Improvement of the Catalytic Ability of a Thermostable and Acidophilic β-Mannanase Using a Consensus Sequence Design Strategy.

ABSTRACT: In order to improve the catalytic efficiency of a thermostable and acidophilic β-mannanase (ManAK; derived from marine Aspergillus kawachii IFO 4308), three mutants were designed by amino acid sequence consensus analysis with a second β-mannanase (ManCbs), which also belongs to the glycoside hydrolase family 5 (GH5) and has excellent catalytic efficiency. Three mutants were constructed and their biochemical characteristics were measured after heterologous expression in Pichia pastoris. The results revealed that the k_cat/K_m values of the three recombinant mannanases ManAK^C292V, ManAK^L293V, and ManAK^L294H were enhanced by 303.0, 280.4, and 210.1%, respectively. Furthermore, ManAK^L293V showed greater thermostability than ManAK, retaining 36.5% of the initial enzyme activity after incubation at 80°C for 5min. This study therefore provides a rational design strategy based on consensus sequence analysis to develop industrially valuable β-mannanase for future applications in marine aquafeed.

SUBMITTER: Liang Q 

PROVIDER: S-EPMC8440911 | biostudies-literature |

REPOSITORIES: biostudies-literature