Microorganism including gene encoding protein having dehalogenase activity, and method of reducing concentration of fluorine-containing compound in sample using the same

Patent metadata

Field	Value
Publication number	US-2019032034-A1
Application number	US-201816046685-A
Country	US
Kind code	A1
Filing date	Jul 26, 2018
Priority date	Jul 28, 2017
Publication date	Jan 31, 2019
Grant date	—

Abstract

Provided are a microorganism including a gene encoding a protein having a dehalogenase activity, a composition including the microorganism for use in reducing a concentration of a fluorine-containing compound in a sample, and a method of reducing the concentration of the fluorine-containing compound in the sample by using the microorganism.

First claim

What is claimed is:

1 . A recombinant Pseudomonas saitens microorganism comprising a genetic modification that increases dehalogenase activity as compared to a parent strain Pseudomonas saitens KCTC 13107BP, wherein the dehalogenase activity comprises the activity of at least one dehalogenase selected from the group consisting of a haloalkane dehalogenase, a polypeptide having a sequence identity of 85% or more with respect to an amino acid sequence of SEQ ID NO: 3, and a polypeptide having a sequence identity of 85% or more with respect to an amino acid sequence of SEQ ID NO: 5.

2 . The recombinant microorganism of claim 1 , wherein the genetic modification increases expression of a gene encoding the dehalogenase.

3 . The recombinant microorganism of claim 1 , wherein the genetic modification increases a copy number of a gene encoding the dehalogenase.

4 . The recombinant microorganism of claim 1 , wherein the haloalkane dehalogenase is an enzyme belonging to EC 3.8.1.5.

5 . The recombinant microorganism of claim 1 , wherein haloalkane dehalogenase has a sequence identity of 85% or more with respect to an amino acid sequence of SEQ ID NO: 1.

6 . A method of reducing a concentration of a fluorine-containing compound in a sample, the method comprising:
contacting a recombinant microorganism of claim 1 with a sample comprising a fluorine-containing compound represented by Formula 1 or Formula 2, so as to reduce the concentration of the fluorine-containing compound in the sample:
C(R 1 )(R 2 )(R 3 )(R 4 )  <Formula 1>

(R 5 )(R 6 )(R 7 )C—[C(R 11 )(R 12 )] n -C(R 8 )(R 9 )(R 10 )  <Formula 2>

wherein, in Formulae 1 and 2, n is an integer from 0 to 10, R 1 , R 2 , R 3 , and R 4 are each independently fluorine (F), chlorine (CI), bromine (Br), iodine (I), or hydrogen (H), and at least one selected from R 1 , R 2 , R 3 , and R 4 is F, and R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , and R 12 are each independently F, Cl, Br, I, or H, and at least one selected from R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , and R 12 is F.

7 . The method of claim 6 , wherein the genetic modification increases expression of a gene encoding the dehalogenase.

8 . The method of claim 6 , wherein the genetic modification increases a copy number of a gene encoding the dehalogenase.

9 . The method of claim 6 , wherein the haloalkane dehalogenase is an enzyme belonging to EC 3.8.1.5.

10 . The method of claim 6 , wherein the haloalkane dehalogenase has a sequence identity of 85% or more with respect to an amino acid sequence of SEQ ID NO: 1.

11 . The method of claim 6 , wherein the contacting is performed in a closed container.

12 . The method of claim 6 , wherein the contacting comprises culturing or incubating the recombinant microorganism in contact with the sample.

13 . The method of claim 6 , wherein the contacting comprises culturing the recombinant microorganism under conditions in which the recombinant microorganism proliferates in a closed container.

14 . The method of claim 6 , wherein the fluorine-containing compound is CF 4 , CHF 3 , or CH 2 F 2 .

15 . The method of claim 6 , wherein the recombinant microorganism cleaves a C—F bond of the fluorine-containing compound, converts the fluorine-containing compound into a different substance, or accumulates the fluorine-containing compound in the recombinant microorganism, to reduce the concentration of the fluorine-containing compound in the sample.

16 . The method of claim 6 , wherein the sample is a liquid or gas.

17 . A method of preparing a recombinant microorganism of claim 1 having an increased dehalogenase activity, the method comprising
introducing into a Pseudomonas saitens KCTC 13107BP microorganisma gene encoding a haloalkane dehalogenase, a gene encoding a polypeptide (BC3334 N122Y) having a sequence identity of 85% or more with respect to an amino acid sequence of SEQ ID NO: 3, a gene encoding a polypeptide (hdl4a) having a sequence identity of 85% or more with respect to an amino acid sequence of SEQ ID NO: 5, or a combination thereof.

Assignees

• Samsung Electronics Co Ltd

Inventors

• Kim Jinha
• Byun Jongwon
• Park Jinhwan

Classifications

• C02F2101/36

  containing halogen · CPC title

• B01D53/70

  Organic halogen compounds · CPC title

• B01D53/84

  Biological processes · CPC title

• C02F3/342

  characterised by the enzymes used · CPC title

• C12Y308/01002

  (S)-2-Haloacid dehalogenase (3.8.1.2) · CPC title