	US 11,866,713 B2
	Compositions and methods for increased protein production in bacillus licheniformis
Marc Anton Bernhard Kolkman, Oegstgeest (NL); Steven D. Doig, Wilmington, DE (US); Ryan L. Frisch, Newark, DE (US); Hongxian He, Wilmington, DE (US); Frank Wouter Koopman, Utrecht (NL); and Chris Leeflang, Twisk (NL)
Assigned to DANISCO US INC., Palo Alto, CA (US)
Appl. No. 16/486,892
Filed by DANISCO US INC, Palo Alto, CA (US)
PCT Filed Feb. 22, 2018, PCT No. PCT/US2018/019140 § 371(c)(1), (2) Date Aug. 19, 2019, PCT Pub. No. WO2018/156705, PCT Pub. Date Aug. 30, 2018.
Claims priority of provisional application 62/463,268, filed on Feb. 24, 2017.
Prior Publication US 2020/0056193 A1, Feb. 20, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/75 (2006.01); C07K 14/32 (2006.01); C12P 21/02 (2006.01)

CPC C12N 15/75 (2013.01) [C07K 14/32 (2013.01); C12P 21/02 (2013.01)]

4 Claims

1. A Bacillus licheniformis cell comprising:

1) A rghR2 gene encoding a RghR2 protein comprising at least 90% sequence identity to SEQ ID NO: 4, wherein the rghR2 gene contains an 18-nucleotide duplication encoding a repeat of amino acids AAAISR at amino acid positions 38-43 of the RghR2 protein of SEO ID NO: 4, and

2) A targeting vector configured to delete the 18-nucleotide duplication in the rghR2 gene, which upon deletion of the 18-nt duplication results in a modified cell comprising a modified rghR2 gene encoding a RghR2 protein comprising at least 90% sequence identity to SEQ ID NO: 2, wherein the modified cell produces an increased amount of a protein of interest relative to an unmodified cell.