Open Access. Powered by Scholars. Published by Universities.®

Biomedical Engineering and Bioengineering Commons

Open Access. Powered by Scholars. Published by Universities.®

Chemical and Biological Engineering Publications

2000

Articles 1 - 1 of 1

Full-Text Articles in Biomedical Engineering and Bioengineering

Replacement And Deletion Mutations In The Catalytic Domain And Belt Region Of Aspergillus Awamori Glucoamylase To Enhance Thermostability, Hsuan-Liang Liu, Yann Doleyres, Pedro M. Coutinho, Clark Ford, Peter J. Reilly Jan 2000

Replacement And Deletion Mutations In The Catalytic Domain And Belt Region Of Aspergillus Awamori Glucoamylase To Enhance Thermostability, Hsuan-Liang Liu, Yann Doleyres, Pedro M. Coutinho, Clark Ford, Peter J. Reilly

Chemical and Biological Engineering Publications

Three single-residue mutations, Asp71→Asn, Gln409→Pro and Gly447→Ser, two long-to-short loop replacement mutations, Gly23-Ala24-Asp25-Gly26-Ala27-Trp28-Val29-Ser30→Asn-Pro-Pro (23–30 replacement) and Asp297-Ser298-Glu299-Ala300-Val301→Ala-Gly-Ala (297–301 replacement) and one deletion mutation removing Glu439, Thr440 and Ser441 (Δ439–441), all based on amino acid sequence alignments, were made to improve Aspergillus awamori glucoamylase thermostability. The first and second single-residue mutations were designed to introduce a potential N-glycosylation site and to restrict backbone bond rotation, respectively, and therefore to decrease entropy during protein unfolding. The third single-residue mutation was made to decrease flexibility and increase O-glycosylation in the already highly O ...