	US 11,926,853 B2
	Botulinum toxin producing method
Daisuke Yokouchi, Kanonji (JP); Akiko Miura, Kanonji (JP); Ryu Okada, Kanonji (JP); Yuzo Yamashita, Kanonji (JP); Yuya Sasaki, Kanonji (JP); and Shogo Nishihata, Kanonji (JP)
Assigned to The Research Foundation For Microbial Diseases of Osaka University, Suita (JP)
Appl. No. 17/416,342
Filed by The Research Foundation For Microbial Diseases of Osaka University, Suita (JP)
PCT Filed Dec. 17, 2019, PCT No. PCT/JP2019/049436 § 371(c)(1), (2) Date Jun. 18, 2021, PCT Pub. No. WO2020/129986, PCT Pub. Date Jun. 25, 2020.
Claims priority of application No. 2018-239214 (JP), filed on Dec. 21, 2018.
Prior Publication US 2022/0081682 A1, Mar. 17, 2022
Int. Cl. C12P 21/06 (2006.01); C12N 9/52 (2006.01)

CPC C12N 9/52 (2013.01) [C12Y 304/24069 (2013.01)]

14 Claims

1. A method for producing a botulinum toxin, comprising steps of:

(A) producing a botulinum toxin from a botulinum toxin-producing bacterium in a culture medium to produce a mixture a containing a bacterial component and a nucleic acid component both derived from the botulinum toxin and the botulinum toxin;

(B) subjecting the mixture a to a removal of the bacterial component to produce a mixture b containing the nucleic acid component and the botulinum toxin;

(C) adding an endonuclease to the mixture b to produce a mixture c containing a nucleic acid degradation product and the botulinum toxin; and

(D) subjecting the mixture c to a removal of the nucleic acid degradation product to produce an isolated botulinum toxin liquid d,

wherein the method excludes any acid precipitation step.