A strain of Bacillus paramycoides and its application in preparing palmitoleic acid

Abstract

The present application belongs to the field of microbial fermentation technology, and specifically relates to a strain of Bacillus paramycoides and its application in the preparation of palmitoleic acid. The Bacillus paramycoides SS6 is a Bacillus paramycoides strain. Bacillus paramycoides , deposited in the China General Microbiological Culture Collection on October 28, 2024, with the deposit number: CGMCC No.32390; the address of the deposit unit: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; palmitic acid can be obtained by fermenting sugarcane juice with it; after subsequent extraction, it can be used for industrial production.

Description

Technical Field

[0001] The present application belongs to the field of microbial fermentation technology, and specifically relates to a strain of Bacillus paramycoides and its application in the preparation of palmitoleic acid. Background Art

[0002] Palmitoleic acid contains 16 carbon atoms and a double bond (CH=CH, also known as palmitoleic acid), and its molecular formula is CH3(CH2)5CH=CH(CH2)7COOH. It is a monobasic cis fatty acid of the OMEGA-7 family (w-7), with a carbon number of 16, and can be extracted from palm. Palmitoleic acid is mainly derived from whale oil and cod liver oil, but due to the international ban on whaling and the scarcity of fishery resources, the production of palmitoleic acid is greatly restricted, making it difficult to meet social needs. The content of palmitoleic acid is relatively low in general oil crops, but it is also very high in some special plants. Therefore, finding plant sources and extracting them can better make up for the lack of palmitoleic acid production. In addition, palmitoleic acid is also found in some microbial extracts.

[0003] Palmitoleic acid, as a functional fatty acid, has multiple biological functions such as promoting cell metabolism, protecting skin moisture, improving patient metabolic syndrome and anti-inflammation (Tu Xinghao, Du Liqing, Wei Fang, et al. Research progress on the physical and chemical properties and efficacy of palmitoleic acid in macadamia nuts [J]. Agricultural Research and Application, 2021, 34(4): 35-43.); Souza et al. (SOUZA CO, TEIXEIRA AAS, BIONDO LA, et al. Palmitoleic acid reduces high fat diet-induced liver inflammation by promoting PPAR-γ-independent M2apolarization of myeloid cells [J]. Biochi-mica et Biophysica Acta Molecularand Cell Biology of Lipids, 2020, 1865(10): 158776.) studied POA The effect of palmitoleic acid on high-fat diet-induced hepatitis in mice showed that the inflammatory cytokines in the liver of mice treated with palmitoleic acid were significantly lower than those in the oleic acid-treated group. The reason is that palmitoleic acid inhibits the activity of macrophages by activating the expression of the fat biological transcription factor peroxisome proliferator-activated receptor γ (PPAR-γ) in hepatitis mice, thereby reducing the number of macrophages and alleviating the inflammatory response. Related studies have also found that seabuckthorn fruit oil can alleviate the clinical symptoms of ulcerative colitis in rats (Li Xinping. Study on the effects and mechanisms of seabuckthorn seed oil and fruit oil on ulcerative colitis model rats [D]. Taiyuan: Shanxi Medical University, 2012.). Summary of the invention

[0004] In view of the problem of low palmitoleic acid production at present, the present invention provides a strain of Bacillus paramycoides, which can be used to ferment sugarcane juice to obtain palmitoleic acid; the fermentation process is simple and easy to industrialize.

[0005] The technical solution of this application is as follows:

[0006] In a first aspect of the present invention, a strain of Bacillus paramycoides SS6 is protected, wherein the Bacillus paramycoides SS6 is Bacillus paramycoides , deposited in the China General Microbiological Culture Collection on October 28, 2024, with the deposit number: CGMCC No. 32390; the address of the deposit unit is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0007] Another aspect of the present invention is to protect the use of the above-mentioned Bacillus paramycoides in the preparation of palmitoleic acid.

[0008] Preferably, palmitoleic acid is obtained by fermenting sugarcane juice with Bacillus paramycoides.

[0009] Preferably, yeast EC1118 is also included. The yeast EC1118 is a commercial yeast.

[0010] Bacillus paramycoides Bacillus paramycoides Originated from the surface of tobacco leaves; the preservation information is as follows:

[0011] Storage time: October 28, 2024

[0012] Deposit number: CGMCC No. 32390;

[0013] Address of the depository: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing;

[0014] Classification name: Bacillus paramycoides Bacillus paramycoides .

[0015] Beneficial Effects of the Invention

[0016] A strain of Bacillus paramycoides was isolated from the surface of tobacco leaves Bacillus halotolerans ; It is used in the fermentation of sugarcane juice, and by controlling the reaction conditions, palmitoleic acid and / or phenylethanol and / or β-damascenone can be obtained.

[0017] The relationship between β-damascone and fermentation temperature is not a simple positive correlation. At the lower temperature stage (20°C), it is the suitable temperature for EC1118 yeast. At 25°C, the increase in temperature and the mutual inhibition of the two strains lead to a decrease in the production of β-damascone. At 30°C, the production of β-damascone (norisoprene compounds) by the SS6 strain increases and becomes the main source of β-damascone due to the increase in temperature. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 is the content of β-damascenone, phenylethanol and palmitoleic acid at different temperatures (μg / L). DETAILED DESCRIPTION

[0019] The yeast EC1118 used in the examples was purchased from Raman Yeast Company.

[0020] Example 1

[0021] A method for preparing palmitoleic acid comprises the following steps:

[0022] Adjust the sugarcane juice to 20 °Bx (20°C), sterilize, and ferment with yeast EC1118 at 20°C, 25°C, and 30°C, and add Bacillus paramycoides SS6 inoculant at 2% of the total fermentation liquid volume; when the sugar content no longer decreases, the fermentation ends;

[0023] Yeast EC1118 was activated by adding warm water three times at a dosage of 1 mg / L; then SS6 bacterial agent was added.

[0024] The Bacillus paramycoides SS6 inoculum is prepared by the following steps: inoculating the Bacillus paramycoides SS6 in an inoculum amount of 1% into an LB liquid culture medium, and culturing at 35° C. and 200 rpm.

[0025] When the concentration of Bacillus paramycoides SS6 was 1×10 9 / mL, stop culturing and obtain the bacterial agent.

[0026] The results of fermentation at different temperatures are shown in Figure 1 .from Figure 1 It can be seen that the production of the three compounds is the highest at a higher temperature (30°C). Among them, the relationship between β-damascone and fermentation temperature is not simply positively correlated. At a lower temperature stage (20°C), it is the suitable temperature for EC1118 yeast, and at 25°C, the production of β-damascone decreases due to the increase in temperature and the mutual inhibition of the two strains. At a temperature of 30°C, the production of β-damascone (norisoprene compounds) by the SS6 strain increases and becomes the main source of β-damascone due to the increase in temperature.

[0027] Example 2

[0028] A method for preparing palmitoleic acid comprises the following steps:

[0029] Adjust the sugarcane juice to 20 °Bx (20°C), sterilize, and ferment at 30°C using yeast EC1118, and add Bacillus paramycoides SS6 inoculant at 1% of the total fermentation liquid volume; when the sugar content no longer decreases, the fermentation ends;

[0030] Yeast EC1118 was activated by adding warm water three times at a dosage of 1 mg / L; then SS6 bacterial agent was added.

[0031] The Bacillus paramycoides SS6 inoculum is prepared by the following steps: inoculating the Bacillus paramycoides SS6 in an inoculum amount of 1% into an LB liquid culture medium, and culturing at 35° C. and 200 rpm; when the concentration of Bacillus paramycoides SS6 is 1×10 9 / mL, stop culturing and obtain the bacterial agent.

[0032] Example 3

[0033] A method for preparing palmitoleic acid comprises the following steps:

[0034] Adjust the sugarcane juice to 20 °Bx (20°C), sterilize, and ferment at 30°C using yeast EC1118, and add Bacillus paramycoides SS6 inoculant at 4% of the total fermentation liquid volume; when the sugar content no longer decreases, the fermentation ends;

[0035] Yeast EC1118 was activated by adding warm water three times at a dosage of 1 mg / L; then SS6 bacterial agent was added.

[0036] The Bacillus paramycoides SS6 inoculum is prepared by the following steps: inoculating the Bacillus paramycoides SS6 in an inoculum amount of 1% into an LB liquid culture medium, and culturing at 35° C. and 200 rpm; when the concentration of Bacillus paramycoides SS6 is 1×10 9 / mL, stop culturing and obtain the bacterial agent.

[0037] Example 4

[0038] The sugar content of sugarcane juice was adjusted to 20 °Bx (20℃), and after sterilization, Bacillus paramycoides SS6 inoculant was added according to 2% of the total fermentation liquid volume. The fermentation temperature was 30℃. Fermentation ended when the sugar content no longer decreased. The fermentation ended about 3-4 days earlier than the fermentation group containing yeast EC1118 (the sugar content of the fermentation liquid with yeast EC1118 stopped decreasing after about 10 days, and the sugar content of the fermentation liquid without yeast EC1118 stopped decreasing after about 7 days).

[0039] The Bacillus paramycoides SS6 inoculum is prepared by the following steps: inoculating the Bacillus paramycoides SS6 in an inoculum amount of 1% into an LB liquid culture medium, and culturing at 35° C. and 200 rpm; when the concentration of Bacillus paramycoides SS6 is 1×10 9 / mL, stop culturing and obtain the bacterial agent.

[0040] Comparative Example 1: No SS6 inoculant was added to assist fermentation

[0041] A method for preparing palmitoleic acid comprises the following steps:

[0042] S1 adjusts the sugar content of sugarcane juice to 20 °Bx (20℃). After sterilization, only yeast EC1118 is used for fermentation. Yeast EC1118 is added at a dosage of 1 mg / L and activated by adding warm water three times. The fermentation temperature is 30℃. Fermentation ends when the sugar content no longer decreases.

[0043] Implementation effect example

[0044] The fermentation broths of Examples 1-4 and Comparative Example 1 were taken out and analyzed. The contents of β-damascone, phenylethyl alcohol and palmitoleic acid were compared in Table 1.

[0045] Table 1 Product analysis of examples and comparative examples (μg / L)

[0046]

[0047] It should be noted that the embodiments described above are only used to explain the present invention and do not constitute any limitation to the present invention. The present invention has been described with reference to typical embodiments, but it should be understood that the words used therein are descriptive and explanatory words, rather than restrictive words. The present invention may be modified as specified within the scope of the claims of the present invention, and the present invention may be revised without departing from the scope and spirit of the present invention. Although the present invention described therein relates to specific methods, materials and embodiments, it does not mean that the present invention is limited to the specific examples disclosed therein, on the contrary, the present invention can be extended to all other methods and applications with the same functions.

Claims

1. A strain of Bacillus paramycoides, characterized in that: The paramycoides Bacillus is paramycoides Bacillus paramycoides , deposited in the China General Microbiological Culture Collection on October 28, 2024, with the deposit number: CGMCC No.32390; the address of the deposit unit: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

2. The use of Bacillus paramycoides according to claim 1 in the preparation of palmitoleic acid, characterized in that: Palmitoleic acid is prepared by using the Bacillus paramycoides described in claim 1 and adding yeast EC1118 to ferment sugarcane juice.

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