solution, fermentor tank rotating speed is 220r/min, and 30 DEG C ferment 3 days.
Embodiment 10
Cultivate rhodotorula mucilaginosa seed liquor: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in and be equipped with in the 250mL triangular flask of 50mL seed culture medium, 28 DEG C, 180r/min condition shaking table cultivate 24h obtain primary seed solution, being inoculated in this seed liquor 5mL is equipped with in the 500mL triangular flask of 100mL seed culture medium, 28 DEG C, 180r/min condition shaking table cultivate 24h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
Above-mentioned seed culture medium is glucose 25g/L, yeast powder 3.7g/L, sodium-chlor 4.5g/L, pH value 6.2.
The nutrient solution of 3.5L is added in 5L fermentor tank, nutrient solution comprises: glycerine 50g/L, yeast powder 4g/L, potassium primary phosphate 0.2g/L, dipotassium hydrogen phosphate 0.2g/L, urea 5g/L, sodium-chlor 4g/L, medium pH value 6, accesses the rhodotorula mucilaginosa seed liquor of 3% in described nutrient solution, fermentor tank rotating speed is 220r/min, and 30 DEG C ferment 3 days.
As can be seen from Figure 1, in different carbon source, sucrose and glucose are comparatively conducive to the synthesis of astaxanthin, and wherein glucose is the most favourable, and extra large red astaxanthin yield is the highest.
Embodiment 11
On the basis of embodiment 6, the implementation case illustrates the impact of different glucose concn on the red astaxanthin yield in sea, adding concentration respectively is in the fermentation medium the glucose of 0g/L, 40g/L, 50g/L, 60g/L, 70g/L, 80g/L and 90g/L, leavening temperature is 30 DEG C, rotating speed is 180r/min, fermentation 3d.The accumulation volume of the red astaxanthin in sea is proportionate along with the increase of glucose concn, but the increase of later stage addition, the output of astaxanthin increases little.
Embodiment 12
On the basis of embodiment 6, ferment, fermentation condition except nitrogenous source is all identical, the implementation case illustrates that different nitrogen sources produces the impact of extra large red astaxanthin yield to rhodotorula mucilaginosa, nitrogenous source is the main source of the nitrogenous substances such as protein and nucleic acid in synthesis thalline, and therefore in microbial cultivation process, its importance is only second to carbon source.Produce in extra large red astaxanthin process in rhodotorula mucilaginosa fermentation, yeast powder and urea are be beneficial to the organic nitrogen source of extra large red astaxanthin accumulation and inorganic nitrogen-sourced respectively, in order to ensure in whole fermenting process, what can continue provides nitrogenous source, this test have chosen the mixture of the inorganic nitrogen-sourced of different concns and organic nitrogen source as fermentation nitrogen source, and have studied its impact on the red astaxanthin yield in sea.
As shown in Table 1, when the urea of the yeast powder and 2g/L that add 5g/L in fermention medium is as fermentation nitrogen source, the output of extra large red astaxanthin is the highest for result.
Table 1: different organic nitrogen source and inorganic nitrogen-sourced proportioning are on the impact of the red astaxanthin yield in sea
| Nitrogenous source kind and concentration/(g/L) | The red astaxanthin productive rate (%) in sea | Thalline productive rate (%) |
| Yeast powder 7 | 0.66 | 11.3 |
| Yeast powder 6+ urea 1 | 1.06 | 14.8 |
| Yeast powder 5+ urea 2 | 1.14 | 18.25 |
| Yeast powder 4+ urea 3 | 1.09 | 16.7 |
| Yeast powder 3+ urea 4 | 0.92 | 14.1 |
| Yeast powder 2+ urea 5 | 0.75 | 10.9 |
| Urea 7 | 0.38 | 9.6 |
Above-mentioned explanation illustrate and describes some preferred embodiments of the present invention, but as previously mentioned, be to be understood that the present invention is not limited to the form disclosed by this paper, should not regard the eliminating to other embodiments as, and can be used for other combinations various, amendment and environment, and can in invention contemplated scope described herein, changed by the technology of above-mentioned instruction or association area or knowledge.And the change that those skilled in the art carry out and change do not depart from the spirit and scope of the present invention, then all should in the protection domain of claims of the present invention.
Claims (4)
1. a strain rhodotorula mucilaginosa (Rhodotorula mucilaginosa), deposit number: CGMCC No.10123.
2. the application of rhodotorula mucilaginosa according to claim 1 in the red astaxanthin in fermentative production sea.
3. the application of rhodotorula mucilaginosa according to claim 2 in the red astaxanthin in fermentative production sea, it is characterized in that, realized by following steps: cultivate rhodotorula mucilaginosa seed liquor, the nutrient solution of 2.5-4L is added in fermentor tank, nutrient solution comprises: glucose, sucrose, fructose, a kind of 6-120g/L in ethanol or glycerine, yeast powder 1-5g/L, potassium primary phosphate 0.1-0.5g/L, dipotassium hydrogen phosphate 0.1-0.5g/L, urea 2-5g/L, sodium-chlor 2-9g/L, medium pH value is between 5.5-7.5, the rhodotorula mucilaginosa seed liquor of 2-10% is accessed in described nutrient solution, fermentor tank rotating speed is 160-300r/min, ferment 2-4 days in the temperature range of 28-33 DEG C,
Described cultivation rhodotorula mucilaginosa seed liquor is realized by following steps: from test tube slant, provoke rhodotorula mucilaginosa thalline one ring be inoculated in the triangular flask that 50mL seed culture medium is housed, 25-33 DEG C, shaking table is cultivated 22-24h and is obtained primary seed solution under 150-300r/min condition, getting this seed liquor 5mL is inoculated in the triangular flask that 100mL seed culture medium is housed, 25-33 DEG C, 150-300r/min condition shaking table cultivate 22-24h obtain secondary seed solution, this secondary seed solution is described rhodotorula mucilaginosa seed liquor.
4. the application of the rhodotorula mucilaginosa according to Claims 2 or 3 in the red astaxanthin in fermentative production sea, it is characterized in that, described seed culture medium comprises: glucose 6-48g/L, yeast powder 1-5g/L, urea 2-5g/L, sodium-chlor 2-9g/L, pH value 5.5-7.5.
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