Improving Thermostability of Chimeric Enzymes Generated by Domain Shuffling Between Two Different Original Glucoamylases

Frontiers in Bioengineering and Biotechnology

Zhongxiu Chen¹  Yuyu Shen¹  Dunji Hu¹  Longbin Wang¹  Liying Zhou¹  Fuping Lu³  Ming Li³ 

[1] College of Biotechnology, Tianjin University of Science and Technology, Tianjin, China;Key Laboratory of Industrial Fermentation Microbiology (Tianjin University of Science and Technology), Ministry of Education, Tianjin, China;Tianjin Key Laboratory of Industrial Microbiology, Tianjin, China;

关键词: glucoamylase;    domain shuffling;    chimeric enzyme;    thermostability;    enzymatic properties;   

DOI  :  10.3389/fbioe.2022.881421

来源: DOAJ

【 摘 要 】

In order to improve enzymatic properties of glucoamylases, six recombinant genes GA1–GA6 were created by domain shuffling of glucoamylase genes GAA1 from Aspergillus niger Ld418AI and GATE from Talaromyces emersonii Ld418 TE using overlap extension PCR and were expressed in Saccharomyces cerevisiae W303-1B; only activities of GA1 and GA2 in the fermentation broth were higher than those of GAA1 but less than those of GATE. Further research results of GA1 and GA2 indicated that chimeric glucoamylases GA1 and GA2 revealed increased thermostability compared with GAA1 and GATE, although with a slight change in the activity and optimal temperature. However, GA1 had almost the same catalytic efficiency as GATE, whereas the catalytic efficiency of GA2 was slightly less than that of GATE, but still higher than that of GAA1. The structural analysis showed that the change of enzymatic properties could be caused by the increased and extended α-helix and β-sheet, which change the secondary and tertiary structures of chimeric glucoamylases. These results demonstrated that domain shuffling was feasible to generate a chimeric enzyme with novel properties.

【 授权许可】

Unknown