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Method for increasing yield of yeast astaxanthin by inactivating ubiquitin ligase and recombinant strain

A ubiquitin ligase, recombinant strain technology, applied in the direction of ligase, microorganism-based methods, other methods of inserting foreign genetic materials, etc., can solve problems such as unclear gene function

Active Publication Date: 2022-06-21
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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Problems solved by technology

However, the functions of many genes in Phaffia rhodozyme are still unclear, especially the metabolic regulation mechanism of astaxanthin synthesis

Method used

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  • Method for increasing yield of yeast astaxanthin by inactivating ubiquitin ligase and recombinant strain
  • Method for increasing yield of yeast astaxanthin by inactivating ubiquitin ligase and recombinant strain
  • Method for increasing yield of yeast astaxanthin by inactivating ubiquitin ligase and recombinant strain

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Embodiment

[0041] 1. Materials and Methods

[0042] 1.1 Strains and culture conditions

[0043] The Phaffia rhodozyme strain CBS 6938 (purchased from the Dutch CBS Culture Collection) was used as the experimental material. The strain was cultured in YPD (glucose 20g / L, peptone 20g / L and yeast extract 10g / L) liquid medium at a culture temperature of 22°C and a rotational speed of 250rpm. After all the media were subpackaged, they were sterilized at 115°C for 30 minutes.

[0044] 1.2 Construction of gene knockout vector

[0045] The knockout of E3 ubiquitin ligase gene adopts homologous recombination method, and the knocked out E3 ubiquitin ligase gene has been annotated in the GenBank genome database. In this example, knockout vectors for six genes as shown in Table 1 were constructed.

[0046] Table 1. GenBank ID of E3 ubiquitin ligase gene and Scaffold

[0047] serial number E3 ubiquitin ligase gene GenBankID 1 RIN1 CDZ97817.1 2 RIN2 CDZ97878.1 3 ...

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Abstract

The invention relates to the technical field of biology, in particular to a method for increasing the yield of yeast astaxanthin by inactivating ubiquitin ligase and a recombinant strain. The recombinant strain is obtained by genetic modification and knockout of at least one E3 ubiquitin ligase of an original strain phaffia rhodozyma, and the yield of astaxanthin of phaffia rhodozyma is increased by the obtained recombinant strain. The phaffia rhodozyma E3 ubiquitin ligase coding gene is knocked out, and the negative regulation effect of E3 ubiquitin ligase astaxanthin synthesis is eliminated, so that the yield of astaxanthin is increased. Compared with a control strain, the astaxanthin cell content of the E3 ubiquitin ligase gene knockout strain can be obviously improved by 8.3 times. The method can be applied to construction of astaxanthin high-yield strains and increase of astaxanthin yield.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for increasing the yield of yeast astaxanthin by inactivating ubiquitin ligase and a recombinant strain. Background technique [0002] Astaxanthin is a carotenoid that usually exists in organisms such as bacteria, algae, yeast and plants (Dhankhar J, et al. International Journal of Pharmaceutical sciences & Research, 2012, 3: 1246-1259), has coloring, strong antioxidant , anti-inflammatory and anti-cancer functions (Park J, Chyun J, Kim Y, et al. Nutr Metab, 2010, 7:18). The global astaxanthin market size (2016) was valued at USD 555 million and is projected to exceed USD 800 million by 2022, according to a May 2017 Foresight Market Analysis Research Report (http: / / www.marketsandmarkets.com / Market- Reports / astaxanthin-mark et-162119410.html, as accessed on 14th September, 2017). And, as the demand for astaxanthin continues to increase, its market will further expand. [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N15/81C12N15/90C12N15/64C12P23/00C12R1/645
CPCC12N9/93C12N15/815C12N15/905C12P23/00
Inventor 王士安丁瑞瑞刘瑜黄睿林李福利
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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