US 11,674,161 B2
Modified monooxygenases for the manufacture of hydroxylated hydrocarbons
Ruijing Guo, Shanghai (CN); Jen-Chieh Lin, Singapur (SG); Sha Tao, Nanjing (CN); Ying Qian, Nanjing (CN); Chenggang Qiu, Nanjing (CN); Kequan Chen, Nanjing (CN); and Kang Li, Nanjing (CN)
Assigned to Covestro Intellectual Property GmbH & Co. KG, Leverkusen (DE)
Appl. No. 17/607,922
Filed by Covestro Intellectual Property GmbH & Co. KG, Leverkusen (DE)
PCT Filed Jun. 18, 2020, PCT No. PCT/EP2020/066892
§ 371(c)(1), (2) Date Nov. 1, 2021,
PCT Pub. No. WO2020/260119, PCT Pub. Date Dec. 30, 2020.
Claims priority of application No. PCT/CN2019/093275 (WO), filed on Jun. 27, 2019; application No. PCT/CN2019/093326 (WO), filed on Jun. 27, 2019; application No. 19192042 (EP), filed on Aug. 16, 2019; and application No. 19192044 (EP), filed on Aug. 16, 2019.
Prior Publication US 2022/0220513 A1, Jul. 14, 2022
Int. Cl. C12N 9/00 (2006.01); C12P 7/22 (2006.01); C12N 9/02 (2006.01); C12P 17/06 (2006.01)
CPC C12P 7/22 (2013.01) [C12N 9/0077 (2013.01); C12N 9/0079 (2013.01); C12P 17/06 (2013.01); C12Y 114/15004 (2013.01)] 11 Claims
 
1. A modified P450 monooxygenase having an amino acid sequence with at least 90% sequence identity with SEQ ID NO: 1, wherein the asparagine at position 199 of the amino acid sequence of SEQ ID NO. 1 is substituted by an amino acid selected from the group consisting of glutamine, isoleucine, leucine, phenylalanine, histidine, methionine, arginine, serine, threonine, tyrosine, tryptophan, alanine, valine, and lysine to provide a functional mutation, wherein said functional mutation leads to an improved reactivity on hydroxylation of aromatic hydrocarbons as compared to a non-modified P450 monooxygenase.