CN111494355A

CN111494355A - Preparation method and application of Xanthocillin compound

Info

Publication number: CN111494355A
Application number: CN202010265252.2A
Authority: CN
Inventors: 张长生; 张海波; 伊穆然可汗; 彭方; 张丽萍; 张庆波; 刘威; 张光涛
Original assignee: South China Sea Institute of Oceanology of CAS
Current assignee: Wuhan University WHU; South China Sea Institute of Oceanology of CAS; Southern Marine Science and Engineering Guangdong Laboratory Guangzhou
Priority date: 2020-04-07
Filing date: 2020-04-07
Publication date: 2020-08-07
Anticipated expiration: 2040-04-07
Also published as: CN111494355B

Abstract

The invention discloses a preparation method and application of a Xanthocillin compound. The application of the compound 1 or the compound 2 shown in the formula I in the preparation of antibacterial drugs or tyrosinase inhibitors. The compound 1 and the compound 2 with antibacterial and tyrosinase inhibitory activities are separated from Penicillium chrysogenum Y-S6-1 CCTCC M2020019, so that the compounds can be applied to antibacterial drugs and tyrosinase inhibitors.

Description

Preparation method and application of Xanthocillin compound

Technical Field

The invention relates to the technical field of biology, in particular to a preparation method and application of a Xanthocillin compound.

Background

Gram-negative bacteria generally refer to bacteria that show a red gram stain, and are represented by Escherichia coli. Gram-positive bacteria generally refer to bacteria that exhibit purple gram staining, as typified by staphylococcus aureus. Gram-negative bacteria have an outer membrane (outer wall) consisting of lipopolysaccharide, phospholipid, lipoprotein and the like outside a cell membrane, and the lipopolysaccharide can prevent larger molecules such as lysozyme, antibiotics (penicillin and the like), detergents, certain dyes and the like from entering the cell membrane, so that most antibiotics can not effectively treat gram-negative bacterial infection. The data statistics of china CHINET for various hospitals in 2014 show that: the pathogenic bacteria responsible for patient infection are primarily drug resistant gram negative bacteria. Among 78955 pathogenic bacteria isolated from clinical specimens, 57320 gram-negative bacteria were found, accounting for 72.6% of the total isolated strains. Clinically common gram-negative pathogenic bacteria include Escherichia coli (Escherichia coli), Acinetobacter baumannii (Acinetobacter baumannii), Pseudomonas aeruginosa (Pseudomonas aeruginosa) and the like, and gram-positive pathogenic bacteria include Staphylococcus aureus (MRSA). In the aspect of aquaculture, gram-negative bacteria Vibrio alginolyticus (Vibrio algicidalus) are common pathogenic bacteria of marine culture animals, and shell animals such as cultured shellfish and shrimps are more easily infected with the bacteria than cultured fishes. The vibrio alginolyticus is pathogenic bacteria causing the black gill and brown spot syndrome of the prawns. The bacteria are also pathogenic bacteria in Meretrix meretrix Linnaeus and Pinctada martensii, and can also cause death of sea bream, Epinephelus akaara, Pagrus major, etc.

Disclosure of Invention

The invention has the first purpose of providing the application of the compound 1 or the compound 2 shown in the formula I or the medicinal salt thereof in preparing antibacterial drugs or tyrosinase inhibitors;

the inventor isolated a fungus Penicillium chrysogenum Y-S6-1 CCTCC M2020019 from a sample of accumulated dung of Antarctic Allandica penguin, found that a DMSO solution of a crude extract produced by the fungus in a culture medium of Curchia maxima has strong inhibitory activity to Acinetobacter baumannii (gram negative pathogenic bacteria common to hospital infection), Pseudomonas aeruginosa (gram negative pathogenic bacteria common to hospital infection), Vibrio alginolyticus (gram negative pathogenic bacteria common to marine culture animals) and the like, and also has strong inhibitory activity to Multi-drug resistant Staphylococcus aureus (Multi-drug resistant Staphylococcus aureus, MRSA), Staphylococcus aureus (Staphylococcus aureus), and the like, and subsequently found that the culture medium of Penicillium chrysogenum Y-S6-1 CCTCC M2020019 has strong inhibitory activity to tyrosinase production (X1) and also has strong inhibitory activity to culture medium of tyrosinase X1 (gram negative compounds) and high yield of Xpenicillin X1.

The antibacterial drug is preferably a drug for resisting acinetobacter baumannii, escherichia coli, multi-drug-resistant staphylococcus aureus, micrococcus luteus, staphylococcus aureus, apple ring rot, wheat sheath blight or tomato early blight.

The tyrosinase inhibitor is preferably whitening cosmetics.

The second purpose of the invention is to provide a preparation method for preparing the compound 1 or the compound 2, wherein the compound 1 and the compound 2 are prepared and separated from a fermentation culture of Penicillium chrysogenum Y-S6-1 CCTCC M2020019.

Preferably, the specific steps are as follows:

preparing a fermentation culture of Penicillium chrysogenum Y-S6-1 CCTCC M2020019, separating a supernatant and a mycelium, extracting the supernatant with ethyl acetate, and concentrating an ethyl acetate extract to remove ethyl acetate to obtain a crude extract of the supernatant; extracting mycelium with acetone-water solution, concentrating the extract to remove acetone, extracting the residual water solution with ethyl acetate, evaporating ethyl acetate from ethyl acetate extract to obtain mycelium crude extract, and mixing the supernatant crude extract and the mycelium crude extract to obtain extract;

performing gradient elution on the extract by medium-pressure reversed-phase column chromatography, and performing gradient elution on the extract by using water in a volume ratio of: methanol, 100: 0 to 0: performing 100-gradient elution, and collecting a fraction F1 obtained by eluting with 70% methanol-water by volume fraction, a fraction F2 obtained by eluting with 75% methanol-water by volume fraction, and a fraction F3 obtained by eluting with 80% methanol-water by volume fraction;

fraction F2 precipitated Compound 1 in a 72% volume fraction methanol-water solution at room temperature, fraction F3 being Compound 1;

fraction F1 was purified to give compound 2.

The purification was done with HP L C.

The fermentation culture for preparing the Penicillium chrysogenum Y-S6-1 CCTCC M2020019 is obtained by performing fermentation culture by taking a Chaudhur liquid culture medium as a fermentation culture medium.

The third purpose of the invention is to provide the application of Penicillium chrysogenum Y-S6-1 CCTCC M2020019 in preparing the compound 1 or the compound 2.

The compound 1 and the compound 2 with antibacterial or tyrosinase inhibitory activity are separated from Penicillium chrysogenum Y-S6-1 CCTCC M2020019, so that the compounds can be applied to antibacterial drugs and tyrosinase inhibitors.

The Penicillium chrysogenum (Penicillium chrysogenum) Y-S6-1 is preserved in China Center for Type Culture Collection (CCTCC) at 1 month and 7 days of 2020, with the address: wuhan and Wuhan university in Hubei province, postcode: 430072 with preservation number of CCTCC NO: m2020019.

Description of the drawings:

FIG. 1 is a strain morphology diagram of Penicillium chrysogenum Y-S6-1 CCTCC M2020019;

FIG. 2 is a Penicillium chrysogenum Y-S6-1 CCTCC M2020019 phylogenetic tree;

figure 3 is a nmr hydrogen spectrum (deuterated DMSO, 700 mhz) of compound 1(Xanthocillin X);

figure 4 is a nuclear magnetic resonance carbon spectrum (deuterated DMSO, 175 mhz) of compound 1(Xanthocillin X);

FIG. 5 is a drawing showing the analysis of Compound 1 (Xanthocilin X) L R-ESI-MS (positive ion mode, [ M + H ]]⁺)；

FIG. 6 is an X-ray single crystal diffractogram of Compound 1 (Xanthocilin X);

figure 7 is a nmr hydrogen spectrum (deuterated DMSO, 700 mhz) of compound 2(Xanthocillin Y1);

FIG. 8 is a carbon nuclear magnetic resonance spectrum (deuterated DMSO, 175 MHz) of Compound 2 (Xanthocilin Y1);

FIG. 9 is a drawing showing the analysis of Compound 2 (Xanthocilin Y1) L R-ESI-MS (positive ion mode, [ M + H ])]⁺)；

FIG. 10 shows that Penicillium chrysogenum Y-S6-1 CCTCC M2020019 extract (sample No. 32) has strong inhibitory effect on Acinetobacter baumannii, Pseudomonas aeruginosa, Vibrio alginolyticus, multidrug-resistant Staphylococcus aureus (MRSA), Staphylococcus aureus (Staphylococcus aureus);

FIG. 11 shows that compounds 1 and 2 have strong inhibitory activity against Rhizoctonia cerealis (Ceratopsis cornigerum) and Physalospora Mali Aphylla (Physalospora piricola Nose), and that sample No. 27 (2) also has inhibitory activity against Altemiarosa (Altemianella solani) (positive control:. fluconazole, P1: nystatin, P2: amphotericin).

The specific implementation mode is as follows:

the following examples are further illustrative of the present invention and are not intended to be limiting thereof.

Example 1:

1. the inventor separates a fungus Penicillium chrysogenum Y-S6-1 CCTCC M2020019 (figure 1) from a sample of accumulated excrement of a penguin in Antarctic Adelia, extracts genome DNA of the strain, obtains a 28S rDNA sequence (the sequence of which is shown as SEQ ID NO. 1) through Polymerase Chain Reaction (PCR) amplification, and establishes a phylogenetic tree through sequence comparison. The results show that: the similarity of the strain and Penicillium chrysogenum reaches 100% (figure 2), which indicates that the strain Y-S6-1 is Penicillium, named as Penicillium chrysogenum Y-S6-1, which is preserved in China Center for Type Culture Collection (CCTCC) at 1-7 days of 2020, addresses: wuhan and Wuhan university in Hubei province, postcode: 430072 with preservation number of CCTCC NO: m2020019.

2. Penicillium chrysogenum Y-S6-1 CCTCC M2020019 was inoculated from a slant storage tube to a PDA plate for activation, spores were picked and inoculated into 1L Erlenmeyer flasks, each flask containing 200 mL of a Czochralski liquid medium (each 1L of the medium contains 3 g of sodium nitrate, 1 g of dipotassium hydrogen phosphate, 0.5 g of magnesium sulfate, 1 g of potassium chloride, 0.01 g of ferrous sulfate, 20g of sucrose, and the balance water, the components were mixed well, adjusted to pH7.0, sterilized at 115 ℃ for 30 minutes), and 55 bottles (11L) were used. The mixture was placed on a shaker at 28 ℃ and cultured with shaking at 200rpm for 20 days to obtain a fermentation culture. The supernatant and the mycelia of the fermentation culture were separated by centrifugation at 4000 rpm. The supernatant was extracted three times with ethyl acetate, and the ethyl acetate was distilled off to obtain a crude extract of the supernatant. The mycelia were extracted three times with 70% by volume acetone-water, and the acetone was recovered from the extract by rotary evaporator. The remaining aqueous solution was extracted three times with ethyl acetate, and ethyl acetate was distilled off to obtain a crude extract of mycelia. And combining the crude extract of the supernatant and the crude extract of the mycelium to obtain 8.72g of crude extract of Penicillium chrysogenum Y-S6-1 CCTCC M2020019.

Subjecting a crude extract (8.72g) of Penicillium chrysogenum Y-S6-1 CCTCC M2020019 to medium-pressure reverse-phase column chromatography gradient elution (gradient elution is carried out from water to methanol in a volume ratio of 100: 0-0: 100), and collecting a fraction F1 obtained by eluting with 70% methanol-water in volume fraction, a fraction F2 obtained by eluting with 75% methanol-water in volume fraction and a fraction F3 obtained by eluting with 80% methanol-water in volume fraction.

Stream F2 was left to stand at room temperature for two days, and then precipitated pale yellow crystals (compound 1, 181 mg) in a 72% methanol-water volume fraction solution, and after recrystallization from DMSO, the crystals were collected and subjected to X-ray single crystal analysis, and found to be Xanthocillin X having an isonitrile group (fig. 6, compound 1). stream F3 weighed 4.1 g, and was found to be almost all of compound 1 by comparative analysis with high performance liquid chromatography (HP L C).

Fraction F1 was further subjected to the semi-preparative HP L C method (acetonitrile B phase-water A phase; gradient elution (elution procedure: 0-23min, A: 95% -0%, B: 5% -100%, 23-26min, A: 0%, B: 100%, 26-28min, A: 0% -95%, B: 100% -5%, 28-30min, A: 95%, B: 5%, flow rate of 2.5ml/min), Phenomex TitankC-18 column, 250 × 10mm, flow rate of 2.5ml/mim) to yield Compound 2(8.7 mg, retention time of 15.5 min).

The NMR spectrum (deuterated DMSO, 700 MHz) of Compound 1 is shown in FIG. 3, the NMR spectrum (deuterated DMSO, 175 MHz) is shown in FIG. 4, and the analysis chart of L R-ESI-MS (positive ion mode, [ M + H ], [ M ] is shown in FIG. 4]⁺) As shown in fig. 5. The X-ray single crystal diffractogram of Compound 1 (Xanthocilin X) is shown in FIG. 6. Compound 1: the brown crystals are obtained by crystallization of a brown crystal,¹H-NMR、¹³C-NMR and reported XanthocillinX data are in agreement, and low resolution mass spectrometry (L R-ESIMS) gives an excimerIon Peak M/z289.1([ M + H)]⁺) The data relate to the formula C of Xanthocillin X₁₈H₁₂N₂O₂(molecular weight 288.1). Thus identified as XanthocillinX.

The NMR spectrum (deuterated DMSO, 700 MHz) of Compound 2 is shown in FIG. 7, the NMR spectrum (deuterated DMSO, 175 MHz) is shown in FIG. 8, and the analysis chart of L R-ESI-MS (positive ion mode, [ M + H ], [ M ] is shown in FIG. 8]⁺) As shown in fig. 9. Compound 2, a dark brown amorphous powder,¹H-NMR、¹³C-NMR and reported Xanthocillin Y1 data are in agreement, L R-ESIMS gives the excimer ion peak M/z 305.1([ M + H ]]⁺) The data relate to the formula C of Xanthocillin Y1₁₈H₁₂N₂O₃(molecular weight 304.1). Thus identified as Xanthocilin Y1.

TABLE 1 NMR data for Compounds 1 and 2

Table1.NMR data of compounds 1 and2(¹H at 700MHz,¹³C at 175MHz)

^aRecorded in DMSO-d6

Identifying that the compound 1 is Xanthocillin X, and the structural formula of the compound is shown as 1 in the formula I;

identifying that the compound 2 is Xanthocillin Y1, and the structural formula of the compound is shown as 2 in the formula I;

example 2:

paper diffusion method

(1) Test sample solution preparation

About 2 mg of ethyl acetate extract (32: Penicillium chrysogenum Y-S6-1 CCTCCM2020019 crude extract of the invention of example 1) from 51 fermentation cultures (supernatant + mycelium) of Antarctic-origin microorganisms were precisely weighed, and DMSO was filtered through a 0.22 μm microfiltration membrane to prepare a 20mg/ml DMSO solution. The positive control was vancomycin at a concentration of 0.256 mg/ml. Negative control was DMSO solvent. The prepared sample solution should be stored in a refrigerator.

(2) Preparation of bacteria-containing test plate

Acinetobacter baumannii (Acinetobacter baumannii), Pseudomonas aeruginosa (Pseudomonas aeruginosa), Staphylococcus aureus (Staphylococcus aureus), and multidrug-resistant Staphylococcus aureus (Multi-drug resistant Staphylococcus aureus, MRSA) are inoculated in a Mueller-Hinton (MH) broth culture medium, Vibrio alginolyticus (Vibrio algolyticus) is inoculated in a L B culture medium, the above indicator bacteria are all placed on a 37 ℃ and 200rpm constant temperature shaker for overnight culture, the Mueller-Hinton culture medium is prepared by 2.0 g/l of beef extract powder, 17.5 g/l of acid hydrolyzed casein, 1.5 g/l of soluble starch, 17.0 g/l of agar powder (added as required) and water, the components are uniformly mixed, pH is adjusted to 7.3 +/-0.1 (25 ℃ and 0.1 g/l of casein, 25.5 g/l of soluble starch is added in a working Medium (MH) containing 20 g/l of sodium chloride, the components are uniformly mixed in a working medium (28 g/l) containing 20g of agar, the Yeast extract, the Yeast is prepared by a method of cooling a 20 g/l agar plate (28. DEG C, 2.5 g/L) and uniformly mixing the agar medium (2.0 g/L) and adding a working medium (2.7.5 g/L) of agar, the agar, 2.5 g/L) of agar, the mixture is prepared by a working medium (20 g/L) and uniformly mixing the mixture (20 g/L) and uniformly.

(3) Paster piece and application of sample

And (3) attaching sterilized filter paper sheets with the diameter of 5mm to a bacteria-containing test flat plate by using a small forceps, dropwise adding 5 microliters of different samples prepared in the step (1) to each paper sheet according to a pre-marked serial number, and repeating the steps for three times. The bacteria-containing test plate after the sample addition is placed in an incubator at 37 ℃ for culture and observed after 12 hours.

As shown in FIG. 10, it can be seen from FIG. 10 that the crude extract of Penicillium chrysogenum Y-S6-1 CCTCC M2020019 (sample No. 32) had a strong inhibitory effect on Acinetobacter baumannii (Acinetobacter baumannii), Pseudomonas aeruginosa (Pseudomonas aeruginosa), Vibrio alginolyticus (Vibrio algicidal), multidrug-resistant Staphylococcus aureus (MRSA), Staphylococcus aureus (Staphylococcus aureus).

Example 3:

MIC (minimum inhibitory concentration) method

(1) Test sample solution preparation

The pure compounds 1 and 2 were precisely weighed at about 2 mg each, and DMSO was filtered through a 0.22 μm microporous membrane to prepare a 2.56mg/ml solution. The positive control was vancomycin at a concentration of 2.56 mg/ml. Negative control was DMSO solvent. The prepared sample solution should be stored in a refrigerator.

(2) MIC plate preparation

The following experiments required sterile handling in a clean bench. The concentration after overnight incubation corresponded to 1.5X10⁸CFU/ml indicator bacteria (Acinetobacter baumannii, Escherichia coli, Multi-drug resistant Staphylococcus aureus, MRSA, Staphylococcus aureus, Micrococcus luteus) suspension is diluted with MH broth 1: 1000 (OD value 0.04-0.06) as an indicator bacteria solution, and the procedure is carried out according to the following steps that (a) MH broth liquid culture medium is added into a 96-well polystyrene plate, 100 mu L is added into each well, 200 mu L is added into each well as a culture medium control, (2) 100 mu L MH liquid culture medium is added into each well of the No.1, 10 mu L1 of DMSO solution is added into each well, 90 mu L MH liquid culture medium is added into each well of the No. 3, 10 mu 2 of DMSO solution is blown into each well of the pipette 4, 2.56mg/ml sample solution is blown into a lance 4, 5 mu 2 of DMSO solution is blown into each well of the pipette 4, 5 mu 2.5 mu of DMSO solution is blown into each well of the pipette 4, 5 mu 12 mu 2.5 mu, 5 mu of DMSO solution is blown into each well of the pipette 4, 5 mu 2.5 mu 2. 5. mu. 10. 5. mu. 5. of DMSO solution, the pipette 5. 10. 5. mu. 5. 1. 5. 10. of DMSO solution is added into each well of the pipette 5. 10. mu. 10. of DMSO liquid culture medium, the pipette 5. 1. 10. 1. 10. of DMSO liquid culture medium, the pipette 5. of DMSO solution is blown into the pipette, the pipette 5. of the pipette 5^-1. (f) The same bacterial suspension was added to each plate and each suspension was assayed 3 times. Sealing the 96-well polystyrene plate, and then placing the sealed plate in an incubator at 37 ℃ for incubation for 16-20 h.

(3) Result judgment

The test is only meaningful when the bacteria are significantly growing in the negative control wells (i.e., without compound) when viewed visually. The OD value at 560nm was measured with a microplate reader, and the result was judged. The lowest drug concentration that completely inhibited bacterial growth in the wells was the MIC.

The results are shown in table 2:

TABLE 2 test results of the minimum inhibitory concentrations of Compounds 1, 2 against five indicator bacteria (unit: μ g/ml)

From table 2, it can be found that compound 1 and compound 2 have strong inhibitory effect on the above five indicator bacteria.

Example 4:

the pathogenic fungus experiment step comprises (1) preparing a potato glucose agar (PDA) plate, wherein a PDA culture medium formula comprises 200g of potatoes, 20g of glucose, 20g of agar and 1000 ml of distilled water, and the pH value is 7.0 +/-0.1 (25 ℃), firstly cleaning and peeling the potatoes, then weighing 200g of potatoes, cutting the potatoes into small blocks, adding water, boiling the small blocks for 20-30 minutes, puncturing the small blocks by a glass rod, filtering the small blocks by eight layers of gauze, discarding filter residues, heating the filtrate, adding 20g of agar, continuously heating and uniformly stirring, adding 20g of glucose after the agar is dissolved, uniformly stirring, slightly cooling, then adding water to 1000 ml, subpackaging the obtained mixture into a conical flask, sealing and binding, sterilizing at 115 ℃ for 30 minutes, putting the sterilized potato glucose agar (PDA) culture medium into a plastic culture dish with the diameter of 90mm, inversely cooling the obtained product, and then respectively inoculating wheat rot fungi (Ceramium), apple physa spore paper and a sample to a plate in a super clean bench, and inoculating a sample to a sample containing no-agar, wherein the sample is prepared by a method comprising the steps of adding a 3-48 g of potato glucose, and sterilizing the same as a sample, and inoculating a sample to a biochemical strain.

As shown in FIG. 11, it can be seen from FIG. 11 that compounds 1 and 2 both had potent inhibitory activity against Rhizoctonia cerealis (Ceratopsis cornigerum) and Physalospora piricola (Physalospora Nose), and sample No. 27 (compound 2) also had inhibitory activity against Altemaria solani tomato early blight (positive control: +: fluconazole, P1: nystatin, P2: amphotericin; the above control was also a chloroform-methanol solution at a volume ratio of 1:1 and a concentration of 2. mu.g/. mu.l).

Example 5: tyrosinase Activity study

Tyrosinase, also known as polyphenol oxidase, is the rate-limiting enzyme for melanin synthesis and widely exists in microorganisms, animals, plants and human bodies. Tyrosinase is a determinant factor for generating melanin, and the synthesis amount of the melanin is in positive correlation with the activity of the tyrosinase, so that the activity of the tyrosinase is inhibited, the generation of the melanin is reduced, and the tyrosinase has close relation with human aging, insect wound healing and development and browning of vegetables and fruits. Tyrosinase activity determination principle: during the process of generating melanin by tyrosinase reaction, tyrosinase catalyzes tyrosine to be dopa, and dopa is converted into dopaquinone. Dopaquinone is a coloured substance with a characteristic absorption at 475nm that can be quantitatively measured spectrophotometrically.

Preparation of enzyme solution: the tyrosinase dry powder was made into a stock solution with a concentration of 10000U/ml using 0.05mol/l PBS pH6.8, and stored at-20 ℃ for further use. For the experiments, the samples were diluted to a concentration of 100U/ml with 0.05mol/l PBS, pH 6.8.

Preparation of substrate, accurately weighing appropriate amount of levodopa (L-dopa) powder, and preparing into 0.01 mol/l. L-dopa solution with 0.05mol/l PBS (pH 6.8) to be unstable.

Sample preparation: a certain amount of sample is accurately weighed, and is prepared into mother liquor with the concentration of 100mg/ml by DMSO for later use, and the mother liquor is diluted into the required concentration by PBS during the experiment.

Accurately absorbing reaction liquid of a blank control group, a negative control group, a sample group (taking kojic acid as a positive control) and a sample background group in a 96-hole enzyme label plate according to the following table 4, uniformly mixing, reacting at constant temperature of 30 ℃ for 30min, and measuring light absorption at wavelength of 475nmDegree a, 3 replicates per sample and 3 replicates were taken and averaged. The action concentration of the sample during primary screening is 1mg/ml, and at least 5 action concentrations are set for calculating IC for the sample with clearance rate of more than 50% during primary screening₅₀. The inhibition rate of the sample on tyrosinase was calculated according to the following formula:

inhibition rate (%) ([ 1- (A) ]_{Sample (I)}-A_{Sample background})/(A_{Negative control}-A_{Blank control})]×100％(3-4)

TABLE 4 composition of the reaction solution

Tyrosinase (Tyrosinase) is a copper-containing metalloenzyme and is widely distributed in microorganisms, animals, plants and human bodies, wherein the Tyrosinase mainly participates in two reaction processes, namely L-tyrosine hydroxylation is catalyzed and converted into L-dopa and oxidation L-dopa to form dopaquinone, the dopaquinone forms melanin after a series of reactions, and the Tyrosinase has important physiological functions in organisms, and meanwhile, the Tyrosinase is related to the occurrence of diseases such as melanin excessive deposition of human freckles, chloasma and the like, and has great relation with the molting of insects and the browning of fruits and vegetables.

The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.

Sequence listing

<110> Nanhai ocean institute of Chinese academy of sciences

Preparation method and application of <120> Xanthocillin compound

<160>1

<170>SIPOSequenceListing 1.0

<210>1

<211>561

<212>DNA

<213> Penicillium chrysogenum Y-S6-1(Penicillium chrysogenum)

<400>1

agtaacggcg agtgaagcgg caagagctca aatttgaaag ctggctcctt cggggtccgc 60

attgtaattt gtagaggatg cttcgggagc ggtccccatc taagtgccct ggaacgggac 120

gtcatagagg gtgagaatcc cgtatgggat ggggtgtccg cgcccgtgtg aagctccttc 180

gacgagtcga gttgtttggg aatgcagctc taaatgggtg gtaaatttca tctaaagcta 240

aatattggcc ggagaccgat agcgcacaag tagagtgatc gaaagatgaa aagcactttg 300

aaaagagagt taaaaagcac gtgaaattgt tgaaagggaa gcgcttgcga ccagactcgc 360

tcgcggggtt cagccggcat tcgtgccggt gtacttcccc gcgggcgggc cagcgtcggt 420

ttgggcggtc ggtcaaaggc cctcggaagg taacgcccct aggggcgtct tatagccgag 480

ggtgcaatgc gacctgccta gaccgaggaa cgcgcttcgg ctcggacgct ggcataatgg 540

tcgtaaacga cccgtcttga a 561

Claims

1. The application of the compound 1 or the compound 2 shown in the formula I or the medicinal salt thereof in preparing antibacterial drugs and tyrosinase inhibitors;

2. the use of claim 1, wherein the antibacterial agent is against acinetobacter baumannii, escherichia coli, multidrug-resistant staphylococcus aureus, micrococcus luteus, staphylococcus aureus, ring rot of apple, rhizoctonia cerealis, or early blight of tomato.

3. The use according to claim 1, wherein the tyrosinase inhibitor is a whitening cosmetic.

4. A process for the preparation of compound 1 or compound 2 according to claim 1, wherein compound 1 or compound 2 is isolated from a fermentation culture of Penicillium chrysogenum Y-S6-1 CCTCC M2020019.

5. The preparation method according to claim 4, comprising the following steps:

fraction F1 was purified to give compound 2.

6. The method of claim 5, wherein the purification is HP L C.

7. The method according to claim 5, wherein the Penicillium chrysogenum Y-S6-1 CCTCC M2020019 is prepared by performing fermentation culture in a Chao' S broth as a fermentation medium.

The application of Penicillium chrysogenum Y-S6-1 CCTCC M2020019 in preparing compound 1 or compound 2.