Method for immobilizing and releasing microorganism

Patent metadata

Field	Value
Publication number	US-9631189-B2
Application number	US-201414779717-A
Country	US
Kind code	B2
Filing date	Mar 14, 2014
Priority date	Mar 26, 2013
Publication date	Apr 25, 2017
Grant date	Apr 25, 2017

Abstract

Intended is to provide a more practical technique for immobilizing a microorganism using an adhesive protein AtaA derived from Acinetobacter sp. Tol 5. Provided is a method for attaching and releasing a microorganism, including (1) a step of contacting a microorganism, into which DNA encoding autotransporter adhesin derived from a microorganism belonging to the genus Acinetobacter has been introduced to impart or enhance non-specific adhesiveness, with a carrier under a high ionic strength and thus attaching the microorganism to the carrier; and (2) a step of releasing the microorganism from the carrier by washing under a low ionic strength.

First claim

The invention claimed is:

1. A method for attaching and releasing a microorganism comprising the following steps (1) and (2):
(1) contacting a microorganism, into which DNA encoding an autotransporter adhesion derived from the ataA gene of a microorganism belonging to the genus Acinetobacter has been introduced to impart or enhance non-specific adhesiveness, with a carrier under a high ionic strength and thus attaching the microorganism to the carrier; and
(2) releasing the microorganism from the carrier by washing under a low ionic strength,
wherein the high ionic strength is from 20 mM to 200 mM, and the low ionic strength is less than 10 mM.

2. The method of claim 1 , wherein the Acinetobacter is Acinetobacter sp. Tol 5.

3. The method of claim 1 , wherein the DNA is any of (a), (b) or (c):
(a) DNA composed of the base sequence of SEQ ID NO: 1;
(b) DNA composed of the base sequence having 90% or more homology with the base sequence of SEQ ID NO: 1, and encoding the protein having activity of imparting or enhancing non-specific adhesiveness to a microorganism;
(c) DNA composed of a part of the base sequence of SEQ ID NO: 1, and encoding the protein having activity of imparting or enhancing non-specific adhesiveness to a microorganism.

4. The method of claim 3 , wherein the DNA of the following (a) or (b) is introduced into the microorganism together with the DNA encoding an autotransporter adhesin:
(a) DNA composed of the base sequence of SEQ ID NO: 3;
(b) DNA composed of the base sequence having 90% or more homology with the base sequence of SEQ ID NO: 3.

5. The method of claim 1 , wherein the DNA of the following (a) or (b) including the above-described DNA has been introduced into the microorganism:
(a) DNA composed of the base sequence of SEQ ID NO: 5;
(b) DNA composed of the base sequence having 90% or more homology with the base sequence of SEQ ID NO: 5, and having activity of imparting or enhancing non-specific adhesiveness to a microorganism.

6. The method of claim 1 , wherein the following step (3-1) is carried out after the step (2):
(3-1) recovering the released microorganism.

7. The method of claim 1 , wherein the following step (3-2) is carried out after the step (2):
(3-2) recovering the carrier.

8. The method of claim 1 , wherein the following step (i) is carried out between the steps (1) and (2):
(i) contacting the microorganism attached to the carrier with a liquid to be treated, and keeping the contact state.

9. The method of claim 8 , wherein the microorganism produces a specific enzyme, and the liquid to be treated comprises a substrate of the specific enzyme that is treated by the specific enzyme.

10. The method of claim 1 , wherein the microorganism is a microorganism belonging to the genus Escherichia.

Assignees

• Nat Univ Corp Nagoya Univ

Inventors

• Hori Katsutoshi

Classifications

• C12N11/00 Primary

  Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof · CPC title

• C12N11/08

  the carrier being a synthetic polymer · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• C12N11/02

  Enzymes or microbial cells immobilised on or in an organic carrier · CPC title

• C12N11/14

  Enzymes or microbial cells immobilised on or in an inorganic carrier · CPC title