	US 12,234,491 B2
	Methods of recombinantly producing neutral protease originating from Paenibacillus polymxa
Thomas Greiner-Stoeffele, Soemmerda (DE); and Stefan Schoenert, Leipzig (DE)
Assigned to Roche Diagnostics Operations, Inc., Indianapolis, IN (US)
Filed by Roche Diagnostics Operations, Inc., Indianapolis, IN (US)
Filed on Nov. 22, 2022, as Appl. No. 18/057,800.
Application 18/057,800 is a continuation of application No. 17/386,106, filed on Jul. 27, 2021, granted, now 11,542,489.
Application 17/386,106 is a continuation of application No. 16/355,259, filed on Mar. 15, 2019, abandoned.
Application 16/355,259 is a continuation of application No. 15/334,658, filed on Oct. 26, 2016, granted, now 10,526,594, issued on Jan. 7, 2020.
Application 15/334,658 is a continuation of application No. 14/592,969, filed on Jan. 9, 2015, abandoned.
Application 14/592,969 is a continuation of application No. PCT/EP2013/064271, filed on Jul. 5, 2013.
Claims priority of application No. 12175563 (EP), filed on Jul. 9, 2012.
Prior Publication US 2024/0132866 A1, Apr. 25, 2024 Prior Publication US 2024/0228999 A9, Jul. 11, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/54 (2006.01); C07K 14/195 (2006.01); C12N 5/071 (2010.01); C12N 9/52 (2006.01); C12N 15/57 (2006.01); C12N 15/75 (2006.01)

CPC C12N 9/52 (2013.01) [C07K 14/195 (2013.01); C12N 5/0602 (2013.01); C12Y 304/24 (2013.01); C12Y 304/24028 (2013.01); C12N 2509/00 (2013.01)]

3 Claims

1. A method for recombinantly producing a neutral protease, the method comprising the steps of

(a) providing in an expression vector a DNA with a sequence encoding a preproenzyme comprising the amino acid sequence of SEQ ID NO:5, and transforming a host organism with the expression vector, thereby obtaining a transformed host organism, wherein the host organism is a gram-positive prokaryotic species; followed by

(b) expressing the DNA in the transformed host organism, wherein the transformed host organism secretes the neutral protease as a proteolytic processing product of said preproenzyme; followed by

thereby recombinantly producing the neutral protease, and

wherein the gram-positive prokaryotic species is Bacillus amyloliquefaciens.