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Phaffia rhodozyma strain with high yield of natural astaxanthin as well as breeding method and application thereof

A technology of Phaffia rhodozyma and yeast strains, which is applied in the field of high-yield natural astaxanthin-producing Phaffia rhodozyme strains and its breeding field, achieving quick results, cost-effective effects, and easy promotion

Active Publication Date: 2013-01-09
浙江皇冠科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no astaxanthin production enterprise in China, so it is of great research significance and great development space to study a production method of astaxanthin suitable for industrial production

Method used

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  • Phaffia rhodozyma strain with high yield of natural astaxanthin as well as breeding method and application thereof
  • Phaffia rhodozyma strain with high yield of natural astaxanthin as well as breeding method and application thereof
  • Phaffia rhodozyma strain with high yield of natural astaxanthin as well as breeding method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1) LiCl-ultraviolet compound mutagenesis breeding

[0042] The parental strain is Phaffia rhodozyma 02, which was collected from the School of Food and Bioengineering of Zhejiang Gongshang University and donated by Liang Xinle (Hangzhou, Zhejiang). Dilute to 10 with sterile saline 6 cells / mL, take 5 mL of bacterial suspension in a petri dish with a magnetic rotor, add 300 μL of 10% LiCl, mix well, and then open the cover for ultraviolet irradiation. Mutagenesis conditions: 30cm, 15W ultraviolet lamp. Irradiation time is 7min, after dark treatment, dilute 10 3 times, the YM medium was coated on the plate, and cultured in the dark for 4-6 days in a constant temperature incubator at 22°C. At the same time, the unmutated bacterial suspension was operated in the same manner as a control, and the calculated lethality was 83%.

[0043] (2) 60 Co-γ-ray mutagenic breeding

[0044] Adjust the concentration of the prepared bacterial suspension to 10 6 cells / mL, take the ba...

Embodiment 2

[0073] Phaffia rhodozyma CZ10 bred in Example 1 was used for fermentation and culture to produce astaxanthin.

[0074] Get the Phaffia rhodozyma CZ10 selected and bred in Example 1 and inoculate it into a Erlenmeyer flask equipped with a fermentation medium for continuous shaking flask fermentation 3 times at 22 ± 1° C., 150 r / m, cultivate for 48 hours, and extract the fermentation broth Astaxanthin in, measure the content of astaxanthin, repeat test, its astaxanthin production is all stable at 68mg / L (the amount of astaxanthin produced per L fermented liquid).

[0075] The extraction and determination methods of astaxanthin in the fermentation broth are the same as in Example 1.

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Abstract

The invention discloses a phaffia rhodozyma strain with high yield of natural astaxanthin as well as a breeding method and application thereof. The strain is named phaffia rhodozyma CZ10, and is collected with the collection number CGMCC No.6355. The breeding method comprises the following steps of: (1) selecting parent strains, classifying the parent strains into two groups, performing LiCl-ultraviolet compound mutation and 60Co-gamma ray mutation respectively, and screening to obtain a plurality of mutated strains I and a plurality of mutated strains II; (2) performing protoplast fusion, regeneration and screening on the mutated strains I and the mutated strains II to obtain a plurality of fusion strains with higher yield of the natural astaxanthin than the mutated strains I and the mutated strains II; and (3) performing recursive protoplast fusion on the obtained fusion strains obtained in the step (2) at least five times to obtain the phaffia rhodozyma strain. The strain disclosedby the invention is applied to industrial production of astaxanthin; and the astaxanthin yield is up to 68mg / L, which is increased by 14.3 times than that of the parent strains before breeding.

Description

technical field [0001] The invention relates to the technical field of biological fermentation engineering, in particular to a Phaffia rhodozyme strain with high yield of natural astaxanthin, a breeding method and application thereof. Background technique [0002] Astaxanthin (3,3'-dihydroxy-4,4'diketo-β,β'carotene) is a keto carotenoid with a natural red color. Astaxanthin was originally used as a feed additive in aquaculture. Later pharmacological and physiological studies showed that astaxanthin has strong anti-oxidation and free radical quenching functions, and can promote the production of antibodies and enhance the immune function of the host. It has broad application prospects in the fields of food, medicine and cosmetics, and has high economic value. [0003] Phaffia rhodozyma is the only yeast that can naturally produce astaxanthin, and its trans-astaxanthin was approved by FDA in 2000 for food additives. It is the only species in the kingdom Fungi, Phylum Fungi, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12P23/00C12N15/01C12N15/04C12R1/645
Inventor 胡向东朱静梁新乐
Owner 浙江皇冠科技有限公司
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