A kind of astaxanthin high-yield bacterial strain and its application

A high-yield strain, astaxanthin technology, applied in fungi, microorganisms, biochemical equipment and methods, etc., can solve the problems of inability to meet industrial production, low astaxanthin yield, etc., and achieve the effect of great social significance and economic value.

Inactive Publication Date: 2017-02-15
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem that the current astaxanthin yield of Phaffia yeast is low and cannot meet the needs of industrial production, the present invention provides a high-yielding astaxanthin strain and its application

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  • A kind of astaxanthin high-yield bacterial strain and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Example 1 This example illustrates the Phaffia yeast ( Xanthophyllomyces dendrorhous ) method for mutagenesis of vr01.

[0036] Phaffia yeast preserved in the laboratory ( Xanthophyllomyces dendrorhous )vr01 is the starting strain for mutagenesis, and the specific steps are as follows:

[0037] (a) In order to obtain a high-yielding strain of astaxanthin, after comparing various mutagenesis methods, the method of mutagenizing the strain with nitrosoguanidine (NTG) was selected. from Phaffia yeast in liquid medium ( Xanthophyllomyces dendrorhous ) The cell suspension to be mutated was obtained from the vr01 culture. At 20°C and 100rpm, the 10 8 Cells / mL cell suspension in 0.1M sodium citrate buffer of 250ug / Ml NTG solution, pH 5.0, warm bath for 60-120min. (b) The mutagenized cells were then cultured in liquid intermediate medium for 10 h at 23 °C and 100 rpm.

[0038](c), screening of mutagenic strains:

[0039] Screening: Introduce the culture solution obtain...

Embodiment 2

[0043] Example 2 This example illustrates the high-yield strain Phaffia ( Xanthophyllomyces dendrorhous ) VR-032 fermentation process for producing astaxanthin.

[0044] The medium formula used in this embodiment:

[0045] The solid medium is: 3.0% sucrose, 1.0% malt extract powder, 1.5% agar powder, pH 5.8-6.0, and the rest is water.

[0046] The seed medium is: 4.0% sucrose, 2.0% peanut powder, 0.2% potassium dihydrogen phosphate, 0.05% magnesium sulfate, pH 5.8~6.0, and the rest is water.

[0047] The fermentation medium is: 2.0% sucrose, 4.0% peanut cake powder, 0.2% potassium dihydrogen phosphate, 0.05% magnesium sulfate, vitamin B 1 0.0002%, pH 5.8~6.0, the rest is water.

[0048] Inoculate Phaffia yeast VR-032 on solid medium, culture temperature is 21°C, and culture time is 48h; inoculate the solid-cultured Phaffia yeast VR-032 culture into seed medium, and fill the liquid in a 250mL culture bottle The volume is 30mL, the culture temperature is 21°C, and the rota...

Embodiment 3

[0049] Example 3 This example illustrates the morphological, physiological and biochemical characteristics and passage stability of mutant strains

[0050] Phaffia yeast ( Xanthophyllomyces dendrorhous ) VR-032, the morphological, physiological and biochemical characteristics are as follows: colony color: off-white; aerobic mode: aerobic growth; colony size: 4~7 mm; optimum growth temperature: 22°C; optimum growth pH: 7.0~ 7.2; Bacterial morphology: round colony, dull; Gram stain: positive.

[0051] In the fermentation medium using sucrose as a carbon source (fermentation culture conditions are the same as in Example 2), the mutant strain Phaffia ( Xanthophyllomyces dendrorhous ) The output of VR-032, the passage stability of the observation period, and the strain passage fermentation test results are shown in Table 1:

[0052] Table 1 High-yielding mutant strain Phaffia ( Xanthophyllomyces dendrorhous ) VR-032 stability test

[0053] Number of passages Asta...

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Abstract

The invention discloses an astaxanthin high-yield strain and application thereof. The astaxanthin high-yield strain is classified to be named as Xanthophyllomyces dendrorhous VR-032 and collected in CCTCC (China Center For Type Culture Collection), and the collection number is CCTCCM2014190. According to the invention, a target strain VR-032 is finally screened by virtue of nitrosoguanidine mutagenesis by adopting a strain which is obtained by fermentation and screening based on a shake flask and has high astaxanthin yield as an initial strain of a next round of mutagenesis, and the strain can be used for more greatly increasing astaxanthin components contained in a fermentation product and decreasing other components and has good hereditary stability. According to the astaxanthin high-yield strain disclosed by the invention, the sugar cane is utilized as a fermentation carbon source, the yield of the astaxanthin obtained by fermentation by utilizing the low-cost sugar cane as the carbon source in a 5L fermentation tank is up to 68.7mg / L, the yield of the astaxanthin is increased by 20.8% compared with that of an original initial strain, and the astaxanthin high-yield strain can be applied to industrial production, greatly improving the fermentation unit and achieving significant economic application value.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to an astaxanthin high-yielding bacterial strain and application thereof. Background technique [0002] Astaxanthin (3,3'-dihydroxy-4,4'diketo-beta, beta' carotene) is a keto carotenoid with a natural red color. Astaxanthin was originally used as a feed additive in aquaculture, and later pharmacological and physiological studies showed that astaxanthin has strong anti-oxidation and quenching free radical functions, and can promote the production of antibodies and enhance the immune function of the host , has broad application prospects in the fields of food, medicine and cosmetics, and has high economic value. Phaffiarhodozyma 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, Subphylum Deuteromycota, Cryptococcus family, Phaffia rhodozym...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16C12P23/00C12R1/645
Inventor 虞龙张旭琛陈晓双黄思思
Owner NANJING TECH UNIV
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