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Astaxanthin high-yield strain and application thereof

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: 2014-12-03
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

Method used

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  • Astaxanthin high-yield strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

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

[0037] (a) In order to obtain a high-yield astaxanthin strain, after comparing various mutagenesis methods, the method of nitrosoguanidine (NTG) mutagenesis strain was selected. From Phaffia in liquid medium ( Xanthophyllomyces dendrorhous ) Obtain the cell suspension to be mutagenized in the vr01 culture. At 20 ℃ and 100 rpm, 10 8 The cell suspension of cells / mL was bathed in 250ug / Ml NTG solution 0.1M sodium citrate buffer, pH 5.0 for 60-120min. (B) Then the mutagenized cells were cultured in a liquid intermediate medium at 23°C and 100 rpm for 10 hours.

[0038] (c) Screening of mutagenic strains:

[0039] Screening: Connect the culture solution induced in step (b) to the seed culture medium, expand t...

Embodiment 2

[0043] Example 2 This example illustrates the high-yielding strain Phaffia ( Xanthophyllomyces dendrorhous ) VR-032 fermented production process of astaxanthin.

[0044] The medium formula used in this example:

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

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

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

[0048] Inoculate Phaffia VR-032 on a solid medium at a temperature of 21°C and a culture time of 48h; inoculate the solid culture of Phaffia VR-032 into a seed medium, and fill the liquid in a 250mL culture bottle The amount is 30mL, the culture temperature is 21℃, and the rotation speed is 200rpm for 48h; the seed cul...

Embodiment 3

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

[0050] Phaffia ( 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℃; suitable growth pH: 7.0~ 7.2; Cell morphology: round colony, dull; Gram stain: positive.

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

[0052] Passages Astaxanthin production (mg / L)vr01 Astaxanthin production (mg / L) VR-032 F156.469.8 F255.870.0 F356.969.5 F456.769.3 F555.969.5

[0053] It can be...

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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. Phaffia rhodozyma is the only yeast that can naturally produce astaxanthin, and its trans-astaxanthin has been approved by FDA in 2000 for food additives. It is the only species in the kingdom Fungi, Phylum Fungi, Subphylum Deuteromycota, Cryptococcus family, Phaffia ...

Claims

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Application Information

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Patent Type & Authority Applications(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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