Heterologous expression and biophysical characterization of a mesophilic tannase following manganese nanoparticle immobilization

Nalok Dutta; Shahriar Md Miraz; Muhammad Usman Khan; Supriya Charuhas Karekar; Muhammad Usman; Shahbaz Manzoor Khan; Usman Amin; Maksim Rebezov; Mohammad Ali Shariati; Muthu Thiruvengadam

doi:10.1016/j.colsurfb.2021.112011

Heterologous expression and biophysical characterization of a mesophilic tannase following manganese nanoparticle immobilization

Colloids Surf B Biointerfaces. 2021 Nov:207:112011. doi: 10.1016/j.colsurfb.2021.112011. Epub 2021 Jul 29.

Affiliations

¹ Bioproducts Science & Engineering Laboratory, Department of Biological Systems Engineering, Washington State University Tri-Cities Campus, 2710 Crimson Way East 229, Richland, WA 99354, United States. Electronic address: [email protected].
² Department of Chemical Engineering, Konkuk University, Seoul, 05029, South Korea.
³ Department of Energy Systems Engineering, Faculty of Agricultural Engineering and Technology, University of Agriculture, Faisalabad, Pakistan.
⁴ Bioproducts Science & Engineering Laboratory, Department of Biological Systems Engineering, Washington State University Tri-Cities Campus, 2710 Crimson Way East 229, Richland, WA 99354, United States.
⁵ Department of Pathobiology, University of Illinois, 2522 Veterinary Medicine Basic Sciences Bldg. 2001 South Lincoln Avenue, Urbana, IL 61802, United States.
⁶ National Institute of Food Science and Technology, University of Agriculture, Faisalabad, Pakistan.
⁷ V M Gorbatov Federal Research Center for Food Systems of Russian Academy of Sciences, 26 Talalikhina St., Moscow, 109316, Russian Federation; Prokhorov General Physics Institute of the Russian Academy of Science, 38 Vavilova Str., Moscow, 119991, Russian Federation.
⁸ K.G. Razumovsky Moscow State University of Technologies and Management (the First Cossack University), 73 Zemlyanoy Val, 109004, Moscow, Russian Federation.
⁹ Department of Crop Science, College of Sanghuh Life Science, Konkuk University, Seoul, 05029, Republic of Korea. Electronic address: [email protected].

PMID: 34339969
DOI: 10.1016/j.colsurfb.2021.112011

Abstract

In the current study, we analyzed the efficacy of manganese oxide nanoparticle (MnNP)-water dispersion as an immobilization matrix for bacterial tannase. The tannase-secreting Bacillus subtilis strain NJKL.tan.2 obtained from tannery effluent soil was subsequently purified and cloned in pET20b vector. The activity of MnNP-tan (tannase activated by manganese nanoparticles) was 1.51- and 3.5-fold higher at 20 °C and 80 °C, respectively, compared with the free enzyme. MnNP-tan decreased K_m by 41.66 % and 3-fold, whereas free tannase showed two-fold and six-fold improvement in K_cat at 37 °C and 80 °C, respectively. MnNP-tan showed an increase in (half-life)t_1/2and E_d by 13-fold and 50.05 units, respectively, at 80 °C, in contrast to the native enzyme. MnNP-tan retained its residual activity by 78.2 % at 37 °C and 34.24 % at 80 °C after 180 min of incubation when compared with untreated set. MnNP-tan retained 51 % of its activity after 120 days with the native enzyme losing ∼50 % functionality following 40 days of incubation. The MnNP-mediated tannase immobilization technique is being reported for the first time. The technique has numerous advantages due to the use of MnNP as a potential matrix for biomolecule immobilization, which can be further extended to immobilize other biocatalysts used in agro-industrial and lab-based applications.

Keywords: Biophysical parameters; Heterologous expression; Manganese oxide nanoparticle.

MeSH terms

Bacillus subtilis
Carboxylic Ester Hydrolases* / chemistry
Carboxylic Ester Hydrolases* / genetics
Enzymes, Immobilized
Hydrogen-Ion Concentration
Ions
Manganese*
Nanoparticles*

Substances

Enzymes, Immobilized
Ions
Manganese
Carboxylic Ester Hydrolases
tannase