Mutant of Bacillus thuringiensis and application thereof

• US 10,927,339 B2
• Filed: 03/16/2018
• Issued: 02/23/2021
• Est. Priority Date: 03/17/2017
• Status: Active Grant

• Current Assignee: Industrial Technology Research Institute
• Original Assignee: Industrial Technology Research Institute
• Inventors: Chen, Chang-Chieh, Bai, Ming-Der, Wu, Shao-I, Kuo, Hsin-Tzu, Lu, Wen-Chang, Wan, Hou-Peng
• Primary Examiner(s): Prouty, Rebecca E
• Application Number:
  US15/923,465
• Publication Number:
  US 20180265836A1
• Time in Patent Office: 1,075 Days
• Field of Search: None
• CPC Class Codes: C12N 1/02 Separating microorganisms from their culture media
• CPC Class Codes: C12N 1/12 Unicellular algae; Culture media therefor as new plants A01H13/00
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12N 1/38 Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound C12N1/34 takes precedence
• CPC Class Codes: C12N 15/75 for Bacillus
• CPC Class Codes: C12R 2001/075 Bacillus thuringiensis

Abstract

The present disclosure provides a new mutant of Bacillus thuringiensis which is deposited under Accession number DSM 32419. The present disclosure also provides a method for manufacturing a growth promoter for promoting microalgal cell growth, including: (a) inoculating a mutant of Bacillus thuringiensis into a culturing medium to obtain a bacterial suspension, wherein the mutant of Bacillus thuringiensis is deposited under Accession number DSM 32419; (b) culturing the mutant of Bacillus thuringiensis in the bacterial suspension at least to a stationary phase to obtain a cultured medium of the mutant of Bacillus thuringiensis; and (c) performing a vacuum heating procedure on the cultured medium of the mutant of Bacillus thuringiensis to obtain the growth promoter for promoting microalgal cell growth, wherein the growth promoter for promoting microalgal cell growth contains active substances for promoting microalgal cell growth.

Claims

1. A method for manufacturing a growth promoter for promoting microalgal cell growth, comprising:

(a) inoculating a mutant of Bacillus thuringiensis into a culturing medium to obtain a bacterial suspension, wherein the mutant of Bacillus thuringiensis is deposited under Accession number DSM 32419;

(b) culturing the mutant of Bacillus thuringiensis in the bacterial suspension at least to a stationary phase to obtain a cultured medium of the mutant of Bacillus thuringiensis, wherein a temperature for culturing the mutant of Bacillus thuringiensis is ranging from 20°

C. to 40°

C.; and

(c) performing a vacuum heating procedure on the cultured medium of the mutant of Bacillus thuringiensis to obtain the growth promoter for promoting microalgal cell growth, wherein the growth promoter for promoting microalgal cell growth contains active substances for promoting microalgal cell growth, and wherein a heating temperature for the vacuum heating procedure is ranging from 40°

C. to 50°

C. and a pressure of the vacuum heating procedure is ranging from 50 mmHg to 100 mmHg.

2. The method as claimed in claim 1, further comprising a step of removing the bacterial cells from the cultured medium of the mutant of Bacillus thuringiensis between step (b) and step (c).

3. The method as claimed in claim 2, wherein a way for removing the bacterial cells from the cultured medium of the mutant of Bacillus thuringiensis comprises performing centrifugation or filtration on the cultured medium of the mutant of Bacillus thuringiensis.

4. The method as claimed in claim 1, wherein a composition of the culturing medium comprises peptone and yeast extract.

5. The method as claimed in claim 1, wherein the mutant of Bacillus thuringiensis in the bacterial suspension is cultured for 12-72 hours at least to a stationary phase.

6. The method as claimed in claim 1, wherein the heating temperature for the vacuum heating procedure is 50°

C. and the pressure of the vacuum heating procedure is 80 mmHg.

7. A growth promoter for promoting microalgal cell growth, which is manufactured by the method for manufacturing a growth promoter for promoting microalgal cell growth as claimed in claim 1.

8. The growth promoter as claimed in claim 7, wherein the method further comprises a step of removing the bacterial cells from the cultured medium of the mutant of Bacillus thuringiensis between step (b) and step (c).

9. The growth promoter as claimed in claim 8, wherein a way for removing the bacterial cells from the cultured medium of the mutant of Bacillus thuringiensis comprises performing centrifugation or filtration on the cultured medium of the mutant of Bacillus thuringiensis.

10. The growth promoter as claimed in claim 7, wherein a composition of the culturing medium comprises peptone and yeast extract.

11. The growth promoter as claimed in claim 7, wherein the mutant of Bacillus thuringiensis in the bacterial suspension is cultured for 12-72 hours at least to a stationary phase.

12. The growth promoter as claimed in claim 7, wherein the heating temperature for the vacuum heating procedure is 50°

C. and the pressure of the vacuum heating procedure is 80 mmHg.

13. A method for promoting microalgal cell growth, comprising:

culturing a microalgal cell in the presence of the growth promoter for promoting microalgal cell growth as claimed in claim 7 to promote microalgal cell growth.

14. The method as claimed in claim 13, wherein a way for culturing a microalgal cell in the presence of the growth promoter for promoting microalgal cell growth comprises adding the growth promoter for promoting microalgal cell growth to a solution or medium containing a microalgal cell to form a mixture solution and culturing the mixture solution.

15. The method as claimed in claim 14, wherein the growth promoter for promoting microalgal cell growth accounts for 1-60% (v/v) of the mixture solution.

16. The method as claimed in claim 13, wherein the microalgal cell is a eukaryotic or prokaryotic unicellular or multicellular microalgae, and the microalgal cell has a cell wall and is capable of performing photosynthesis.

17. The method as claimed in claim 16, wherein the microalgal cell comprises Haematococcus pluvialis, Nannochloropsis sp., Chlorella sp., Tetraselmis sp., Isochrysis sp. or Dunaliella sp.

18. The method as claimed in claim 17, wherein the Tetraselmis sp. is Tetraselmis chuff.

19. The method as claimed in claim 17, wherein the Isochrysis sp. is Isochrysis galbana.