Cell-wall degrading enzyme variants

US6808915B2 · kind B2 · utility

Assignee

• Novozymes A/S · DK

Inventors

• Sanne O. Schroder Glad · Rang Taché Est, DK
• Carsten Andersen
• Martin Schulein · Copenhagen, DK
• Torben Peter · Paseo Los Muchachos, DK

Key dates

• Filing date: Mar 31, 2003
• Grant date: Oct 26, 2004
• Expiry date: Mar 31, 2023

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N9/88
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention relates to variants of a cell-wall degrading enzyme having a beta-helix structure, which variant has at least one substituent in a position determined by identifying all residues potentially belonging to a stack; characterizing the stack as interior or exterior; characterizing the stack as polar, hydrophobic or aromatic/heteroaromatic based on the dominating characteristics of the parent or wild-type enzyme stack residues and/or its orientation relative to the beta-helix (interior or exterior); optimizing all stack positions of a stack either to hydrophobic aliphatic amino acids, hydrophobic aromatic or polar amino acids by allowing mutations within one or all positions to amino acids belonging to one of these groups; measuring thermostability of the variants by DSC or an application-related assay such as a Pad-Steam application test; and selecting the stabilized variants. Variant of a wild-type parent pectate lyase (EC 4.2.2.2) having the conserved amino acid residues D111, D141 or E141, D145, K165, R194 and R199 when aligned with the pectate lyase comprising the amino acid sequence of SEQ ID NO: 2 are preferred.