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Application of M. frugosa in the biotransformation of 3-indoleacetonitrile to synthesize 3-indoleacetamide

A technology of sword rhinoceros oleracea and indoleacetamide is applied in the field of microorganisms to achieve the effects of simple process, easy cultivation and high transformation rate

Inactive Publication Date: 2016-09-14
NANJING NORMAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no literature report on the synthesis of 3-indoleacetamide from 3-indoleacetonitrile catalyzed by bacteria of the genus Ensifer

Method used

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  • Application of M. frugosa in the biotransformation of 3-indoleacetonitrile to synthesize 3-indoleacetamide
  • Application of M. frugosa in the biotransformation of 3-indoleacetonitrile to synthesize 3-indoleacetamide
  • Application of M. frugosa in the biotransformation of 3-indoleacetonitrile to synthesize 3-indoleacetamide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1: Streak Mushroom CGMCC7333 on LB solid agar medium. The composition of LB medium is: peptone 10g / L, yeast extract 5g / L, NaCl 10g / L, pH adjusted to 7.0, and 20g / L agar powder is added to prepare solid medium. Cultivation was based on autoclaving at 115°C for 20 minutes. After a single colony grows on the solid plate, pick a single colony and inoculate it into 10mL LB liquid medium with a final concentration of 0.1mmol / L cobalt chloride, culture at 30°C, shake at 220rpm for 24h, then centrifuge at 8000rpm for 10min to collect The cells were suspended in 0.05 mol / L potassium phosphate buffer at pH 7.5, and the cell concentration (OD600) was adjusted to 5.0. Take 5 mL of the above cell suspension in a 50 mL centrifuge tube, and add 0.5 mg of 3-indoleacetonitrile (concentration: 0.5 g / L). The centrifuge tube was reacted in a shaker at 30°C and 220rpm. After 12 hours of reaction, samples were taken, centrifuged at 8000rpm for 10 minutes, and the supernatant was ta...

Embodiment 2

[0013] Example 2: Use a sterile toothpick to pick up a ring of activated Sword Mushroom CGMCC7333 and place it in a 100mL Erlenmeyer flask containing 20mL of liquid LB medium. After culturing for 16 hours, take 1 mL and inoculate it in 100 mL of LB liquid medium containing 0.1 mmol / L cobalt chloride at 30°C, shake at 220 rpm for 12 hours, centrifuge at 10,000 rpm for 10 min, collect the cells and suspend them in 0.05 mol / L of pH 7.5 In potassium phosphate buffer, the cell concentration (OD600) was adjusted to 5.0. The content of 3-indoleacetamide was sampled and determined after 12 hours of conversion, and the result showed that under the above conditions, the concentration of 3-indoleacetamide generated was 0.76 g / L.

Embodiment 3

[0014] Example three: basically the same as Example two, the conversion time is 24h, and the HPLC quantitative analysis results show that the concentration of the generated 3-indoleacetamide is 1.03g / L.

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Abstract

The invention belongs to the technical field of microbes, and discloses the application of the sword fungus Ensifer meliloti CGMCC7333 in the biotransformation of 3-indole acetonitrile to generate 3-indole acetamide. The bacterial strain used in the present invention is easy to cultivate, and is applied to biocatalyze 3-indoleacetonitrile to synthesize 3-indoleacetamide, which has the characteristics of simple process, high conversion rate and the like. The product 3‑indoleacetamide can be used as a pharmaceutical intermediate and for the synthesis of plant hormones such as 3‑indoleacetic acid.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to the application of C. meliloti CGMCC7333 to catalyze 3-indoleacetonitrile to generate 3-indoleacetamide. Background technique [0002] Biocatalysis refers to the process of chemical transformation using enzymes or whole cells as catalysts. Since the nature of biocatalytic reactions is enzyme-catalyzed reactions, it has the characteristics of high efficiency, diversity, substrate specificity, regioselectivity, chemoselectivity, enantioselectivity and mild reaction conditions. At present, biocatalytic reactions have been widely used in chemical, pharmaceutical, pesticide and food industries. [0003] Nitrile compounds are an important class of chemical synthesis raw materials. Starting from nitrile compounds, a class of compounds with higher value and wider application range such as amides and acids can be obtained. Among them, the enzyme that catalyzes nitrile comp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/10C12R1/01
Inventor 戴亦军周玲艳王鑫孙仕磊
Owner NANJING NORMAL UNIVERSITY
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