Strain metabolism to obtain compounds and preparation and application thereof

Abstract

The present application relates to the fields of biological medicine and pesticide, and particularly relates to a compound (hexadecyl macrocyclic lactone and peniorcinol compound) obtained by metabolism of a strain, and preparation and application thereof.The compound is a hexadecyl macrocyclic lactone compound with a theta-keto-alpha, beta, gamma, delta-unsaturated lactone structure fragment as shown in compound 1, and a peniorcinol compound as shown in compound 2.The secondary metabolite of the fungus has hypoglycemic, seed germination promoting or antioxidant activity, and can be used as a lead compound for hypoglycemic, seed germination promoting or antioxidant activity.The compound can be produced by fermentation of the fungus, and the preparation method is simple, efficient, and the obtained product has high purity.The present application has good application prospect.

Description

Technical Field

[0001] This invention relates to the fields of biomedicine and pesticides, specifically to compounds (sixteen-membered macrocyclic lactones and peniorcinol compounds) obtained by the metabolism of a bacterial strain, their preparation and application. Background Technology

[0002] Diabetes mellitus is a chronic, common metabolic disease, classified into type 1 and type 2 diabetes. Type 2 diabetes (T2DM), also known as non-insulin-dependent diabetes mellitus, accounts for 90% of all diabetes cases worldwide. The World Health Organization reports that over 400 million people have diabetes, a number projected to exceed 592 million by 2035. Alpha-glucosidase is a key target enzyme associated with T2DM. After a meal, alpha-glucosidase, present on the brush border of the small intestinal mucosa, hydrolyzes carbohydrates in food into glucose. Glucose is absorbed into the bloodstream, raising blood glucose levels. Therefore, alpha-glucosidase is one of the main target enzymes controlling postprandial blood glucose. Alpha-glucosidase inhibitors reversibly inhibit small intestinal alpha-glucosidase, reducing carbohydrate hydrolysis, delaying glucose production and absorption, and thus lowering postprandial blood glucose levels. They are symptomatic treatments for controlling postprandial blood glucose. To date, there are three main naturally occurring alpha-glucosidase inhibitors: voglibose, miglitol, and acarbose.

[0003] In recent years, with the rapid development of the free radical theory, aging and the occurrence of various diseases have been proven to be closely related to the excessive production of free radicals. Supplementing the body with exogenous antioxidants or administering compounds that help restore the body's endogenous antioxidants plays an auxiliary role in the treatment of aging and some diseases. Traditional antioxidants are generally derived from chemical synthesis, which has disadvantages such as poor stability and strong toxic side effects. Natural antioxidants, on the other hand, have advantages such as safety, non-toxicity, high stability, and naturalness, making them more in line with consumer requirements, and some have already been well developed and utilized. Therefore, the research and development of natural antioxidants has become a research hotspot in the food, traditional Chinese medicine, and cosmetic fields.

[0004] Seed germination is the most important stage in the plant life cycle, and in agricultural production, rapid and uniform seed germination is also a necessary condition for high yields. The vast diversity of plants has shaped a rich and unique diversity of endophytic fungi, which produce a wide variety of secondary metabolites that promote seed germination. Summary of the Invention

[0005] The purpose of this invention is to provide a compound (a sixteen-membered macrocyclic lactone and peniorcinol-like compound) obtained by bacterial metabolism, as well as its preparation and application.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] A strain of bacteria metabolized to obtain compounds, namely, compound 1, a sixteen-membered macrocyclic lactone compound with an θ-keto-α,β,γ,δ-unsaturated lactone structural fragment, and compound 2, a peniorcinol-like compound.

[0008]

[0009] A method for preparing the compound obtained by the metabolism of the strain described above, wherein the compound is obtained by separating and purifying the fermentation broth of plant endophytic fungus F4a after cultivation;

[0010] The plant endophytic fungus F4a is a strain with accession number CCTCC NO: M 2012531, deposited at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, China, on December 18, 2012, and its Latin name is Eupenicillium brefeldianum F4a.

[0011] Specifically:

[0012] 1) Plant endophytic fungus F4a was inoculated on PSA solid medium and cultured for 48-96 h to activate it. After activation, it was inoculated on liquid medium and cultured at 28-32℃ for 48-72 h. The resulting culture was used as seed culture. The seed culture was mixed with liquid medium at a volume ratio of 1:27-1:13 and cultured at 26-30℃ with shaking for 7-9 days.

[0013] 2) Centrifuge the fermentation broth to remove the bacterial cells and obtain the fermentation supernatant. Use HP20 macroporous adsorption resin to adsorb the active ingredients in the fermentation broth on a solid phase, with a resin-to-fermentation broth volume ratio of 1:14-1:26. Adsorb for 1-3 hours, and wash after adsorption to obtain crude extract. Extract the separated bacterial cells with acetone to obtain crude extract. Combine the crude extracts to obtain total extract A. Obtain the compound by separation and purification of total extract A.

[0014] To elaborate further,

[0015] 1) Plant endophytic fungus F4a was inoculated on PSA solid medium and cultured for 48-96 h to activate it. After activation, it was inoculated on liquid medium and cultured at 28-32℃ for 48-72 h. The resulting culture was used as seed culture. The seed culture was mixed with liquid medium at a volume ratio of 1:27-1:13 and cultured at 26-30℃ with shaking for 7-10 days.

[0016] 2) Centrifuge the fermentation broth to remove the bacterial cells and obtain the fermentation supernatant. Use HP20 macroporous adsorption resin to adsorb the active ingredients in the fermentation broth on the solid phase. The volume ratio of resin to fermentation broth is 1:14-1:26. Adsorb for 1-3 hours. Wash after adsorption to obtain crude extract. The separated bacterial cells are extracted with acetone to obtain crude extract. Combine the crude extracts to obtain total extract A.

[0017] 3) The total extract A was separated by silica gel column chromatography, eluted with a gradient of dichloromethane:methanol (v / v) ratio of 100:0-0:100, and each component was collected to obtain the target secondary metabolite enrichment.

[0018] 4) The above-mentioned enriched product was separated by silica gel column chromatography, and eluted with pure dichloromethane to obtain fraction F. a Dichloromethane:methanol (v / v) ratio of 100:1-100:3 was used as the elution fraction F. b Dichloromethane:methanol (v / v) ratio of 100:4-100:6 was eluted to fraction F. c Dichloromethane:methanol (v / v) ratio of 100:7-10:9 was eluted to fraction F. d Dichloromethane:methanol (v / v) ratio of 100:10-100:15 eluted to fraction F e .

[0019] Flow segment F b Separated by LH2O gel electrophoresis, the fraction was divided into four subfractions according to the volume of methanol eluted. 50-70 mL of methanol was collected as subfraction F according to the elution order. b4 Receive 80-120 mL of methanol as sub-fraction F b3 Receive 120-140 mL of methanol as sub-fluid F b2 Receive 145-160 mL of methanol as sub-fluid F b1 Sub-flow F b4 After ODS reversed-phase column chromatography and methanol-water gradient separation, a total of 4 subfractions (F) were obtained. b4a -F b4d ), of which 30-40% methanol eluted with water is F. b4a The product eluted with 42-45% methanol and water is F. b4b F was eluted with 46-50% methanol and water. b4c F was obtained by elution with 52-55% methanol and water. b4d F b4d Compounds 1-2 were obtained by semi-preparative HPLC purification, eluting with 50-60% methanol aqueous solution at a flow rate of 2.0 mL / min.

[0020] The liquid culture medium consists of 12-14g starch, 18-22g glucose, 0.5-1.5g yeast extract, 0.3-0.7g soybean meal, 2-3g barley extract, 0.5-1.5g ammonium sulfate, 0.5-1.1g potassium nitrate, 3-5g calcium carbonate, 2-3g magnesium sulfate heptahydrate, 2-4g potassium dihydrogen phosphate, and 0.01-0.03g copper sulfate per liter of water, with a pH of 6.5-7.5.

[0021] An application of the compound obtained by the metabolic metabolism of the strain, wherein the sixteen-membered macrocyclic lactone compound having the θ-keto-α,β,γ,δ-unsaturated lactone structural fragment shown in compound 1 obtained by the metabolic metabolism of the strain is used in the preparation of hypoglycemic agents.

[0022] An application of the compound obtained by the metabolic metabolism of the strain, wherein the sixteen-membered macrocyclic lactone compound having the θ-keto-α,β,γ,δ-unsaturated lactone structural fragment shown in compound 1 obtained by the metabolic metabolism of the strain is used in the preparation of a seed germination promoter.

[0023] An application of the compound obtained by the metabolic process of the strain, specifically the application of the peniocinol-like compound represented by compound 2 in the preparation of antioxidants.

[0024] Advantages of this invention:

[0025] This invention provides a sixteen-membered macrocyclic lactone having an unreported θ-keto-α,β,γ,δ-unsaturated lactone structural fragment and a previously unreported peniorcinol compound.

[0026] The sixteen-membered macrocyclic lactones and peniorcinol compounds described in this invention are new compounds obtained through fungal fermentation and specific separation processes.

[0027] The sixteen-membered macrocyclic lactone compounds obtained in this invention possess α-glucosidase inhibitory activity and seed germination promoting activity, while peniorcinol compounds exhibit ABTS comparable to the positive control drug vitamin C. +· With their scavenging activity, they can serve as ideal candidate compounds for lowering blood sugar, promoting seed germination, or acting as antioxidants. Attached Figure Description

[0028] Figure 1 Compounds 1-2 provided in the embodiments of the present invention 1 H- 1 The important correlation between H COSY and HMBC.

[0029] Figure 2 NMR calculations and DP4+ analysis of compound 1 provided in the embodiments of the present invention.

[0030] Figure 3 The experimental ECD spectrum and calculated ECD spectrum of compound 1 in CH3OH provided in the embodiments of the present invention. Detailed Implementation

[0031] To better understand the content of this invention, further explanation is provided below with reference to specific embodiments, but the scope of protection of this patent is not limited thereto.

[0032] Example 1: Preparation method of sixteen-membered macrocyclic lactones and peniorcinol compounds by fermentation of plant endophytic fungus F4a

[0033] The liquid culture medium used was as follows: 10g starch, 20g glucose, 1g yeast extract, 0.5g soybean meal, 2.5g barley extract, 1g ammonium sulfate, 0.8g potassium nitrate, 4g calcium carbonate, 2.5g magnesium sulfate heptahydrate, 3g potassium dihydrogen phosphate, 0.02g copper sulfate, pH 7.0.

[0034] Plant endophytic fungus F4a (CCTCCM 2012531) was inoculated onto PSA solid medium and cultured for 48 h to activate it. Then, it was inoculated onto the aforementioned liquid medium and cultured at 28°C for 72 h to obtain the seed culture. The seed culture obtained from fermentation was then inoculated onto the same fermentation medium at a volume ratio of 1:18 and cultured at 28°C with shaking for 7 days. The fermentation broth was centrifuged to remove the bacterial cells, obtaining the fermentation supernatant. Macroporous adsorption resin HP20 was added to the fermentation broth for solid-phase adsorption of the active ingredients, with a resin-to-fermentation broth volume ratio of 1:25. After adsorption for 2 h, the resin was washed successively with distilled water and 15% methanol aqueous solution, followed by desorption with methanol. The crude extract was obtained after recovering the methanol solvent. The bacterial cells were sonicated with acetone for 30 min, and the acetone solvent was recovered to obtain the crude extract. The two crude extracts were combined to obtain total extract A.

[0035] Extract A was separated by rapid silica gel column chromatography, using a gradient elution of dichloromethane:methanol (v / v) at a ratio of 100:0 to 0:100, to obtain 5 fractions (F... a -F e ), in which pure dichloromethane was eluted to fraction F. a Dichloromethane:methanol (v / v) ratio of 100:1-100:3 was used as the elution fraction F. b Dichloromethane:methanol (v / v) ratio of 100:4-100:6 is the flow fraction F. c The dichloromethane:methanol (v / v) ratio is 100:7-10:9. The flow fraction F... d Dichloromethane:methanol (v / v) ratio of 100:10-100:15 is the fraction F. e .

[0036] The above collection flow is divided into F b Separated by LH2O gel electrophoresis, the fraction was divided into four subfractions according to the volume of methanol eluted. 40-60 mL of methanol was collected as subfraction F according to the elution order. b4 Receive 62-100 mL of methanol as sub-fraction F b3Receive 102-120 mL of methanol as sub-fluid F b2 Receive 125-140 mL of methanol as sub-fluid F b1 Sub-flow F b4 After ODS reversed-phase column chromatography and methanol-water gradient separation, a total of 4 subfractions (F) were obtained. b4a -F b4d ), of which 30-40% methanol eluted with water is F. b4a The product eluted with 42-45% methanol and water is F. b4b F was eluted with 46-50% methanol and water. b4c F was obtained by elution with 52-55% methanol and water. b4d F b4d Penidione A (1, 4.3 mg, t) was obtained by semi-preparative HPLC purification, eluting with 50-60% methanol-water solution at a flow rate of 2.0 mL / min. R =21.8min) and peniorcinol D (2, 3.3mg, t) R =19.8min).

[0037] The above provides structural analyses of sixteen-membered macrocyclic lactones and peniorcinol compounds (see Table 1 and...). Figure 1-3 ):

[0038] Compound 1 is a white powder, and HRESIMS data show an m / z of 303.1560 [M+Na]. + (calcd forC 16 H 24 O4Na, 303.1572), its molecular formula is presumed to be C 16 H 24 O4 has an unsaturation degree of 5. 1 The 1H NMR (600MHz, DMSO-d6) spectrum showed the presence of four olefinic proton signals δ. H The two oxygen-containing methine proton signals δ were 7.03 (1H, dd, J = 15.4, 11.3 Hz), 6.37 (1H, ddd, J = 15.2, 11.3, 1.3 Hz), 5.81 (1H, d, J = 15.4 Hz), and 5.77 (1H, dd, J = 15.2, 3.7 Hz). H 4.30 (1H, m) and 4.84 (1H, m), a set of aliphatic methyl proton signals δ H 1.20 (3H, d, J = 6.4Hz) (Table 1). In 13The C10 NMR (150 MHz, DMSO-d6) and HSQC spectra showed the presence of one ketone carbonyl group, one ester carbonyl group, four olefinic hydrogen carbons, two oxygen-containing methine carbons, seven methylene carbons, and one methyl carbon (Table 1). Based on the unsaturation degree of 5 of compound 1, sp... 2 Besides the hybridized alkene-hydrocarbon group (Ω=2), a ketone carbonyl group (Ω=1), and an ester carbonyl group (Ω=1), compound 1 has one remaining degree of unsaturation, thus indicating that it is a sixteen-membered macrocyclic lactone. 1 H- 1 The correlation signals of H-2 / H-3 / H-4 / H-5 / H-6 / H-7 / H-8 in H COSY and the long-range correlations of H-2 and H-3 with C-1 in the HMBC spectrum suggest that compound 1 contains a 6-hydroxy-2,4-dioctenic acid fragment. Figure 1 ).Depend on 1 H- 1 The H-10 / H-11 / H-12 / H-13 / H-14 / H-15 / H-16 related signals in H COSY suggest the presence of a 2-heptanol fragment. Figure 1 The HMBC spectrum shows that H-7, H-8, and H-10 exhibit long-range correlation signals with C-9, and H-15 exhibits a long-range correlation signal with C-1. It is speculated that these two fragments are linked by an ester bond and a ketone carbonyl group. Figure 1 The coupling constants of the two double bonds are 15.4 Hz and 15.2 Hz, respectively, suggesting that both double bonds are in the trans configuration.

[0039] To determine the relative configuration of compound 1, NMR calculations were performed in conjunction with DP4+ analysis. Figure 2 Two configurations were calculated: (2E, 4E, 6S*, 15R*)-1a and (2E, 4E, 6S*, 15S*)-1b. Combined with DP4+ analysis, the relative configuration of compound 1 is 1a (99.99%). Figure 2 By fitting the calculated ECD curves to the experimental ECD curves, the absolute configuration of compound 1 was determined to be 2E, 4E, 6S, 15R. Figure 3 Therefore, compound 1 can be identified as a previously unreported sixteen-membered macrocyclic lactone with an θ-keto-α,β,γ,δ-unsaturated lactone structural fragment, named penidione A.

[0040] Compound 2 is a white powder, and HRESIMS data show an m / z of 247.1325 [M+H]. + (calcd forC 15 H 19 O3 (247.1334), its molecular formula is inferred to be C. 15 H18 O3, with an unsaturation degree of 7. Combined with 1D NMR and HSQC spectral analysis (Table 1), compound 2 shares the same 4-hydroxy-2-methoxy-6-methylbenzyl fragment as peniorcinols AC (J. Fungi 2021, 7, 913). 1 H- 1 The presence of correlation signals between H-10 / H-11 in the H COSY spectrum, and the long-range correlation signals between H-11 and C-8, C-9, and C-12 in the HMBC spectrum, along with the long-range correlation signals between H-13 and C-11 and C-12, suggest that compound 2 contains a 3-methylcyclopent-2-en-1-one fragment. The HMBC spectrum also shows long-range correlation signals between H-7 and C-2, C-9, and C-12, indicating that these two fragments are linked by a methylene group. Figure 1 Therefore, compound 2 can be identified as a previously unreported peniorcinol class compound, named peniorcinol D.

[0041] Table 1. 1H and 1C spectra of compounds 1-2 (DMSO-d6, 600 / 150MHz, δin ppm)

[0042]

[0043]

[0044] Example 2: α-glucosidase inhibitory activity of compounds 1-2

[0045] Alpha-glucosidase plays a crucial role in food absorption; food can only be digested and absorbed after binding with it. The hypoglycemic mechanism of alpha-glucosidase inhibitors is to inhibit alpha-glucosidase on the intestinal mucosa, slowing the rate at which starch is broken down into glucose, reducing and delaying glucose absorption in the small intestine, thereby lowering blood glucose levels. This effect is particularly pronounced in treating postprandial hyperglycemia.

[0046] Assay method: Accurately pipette 20 μL of each of the test sample series solutions, add 30 μL of α-glucosidase solution, then add 800 μL of 0.1 M phosphate buffer solution (pH 6.8), and heat at 37 °C for 5 min; add 150 μL of pNPG solution (10 mM), mix well, and heat at 37 °C for 30 min; stop the reaction by adding 650 μL of Na2CO3 stop solution (1 M), and measure the absorbance at 405 nm using an enzyme-linked immunosorbent assay (ELISA) reader. The positive control is acarbose, and the negative control is 20 μL of DMSO. Each test sample (compounds 1-2, positive control, and negative control) should be measured three times.

[0047] α-glucosidase inhibition rate % = (1-A samp / A cont )×100%, where A samp A represents the absorbance value of the test solution. cont The absorbance value represents the negative control solution; the final experimental results are expressed as EC. 50 Value (μM) is used to represent this.

[0048] Experimental Results: The results of the α-glucosidase inhibition experiments of compounds 1-2 are shown in Table 2. The results indicate that only compound 1 showed inhibitory effects compared to the positive control acarbose (EC). 50 Compared to 2.5 μM, it exhibits weak α-glucosidase inhibitory activity (EC). 50 =52.3 μM). Notably, the hypoglycemic activity of a sixteen-membered macrocyclic lactone compound with the θ-keto-α,β,γ,δ-unsaturated lactone structural fragment is reported for the first time. Compound 2 did not exhibit α-glucosidase inhibitory activity.

[0049] Example 3 Antioxidant activity of compounds 1-2

[0050] There are many methods for evaluating and screening the antioxidant activity of substances in vitro. The 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl (DPPH) method and the 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)ammonium salt (ABTS) method have advantages such as good stability, high sensitivity, and simple operation. Both methods have been widely used worldwide for the quantitative analysis of free radical scavenging ability.

[0051] (1)DPPH · Scavenging activity

[0052] Assay method: Accurately pipette 100 μL of each of the above-mentioned test solutions and 100 μL of DPPH ethanol solution into a 96-well plate, mix well, react at room temperature in the dark for 30 min, then place in a microplate reader and measure the absorbance at 517 nm. Vitamin C is used as a positive control, and 100 μL of anhydrous ethanol is used as a negative control. Each test sample (compounds 1-2, positive control, and negative control) is measured three times. · The formula for calculating the clearance rate is as follows: Clearance rate % = (1 - A) samp / A cont )×100%, where Asamp A represents the absorbance value of the test solution. cont The absorbance value represents the negative control solution; the final experimental results are expressed as EC. 50 Value (μM) is used to represent this.

[0053] Experimental results: DPPH of compounds 1-2 · The results of the scavenging activity assay are shown in Table 2. The results indicate that neither compound 1 nor compound 2 exhibited DPPH. · Eliminate activity.

[0054] (2) ABTS +· Scavenging activity

[0055] Test method: Prepare ABTS + The solution was diluted approximately 32 times with 0.01M phosphate buffer. After dilution, it was incubated in the dark for 30 minutes (to ensure accurate absorbance values). The absorbance was measured using a UV spectrophotometer; a value within 0.7 ± 0.02 was considered acceptable and usable. Accurately pipette 100 μL of each of the above test solutions and ABTS. +· Add 3 mL of solution to a glass test tube, mix well, and react at room temperature in the dark for 6 min. Then, place the tube in a microplate reader and measure the absorbance at 734 nm. The positive control is vitamin C, and the negative control is 100 μL of anhydrous ethanol. Each test sample (compounds 1-2, positive control, and negative control) should be measured three times. (ABTS) +· The formula for calculating clearance rate is the same as that for DPPH. · The formula for calculating the clearance rate, and the final experimental results using EC 50 Value (μM) is used to represent this.

[0056] Experimental results: ABTS of compounds 1-2 +· The results of the scavenging activity experiment are shown in Table 2. The results indicate that compound 2 scavenges ABTS... +· Scavenging activity (EC) 50 =29.8μM) and positive control vitamin C (EC) 50 =22.3 μM) equivalent, compound 1 did not exhibit ABTS +· Eliminate activity.

[0057] Table 2. α-glucosidase inhibitory and antioxidant activities of compounds 1-2

[0058]

[0059] ND means no detection.

[0060] Example 4: Seed germination promoting activity of compound 1

[0061] First, monomer compound 1 and the positive control 3-indoleacetic acid were dissolved in methanol solution, and a 0.01 μM test solution (1% methanol) was prepared with sterile water. Next, pakchoi seeds and rice seeds were sterilized with 75% alcohol for 5 min and washed 5 times with distilled water. Then, the treated seeds (N = 30–60) were placed in glass petri dishes containing 9 cm cellulose filter paper, and 5 mL of the test solution was added to each dish. After incubation at 24°C in the dark for 48 h, the seed germination rate was calculated. Seeds were considered to have germinated when the radicle protruded more than 2 mm.

[0062] Seed germination experiments showed that compound 1, at a concentration of 0.01 μM, promoted the germination rate of pakchoi seeds by 7.4%, which was stronger than the positive control 3-indoleacetic acid (germination rate of 5.6%). Furthermore, compound 1, at a concentration of 0.01 μM, promoted the germination rate of rice seeds by 14.3%, the same as the positive control 3-indoleacetic acid. Therefore, compound 1 exhibits significant seed germination-promoting activity.

Claims

1. A compound obtained by the metabolism of a bacterial strain, characterized in that: The compound shown is compound 1, which has θ -keto- α , β , γ , δ - Sixteen-membered macrocyclic lactone compounds with unsaturated lactone structural fragments, and peniorcinol-like compounds as shown in compound 2. 。 2. A method for preparing a compound obtained by the metabolism of the strain according to claim 1, characterized in that: The compound of claim 1 was obtained by separating and purifying the fermentation broth of plant endophytic fungus F4a after cultivation; The plant endophytic fungus F4a is a strain with the preservation number CCTCC NO: M 2012531; The specific steps are as follows: 1) Plant endophytic fungus F4a was inoculated on PSA solid medium and cultured for 48-96 h to activate it. After activation, it was inoculated on liquid medium and cultured at 28-32°C for 48-72 h. The resulting culture was used as seed culture. The seed culture was mixed with liquid medium at a volume ratio of 1:27-1:13 and cultured at 26-30°C with shaking for 7-9 days. 2) Centrifuge the fermentation broth to remove bacterial cells and obtain the fermentation supernatant. Use HP20 macroporous adsorption resin to adsorb the active ingredients in the fermentation broth on a solid phase, with a resin-to-fermentation broth volume ratio of 1:14-1:

26. Adsorb for 1-3 h, and wash after adsorption to obtain a crude extract. Extract the separated bacterial cells with acetone to obtain a crude extract. Combine the crude extracts to obtain total extract A. Separate and purify total extract A to obtain the compound described in claim 1. The liquid culture medium consists of 12-14 g starch, 18-22 g glucose, 0.5-1.5 g yeast extract, 0.3-0.7 g soybean meal, 2-3 g barley extract, 0.5-1.5 g ammonium sulfate, 0.5-1.1 g potassium nitrate, 3-5 g calcium carbonate, 2-3 g magnesium sulfate heptahydrate, 2-4 g potassium dihydrogen phosphate, and 0.01-0.03 g copper sulfate per liter of water, with a pH of 6.5-7.

5.

3. The application of the compound obtained by the strain according to claim 1 through metabolism, characterized in that: The strain metabolized to obtain compound 1, which has the characteristics shown in compound 1. θ -keto- α , β , γ , δ Application of sixteen-membered macrocyclic lactone compounds with unsaturated lactone structural fragments in the preparation of hypoglycemic agents.

4. The application of the compound obtained by the strain according to claim 1 through metabolism, characterized in that: The strain metabolized to obtain compound 1, which has the characteristics shown in compound 1. θ -keto- α , β , γ , δ Application of sixteen-membered macrocyclic lactone compounds with unsaturated lactone structural fragments in the preparation of seed germination promoters.

5. The application of the compound obtained by the strain according to claim 1 through metabolism, characterized in that: The application of the peniocinol-like compounds, represented by compound 2, obtained by the metabolism of the strain in the preparation of antioxidants.

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