Details

Autor(en) / Beteiligte

• Sahnoun, Mouna
• Jaoua, Mouna
• Bejar, Samir
• Jemli, Sonia

Titel

Highlight on mutations affecting the US132 cyclodextrin glucanotransferase binding specificity, thermal stability, and anti-staling activity

Ist Teil von

• Colloids and surfaces, B, Biointerfaces, 2022-04, Vol.212, p.112375-112375, Article 112375

Ort / Verlag

Netherlands: Elsevier B.V

Erscheinungsjahr

2022

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• We have already reported that the triple mutant (K47E-S382P-N655S of Paenibacillus pabuli US132 cyclodextrin glucanotransferase US132 (CGTase)) altered the CGTase specificity. In the current study, the single (K47E, S382P and N655S) and double (K47E+S382P, K47E+N655S, and S382P+N655S) mutants were constructed to elucidate the synergic or antagonist substitutions effect on the enzyme behavior. For the six generated mutants, an improvement of the dextrinization/cyclization ratio from 4.4 to 6-fold was observed when compared to the wild-type enzyme. The mutations effect on enzyme specificity was not attributed to synergy modulation since the single mutant N655S had the highest ratio enhancement. Moreover, the mutant N655S revealed the highest β-cyclodextrin binding affinity with a high amount of hydrophobic bonds which might be contributed to the apparent decrease in the cyclization activity. On the other hand, mutations N655S, K47E, and (K47E-N655S) showed the same positive effect on thermal activity. The highest stability was attained at 70 °C by N655S to be 3.6-fold higher than the wild-type. The addition of N655S to wheat flour induced a decrease of dough and bread hardness and led to an increase in dough and bread cohesiveness and a rise in bread masticability values compared to the control. This mutant addition also corrected the dough elasticity decrease engendered by the wild-type CGTase indicating that N655S-CGTase could be an alternative anti-staling agent. [Display omitted] •Six mutants were constructed to elucidate the substitutions effect.•The dextrinization/cyclization ratio was improved from 4.4 to 6-fold.•N655S had the highest D/C ratio and thermal stability.•N655S-CGTase could be an alternative anti-staling agent.