Phenolic acid amide derivative and preparation method and application thereof
阅读说明：本技术 一种酚酸酰胺衍生物及其制备方法和应用 (Phenolic acid amide derivative and preparation method and application thereof ) 是由 杜丰玉 王兆富 肖�琳 张相飞 于 2019-09-23 设计创作，主要内容包括：本发明涉及微生物农药技术领域,具体的说是一种从菌株<I>Penicillium</I>sp.TR85(保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏日期：2019年8月2日,保藏号：CGMCC No.18116)液体发酵产物中得到酚酸酰胺衍生物及其分离纯化方法和在除草活性方面的应用。酚酸酰胺衍生物为式(一)所示化合物,在20μg/mL浓度时,式(一)所示化合物对马唐[<I>Digitaria </I><I>sanguinalis</I>]胚芽鞘的生长抑制率为68.96%,优于商品化除草剂乙草胺；对马唐胚根的生长抑制率为64.74%,与乙草胺接近。因此,式(一)所示酚酸酰胺衍生物可作为具有除草活性的先导化合物,进而开发微生物源除草剂,<Image he="147" wi="272" file="RE-675300DEST_PATH_IMAGE001.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>。(The invention relates to the technical field of microbial pesticides, in particular to a slave strain Penicillium sp.TR85 (preserved in China general microbiological culture Collection center, preservation date: 2019, 8 and 2 months, preservation number: CGMCC No. 18116) liquid fermentation product to obtain phenolic acid amide derivatives, a separation and purification method thereof and application thereof in the aspect of herbicidal activity. The phenolic acid amide derivative is a compound represented by the formula (I), and the compound represented by the formula (I) is used for the digitaria sanguinalis [ alpha ], [ alpha Digitaria sanguinalis ]The growth inhibition rate of the coleoptile is 68.96%, which is superior to that of the commercial herbicide acetochlor; the growth inhibition rate of the crabgrass radicle is 64.74 percent, which is close to that of acetochlor. Therefore, the temperature of the molten metal is controlled,the phenolic acid amide derivative shown in the formula I can be used as a lead compound with herbicidal activity to further develop a microbial source herbicide, 。)
1. A phenolic acid amide derivative characterized by: the phenolic acid amide derivative is a compound shown as a formula (I), and the molecular formula is C15H13NO4，
。
2. A process for the preparation of a phenolic acid amide derivative according to claim 1, wherein:
1) the strain isPenicilliumsp, TR85 (preserved in the general microbiological culture Collection center of China Committee for culture Collection of microorganisms, the preservation date is 8 and 2 days in 2019, the preservation number is CGMCC number 18116) is placed in a liquid culture medium for standing and fermentation for 30 days, the fermentation product is repeatedly soaked and extracted by ethyl acetate, and the extracting solution is combined and concentrated to obtain a fermentation crude extract;
2) subjecting the crude fermentation extract to silica gel column chromatography, and gradient eluting with petroleum ether-ethyl acetate as solvent at gradient ratio of 40:1 to 1:1 (v/v, the same below);
3) performing reverse phase silica gel column chromatography on the petroleum ether-ethyl acetate =1:1 elution component in the step 2), and performing gradient elution by using methanol-water with gradient of 2:8 to 1:0 as a solvent;
4) purifying the eluted fraction with methanol-water =5:5 in step 3) by semi-preparative high performance liquid chromatography, wherein the mobile phase is 45% acetonitrile-water, the detection wavelength is 230 nm, the flow rate is 3 mL/min, and the collection retention timet RThe component with the value of 13.8 min is the phenolic acid amide derivative shown in the formula (I).
3. A process for the preparation of a phenolic acid amide derivative according to claim 2, wherein: the liquid culture medium comprises 40 g of glucose, 10 g of peptone, 5 g of yeast powder, 5 g of malt extract and 1000 mL of natural seawater.
4. The use of a phenolic acid amide derivative according to claim 1, wherein: the phenolic acid amide derivative shown in the formula I can be used for preparing a lead compound with herbicidal activity.
5. The use of a phenolic acid amide derivative according to claim 4, wherein: the weed is digitaria sanguinalis [ 2]Digitaria sanguinalis]。
Technical Field
The invention relates to the technical field of microbial pesticides, in particular to a slave strainPenicilliumsp, TR85 liquid fermentation product to obtain phenolic acid amide derivative, its separation and purification method and application in weeding activity.
Background
Tang 2Digitaria sanguinalis]Is one of the most main malignant weeds in farmlands, and because seedlings of large crabgrass are similar to rice seedlings, rice production in China is seriously damaged. At present, the crabgrass is mainly prevented and removed by adopting a chemical weeding technology, and the defects are that the environment is easily polluted and resistant weeds are generated. The substitution of the chemical weeding technique by the weed biological control technique can alleviate a series of problems caused by the use of chemical herbicides, so that the search for lead compounds with herbicidal activity from plants or microorganisms and the further development of biogenic herbicides have become important directions for the development of herbicides.
According to literature research, the phenolic acid amide derivative is a novel compound, and no related report on herbicidal activity exists.
Disclosure of Invention
The invention aims to provide a phenolic acid amide derivative, and a preparation method and application thereof.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a phenolic acid amide derivative characterized by: the phenolic acid amide derivative is a compound shown as a formula (I), and the molecular formula is C15H13NO4。
A preparation method of phenolic acid amide derivatives comprises the following steps:
1) the strain isPenicilliumsp, TR85 (deposited in China general microbiological culture Collection center, CGMCC for short, with the address of No. 3 West Lu No.1, North Sungyang district, Beijing, China academy of sciences microbiological research, preservation date of 8 months and 2 days in 2019, preservation number of CGMCC No. 18116) in a liquid culture medium, standing for fermentation, repeatedly soaking and extracting the fermentation product with ethyl acetate, combining the extracting solutions and concentrating to obtain a fermentation crude extract;
2) subjecting the crude fermentation extract to silica gel column chromatography, and gradient eluting with petroleum ether-ethyl acetate as solvent at gradient ratio of 40:1 to 1:1 (v/v, the same below);
3) performing reverse phase silica gel column chromatography on the petroleum ether-ethyl acetate =1:1 elution component in the step 2), and performing gradient elution by using methanol-water with gradient of 2:8 to 1:0 as a solvent;
4) purifying the eluted fraction with methanol-water =5:5 in step 3) by semi-preparative high performance liquid chromatography, wherein the mobile phase is 45% acetonitrile-water, the detection wavelength is 230 nm, the flow rate is 3 mL/min, and the collection retention timet RThe component with the value of 13.8 min is the phenolic acid amide derivative shown in the formula (I).
The application of the phenolic acid amide derivative shown in the formula (I) can be used for preparing a lead compound with herbicidal activity.
The weed is digitaria sanguinalis [ 2]Digitaria sanguinalis]。
The invention has the advantages that:
1) the phenolic acid amide derivative shown as the formula (I) is a novel compound, and when the concentration of the phenolic acid amide derivative is 20 mu g/mL, the compound shown as the formula (I) is used for the digitaria sanguinalis [ digitalis ], [ solution of ] ADigitaria sanguinalis]The growth inhibition rate of the coleoptile is 68.96%, which is superior to that of the commercial herbicide acetochlor; the growth inhibition rate of the crabgrass radicle is 64.74 percent, which is close to that of acetochlor.
2) The phenolic acid amide derivative shown as the formula (I) consists of bacterial strainsPenicilliumsp, TR85, facilitating scale fermentation production; and secondly, the phenolic acid amide derivative shown in the formula (I) is a natural product, and has the advantages of good environmental compatibility, difficult generation of drug resistance and the like compared with a chemical synthetic pesticide.
3) The phenolic acid amide derivative shown in the formula (I) related by the invention is convenient to reform, and can be used as a lead compound with herbicidal activity to further develop a microbial source herbicide.
Drawings
FIG. 1 is a diagram showing a process for producing a phenolic acid amide derivative represented by the formula (I)1H NMR spectrum.
FIG. 2 is a diagram showing a process for producing a phenolic acid amide derivative represented by the formula (I)13C NMR spectrum.
FIG. 3 shows the inhibitory effect of the amide phenolate derivative represented by formula (I) on the germination of seeds of digitaria sanguinalis.
Detailed Description
The invention will now be further described with reference to some non-limiting examples of embodiments in order to clarify the understanding of the characteristics of the invention.
The invention relates to a strainPenicilliumsp, TR85 (deposited in China general microbiological culture Collection center, CGMCC for short, with the address: No. 3, West Siway 1, Kyoho, Beijing, Chaoyang, China academy of sciences microbiological research, date of deposit: 2019, 8, 2 days, and number of CGMCC 18116) to obtain the compound indicated in the following examples, wherein a phenolic acid amide derivative has the structure shown in formula (one) (the Arabic number in the structure is the index position of carbon atom):