Nile strain JL1B1071 for producing aryl polyene and application of Nile strain JL1B1071

Abstract

The invention relates to the technical field of microorganisms, in particular to a Niesia strain JL1B1071 for producing aryl polyene and application of the Niesia strain JL1B1071. The preservation number of the bacterial strain JL1B1071 is as follows: GDMCC (China General Microbiological Culture Collection Center) No.64459. According to the invention, a Niele strain JL1B1071 is separated from the surface of spacecraft equipment, and the strain has adaptive capacity to extreme space environment. Microbial genomics and metabolic pathway analysis show that the strain JL1B1071 can be used for generating bacterial pigment aryl polyene, and the discovery of the strain has important value in the field of microorganisms.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a strain JL1B1071 of the genus Neel for producing aromatic polyenes and an application thereof. Background Art

[0002] Niallia microorganisms belong to the Bacillaceae family and are a class of Gram-positive, aerobic, rod-shaped bacteria. Niallia microorganisms are considered to be one of the most widely used genera in the Bacillaceae family and one of the microorganisms with the greatest biotechnological potential, producing a range of enzymes, antimicrobial metabolites and bioactive antioxidant compounds. The model species of this genus is Niallia cyclophane Niallia circulans , is the main industrial producer of β-galactosidase. At the same time, microorganisms of this genus are considered to have good application prospects in the fields of lignin hydrolysis, biofertilizer, and bioremediation.

[0003] Secondary metabolites of microorganisms are often active compounds with potentially advantageous properties. Arylpolyene is a class of abundant bacterial natural products that function similarly to antioxidant carotenoids and can protect bacteria from photooxidative damage and lipid peroxidation. Bacterial natural products with such properties play an important role in the development of bioactive molecules related to human health, such as effective UV protection, prevention or treatment of degenerative diseases caused by oxidative stress, and antagonism against food pathogens. Summary of the invention

[0004] In order to solve the problems existing in the prior art, the present invention provides a strain of the genus Neel JL1B1071 for producing aromatic polyenes and application thereof.

[0005] The strain JL1B1071 was isolated from the surface of a Chinese spacecraft and has the ability to adapt to the extreme space environment. Through microbial genomics and metabolic pathway analysis, the present invention found that the patented strain JL1B1071 can be used as a strain for the production of bacterial pigment aromatic polyenes, and the synthesis of this natural product is also a potential mechanism for the strain to adapt to the space environment. The discovery of this strain has important technical, economic and social value.

[0006] The present invention has made a biological deposit for the strain JL1B1071, and the deposit information is as follows: Deposit number GDMCC No: 64459; Classification name: Niallia sp.; Depository unit: Guangdong Microbial Culture Collection Center; Deposit address: Institute of Microbiology, Guangdong Academy of Sciences, 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou; Deposit date: March 28, 2024.

[0007] In a first aspect, the present invention provides a bacterial agent, which includes the strain JL1B1071 or its fermentation product.

[0008] The fermentation product of the present invention can be the supernatant obtained after the strain JL1B1071 is fermented, or it can be the whole fermentation product (including the strain JL1B1071).

[0009] Furthermore, the bacterial agent is a solid bacterial agent, a liquid bacterial agent or a microbial bacterial agent, and the total viable count of the strain JL1B1071 in the bacterial agent is 1×10 7-10 cfu / g.

[0010] In a second aspect, the present invention provides the strain JL1B1071 or the use of the bacterial agent in the production of aromatic polyenes, terpenes or ribosomal peptide analogs.

[0011] Furthermore, the strain JL1B1071 synthesizes aromatic polyenes through genes JL1B1071GL003657, JL1B1071GL003664 and JL1B1071GL003666.

[0012] The present invention provides the strain JL1B1071 or the use of the bacterial agent in preparing antioxidant products.

[0013] Furthermore, the product is a medicine, an antibacterial agent, a food or a cosmetic.

[0014] The present invention has the following beneficial effects: The present invention screened and obtained a strain of the genus Neele ( Niallia sp. ) strain JL1B1071, which is identified as a new species. The strain JL1B1071 screened by the present invention has the ability to produce aromatic polyenes, terpenes or ribosomal peptide analogs, and can be used in the preparation of functional products (such as food, cosmetics or medicines). The strain JL1B1071 provided by the present invention is a new source of biologically active compounds and has important application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0016] Figure 1 This is the phylogenetic tree of strain JL1B1071 provided in Example 1 of the present invention.

[0017] Figure 2 This is the location map of the aromatic polyene biosynthesis gene cluster of strain JL1B1071 provided in Example 1 of the present invention. DETAILED DESCRIPTION

[0018] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the drawings of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0019] The experimental methods involved in the following examples, unless otherwise mentioned, are all conventional methods in the art, for example, reference may be made to experimental manuals in the art, or the conditions recommended in the manufacturer's instructions.

[0020] Unless otherwise specified, the experimental materials and reagents involved in the following examples can be obtained from commercial sources.

[0021] Example 1

[0022] The present invention screened and obtained a strain of the genus Neel JL1B1071, which includes the following process: 1. Separation

[0023] Strain JL1B1071 was isolated from the surface of spacecraft equipment. Microorganisms were collected using pre-moistened sterile wipes, which were then aseptically transferred into 200 mL of elution solution. Ultrasonic elution was performed, and then the bacteria were cultured overnight by pouring the plate. The culture medium was TSA medium at 37°C. The single colony produced was picked and streaked to pure culture. Microscopic examination results showed that the cells of strain JL1B1071 were Gram-positive, aerobic, rod-shaped, and could produce endospores with a size of (0.3-0.8) μm×(1.5-5.6) μm. After 24 hours of culture on TSA medium, the strain presented white colonies with a smooth surface.

[0024] 2. Identification

[0025] The present invention uses 16SrRNA gene primer 27F / 1492R for identification. The software CLUSTALW is used to compare the strain JL1B1071 and other closely related species. 16S rRNA Gene sequence; phylogenetic tree of strains was constructed using MEGA11 software. Bootstrap was used for support rate verification during construction, and the parameter was set to bootstrap=1000.

[0026] The 16sRNA sequence of strain JL1B1071 (SEQ ID NO.1) is as follows:

[0027] The results are as follows Figure 1 As shown, the phylogenetic tree shows that strain JL1B1071 is a separate branch and is a new species of microorganism.

[0028] 3. Genomic characteristics

[0029] The genome sequence of strain JL1B1071 is 5166230bp in length, with a G+C content of 35.63mol%. The results of gene composition prediction show that strain JL1B1071 contains 5020 genes, 33 rRNAs, 83 tRNAs, 26 sRNAs, and 4 CRISPR elements. The functional annotation of the genome of strain JL1B1071 shows that this strain is a new source of bioactive compounds, including industrial enzymes, antibiotics, antibacterial agents, etc.

[0030] 4. Analysis of secondary metabolites of strains

[0031] The present invention predicts the secondary metabolite synthesis gene cluster in the genome of strain JL1B1071 by antiSMASH (v7.1.0) software and 2nd find online software, and as a result, a total of 3 secondary metabolite gene clusters are annotated, namely: aromatic polyene synthesis gene cluster, terpene synthesis gene cluster, and ribosomal peptide analog synthesis gene cluster. Among them, the aromatic polyene biosynthesis gene cluster has the highest similarity with the published gene cluster, indicating that the main secondary metabolites of strain JL1B1071 are aromatic polyenes and terpenes.

[0032] The present invention further compares the composition of secondary metabolic biosynthetic gene clusters of strain JL1B1071 and its closest relative, N. cyclophila. The results show that the main secondary metabolites of N. cyclophila are terpenes, lasso peptides and lanthipeptides. Compared with N. cyclophila, strain JL1B1071 has undergone significant differentiation in the synthesis of secondary metabolites, and the synthesis of aromatic polyene compounds is its main feature, indicating that strain JL1B1071 is a microbial strain with unique genetic characteristics.

[0033] Table 1 Secondary metabolism gene clusters of strain JL1B1071 and its related bacteria

[0034] “√” indicates that the genome has the corresponding biosynthetic gene cluster.

[0035] 5. Aryl polyene biosynthesis gene cluster of strain JL1B1071

[0036] Aromatic polyenes are a special type of polyunsaturated carboxylic acid, which is the expression product of biosynthetic gene clusters in bacteria. APEs are present in most bacteria throughout the Proteobacteria and Bacteroidetes. Recent studies have classified metabolites containing the core structure of aromatic polyene chains into a family of aromatic polyene metabolites, including xanthomonas produced by Xanthomonas and daunorubicin produced by Flavobacterium, which have relatively conserved biosynthetic gene clusters.

[0037] The aryl polyene biosynthesis gene cluster of strain JL1B1071 is located at 3712253-3753404 of the chromosomal DNA, containing a total of 41151 bp, including 33 genes (e.g. Figure 2 BLAST was used to compare and annotate each gene in the NCBI database. The specific annotation results are shown in Table 2.

[0038] Table 2 Gene composition and function prediction of aromatic polyene biosynthesis gene cluster

[0039] Table 3 Gene composition and function prediction of aromatic polyene biosynthesis gene cluster

[0040] Among the 33 genes, JL1B1071GL003657, JL1B1071GL003664, and JL1B1071GL003666 encode acyltransferase, acyl carrier protein, and acylreductase, respectively, and are essential genes for strain JL1B1071 to synthesize aromatic polyene compounds. Among them, the JL1B1071GL003664 gene is involved in the synthesis of aromatic PKS (polyketide synthase) and contains a ketone synthase domain, which mainly synthesizes aromatic compounds. Phylogeny shows that the synthesis of aromatic polyenes is similar to the biosynthesis mechanism of fatty acids and type II polyketides. The JL1B1071GL003664 gene encodes the N-terminal domain protein of β-ketoacyl synthase, which participates in the polymerization reaction in acyl chain elongation and plays an important role in the synthesis pathway of natural products such as polyketides. The JL1B1071GL003666 gene is also highly involved in the synthesis of aromatic PKS (polyketide synthase) and contains an enoyl reductase domain, which is responsible for catalyzing the final reduction reaction of fatty acids, reducing enoyl-ACP to saturated acyl-ACP. Under certain circumstances, the enzyme can also catalyze the alkylation reaction of the substrate.

[0041] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A species of Neelid Niallia sp. ) strain JL1B1071, characterized in that The deposit number of the strain JL1B1071 is: GDMCC No: 64459.

2. A bacterial agent, characterized in that The invention comprises the strain JL1B1071 or a fermentation product thereof according to claim 1.

3. The bacterial agent according to claim 2, characterized in that The bacterial agent is a solid bacterial agent, a liquid bacterial agent or a microbial bacterial agent. In the bacterial agent, the total viable count of the strain JL1B1071 is 1×10 7-10 cfu / g.

4. Use of the strain JL1B1071 according to claim 1, or the bacterial agent according to claim 2 or 3 in the production of aromatic polyenes, terpenes or ribosomal peptide analogs.

5. The use according to claim 4, characterized in that: The strain JL1B1071 synthesizes aromatic polyenes through genes JL1B1071GL003657, JL1B1071GL003664 and JL1B1071GL003666.

6. Use of the strain JL1B1071 according to claim 1, or the bacterial agent according to claim 2 or 3 in the preparation of antioxidant products.

7. The use according to claim 6, characterized in that: The product is a medicine, an antibacterial agent, a food or a cosmetic.

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