Modifing Aspergillus Oryzae S2 amylase substrate specificity and thermostability through its tetramerisation using biochemical and in silico studies and stabilization

Sahnoun, Mouna; Jemli, Sonia; Trabelsi, Sahar; Béjar, Samír

doi:10.1016/j.ijbiomac.2018.05.136

Cited by 6 publications

(2 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Notably, GH13 enzymes share a common structural arrangement composed of conserved domains A, B, and C . In the GH13 family, α-amylase, trehalose synthase, and cyclodextrin-hydrolyzing enzyme are also present as homotetramers in solution. However, to the best of our knowledge, CT-ASase is the first homotetrameric ASase reported to date.…”

Section: Resultsmentioning

confidence: 99%

“…As shown in Table 1, TR-ASase, DG-ASase, DRd-ASase, DRp-ASase, MF-ASase, and MA-ASase exist as dimers in solution, while other ASases from mesophilic micro-organisms usually exist in a monomeric conformation. Notably, GH13 enzymes share a common structural arrangement composed of conserved domains A, B, and C. 32 In the GH13 family, αamylase, 33 trehalose synthase, 34 and cyclodextrin-hydrolyzing enzyme 20 are also present as homotetramers in solution.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Thermostable Amylosucrase from Calidithermus timidus DSM 17022: Insight into Its Characteristics and Tetrameric Conformation

Tian

Guang

et al. 2019

J. Agric. Food Chem.

View full text Add to dashboard Cite

Amylosucrase (EC 2.4.1.4, ASase), a typical carbohydrate-active enzyme, can catalyze 5 types of reactions and recognize more than 50 types of glycosyl acceptors. However, most ASases are unstable even at 50 °C, which limits their practical industrial applications. In this study, an extremely thermostable ASase was discovered from Calidithermus timidus DSM 17022 (CT-ASase) with an optimal activity temperature of 55 °C, half-life of 1.09 h at 70 °C, and melting temperature of 74.47 °C. The recombinant CT-ASase was characterized as the first tetrameric ASase, and a structure-based truncation mutation was conducted to confirm the effect of tetrameric conformation on its thermostability. In addition, α-1,4-glucan was found to be the predominant product of CT-ASase at pH 6.0−8.0 and 30−60 °C.

show abstract

Section: Resultsmentioning

confidence: 99%