US 9,404,139 B2 · Mutated cephalosporin hydroxylase and its application in deacetylcephalosporanic acid synthesis

Mutated cephalosporin hydroxylase and its application in deacetylcephalosporanic acid synthesis

AssigneeOrchid Chemicals & Pharmaceuticals Limited

InventorsMicheal Durairaaj, Ramanan Thirumoorthy, Kanhu Charan Mishra, Thangadurai Chinnathambi, Cavery Manian Krishnan, Padma Rajasekaran, Sugumar Subramani, Kavitha Daffrose Selvaraj, Nataraj Balakrishnan, Sathish Ravikumar Chakravarthy, Arulmozhi Natrajan Madhiyazhagan

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Abstract

A mutant hydroxylase with increased activity and greater substrate specificity towards phenylacetyl-7-ADCA derivatives for the production of phenylacetyl-7-HACA derivatives, which carries one or more amino acid modification at residue positions when compared with certain wild type hydroxylase from certain groups of residues. Also disclosed is a process for the preparation of deacetyl cephalosporanic acid from the corresponding deacetoxy cephalosporanic acid that includes an enzyme of the present invention. Also provided is a method for the production and processing of such enzymes.

Claims9

Dependent claims

A mutant hydroxylase of a wild type hydroxylase having the amino acid sequence set forth in SEQ ID NO:

1, the mutant hydroxylase comprising an amino acid modification of at least one amino acid residue selected from the group consisting of P7, A40, T51, M53, E82, R91, T96, G108, I149, V171, A177, R182, M184, I193, F195, Q209, A210, V226, M233, L236, A237, A241, V249, R250, S251, G255, E258, 5260, F267, A280, V307, and with N313, wherein the said mutant hydroxylase comprises increased hydroxylase activity than that of the wild type hydroxylase of the amino acid sequence of SEQ ID NO:
1.

Dependencies are inferred from the claim text.

Facts

Application

14/371,402Filed 9 Jan 2013

Priority

IN 105/CHE/201210 Jan 2012

Published

US 2015/0240282 A127 Aug 2015

Granted

2 Aug 2016

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Classification7 codes

C12P 35/00
Preparation of compounds having a 5-thia-1-azabicyclo [4.2.0] octane ring system
C12N 9/0071
Oxidoreductases: acting on paired donors with incorporation of molecular oxygen
C12P 35/06
Preparation of compounds having a 5-thia-1-azabicyclo [4.2.0] octane ring system, e.g. cephalosporin: cephalosporin C; Derivatives thereof
Y02P 20/52
Improvements relating to the production of bulk chemicals: using catalysts, e.g. selective catalysts