Purification, characterization and anticancer evaluation of l-methioninase from Trichoderma harzianum.

Abstract:

:The Trichoderma harzianum l-methioninase was purified 7.15-fold with a recovery of 47.9% and the specific activity of 74.4 U/mg of protein. The purified enzyme has an apparent molecular mass of 48 kDa on SDS-PAGE and exhibited maximum activity at pH 8 and 35 °C. The enzyme was catalytically stable below 50 °C and at a pH range of 6.0-8.5. The thermal inactivation of l-methioninase exhibited first-order kinetics with the k value between 5.71 × 10-4 min-1 and 1.83 × 10-2 min-1. The studies on thermodynamic parameters of l-methioninase indicated the compaction and aggregation of the enzyme molecule during denaturation. This is the first report of thermodynamic analysis of thermal inactivation in l-methioninase. The enzyme activity was enhanced by Li+ and inhibited by Cu2+, Co2+, Fe2+, Hydroxylamine and PMSF. The purified enzyme showed K m , Vmax and kcat value of 1.19 mM, 21.27 U/mg/min and 16.11 s-1, respectively. The l-methioninase inhibited the growth of human cell lines hepatocellular carcinoma (Hep-G2) and breast carcinoma (MCF-7) with IC50 values of 14.12 μg/ml and 20.07 μg/ml, respectively. The in vivo antitumor activity of l-methioninase was evaluated against DAL cell lines bearing in Swiss albino mice. The enzyme effectively reduced tumor volume, packed cell volume, viable cell count and restored hematological parameters, serum enzyme and lipid profile to normal levels compared to DAL control mice. The present study has demonstrated the high efficacy of Trichoderma harzianum l-methioninase against cancer cell lines in vitro and in vivo conditions. The purified l-methioninase has significant thermal stability and better catalytic properties than the enzyme purified from other sources.

journal_name

3 Biotech

journal_title

3 Biotech

authors

Salim N,Santhiagu A,Joji K

doi

10.1007/s13205-020-02494-w

subject

Has Abstract

pub_date

2020-11-01 00:00:00

pages

501

issue

11

eissn

2190-572X

issn

2190-5738

pii

2494

journal_volume

10

pub_type

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