A type of alpine spore fungus and its application

Abstract

This invention proposes an alpine Mortierella and its applications, belonging to the field of microbial technology. The invention aims to address the technical problem of increasing arachidonic acid production by providing an alpine Mortierella and its applications. This invention provides an alpine Mortierella, named LSZJ001, which was deposited on June 17, 2024, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 41340. This provides a theoretical basis for arachidonic acid production.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, and in particular relates to an alpine spore mold and its applications. Background Technology

[0002] Currently, the sources of arachidonic acid (ARA) are mainly divided into two categories: traditional extraction methods and microbial synthesis methods. However, both methods have limitations to varying degrees, hindering the sustainable development of the industry and the satisfaction of market demand. Traditionally, ARA is mainly extracted from deep-sea fish oil. This technical route suffers from problems such as unstable raw material supply, product purity and quality defects, and high extraction costs. Although microbial synthesis has been recognized as a green alternative to fish oil extraction, the strains currently relied upon for commercial production still face bottlenecks such as limited unit capacity, long fermentation cycles, and insufficient strain stability and adaptability, which are not conducive to stable industrial production. Summary of the Invention

[0003] In view of this, the present invention aims to provide a type of alpine argentea and its application to solve the technical problem of increasing the yield of arachidonic acid.

[0004] To achieve the above objectives, the present invention adopts the following technical solution: a type of alpine spores ( Mortierella alpina The alpine spores, named LSZJ001, were deposited on June 17, 2024, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 41340.

[0005] This invention provides a microbial preparation containing the above-mentioned alpine spores.

[0006] This invention provides the application of the above-mentioned *Morchella alpina* or the above-mentioned microbial preparation in the production of arachidonic acid.

[0007] This invention provides a method for producing arachidonic acid, obtained by fermentation of the aforementioned *Morchella alpina*.

[0008] Furthermore, the carbon source utilized by *Morchella alpineensis* is potato.

[0009] Furthermore, the nitrogen source utilized by Alpine Flooding is glucose.

[0010] Furthermore, the fermentation medium is PDA medium.

[0011] Furthermore, the fermentation temperature is specified as 25°C.

[0012] Furthermore, the fermentation time is limited to 18 days.

[0013] Compared with the prior art, the beneficial effects of the present invention are as follows: the alpine spore strain is inoculated into a culture box, cultured at 25°C, fermented for 18 days, and metabolites are obtained. After extraction, oil is obtained, and the arachidonic acid yield can reach up to 16 g / L. According to GB5009.168-2016, fatty acid composition analysis shows that the arachidonic acid content in the oil can reach up to 62.5%.

[0014] [Biological Preservation Information]: A type of alpine spore fungus ( Mortierella alpina The alpine spores, named LSZJ001, were deposited on June 17, 2024, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 41340. The deposit address is No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences. Attached Figure Description

[0015] Figure 1 Image of a colony of *Morchella alpinea*. Detailed Implementation

[0016] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. It should be noted that, unless otherwise specified, the embodiments and features in the embodiments of the present invention can be combined with each other, and the described embodiments are only some embodiments of the present invention, not all embodiments.

[0017] Example 1. Obtaining Alpine Monascus This strain was isolated from the soil of the wheat experimental field of Leshi Company and obtained through strain screening.

[0018] Isolation method: Soil was placed in an Erlenmeyer flask, 50 mL of distilled water was added, and the soil was spread onto PDA medium using a spreader. After incubation at 25°C for 3 days, the morphology of the bacterial strain was observed under a microscope. The colony was a rosette shape composed of small leaves, white and cottony, and grew vigorously. Figure 1 As shown.

[0019] Screening method: All obtained alpine molluscs were inoculated into PDA medium containing aspirin, and non-growing strains were selected for 18S RNA identification to obtain the alpine mollusc strain used in this patent.

[0020] 18sRNA sequence: (SEQ ID NO.1) The comparison results are shown in Table 1: Table 1

[0021] The species with the highest similarity to the NCBI database was Mortierella, whose taxonomic unit is: Fungi (Kingdom); Mucoromycota (Phylum); Mortierellomycetes (Class); Mortierellales (Order); Mortierellaceae (Family); Mortierella (Genus).

[0022] Example 2. A method for producing arachidonic acid Required carbon source: potato; required nitrogen source: glucose; PDA medium was used, containing 200 g / L potato, 20 g / L glucose, and 16 g / L agar. Under laboratory conditions, the strain was inoculated into a culture box and cultured at 25℃ for 18 days to obtain metabolites. After extraction, oil was obtained, with the highest arachidonic acid yield reaching 16 g / L. According to GB 5009.168-2016, fatty acid composition analysis showed that the arachidonic acid content in the oil could reach up to 62.5%. The statistical analysis and identification of arachidonic acid content were performed according to the methods in GB 5009.168-2016.

[0023] Determination of fermentation time: Under laboratory conditions, the strain was inoculated into a culture box and cultured at 25℃. The biomass accumulated on days 3, 5, 7, 9, 11, 13, 15, 19 and 21 of culture was observed. Analysis showed that the content of arachidonic acid in fatty acids reached its peak after 18 days of culture.

[0024] Oil extraction steps: Dry and grind the metabolites obtained from fermentation, soak them in n-hexane for half an hour, filter them using a Buchner funnel, and evaporate the resulting liquid on a rotary evaporator to obtain an orange-yellow oil, which is the target product.

[0025] Table 2

[0026] The specific embodiments of the present invention disclosed above are merely illustrative of the invention. These embodiments do not exhaustively describe all details, nor do they limit the invention to the specific embodiments described. Many modifications and variations can be made based on the content of this specification. This specification selects and specifically describes these embodiments to better explain the principles and practical applications of the invention, thereby enabling those skilled in the art to better understand and utilize the invention.

Claims

1. A type of alpine spore fungus ( Mortierella alpina ), characterized in that, The alpine spore fungus, named LSZJ001, was deposited on June 17, 2024, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 41340.

2. A microbial preparation containing the alpine spores as described in claim 1.

3. The application of the *Morchella alpina* as described in claim 1 in the production of arachidonic acid.

4. The application of the microbial preparation according to claim 2 in the production of arachidonic acid.

5. A method for producing arachidonic acid, characterized in that, The method described in claim 1 is based on the fermentation of *Morchella alpina*.

6. The method according to claim 5, characterized in that, The carbon source utilized by *Morchella alpineensis* is potato.

7. The method according to claim 5, characterized in that, The nitrogen source utilized by alpine spores is glucose.

8. The method according to claim 5, characterized in that, The fermentation medium was PDA medium.

9. The method according to claim 5, characterized in that, The fermentation temperature is 25℃.

10. The method according to claim 5, characterized in that, The fermentation time is 18 days.

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