High-yield carotinoid and copper fermentation culture method by using Rhodotorula benthica

A marine rhodotorula, carotene technology, applied in biochemical equipment and methods, fermentation, microbial-based methods, etc., to achieve high absorption rate, good stability, and increase the effect of carotene and copper production

Active Publication Date: 2015-11-11
QINGDAO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there is no relevant report on the marine rhodotorula that can produce high carotenoids and can also enrich copper. The present invention expects to separate and obtain copper-rich marine rhodotorula with high carotene production from the marine environment, and determine it through single factor experiments and orthogonal experiments. It is suitable for the optimal culture conditions of marine red yeast carotenoids, provides a theoretical basis for industrial production, and explores a new way for the utilization of marine red yeast

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  • High-yield carotinoid and copper fermentation culture method by using Rhodotorula benthica

Examples

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Effect test

Embodiment 1

[0020] Example 1: Acquisition of marine Rhodotorula yeast WYN1 and domestication of copper tolerance

[0021] (1) Fresh sea shrimp, starfish, seaweed and other biological samples collected from the Yellow Sea are placed in sterile YEPD medium, and red or pink round colonies are picked for further purification and culture. According to the accumulation of carotenoids in each strain, the strains with high carotenoid production were screened. Among them, the marine rhodotorula WYN1 with higher carotenoid production has oval cells without pseudohyphae; it forms red colonies on the YEPD medium prepared in seawater, with smooth surface and neat edges; it does not produce eggs on the sporulation medium Cystic spores, multilateral budding, no sporulation; no fermentation of sugar, no formation of starch-like compounds, and no inositol synthesis. Based on the above characteristics, it was preliminarily determined to belong to the genus Rhodotorula. Keep ramps in the refrigerator.

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Embodiment 2

[0038] Example 2: (Comparative Test) Fermentation Conditions for High-yielding Carotenoids of Rhodotorula marina WYN1

[0039] Fermentation medium composition: glucose 2.5%, peptone 2.5%, magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 300mg / L (copper sulfate configuration), sodium chloride 20%, the rest is water; the initial pH is 8 , the inoculum amount was 10%, the shaker speed was 180r / min, and after culturing at 28°C for 24 hours, the temperature was adjusted to 30°C, the pH was adjusted to 5, and the nitrogen source was added once, that is, peptone was added according to the addition amount of 2.5%, and continued Ferment for 24h. The marine Rhodotorula WYN1 was fermented and cultured by the above method, and its biomass, carotenoid production and intracellular copper content were 27.52 g / L, 25.12 mg / L and 6240 μg / g dry bacteria, respectively.

Embodiment 3

[0041] The marine red yeast WYN1 of the present invention can efficiently produce carotenoids by fermenting and cultivating in a copper-containing medium. The specific method is: the preferred fermentation medium is composed of 1% glucose, 2% enzymatically hydrolyzed corn flour, and 2.5% peptone. Magnesium sulfate 0.02%, potassium dihydrogen phosphate 0.15%, copper content 180mg / L (copper sulfate configuration, calculated as copper ions), sodium chloride 20%, the rest is water; initial pH is 8, inoculum size 5%, shaker The rotation speed is 60r / min, after culturing at 26°C for 12 hours, adjust the temperature to 18°C ​​for static culture at low temperature, stop stirring, adjust the pH to 4, and keep it for 3 hours; after that, the temperature is raised to 28°C in a stepwise manner at 2-4°C per hour, and the pH is adjusted. For 5.3, add peptone according to the amount of 2.5%, and add yeast powder according to the amount of 2%; the shaker speed is 80r / min, and the fermentation ...

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Abstract

The invention provides a high-yield carotinoid and copper fermentation culture method by using Rhodotorula benthica, which is implemented by carrying out fermentation culture on the Rhodotorula benthica WYN1 in a copper-containing culture medium to efficiently produce carotinoid, wherein the fermentation culture medium is composed of 1% of glucose, 1-3% of enzymolysis corn flour, 2.5% of peptone, 0.02% of magnesium sulfate, 0.15% of potassium dihydrogen phosphate, 150-200 mg / L copper, 20% of sodium chloride and the balance of water. The method comprises the following steps: culturing at 24-28 DEG C for 10-15 hours, wherein the initial pH value is 8, the inoculum size is 3-8%, and the rotation speed of the shaking table is 50-80 r / min; regulating the temperature to 16-20 DEG C, carrying out low-temperature stationary culture, stopping stirring, regulating the pH value to 3-5, and keeping for 2-4 hours; carrying out gradient heating to 24-30 DEG C, regulating the pH value to 4.6-5.5, adding 2.5% of the peptone, and adding 2% of the yeast powder; and regulating the rotation speed of the shaking table to 50-100 r / min, and continuing fermentation for 10-15 hours.

Description

technical field [0001] The invention relates to a marine rhodotorula strain capable of accumulating carotenoids and enriching copper and a fermentation culture method conducive to carotenoid accumulation, which belongs to the technical field of fermentation and comprehensive utilization of marine microorganisms. Background technique [0002] Carotenoids are a class of polyene compounds that are yellow, orange or red. They are the most ubiquitous and stable natural pigments in nature and widely exist in many animals and plants. As an excellent food additive and nutritional supplement for improving human nutrition, carotenoid has been recognized by countries all over the world for its excellent quality and effect, and is known as the most promising antioxidant. At present, carotenoids are widely used in industries such as food, medicine, cosmetics and feed additives. Due to their good application prospects and excellent functions, they have been favored by people for a long ti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P23/00C12P3/00C12R1/645
Inventor 王亚楠程殿林丛培江孙梦洁涂开杨婷婷刘敏
Owner QINGDAO UNIV
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