CN1181209C - Algae with astaxanthin production and astaxanthin production by yeast mixed culture fermentation - Google Patents
Algae with astaxanthin production and astaxanthin production by yeast mixed culture fermentation Download PDFInfo
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- CN1181209C CN1181209C CNB031053149A CN03105314A CN1181209C CN 1181209 C CN1181209 C CN 1181209C CN B031053149 A CNB031053149 A CN B031053149A CN 03105314 A CN03105314 A CN 03105314A CN 1181209 C CN1181209 C CN 1181209C
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- astaxanthin
- phaffiafhodozyma
- mixed culture
- culture
- yeast
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Abstract
The present invention discloses a method for producing astaxanthin by mixed culture fermentation of algae with astaxanthin production and yeast, which belongs to the technique of astaxanthin production in a biology method. In the method, algae comprising haematococcus pluvialis and yeast comprising carroty yeast are simultaneously inoculated in a reactor with a defined medium for mixed culture fermentation to produce astaxanthin. The method is characterized in that the composition of the culture medium and the culture condition are determined. The method has the advantages that the cost of the culture medium is low, and the conversion rate of sugar, the biomass and the astaxanthin output are enhanced in the cultivation process. The method is suitable for the mixed culture of haematococcus pluvialis, carroty yeast and other kinds of algae with astaxanthin production and yeast.
Description
Technical field
The present invention relates to a kind ofly will produce the algae of astaxanthin and the method that yeast carries out mixed culture, fermentative production astaxanthin, particularly relate to a kind of method of Haematocoocus Pluvialls and phaffiafhodozyma being carried out mixed culture fermentative production astaxanthin.Belong to biological process and produce the astaxanthin technology.
Background technology
Astaxanthin is the extremely strong secondary carotenoid of a kind of oxidation-resistance, has a wide range of applications at aspects such as medicine, food, healthcare products.
Algae is like rain giving birth to all energy synthesizing astaxanthins of haematococcus pulvialis (Haematococcus pluvialis), Chlorococcum (Chlorococcum sp) and yeast such as phaffiafhodozyma (Phaffia rhodozyma), rhodothece rubra (Phodotorula rubra), rhodotorula glutinis (Rhodotorula glutinis), and they are to adopt biotechnology to produce the major microorganisms source of astaxanthin.Haematocoocus Pluvialls and Chlorococcum can carry out photoautotrophy and chemoheterotrophy metabolism simultaneously when illumination, be suitable for little slight alkali environment, and the speed of growth is slow, the content astaxanthin height; Phaffiafhodozyma, rhodothece rubra and rhodotorula glutinis all are unicellular yeasts, are suitable for the slant acidity environment, only carry out the heterotrophism metabolism, and reproduction speed is fast, and content astaxanthin is low.The method of producing at present astaxanthin all is algae or yeast are cultivated individually or to be fermented, in cultivation or fermenting process separately, because nutritional condition, pH and CO
2And O
2The variation of concentration has influenced the growth of cell and the accumulation of astaxanthin, and astaxanthin yield is lower.
Summary of the invention
The object of the present invention is to provide a kind of method of carrying out mixed culture fermentative production astaxanthin with Haematocoocus Pluvialls and phaffiafhodozyma.This procedure condition is easily controlled, and production cost is low, the astaxanthin yield height.
The present invention is realized by following technical proposals: in the reactor of defined medium is housed, inoculates Haematocoocus Pluvialls and phaffiafhodozyma simultaneously and carry out mixed culture fermentative production astaxanthin, it is characterized in that: substratum consists of:
Glucose 1～30g/L K
2HPO
40.01～2.0g/L CuSO
45H
2O 0.001～3.5g/L
NaNO
3 0.01～2.0g/L MgSO
4·7H
2O 0.01～2.5g/L ZnSO
4·7H
2O 0.001～3.0g/L
Co(NO
3)
2 0.01～8.0mg/L CaCl
2·2H
2O 0.01～2g/L KH
2PO
4 0.01～3.6g/L
MnCl
2·4H
2O 0.001～1.5g/L NaCl 0.01～3.0g/L MoO
3 0.01～15mg/L
KOH 0.001～5g/L FeSO
4·7H
2O 0.001～5g/L EDTANa
2 0.001～0.8g/L
H
3BO
3 0.001～1.0g/L
Culture condition is: Haematocoocus Pluvialls and phaffiafhodozyma are inoculated in 1: 0.5～10 ratio, and inoculum size is 1～30% (v/v), and cell density is 1 * 10
2-10
8Cells/ml; Initial pH transfers to 5～9 with hydrochloric acid or sulfuric acid; Culture temperature is 15～30 ℃; Intensity of illumination 1000～6000Lx; Rotating speed is 20～360 rev/mins, cultivates 60～150 hours.
Major advantage of the present invention is that culture medium cost is low, and in the mixed culture process, because the CO that the phaffiafhodozyma fermentation produces
2And organic acid (being neutralized into organic acid salt) can be used as carbon source and is absorbed by Haematocoocus Pluvialls; The O that Haematocoocus Pluvialls photosynthesis is emitted
2Absorbed by yeast again and promote the synthetic of its astaxanthin, thereby improved sugared transformation efficiency, biomass and astaxanthin yield.
Embodiment
Example one substratum is formed (g/l):
glucose 3.5 NaNO
3 0.30 CaCl
2·2H
2O 0.50
MgSO
4·7H
2O 0.075 K
2HPO
4 0.075 NaCl 0.025
KH
2PO
4 0.075 ZnSO
4·7H
2O?0.0082 MnCl
2·4H
2O 0.0014
MoO
3 0.0007.1 CuSO
4·5H
2O?0.00157 Co(NO
3)
2·6H
2O?0.0005
H
3BO
3 0.0114 EDTANa
2 0.05 FeSO
4 0.005
KOH 0.031
PH: transfer to 6.5 with 0.1M hydrochloric acid
In the Erlenmeyer flask of the 250ml of the above-mentioned substratum that 30ml is housed, inoculate Haematocoocus Pluvialls (cell density 2.5 * 10
5Cells/ml) and phaffiafhodozyma (cell density 3.1 * 10
5Cells/ml) each 3ml, and compare with the two single culture.Inoculum size also is respectively 3ml during the two single culture.Erlenmeyer flask is placed the constant temperature bottle swingging machine of band illumination.23.8 ℃ of temperature, rotating speed 100rpm, illumination 35001x cultivated 96 hours.
The extraction of astaxanthin and mensuration: nutrient solution carries out external standard method through centrifugation and extraction with high performance liquid chromatography.
The astaxanthin yield of mixed culture and Haematocoocus Pluvialls and phaffiafhodozyma single culture is respectively: 13.2mg/l, 3.70mg/l, 1.08mg/l.
Example two is with nitrogenous source NaNO in the substratum
3Concentration double (0.6g/l), other condition is constant, the astaxanthin yield that carries out Haematocoocus Pluvialls and phaffiafhodozyma mixed culture, Haematocoocus Pluvialls and phaffiafhodozyma single culture is respectively: 7.80mg/l, 2.05mg/l, 1.69mg/l.
Example three is adjusted to 12g/l with the glucose concn in the substratum, other condition is constant, the astaxanthin yield that carries out Haematocoocus Pluvialls and phaffiafhodozyma mixed culture, Haematocoocus Pluvialls and phaffiafhodozyma single culture is respectively: 6.06mg/l, 2.11mg/l, 1.98mg/l.
Example four is reduced as each 1.5ml with inoculum size, and other condition is constant, and the astaxanthin yield that carries out Haematocoocus Pluvialls and phaffiafhodozyma mixed culture, Haematocoocus Pluvialls and phaffiafhodozyma single culture is respectively: 5.12mg/l, 1.91mg/l, 1.58mg/l.
Example five is adjusted to 2500lx with intensity of illumination, and other condition is constant, and the astaxanthin yield that carries out Haematocoocus Pluvialls and phaffiafhodozyma mixed culture, Haematocoocus Pluvialls and phaffiafhodozyma single culture is respectively: 2.26mg/l, 1.99mg/l, 1.63mg/l.
Example six is adjusted to 200rpm with rotating speed, and other condition is constant, and the astaxanthin yield that carries out Haematocoocus Pluvialls and phaffiafhodozyma mixed culture, Haematocoocus Pluvialls and phaffiafhodozyma single culture is respectively: 8.21mg/l, 5.52mg/l, 3.16mg/l.
Adopt above-mentioned substratum and cultural method, also be applicable to the mixed culture of Haematocoocus Pluvialls and rhodotorula glutinis and rhodothece rubra, and Chlorococcum and above-mentioned phaffiafhodozyma, rhodothece rubra, rhodotorula glutinis three primary yeasts carry out mixed culture.
Claims (1)
1, a kind of method with Haematocoocus Pluvialls and phaffiafhodozyma mixed culture fermentative production astaxanthin, this method is in the reactor of defined medium is housed, inoculate Haematocoocus Pluvialls and phaffiafhodozyma simultaneously and carry out mixed culture fermentative production astaxanthin, it is characterized in that: substratum consists of:
Glucose 1～30g/L K
2HPO
40.01～2.0g/L CuSO
45H
2O 0.001～3.5g/L
NaNO
3 0.01～2.0?g/L MgSO
4·7H
2O?0.01～2.5g/L ZnSO
4·7H
2O?0.001～3.0g/L
Co(NO
3)
2 0.01～8.0mg/L CaCl
2·2H
2O?0.01～2g/L KH
2PO
4 0.01～3.6g/L
MnCl
2·4H
2O?0.001～1.5g/L NaCl 0.01～3.0g/L MoO
3 0.01～15mg/L
KOH 0.001～5g/L FeSO
4·7H
2O?0.001～5g/L EDTANa
2 0.001～0.8g/L
H
3BO
3 0.001～1.0g/L
Culture condition is: Haematocoocus Pluvialls and phaffiafhodozyma are inoculated in 1: 0.5～10 ratio, and inoculum size is 1～30%v/v, and cell density is 1 * 10
2-10
8Cells/ml; Initial pH transfers to 5～9 with hydrochloric acid or sulfuric acid; Culture temperature is 15～30 ℃; Intensity of illumination 1000～6000Lx; Rotating speed is 20～360 rev/mins, cultivates 60～150 hours.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031053149A CN1181209C (en) | 2003-02-24 | 2003-02-24 | Algae with astaxanthin production and astaxanthin production by yeast mixed culture fermentation |
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|---|---|---|---|
| CNB031053149A CN1181209C (en) | 2003-02-24 | 2003-02-24 | Algae with astaxanthin production and astaxanthin production by yeast mixed culture fermentation |
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| CN1181209C true CN1181209C (en) | 2004-12-22 |
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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1298857C (en) * | 2004-03-23 | 2007-02-07 | 中国农业大学 | Synergist for saccharomycetes fermentation |
| CN1324989C (en) * | 2005-10-10 | 2007-07-11 | 华南理工大学 | Nutrition food containing astaxanthin and its preparing method |
| CN100362092C (en) * | 2006-01-05 | 2008-01-16 | 大连轻工业学院 | Method for cultivating phaffiafhodozyma using astaxanthin synthesis accelerant |
| CN101810246B (en) * | 2009-11-13 | 2013-01-16 | 厦门汇盛生物有限公司 | Composite nutrition enhancer and preparation method thereof |
| CN102337215A (en) * | 2011-10-20 | 2012-02-01 | 烟台华融生物科技有限公司 | Methods for culturing haematococcus pluvialis and producing astaxanthin |
| CN102604836B (en) * | 2012-03-01 | 2015-01-21 | 武汉凯迪工程技术研究总院有限公司 | Production method and device for preventing chytrid pollution in haematococcus pluvialis |
| CN103044303B (en) * | 2012-12-19 | 2014-06-04 | 广州优锐生物科技有限公司 | Method for using enzyme to produce astaxanthin |
| CN110408671B (en) * | 2019-07-24 | 2021-07-09 | 嘉必优生物技术（武汉）股份有限公司 | Method for producing DHA and astaxanthin by mixed culture of schizochytrium limacinum and haematococcus pluvialis |
| CN113278557A (en) * | 2021-05-25 | 2021-08-20 | 海南绿藻世界生物科技有限公司 | Symbiotic bacteria composition, preparation method thereof and microalgae culture method |
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