CN111850061B - A kind of preparation method of Jinggang hydroxylamine A ester compound - Google Patents

Abstract

The invention discloses a preparation method of valienamine A ester shown in formula (I), wherein R is C ₁ ‑C ₂₁ Saturated aliphatic radical of (2), C ₁ ‑C ₂₁ Unsaturated aliphatic radical of (A) or (C) ₁ ‑C ₂₁ The halogenated saturated aliphatic group of (a), the process being: the valienamine A and fatty acid RCOOH react in tertiary butanol under the action of a biocatalyst to prepare an esterified product of the valiolamine A shown in a formula (I); the biocatalyst is one of Lipozyme RMIM, lipase CALB, porcine pancreatic Lipase, candida Lipase and Candida rugosa Lipase. The method can accurately control the reaction sites and obtain the validamine A esterified substance with high purity.

Description

A kind of preparation method of Jinggang hydroxylamine A ester compound

technical field

The invention relates to an enzyme-catalyzed method for preparing Jinggang hydroxylamine A ester.

Background technique

Validamycin, a pseudotrisaccharide compound, was isolated in 1970 from the secondary metabolites of Streptomyce hygroscopicus var.lemonesus, and was named validamycin. In 1973, the Shanghai Institute of Pesticides discovered and screened the strain of Streptomyces hygroscopicus var. Jinggangensis (S. hygroscopicus var. jinggangensis) that can inhibit rice sheath blight (Rhizoctonia solani) in the soil of Jinggangshan, Jiangxi and Hangzhou, Zhejiang. After research, it was extracted from it. The chemical structure of the active compound is consistent with validamycin, and its active ingredient is named jinggangmycin. Jinggangmycin can effectively inhibit rice sheath blight, and has a certain bactericidal effect on vegetable seedling wilt. In addition, it has long-term efficacy, low toxicity, low residue, high safety, and low environmental pollution. The advantages make it popular in my country The pollution-free biological source pesticide with the largest area and the lowest cost per mu.

Jinggangmycin A is the most important and main component of Jinggangmycin compounds. Its molecular structure contains Jinggang hydroxylamine A and a D-type glucopyranose group. It is a class of pseudotrisaccharide compounds.

The chemical structure of Jinggang Hydroxylamine A is similar to that of trehalose (as shown below), and it is considered to be a kind of competitive inhibitor of natural trehalase with high activity. It has a novel structure, good bactericidal effect in vitro, and strong selectivity. The inhibitory effect of the enzyme can reach 10 ^-8 -10 ^-9 mol/L, in addition, it has high safety to mammals and higher plants, and has a good application prospect.

Jinggang Hydroxylamine A has a strong trehalase inhibitory activity in vitro, but it is difficult to enter the organism. Therefore, it is necessary to modify the structure of Jinggang Hydroxylamine A to increase its ability to enter the organism, so as to solve the problem that can only be solved by "injection". To exert its insecticidal and bactericidal activity, ester synthesis has become a preferred solution. However, the number of hydroxyl groups on Jinggang hydroxylamine A is relatively large, and it is difficult to accurately control the esterification site by conventional chemical synthesis methods. However, the lipase-catalyzed synthesis method adopted in the present invention can accurately control the reaction sites and finally obtain high-purity derivatives. thing. Further research on its biological activity shows that it has good biological activity.

Contents of the invention

The object of the present invention is to provide an enzyme-catalyzed method for preparing Jinggang hydroxylamine A ester, which can accurately control the reaction site and obtain high-purity Jinggang hydroxylamine A ester.

In order to realize the above-mentioned purpose of the invention, the present invention adopts following technical scheme:

A preparation method of Jinggang hydroxylamine A esterified product shown in formula (I), described method is: Jinggang hydroxylamine A and fatty acid RCOOH are reacted under the effect of biocatalyst in tert-butanol and prepared formula (I) The esterification of Jinggang hydroxylamine A; the biocatalyst is one of Lipozyme RMIM, Lipase CALB, porcine pancreatic lipase, Candida lipase, and Candida rugosa lipase;

R is a C ₁ -C ₂₁ saturated aliphatic group, a C ₁ -C ₂₁ unsaturated aliphatic group or a C ₁ -C ₂₁ halogenated saturated aliphatic group.

Preferably, R=C _n H _2n+1 , where n=1-21. More preferably n=8-18, still more preferably n=10-18, still more preferably 10-17.

Preferably, R=C _n H _2n-1 , where n=1-21. More preferably n=3-18, even more preferably n=5-18, especially preferably n=7-18, such as R=CH ₂ =CHCH ₂ CH ₂ CH ₂ CH ₂ CH ₂ .

Preferably, R=C _n H _2n+1-m X _m , wherein n=1-21, m≤2n+1, X=F, Cl, Br or I. More preferably n=3-18, even more preferably n=5-18, especially preferably n=7-18. More preferably X=Cl. Particular preference is given to R═CCl ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ .

As a preference, an activated molecular sieve is also added to the reaction system, and the amount of the molecular sieve is 0.4-2g/mmoL based on the moles of Jinggang hydroxylamine A, more preferably 1.2-2.0g/mmoL, most preferably 1.2g/mmoL . As a further preference, the molecular sieve is 4A° molecular sieve. As a further preference, the molecular sieve is activated before use. The recommended activation method is: activate at 400°C for 2 hours.

Preferably, the biocatalyst is Lipozyme RMIM, and the conversion rate of Jinggang hydroxylamine A is the highest at this time.

As a preference, the feeding method is: reflux and stir Jinggang hydroxylamine A in tert-butanol at 90-110°C for 4-6h to obtain a saturated solution of Jinggang hydroxylamine A, and then add the saturated solution of Jinggang hydroxylamine A and fatty acid RCOOH to the reaction vessel mix in. This feeding method can significantly increase the solubility of Jinggang hydroxylamine A in tert-butanol, thereby effectively improving the reaction efficiency.

As a further preference, the preparation method is specifically implemented as follows: fully mix the Jinggang hydroxylamine A-tert-butanol saturated solution and the fatty acid RCOOH in a reaction vessel at 25-50°C, then add a biocatalyst and shake the reaction at 25-50°C After fully reacting, the reaction solution is separated and purified to obtain the esterified product of Jinggang Hydroxylamine A; The number of moles is 40-240g/moL.

As a further preference, molecular sieves are also added to the reaction system, the amount of molecular sieves is 0.4-2g/mmoL based on the moles of Jinggang hydroxylamine A, more preferably 1.2-2.0g/mmoL, most preferably 1.2g/mmoL .

As a further preference, the biocatalyst is Lipozyme RMIM, at this time Jinggang hydroxylamine A conversion rate is the highest.

As a further preference, the biocatalyst is Lipozyme RMIM, and the amount of the biocatalyst is 160-240 g/moL, most preferably 160 g/moL, based on the moles of Jinggang Hydroxylamine A.

As a further preference, the biocatalyst is Lipozyme RMIM, and the equilibrium is reached when the reaction time is 48h.

As a further preference, the biocatalyst is Lipozyme RMIM, the reaction temperature is 45-50°C, and the optimum temperature is 45°C.

As a further preference, the biocatalyst is Lipozyme RMIM, and the molar ratio of Jinggang hydroxylamine A to fatty acid RCOOH is 1:2-4, most preferably 1:3.

The reaction condition when the most preferred biocatalyst of the present invention is Lipozyme RMIM is: the molar ratio of Jinggang hydroxylamine A and fatty acid RCOOH is 1:3, and the enzyme dosage is 160g/moL Jinggang hydroxylamine A, and the consumption of molecular sieve is 1.2g/mmoL Jinggang hydroxylamine A, the reaction temperature is 45°C, and the reaction time is 48h.

As a further preference, the specific steps of the separation and purification are: centrifuging to remove the biocatalyst (and molecular sieve), distilling off the solvent, then adding water and adjusting the pH value to neutral with sodium hydroxide, separating through a macroporous adsorption resin column, Gradient elution is carried out with ethanol aqueous solution with a volume concentration of 0%-55% as the elution reagent, and the target liquid is collected, concentrated and dried.

The Jinggang hydroxylamine A ester represented by the formula (I) of the present invention has a good inhibitory effect on rice sheath blight.

Compared with prior art, the beneficial effect of the present invention is:

(1) The present invention provides a method for improving the solubility of Jinggang hydroxylamine A, which can improve the solubility of Jinggang hydroxylamine A in tert-butanol.

(2) The present invention provides an enzyme-catalyzed method for preparing Jinggang hydroxylamine A ester, which can accurately control the reaction site and obtain high-purity Jinggang hydroxylamine A ester.

(3) The present invention can significantly improve the reaction efficiency of Jinggang hydroxylamine A and fatty acid by adding molecular sieves.

detailed description

The present invention will be further described below in conjunction with specific examples, but the protection scope of the present invention is not limited thereto.

Embodiment 1: the fermentation of producing lipase strain

In the present invention, Acinetobacter agarnei zjutfet-1 (preserved in the China Type Culture Collection Center, the preservation date is September 6, 2015, the preservation number CCTCC NO.M2015511, and the preservation address is Wuhan University, Wuhan, China, postcode 430072), Applied to the synthesis of the compound. The culture method of this bacterium is as follows:

Inoculate Acinetobacter agarnei zjutfet-1 into the slant medium and culture at 35° C. for 20 hours to obtain slant colonies; the final concentration of the slant medium is composed of: LB medium + 2% agar powder.

The slant colonies were inoculated on the seed medium, and cultured at 30°C for 6 hours to obtain the seed liquid. The final concentration of the seed medium was composed of peptone 10g/L, yeast extract 5g/L, NaCl 10g/L, the solvent was distilled water, and the pH Value is 7.0;

The seed solution was inoculated on the fermentation medium with an inoculum volume concentration of 0.2%, cultivated at 30°C and 180rpm for 48 hours, centrifuged the fermentation solution, discarded the supernatant to obtain wet bacteria, and then freeze-dried to obtain the following freeze-dried powder of bacteria. The fermentation medium consists of peptone 10g/L, sucrose 5g/L, dipotassium hydrogen phosphate 10g/L, magnesium sulfate 0.5g/L, olive oil 50g/L, solvent is distilled water, and the pH value is 7.0.

Example 2: Preparation of Hydroxylamine Acetate A (Compound 1) (R ₁ =CH ₃ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and acetic acid (0.120g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH = 7, and then pass through a macroporous adsorption resin column (Shanghai Huazhen Technology Co., Ltd. Company, HZ-801) separation, the eluent is 0%-55% ethanol aqueous solution for gradient elution, the target liquid is collected, concentrated and dried to obtain 0.38g Jinggang Hydroxylamine A acetate with a purity of 98.8%. TLC detects the concentrated solution, the developer is n-propanol:water:acetic acid=4:1:1 (volume ratio), and the color developer is iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.58(dd, J=6.2,0.6Hz,1H),4.74(d,J=0.9Hz,2H),4.06(s,1H),3.72(dd,J=2.5 ,1.5Hz,1H),3.57–3.54(m,1H),3.52(dd,J=12.2,8.0Hz,1H),3.35(t,J=8.3Hz,1H),3.27(dd,J=12.4, 8.2Hz, 1H), 3.17(dd, J=7.9, 1.8Hz, 1H), 3.13–3.11(m, 1H), 3.09(dd, J=9.2, 6.3Hz, 1H), 2.08(d, J=1.0 Hz,1H),2.07(dd,J＝6.5,4.1Hz,1H),2.01(s,3H),1.76(s,1H),1.33–1.31(m,1H),1.07–1.05(m,1H) .ESI-MS:[M+H]:378.1719.

Example 3: Preparation of Hydroxylamine Propionate A (Compound 2) (R ₁ =CH ₃ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and propionic acid (0.148g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol was added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with ethanol aqueous solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.42 g of Jinggang Hydroxylamine Propionate A with a purity of 98.5%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.58(dd, J=6.2,0.6Hz,1H),4.72(d,J=0.6Hz,2H),4.06(dd,J=4.2,0.7Hz,1H), 3.85(dt,J=5.5,3.7Hz,1H),3.70(dd,J=9.4,4.3Hz,1H),3.60(dd,J=9.3,4.2Hz,1H),3.52(dd,J=12.4, 7.6Hz, 1H), 3.27(dd, J＝12.4, 7.6Hz, 1H), 3.19–3.14(m, 2H), 3.09(dd, J＝6.2, 4.4Hz, 1H), 2.88–2.81(m, 1H ),2.48(dd,J＝7.5,2.9Hz,1H),2.46–2.42(m,2H),1.68(s,1H),1.37–1.26(m,1H),1.20(t,J＝6.8Hz, 3H), 1.09(dd, J=8.2, 3.6Hz, 1H).

ESI-MS:[M+H] ⁺ :392.1867.

Example 4: Preparation of Hydroxylamine Butyrate A (Compound 3) (R ₁ =CH ₃ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and butyric acid (0.176g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with ethanol aqueous solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.45 g of Jinggang Hydroxylamine A Butyrate, with a purity of 98.2%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ4.73(d, J=13.9Hz, 2H), 4.56(s, 1H), 4.41(dd, J=9.5, 5.8Hz, 1H), 4.31(s, 1H), 3.77(t,J=8.4Hz,1H),3.53(dd,J=10.4,3.8Hz,1H),3.49–3.41(m,1H),3.41–3.23(m,6H),3.09(t,J= 8.7Hz, 1H), 2.13(dt, J＝11.2, 6.9Hz, 1H), 1.89(s, 5H), 1.87–1.80(m, 1H), 1.53(ddd, J＝14.9, 11.7, 5.8Hz, 1H ).

ESI-MS[M+H] ⁺ 406.2067,[M+Na] ⁺ 428.1890

Example 5: Preparation of Jinggang Hydroxylamine A Valerate (Compound 4) (R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and valeric acid (0.204g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with ethanol aqueous solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.46 g of Jinggang Hydroxylamine A Valerate with a purity of 99.1%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.76(s, 1H), 5.44(s, 1H), 4.65(s, 5H), 4.17(dd, J=10.6, 2.8Hz, 1H), 4.09–3.86(m ,3H),3.70(d,J=4.7Hz,1H),2.28(t,J=7.3Hz,2H),2.03–1.93(m,1H),1.77(d,J=14.1Hz,1H),1.51 (dt,J=15.0,7.4Hz,2H),1.38–1.03(m,4H),0.86(t,J=7.3Hz,3H).

ESI-MS[M+H] ⁺ 420.2227,[M+Na] ⁺ 442.2040

Embodiment 6: the preparation of hexanoic acid Jinggang hydroxylamine A ester (compound 5)

(R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and hexanoic acid (0.232g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with ethanol aqueous solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.45 g of hexanoic acid Jinggang hydroxylamine A ester with a purity of 98.4%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ 5.81 (dd, J = 37.3, 3.0Hz, 1H), 4.68 (d, J = 13.1Hz, 2H), 4.35 (d, J = 13.2Hz, 1H), 3.96 ( d,J=17.8Hz,1H),3.69(dd,J=11.3,5.7Hz,1H),3.49(d,J=3.8Hz,31H),3.46–3.17(m,6H),3.11–3.02(m ,1H),2.31(dt,J=22.2,7.3Hz,1H),2.15(t,J=7.4Hz,1H),1.74(dd,J=14.1,3.2Hz,1H),1.56–1.36(m, 2H),1.36–1.03(m,6H),,0.89(t,J＝6.9Hz,3H).

ESI-MS[M+H] ⁺ 434.2381,[M+Na] ⁺ 456.2189

Embodiment 7: the preparation of heptanoic acid Jinggang hydroxylamine A ester (compound 6)

(R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and heptanoic acid (0.260g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with an aqueous ethanol solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.48 g of Jinggang Hydroxylamine A Heptanoate with a purity of 98.5%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.79 (dd, J = 37.3, 3.0Hz, 1H), 4.62 (d, J = 13.1Hz, 2H), 4.33 (d, J = 13.2Hz, 1H), 3.94 ( d,J=17.8Hz,1H),3.71(dd,J=11.3,5.7Hz,1H),3.45(d,J=3.8Hz,31H),3.43–3.17(m,6H),3.11–3.01(m ,1H),2.31(dt,J=22.2,7.3Hz,1H),2.18(t,J=7.4Hz,1H),1.71(dd,J=14.1,3.2Hz,1H),1.56–1.38(m, 2H), 1.36–1.03(m, 8H), 0.91(t, J＝6.9Hz, 3H).

ESI-MS[M+H] ⁺ 448.2532,[M+Na] ⁺ 470.2341

Embodiment 8: the preparation of caprylic acid Jinggang hydroxylamine A ester (compound 7)

(R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang Hydroxylamine A (1.34g, 4.0mmol) and octanoic acid (0.288g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol was added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with ethanol aqueous solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.51 g of octanoic acid Jinggang hydroxylamine A ester with a purity of 98.3%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.81 (dd, J = 37.3, 3.0Hz, 1H), 4.65 (d, J = 13.1Hz, 2H), 4.31 (d, J = 13.2Hz, 1H), 3.94 ( d,J=17.8Hz,1H),3.71(dd,J=11.3,5.7Hz,1H),3.45(d,J=3.8Hz,31H),3.40–3.17(m,6H),3.11–3.01(m ,1H),2.30(dt,J=22.2,7.3Hz,1H),2.18(t,J=7.4Hz,1H),1.74(dd,J=14.1,3.2Hz,1H),1.56–1.40(m, 2H), 1.36–1.01(m, 10H), 0.91(t, J＝6.9Hz, 3H).

ESI-MS[M+H] ⁺ 462.2686,[M+Na] ⁺ 484.2492

Embodiment 9: Preparation of nonanoic acid Jinggang hydroxylamine A ester (compound 8)

(R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang Hydroxylamine A (1.34g, 4.0mmol) and nonanoic acid (0.316g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol was added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with ethanol aqueous solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.49 g of nonanoic acid Jinggang hydroxylamine A ester with a purity of 98.9%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.79 (dd, J=37.3, 3.0Hz, 1H), 4.66 (d, J=13.1Hz, 2H), 4.45 (d, J=13.2Hz, 1H), 3.97( d,J=17.8Hz,1H),3.71(dd,J=11.3,5.7Hz,1H),3.48(d,J=3.8Hz,31H),3.45–3.18(m,6H),3.13–3.04(m ,1H),2.29(dt,J=22.2,7.3Hz,1H),2.17(t,J=7.4Hz,1H),1.76(dd,J=14.1,3.2Hz,1H),1.58–1.40(m, 2H), 1.24(s, 10H), 1.16–1.00(m, 2H), 0.86(t, J＝6.9Hz, 3H).

ESI-MS[M+H] ⁺ 476.2856,[M+Na] ⁺ 498.2616

Embodiment 10: the preparation of decanoic acid Jinggang hydroxylamine A ester (compound 9)

(R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and capric acid (0.344g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with ethanol aqueous solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.55 g of decanoic acid Jinggang hydroxylamine A ester with a purity of 98.4%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.86 (dd, J = 37.6, 3.5Hz, 1H), 4.62 (d, J = 13.0Hz, 1H), 4.48 (d, J = 13.2Hz, 1H), 3.92 ( d,J=18.1Hz,1H),3.65(dd,J=11.0,5.7Hz,1H),3.43(d,J=3.6Hz,1H),3.40–3.13(m,6H),3.11–3.04(m ,1H),2.29(dt,J=22.2,7.1Hz,1H),2.17(t,J=7.7Hz,1H),1.81(dd,J=14.5,3.6Hz,1H),1.58–1.42(m, 2H), 1.24(s, 12H), 1.16–0.98(m, 2H), 0.86(t, J=6.5Hz, 3H).

m/z(ESI-MS):[M ⁺ H]+490.3008,[M ⁺ Na]+512.2746.

Embodiment 11: the preparation of n-undecanoic acid Jinggang hydroxylamine A ester (compound 10)

(R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and n-undecanoic acid (0.372g, 2.0mmol) were successively added into a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with ethanol aqueous solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.52 g of n-undecanoic acid Jinggang Hydroxylamine A ester with a purity of 98.1%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.92 (dd, J = 37.8, 3.7Hz, 1H), 4.71 (d, J = 13.5Hz, 1H), 4.48 (d, J = 13.7Hz, 1H), 3.98 ( d,J=17.2Hz,1H),3.71(dd,J=11.8,5.4Hz,1H),3.46(d,J=3.4Hz,1H),3.42–3.18(m,6H),3.13–3.06(m ,1H),2.30(dt,J=22.2,7.2Hz,1H),2.21(t,J=7.4Hz,1H),1.85(dd,J=14.0,3.2Hz,1H),1.56–1.42(m, 2H), 1.20(s, 14H), 1.16–0.98(m, 2H), 0.86(t, J=6.3Hz, 3H).

m/z(ESI-MS):[M ⁺ H]+504.3161,[M ⁺ Na]+527.1816.

Embodiment 12: the preparation of n-dodecanoic acid Jinggang hydroxylamine A ester (compound 11):

(R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H)

Jinggang hydroxylamine A (1.34g, 4.0mmol) and n-dodecanoic acid (0.400g, 2.0mmol) were successively added into a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with ethanol aqueous solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.54 g of n-dodecanoic acid Jinggang Hydroxylamine A ester with a purity of 98.2%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ 5.95 (dd, J = 36.5, 4.1 Hz, 1H), 4.77 (d, J = 12.8 Hz, 1H), 4.50 (d, J = 12.8 Hz, 1H), 3.41 ( d,J=17.8Hz,1H),3.768(dd,J=11.3,5.7Hz,1H),3.43(d,J=3.8Hz,1H),3.40–3.18(m,6H),3.13–3.06(m ,1H),2.32(dt,J=22.2,7.3Hz,1H),2.25(t,J=7.4Hz,1H),1.86(dd,J=14.1,3.2Hz,1H),1.56–1.42(m, 2H), 1.23(s, 12H), 1.16–0.92(m, 4H), 0.81(t, J＝6.8Hz, 3H).

m/z(ESI-MS):[M ⁺ H]+518.3310,[M ⁺ Na]+540.2876.

Embodiment 13: the preparation of n-tetradecanoic acid Jinggang hydroxylamine A ester (compound 12):

(R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H)

Jinggang hydroxylamine A (1.34g, 4.0mmol) and n-tridecanoic acid (0.428g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with ethanol aqueous solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.49 g of n-tetradecanoic acid Jinggang hydroxylamine A ester with a purity of 98.6%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.93 (dd, J = 37.3, 3.0Hz, 1H), 4.70 (d, J = 13.1Hz, 1H), 4.50 (d, J = 13.2Hz, 1H), 3.94 ( d,J=17.4Hz,1H),3.72(dd,J=11.3,5.7Hz,1H),3.48(d,J=4.2Hz,1H),3.44–3.17(m,6H),3.13–3.02(m ,1H),2.31(dt,J=22.2,7.3Hz,1H),2.24(t,J=7.4Hz,1H),1.87(dd,J=14.1,3.2Hz,1H),1.65–1.40(m, 2H), 1.18(s, 12H), 1.15–0.81(m, 6H), 0.74(t, J=6.4Hz, 3H).

m/z(ESI-MS):[M ⁺ H]+546.3624,[M ⁺ Na]+568.3201

Embodiment 14: Preparation of n-pentadecanoic acid Jinggang hydroxylamine A ester (compound 13)

(R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and n-pentadecanoic acid (0.484g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH = 7, filter to remove the precipitated sodium fatty acid, and then pass through the macroporous adsorption resin Column separation, the eluent is 0%-55% concentration gradient ethanol aqueous solution for gradient elution, the target liquid is collected, concentrated and dried to obtain 0.58 g of n-pentadecanoic acid Jingganghydroxylamine A ester with a purity of 98.4%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.58(dd, J=6.2,0.6Hz,1H),4.76(d,J=0.9Hz,2H),4.06(dd,J=7.3,0.6Hz,1H), 3.75(dd, J=9.2, 7.4Hz, 1H), 3.70(t, J=8.8Hz, 1H), 3.52(dd, J=12.4, 7.1Hz, 1H), 3.27(dd, J=12.4, 7.1Hz ,1H),3.19(t,J=8.9Hz,1H),3.13–3.06(m,2H),2.83(dd,J=10.5,8.7Hz,1H),2.61(dd,J=16.9,7.8Hz, 1H), 2.36(t, J=8.0Hz, 2H), 2.02(t, J=7.9Hz, 1H), 1.97(s, 1H), 1.69(p, J=7.8Hz, 2H), 1.40–1.29( m,23H),1.21–1.16(m,1H),0.99(t,J＝6.4Hz,3H).

m/z(ESI-MS):[M ⁺ H]+560.3833,[M ⁺ Na]+582.3432

Embodiment 15: Preparation of n-hexadecanoic acid Jinggang hydroxylamine A ester (compound 14)

(R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and n-hexadecanoic acid (0.512g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH = 7, filter to remove the precipitated sodium fatty acid, and then pass through the macroporous adsorption resin Column separation, the eluent is 0%-55% ethanol aqueous solution for gradient elution, the target liquid is collected, concentrated and dried to obtain 0.56 g of n-hexadecanoic acid Jingganghydroxylamine A ester with a purity of 98.4%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.58(dd, J=6.2,0.6Hz,1H),4.74(d,J=0.6Hz,2H),4.06(dd,J=4.4,0.9Hz,1H), 3.75(dd,J=9.2,4.4Hz,1H),3.55–3.49(m,2H),3.27(dd,J=12.4,7.0Hz,1H),3.22–3.17(m,1H),3.14–3.07( m,2H),2.83(dd,J=10.3,8.1Hz,1H),2.49(dt,J=9.3,7.6Hz,1H),2.32(t,J=8.2Hz,2H),2.19(t,J ＝8.0Hz, 1H), 1.71(p, J＝8.0Hz, 2H), 1.51(s, 1H), 1.41–1.26(m, 25H), 1.18(t, J＝7.9Hz, 1H), 0.99(t ,J＝6.4Hz,3H).

m/z(ESI-MS):[M ⁺ H]+574.4101,[M ⁺ Na]+596.3614

Embodiment 16: the preparation of n-octadecanoic acid Jinggang hydroxylamine A ester (compound 15)

(R ₁ =CH ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and n-octadecanoic acid (0.568g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. Shake at 45°C for 30 minutes. After the reaction system is fully balanced, add lyophilized bacterial cells (1 g), and shake at 45°C for 48 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH = 7, filter to remove the precipitated sodium fatty acid, and then pass through the macroporous adsorption resin Column separation, the eluent is 0%-55% gradient ethanol solution for gradient elution, the target liquid is collected, concentrated and dried to obtain 0.54 g of n-octadecanoic acid Jingganghydroxylamine A ester with a purity of 98.6%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (500MHz, Chloroform) δ5.58(dd, J=6.2,0.6Hz,1H),4.74(d,J=0.9Hz,2H),4.06(dd,J=4.2,0.6Hz,1H), 3.75(dd, J＝8.9, 4.3Hz, 1H), 3.52(dd, J＝12.4, 7.6Hz, 1H), 3.50–3.46(m, 1H), 3.27(dd, J＝12.4, 7.6Hz, 1H) ,3.18–3.13(m,1H),3.12–3.07(m,2H),2.83(dd,J＝10.4,8.1Hz,1H),2.59(dt,J＝9.2,7.7Hz,1H),2.26(dt ,J=11.2,8.1Hz,3H),1.70(p,J=8.0Hz,2H),1.43–1.30(m,29H),1.17(s,1H),1.11(t,J=8.0Hz,1H) ,0.99(t,J=6.5Hz,3H).

m/z(ESI-MS):[M ⁺ H]+602.4508,[M ⁺ Na]+624.5136

Example 17: Preparation of 7,8-octenoic acid jingganghydroxylamine A ester (compound 16) (R ₁ =CH ₂ =CHCH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and 7,8-octenoic acid (0.28g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol were added. At 45°C, oscillate on a shaking table for 30 minutes. After the reaction system is fully balanced, add bacterial cell freeze-dried powder (1 g), and shake on a shaking table at 45°C for 72 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with ethanol aqueous solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.62 g of 7,8-octenoic acid Jinggang hydroxylamine A ester with a purity of 97.9%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (600MHz, DMSO) δ5.65 (ddt, J＝16.3, 10.0, 6.1Hz, 1H), 5.58 (dd, J＝6.2, 0.6Hz, 1H), 5.02–4.97 (m, 1H), 4.97 –4.93(m,1H),4.77(d,J=0.9Hz,2H),4.06(d,J=1.3Hz,1H),3.77–3.72(m,2H),3.52(dd,J=12.5,7.5 Hz,1H),3.35(s,1H),3.27(dd,J＝12.5,7.5Hz,1H),3.22–3.17(m,1H),3.14–3.11(m,1H),3.09(t,J＝ 4.3Hz, 1H), 2.36(dt, J＝9.8, 7.9Hz, 3H), 2.08(t, J＝8.0Hz, 1H), 2.02(dd, J＝14.2, 7.8Hz, 2H), 1.74–1.66( m,3H),1.38–1.34(m,4H),1.34–1.30(m,1H),1.15(t,J＝7.9Hz,1H).

m/z(ESI-MS):[M+H] ⁺ :460.2506

Example 18: Preparation of Trichlorooctanoic Acid Hydroxylamine A Ester (Compound 17) (R ₁ =CCl ₃ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CO-, R ₂ -R ₈ =H):

Jinggang hydroxylamine A (1.34g, 4.0mmol) and trichlorooctanoic acid (0.50g, 2.0mmol) were successively added to a 50mL Erlenmeyer flask, and 10mL each of cyclohexane and tert-butanol was added. At 45°C, oscillate on a shaking table for 30 minutes. After the reaction system is fully balanced, add bacterial cell freeze-dried powder (1 g), and shake on a shaking table at 45°C for 72 hours. After the reaction is over, add 20 mL of chloroform to stop the reaction, filter the reaction solution, rotary evaporate, add 20 mL of deionized water, then add 1M sodium hydroxide solution to pH=7, then separate through a macroporous adsorption resin column, and the eluent is Gradient elution was carried out with an aqueous ethanol solution with a concentration gradient of 0%-55%, and the target liquid was collected, concentrated and dried to obtain 0.49 g of triclooctanoic acid Jinggang Hydroxylamine A ester with a purity of 98.1%. The concentrated solution was detected by TLC, the developer was n-propanol: water: acetic acid = 4:1:1, and the color developer was iodine vapor and ninhydrin solution.

¹ H NMR (500MHz, Chloroform) δ5.58(dd, J=6.2,0.6Hz,1H),4.74(d,J=0.6Hz,2H),4.06(dd,J=2.6,0.6Hz,1H), 4.00(dd, J=7.0, 3.3Hz, 1H), 3.74(t, J=3.1Hz, 1H), 3.52(dd, J=12.4, 7.1Hz, 1H), 3.35(t, J=8.3Hz, 1H ),3.27(dd,J＝12.4,7.1Hz,1H),3.21–3.15(m,1H),3.11–3.06(m,1H),2.82(dd,J＝10.3,8.3Hz,1H),2.43( dt,J=9.2,7.7Hz,1H),2.37–2.28(m,4H),1.90(t,J=7.9Hz,1H),1.73(p,J=8.0Hz,2H),1.51(s,1H ),1.42–1.32(m,6H),1.31(d,J=4.1Hz,1H),1.24(t,J=7.9Hz,1H).

m/z(ESI-MS):[M+H] ⁺ :565.1462

Embodiment 19: Determination of antibacterial activity

The measurement method used is the mycelial growth rate method. Specific operation: Use a hole puncher with a diameter of 6.0mm to take the activated R. solani bacteria plate on the edge of the potato medium, put the mycelium of the bacteria plate facing down, and inoculate them on the drug-containing agar medium plate and the control group respectively. In the center of the base plate, cultivate in a constant temperature incubator at 25°C. When the diameter of the control colony is between 50mm and 80mm (R.solani generally takes 36h), use a vernier caliper to measure it. Inhibition rate. Mycelium growth inhibition rate calculation formula 4-1 (length unit: mm) is as follows:

During the experiment, first use the inhibition rate obtained by diluting multiple test agents to a concentration of 1000 μg/mL as a reference. If the inhibition rate is 0, discard it. On the contrary, if the inhibition rate is greater than 0, use 500 μg/mL, 250 μg/mL , 125μg/mL, 62.5μg/mL, 31.25μg/mL and other concentration gradients for the determination of antibacterial activity.

Regression equation of drug toxicity and calculation of effective middle concentration EC ₅₀ and EC ₅ : The toxicity of the drug to R.solani was expressed by the rate of inhibition of mycelial growth. The inhibition rate of mycelia growth is converted into the inhibition probability value (y), the concentration of the drug is converted into the concentration logarithm (x), and the regression equation determination coefficient R of the drug to the inhibition probability value of R.solani is obtained by the least square method, according to the toxicity The force regression equation was used to calculate the concentration EC ₅ and EC ₅₀ values of drugs on the inhibition of hyphae growth.

Table 1 Toxicity equation of different drugs to R.solani

It can be seen from the above table that this type of compound has good biological activity. Among them, the inhibitory effect of some compounds on rice sheath blight exceeds that of Jinggangmycin A, and even the lowest effect is compared with Jinggangmycin A or Jinggang Hydroxylamine A. 2-3 orders of magnitude lower. Therefore, this type of compound has great application prospects and commercial value, and is expected to be developed as a new type of high-efficiency and green fungicide to replace Jinggangmycin.

Embodiment 20: Preparation of Jinggang hydroxylamine A-tert-butanol saturated solution

Jinggang hydroxylamine A (5 g) was stirred in tert-butanol (300 mL) for 5 h in an oil bath at 90° C. to prepare a saturated solution of Jinggang hydroxylamine A-tert-butanol.

Jinggang hydroxylamine A (5 g) was stirred in tert-butanol (300 mL) in an oil bath at 90°C for 30 h without reflux to prepare a saturated solution of Jinggang hydroxylamine A-tert-butanol.

The solubility of the two solutions is shown in Table 2:

Table 2

Example 21:

Jinggang hydroxylamine A (5 g) was stirred in tert-butanol (300 mL) for 5 h in an oil bath at 90° C. to prepare a saturated solution of Jinggang hydroxylamine A-tert-butanol. Jinggang hydroxylamine A-tert-butanol saturated solution (21mL, 0.5mmoL) and undecanoic acid (1.0mmoL, 186.33mg) were fully mixed in a reaction vessel at 40°C, then lipase Lipozyme RMIM (100mg) was added and dissolved at 400 4A° molecular sieve (1g) activated for 2h at 40°C was shaken and reacted, sampled after 30h of reaction, diluted with water, passed through a membrane, and analyzed by high performance liquid chromatography; wherein, the mobile phase was a mixture of pH=7.0 PBS solution and methanol ( Wherein the volume fraction of methanol is 3%). Analysis results of high performance liquid chromatography: Jinggang hydroxylamine A conversion rate was 26.87%.

The reaction solution was centrifuged to remove the molecular sieve and catalyst, and the solvent was evaporated, then water was added and the pH value was adjusted to neutrality with sodium hydroxide, separated by a macroporous adsorption resin column (Shanghai Huazhen Technology Co., Ltd., HZ-801), and the volume concentration was 0%-55% ethanol aqueous solution was used as the elution reagent for gradient elution, the target liquid was collected, concentrated and dried to obtain the product, and the ¹ HNMR (600MHz, DMSO) and m/z (ESI-MS) characterization of the product showed that it was compatible with the above compound 10 is the same substance, that is undecanoic acid Jinggang hydroxylamine A ester, and the product purity is 98%.

Example 22: Screening of the most suitable lipase to catalyze the synthesis of undecanoic acid Jinggang Hydroxylamine A

Change lipase, other reaction steps are with embodiment 21, and gained product is undecanoic acid Jinggang hydroxylamine A ester, and wherein high performance liquid chromatography analysis result is as shown in the following table:

Table 3: The conversion rate of Jinggang hydroxylamine A catalyzed by different lipase ^a

The lipases listed in the table ^a were purchased from Xiamen Huijia Biotechnology Co., Ltd.

The results showed that Lipozyme RMIM had the highest catalytic efficiency, and the conversion rate of Jinggang hydroxylamine A reached 26.87%.

Example 23: Lipozyme RMIM Catalyzed Synthesis of Undecanoic Acid Jinggang Hydroxylamine A Condition Optimization—Reaction Time

Jinggang hydroxylamine A-tert-butanol saturated solution (21mL, 0.5mmoL) and undecanoic acid (1.0mmoL, 186.33mg) were thoroughly mixed in a reaction vessel at 40°C, then Lipozyme RMIM (100mg) was added and activated at 400°C The 4A°molecular sieve (1g) was shaken and reacted at 40°C for 2h. During the reaction, samples were taken, diluted with water, passed through a membrane, and analyzed by high-performance liquid chromatography; wherein, the mobile phase was a mixture of pH=7.0 PBS solution and 3% methanol. High performance liquid chromatography analysis result is as shown in table 4:

Jinggang hydroxylamine A transformation rate under the different reaction times of table 4

The results showed that the optimal catalytic time of Lipozyme RMIM was 48 hours, and the conversion rate of Jinggang hydroxylamine A was 26.54%.

Example 24: Lipozyme RMIM Catalyzed Synthesis of Undecanoic Acid Hydroxylamine A Condition Optimization—Temperature

Jinggang hydroxylamine A-tert-butanol saturated solution (21mL, 0.5mmoL) and undecanoic acid (1.0mmoL, 186.33mg) were placed in a reaction vessel (at 25°C, 30°C, 35°C, 40°C, 45°C, 50°C ℃) and mix well, then add lipase (100mg) and 4A° molecular sieve (1g) which has been activated at 400℃ for 2h (at 25℃, 30℃, 35℃, 40℃, 45℃, 50℃) and shake the reaction, the reaction After 48 hours, samples were taken, diluted with water, passed through a membrane, and analyzed by high-performance liquid chromatography; wherein, the mobile phase was a mixture of pH=7.0 PBS solution and 3% methanol. High performance liquid chromatography analysis result is as shown in table 5:

Table 5: Jinggang hydroxylamine A conversion rate at different temperatures

The results showed that the optimal catalytic temperature of Lipozyme RMIM was 45℃.

Example 25: Condition optimization for the synthesis of undecanoic acid hydroxyamine A catalyzed by Lipozyme RMIM—molecular sieve dosage

Jinggang hydroxylamine A-tert-butanol saturated solution (21mL, 0.5mmoL) and undecanoic acid (1.0mmoL, 186.33mg) were thoroughly mixed in a reaction vessel at 45°C, then lipase (100mg) was added and activated at 400°C 4A ° molecular sieves (0g, 0.2g, 0.4g, 0.6g, 0.8g, 1.0g) were shaken and reacted at 45°C for 2 hours, sampled after 48h of reaction, diluted with water, passed through a membrane, and analyzed by high performance liquid chromatography; wherein, The mobile phase is a mixture of pH=7.0 PBS solution and 3% methanol. High performance liquid chromatography analysis result is as shown in table 6:

Table 6 Jinggang hydroxylamine A conversion rate under different molecular sieve additions

The results showed that the optimum molecular sieve content was 1.2g/mmoL.

Example 26: Condition optimization for the synthesis of undecanoic acid hydroxyamine A catalyzed by Lipozyme RMIM - substrate molar ratio

Put Jinggang hydroxylamine A-tert-butanol saturated solution (21mL, 0.5mmoL) and undecanoic acid (0.125mmoL, 0.167mmoL, 0.25mmoL, 0.5mmoL, 1.0mmoL, 1.5mmoL, 2.0mmoL) in a reaction vessel at 45°C Mix well, then add lipase (100mg) and 4A° molecular sieve (0.6g) which has been activated at 400°C for 2h, shake and react at 45°C, take a sample after reacting for 48h, dilute with water, pass through a membrane, and analyze by high performance liquid chromatography; , the mobile phase is a mixture of pH=7.0 PBS solution and 3% methanol. High performance liquid chromatography analysis result is as shown in table 7:

The conversion rate of Jinggang hydroxylamine A under different substrate molar ratios in table 7

The results showed that the optimal molar ratio of Lipozyme RMIM catalyzed reaction was 1:3 considering the materials and conversion rate of the reaction.

The post-treatment method of the reaction solution obtained under different substrate molar ratios is the same as in Example 21, and the obtained products are all undecanoic acid Jinggang Hydroxylamine A esters.

Example 27: Condition optimization for the synthesis of undecanoic acid hydroxyamine A catalyzed by Lipozyme RMIM—enzyme dosage

Jinggang hydroxylamine A-tert-butanol saturated solution (21mL, 0.5mmoL) and undecanoic acid (1.5mmoL, 279.495mg) were thoroughly mixed in a reaction vessel at 45°C, then lipase (20mg, 40mg, 60mg, 80mg , 100mg, 120mg) and 4A° molecular sieve (0.6g) which had been activated at 400°C for 2h were shaken and reacted at 45°C, sampled after 48h of reaction, diluted with water, passed through a membrane, and analyzed by high performance liquid chromatography; wherein, the mobile phase was pH = Mixture of 7.0 PBS solution and 3% methanol. High performance liquid chromatography analysis result is as shown in table 8:

Table 8 The conversion rate of Jinggang hydroxylamine A under different enzyme dosages

The results showed that when the reaction system was 21mL, the optimal amount of enzyme added was 80mg, that is, 160g/moL.

In summary, in Jinggang hydroxylamine A-tert-butanol saturated solution, when using Lipozyme RMIM as catalyst, the molar ratio of Jinggang hydroxylamine A to undecanoic acid is 1:3, the reaction temperature is 45°C, and the amount of molecular sieve is 1.2g /mmoL, when the enzyme dosage was 200g/moL, the conversion rate of Jinggang hydroxylamine A was as high as 43% after 48 hours of reaction.

Claims (13)

1. the preparation method of the Jinggang hydroxylamine A ester compound shown in a kind of formula (I), described method is: Jinggang hydroxylamine A and undecanoic acid react formula (I) under the effect of biocatalyst in tert-butanol The esterified product of Jinggang hydroxylamine A shown; the biocatalyst is Lipozyme RMIM;

R=C _n H _2n+1 , where n=10. 2. the preparation method as claimed in claim 1 is characterized in that: the feeding mode of raw material is: make Jinggang hydroxylamine A in tert-butanol at 90-110 ℃ of reflux stirring 4-6 h and make Jinggang hydroxylamine A saturated solution, Then the Jinggang hydroxylamine A saturated solution and undecanoic acid were mixed in the reaction vessel. 3. The preparation method according to claim 2, characterized in that: the preparation method is specifically implemented as follows: Jinggang hydroxylamine A-tert-butanol saturated solution and undecanoic acid are fully mixed in a reaction vessel at 25-50°C , then add biocatalyst and shake reaction at 25-50 ℃, after fully reacting, reaction solution obtains the esterification product of Jinggang hydroxylamine A through separation and purification; The feeding molar ratio of described Jinggang hydroxylamine A and undecanoic acid is 4:1-1: 4. The amount of the biocatalyst is 40-240 g/moL based on the moles of Jinggang Hydroxylamine A. 4. The preparation method according to claim 3, characterized in that: activated molecular sieves are also added in the reaction system, and the amount of said molecular sieves is 0.4-2 g/mmoL based on the moles of Jinggang hydroxylamine A. 5. The preparation method according to claim 4, characterized in that: the amount of the molecular sieve is 1.2-2.0 g/mmoL in terms of moles of Jinggang hydroxylamine A. 6. preparation method as claimed in claim 5 is characterized in that: the consumption of described molecular sieve is calculated as 1.2g/mmoL with the number of moles of Jinggang hydroxylamine A. 7. preparation method as claimed in claim 3 is characterized in that: the molar ratio of described Jinggang hydroxylamine A and undecanoic acid is 1:2-4; The consumption of described biocatalyst is in the molar number of Jinggang hydroxylamine A It is 160-240 g/moL. 8. The preparation method according to claim 7, characterized in that: the molar ratio of Jinggang hydroxylamine A and undecanoic acid is 1:3. 9. The preparation method according to claim 7, characterized in that: the amount of the biocatalyst is 160 g/moL in terms of moles of Jinggang hydroxylamine A. 10. The preparation method according to claim 3, characterized in that: the reaction temperature is 45-50°C; the reaction time is 48 h. 11. The preparation method according to claim 10, characterized in that: the reaction temperature is 45°C. 12. the preparation method as claimed in claim 3 is characterized in that reaction condition is: the mol ratio of Jinggang hydroxylamine A and undecanoic acid is 1:3, and enzyme dosage is 160 g/moL Jinggang hydroxylamine A, and the consumption of molecular sieve is 1.2g/mmoL Jinggang Hydroxylamine A, the reaction temperature is 45°C, and the reaction time is 48 h. 13. The preparation method as claimed in claim 4, characterized in that: the separation and purification step is specifically: removing the biocatalyst and molecular sieve by centrifugation, removing the solvent by steaming, then adding water and adjusting the pH value to neutral with sodium hydroxide, passing Separation by macroporous adsorption resin column, gradient elution with ethanol aqueous solution with a volume concentration of 0%-55% as the elution reagent, the target liquid is collected, concentrated and dried.