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Nitrile hydratase and a method for producing amides

A technology for nitrile hydratase and nitrile group hydration, which is applied in the fields of nitrile hydratase and amide production, and can solve problems such as impracticality

Inactive Publication Date: 2005-01-26
DAICEL CHEM IND LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Similarly, the method of adding sulfite, acid sulfite, or dithionite requires the addition of ions in an amount equal to or greater than that of the raw material, so this method is not practical

Method used

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  • Nitrile hydratase and a method for producing amides
  • Nitrile hydratase and a method for producing amides
  • Nitrile hydratase and a method for producing amides

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Embodiment Construction

[0247] The present invention is explained in detail below with reference to Examples, but the present invention is not construed as being limited thereto.

[0248] 1. Enzyme Activity Determination Method

[0249] A standard assay for nitrile hydratase activity was employed in the following examples, as follows. The standard composition of the reaction solution used for the enzyme reaction is shown in Tables 1 and 2. The enzymatic reaction is initiated by adding 3-cyanopyridine or 2-hydroxy-4-methylthiobutyronitrile (HMBN) as a substrate compound. Incubate at 20° C. for 10 minutes in the case of 3-cyanopyridine and 15 minutes at 20° C. in the case of HMBN. When using 3-cyanopyridine in the reaction, add 0.1ml of 2N hydrochloric acid to the reaction and shake the mixture vigorously to terminate the reaction; when using HMBN in the reaction, add 0.1ml of the reaction solution to 0.9ml of 0.1% (v / v) phosphoric acid , shake the mixture vigorously to terminate the reaction. The ...

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Abstract

An objective of the present invention is to provide a nitrile hydratase capable of producing 2-hydroxy-4-methylthiobutyroamide. The present invention provides a novel nitrile hydratase producing alpha-hydroxyamide using, alpha-hydroxnitrile as the substrate, and the encoding DNA thereof. The enzyme can be obtained from Rhodococcus sp. Further, the enzymatic activity of the enzyme can he maintained stably during the reaction. The present invention provides a method for producing amide compounds, the method comprising the step of reacting this enzyme to nitrile compounds. According to the present invention, from hydroxy nitrile compounds, corresponding amide compounds can be produced biochemically without reducing the enzyme activity of nitrile hydratase.

Description

technical field [0001] The present invention relates to a novel nitrile hydratase and a method of producing an amide compound from a nitrile compound using the nitrile hydratase. Background technique [0002] In order to increase the expression level of nitrile hydratase, a method of overexpressing nitrile hydratase in microbial cells by genetic engineering and a method of converting nitrile compounds into corresponding amide compounds using these cells have been examined. For example, known nitrile hydratases are derived from the following microorganisms. All nitrile hydratases are enzymes composed of two types of heterogeneous subunits. [0003] Rhodococcus (Examined Japanese Patent Application Publication (JP-B) No. Hei 3-54558) [0004] Rhodococcus (Unexamined Japanese Patent Application Publication (JP-A) No. Hei 2-119778) [0005] Pseudomonas (JP-A Hei 3-251184) [0006] Rhodococcus rhodochrous (European Patent Application No. 455646) [0007] In all cases, howeve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12N1/15C12N1/19C12N1/21C12N5/10C12N9/88C12N15/60C12P13/02C12P17/00C12R1/01
CPCC12N9/88C12P13/02
Inventor 长泽透松山彰收
Owner DAICEL CHEM IND LTD
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