Purification and characterization of β-galactosidase from newly isolated Aspergillus terreus (KUBCF1306) and evaluating its efficacy on breast cancer cell line (MCF-7)

B Vidya; M Palaniswamy; J Angayarkanni; K Ayub Nawaz; M Thandeeswaran; K Krishna Chaithanya; Berihu Tekluu; Karthi Muthusamy; V K Gopalakrishnan

doi:10.1016/j.bioorg.2019.103442

Purification and characterization of β-galactosidase from newly isolated Aspergillus terreus (KUBCF1306) and evaluating its efficacy on breast cancer cell line (MCF-7)

Bioorg Chem. 2020 Jan:94:103442. doi: 10.1016/j.bioorg.2019.103442. Epub 2019 Nov 15.

Authors

B Vidya¹, M Palaniswamy², J Angayarkanni³, K Ayub Nawaz³, M Thandeeswaran³, K Krishna Chaithanya⁴, Berihu Tekluu⁴, Karthi Muthusamy⁵, V K Gopalakrishnan⁶

Affiliations

¹ Department of Biochemistry, Karpagam Academy of Higher Education, Coimbatore, India.
² Department of Microbiology, Karpagam Academy of Higher Education, Coimbatore, India.
³ Department of Microbial Biotechnology, Bharathiar University, Coimbatore, India.
⁴ Department of Chemistry, College of Natural and Computational Sciences, Aksum University, Ethiopia.
⁵ Department of Biochemistry, Bharathiar University, Coimbatore, India.
⁶ Department of Chemistry, College of Natural and Computational Sciences, Aksum University, Ethiopia. Electronic address: vkgopalakrishnan@gmail.com.

PMID: 31761411
DOI: 10.1016/j.bioorg.2019.103442

Abstract

β-galactosidases (EC 3.2.1.23) are able to catalyze two different types of reactions, namely hydrolysis and transgalactosylation. It is a lysosomal exoglycosidase involved in the catabolism of glycoconjugates by sequential release of β-linked terminal galactosyl residues. It has profound significance in cancer cell senescence. It can be derived from microbial sources including bacteria, yeasts, and filamentous fungi. The enzyme was purified from the crude enzyme using ammonium sulfate precipitation, dialysis, ion exchange chromatography using DEAE cellulose, fast protein liquid chromatography and high performance liquid chromatography. The enzyme was purified with 10.78 -fold with specific activity of 62 U/mg of protein and yield of 28.26%. Molecular weight of β -galactosidase as estimated by using SDS-PAGE was 42 kDa. Kinetic parameters K_m and V_max for purified enzyme were 0.48 and 0.96 respectively. Further the characterization and kinetic studies of purified enzyme were carried out. The optimum pH and temperature for maximum β-galactosidase activity were found to be 6, 40 °C, respectively. The present study is aimed to purification, characterization and in vitro efficacy assessment in breast cancer cell line. The β-galactosidase isolated from Aspergillus terreus was found to be effective in the proliferation of MCF-7 breast cancer cells in vitro. The present study is aimed to purification and characterization of enzyme to assess in vitro efficacy of β-galactosidase on MCF-7 cell line to delineate its therapeutic efficacy.

Keywords: Aspergillus terreus; FPLC; HPLC; MCF-7 breast cancer cells; Therapeutic efficacy; β-galactosidase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aspergillus / enzymology*
Breast Neoplasms / metabolism*
Breast Neoplasms / pathology
Cell Proliferation
Female
Humans
Hydrogen-Ion Concentration
MCF-7 Cells
Molecular Structure
Molecular Weight
Temperature
Tumor Cells, Cultured
beta-Galactosidase / chemistry
beta-Galactosidase / isolation & purification
beta-Galactosidase / metabolism*

Substances

beta-Galactosidase

Supplementary concepts

Aspergillus terreus