Cardiff University | Prifysgol Caerdydd ORCA
Online Research @ Cardiff 
WelshClear Cookie - decide language by browser settings

Structure-guided engineering of a family IV cold-adapted esterase expands its substrate range

Noby, Nehad, Johnson, Rachel L., Tyzack, Jonathan D., Embaby, Amira M., Saeed, Hesham, Hussein, Ahmed, Khattab, Sherine N., Rizkallah, Pierre J. ORCID: and Jones, D. Dafydd ORCID: 2022. Structure-guided engineering of a family IV cold-adapted esterase expands its substrate range. International Journal of Molecular Sciences 23 (9) , 4703. 10.3390/ijms23094703

[thumbnail of ijms-23-04703.pdf]
PDF - Published Version
Available under License Creative Commons Attribution.

Download (3MB) | Preview
License URL:
License Start date: 24 April 2022


Cold active esterases have gained great interest in several industries. The recently determined structure of a family IV cold active esterase (EstN7) from Bacillus cohnii strain N1 was used to expand its substrate range and to probe its commercially valuable substrates. Database mining suggested that triacetin was a potential commercially valuable substrate for EstN7, which was subsequently proved experimentally with the final product being a single isomeric product, 1,2-glyceryl diacetate. Enzyme kinetics revealed that EstN7’s activity is restricted to C2 and C4 substrates due to a plug at the end of the acyl binding pocket that blocks access to a buried water-filled cavity. Residues M187, N211 and W206 were identified as key plug forming residues. N211A stabilised EstN7 allowing incorporation of the destabilising M187A mutation. The M187A-N211A double mutant had the broadest substrate range, capable of hydrolysing a C8 substrate. W206A did not appear to have any significant effect on substrate range either alone or when combined with the double mutant. Thus, the enzyme kinetics and engineering together with a recently determined structure of EstN7 provide new insights into substrate specificity and the role of acyl binding pocket plug residues in determining family IV esterase stability and substrate range

Item Type: Article
Date Type: Published Online
Status: Published
Schools: Biosciences
Additional Information: This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (
Publisher: MDPI
ISSN: 1422-0067
Funders: BBSRC
Date of First Compliant Deposit: 3 May 2022
Date of Acceptance: 16 April 2022
Last Modified: 22 May 2023 21:45

Citation Data

Cited 1 time in Scopus. View in Scopus. Powered By Scopus® Data

Actions (repository staff only)

Edit Item Edit Item


Downloads per month over past year

View more statistics