首页|Truncation of C-terminal amino acids of GH26 endo-mannanase (ManB-1601) affects biochemical properties and stability against anionic surfactants

Truncation of C-terminal amino acids of GH26 endo-mannanase (ManB-1601) affects biochemical properties and stability against anionic surfactants

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  • NSTL
  • Elsevier
Hitherto, the contribution of C-terminal amino acids in structure, stability and function of GH26 endomannanases has not been demonstrated. Semi-logarithmic plot of endo-mannanase activity showed a progressive decline with increase in the number of truncated amino acids [ManB-CA5 (129 U/mL), ManB-CA10 (47 U/ mL), ManB-CA15 (0.05 U/mL) and ManB-CA20 (0.02 U/mL)]. ManB-CA5 and ManB-CA10 exhibited similar temperature and pH optima and product profile but biochemical properties (kinetic constants, mannan hydrolysis, response to metal ions and enzyme inhibitors) and stability (in presence of commercial detergents, anionic surfactants and organic solvents and half-life) were markedly affected. Interaction of truncated proteins with anionic surfactants was probed using intrinsic, Nile red, acrylamide quenching, resonance light scattering and synchronous fluorescence spectroscopy studies. Truncation of ten amino acids increased vulnerability to anionic surfactants as conformational changes, exposure of the hydrophobic core and susceptibility to unfolding process were observed. The microenvironment around Trp residues was affected more with surfactants as compared to Tyr residues in truncated proteins. Zn2+ coordination might not play a role in providing stability against SDS. MD simulation studies corroborated that C-terminal amino acids (327-336) helps in structure stabilization, regulating flexibility of loops around the active site and preventing denaturation in the presence of SDS.

GH26 endo-mannanaseC-terminal amino acidsSurfactantsEnzyme stabilityMolecular dynamicsCANDIDA-RUGOSA LIPASEBOVINE SERUM-ALBUMINBACILLUS SP CFR1601THERMAL-STABILITYMOLECULAR DOCKINGFLUORESCENCEPROTEINENDO-BETA-14-MANNANASEFERMENTATIONGUM

Monica, P.、Mutturi, Sarma、Kapoor, Mukesh

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CSIR Cent Food Technol Res Inst

2022

10.1016/j.enzmictec.2022.110031

Enzyme and Microbial Technology

Enzyme and Microbial Technology

EISCI
ISSN：0141-0229
年,卷(期)：2022.157
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