creators_name: Nandanwar, Hemraj S
creators_name: Vohra, Rakesh M
creators_name: Hoondal, Gurinder S
type: article
datestamp: 2019-09-05 14:12:10
lastmod: 2019-09-05 14:12:10
metadata_visibility: show
title: Enhanced stability of newly isolated trimeric l-methionine-N-carbamoylase from Brevibacillus reuszeri HSN1 by covalent immobilization.
ispublished: pub
subjects: QR
note: Copyright of this article belongs to  International Union of Biochemistry and Molecular Biology, Inc./Wiley
abstract: Newly isolated and partially purified trimeric l-methionine-N-carbamoylase from Brevibacillus reuszeri HSN1 was immobilized by covalent coupling to a well-known support material, Eupergit® C. Approximately 80% enzyme activity yield was achieved with ≈61% binding of a soluble protein from a solution containing 5 mg/mL protein. The immobilized preparation was found to be quite unstable due to a poor multisubunit covalent interaction of trimeric enzyme. Additional cross-linking with polyaldehyde-dextran was done to sustain the biotechnological application of immobilized enzyme. The temperature and pH optima of immobilized enzyme were increased by 10°C and 0.5 unit, respectively. The enzyme was significantly stabilized and retained ≈93% enzyme activity when incubated at 60°C for 60 Min, whereas free enzyme lost ≈50% activity. It was recycled nine times with ≈100% conversion efficiency when batch experiments were carried out at 35°C, pH 7.5, for the 180 Min cycle, using 5% N-carbamoyl-l-methionine as the substrate. The half-life of the immobilized preparation was determined as 23 cycles and is significant. Approximately 50% of enzyme activity was retained even after 5 months of storage at 4°C, whereas free enzyme lost complete enzyme activity. Hence, we could enhance the stability of l-methionine-N-carbamoylase to make it a potential biocatalyst for biotechnological production of α-amino acids.
date: 2013-05-18
publication: Biotechnology and applied biochemistry
volume: 60
number: 3
publisher: Wiley
pagerange: 305-15
refereed: TRUE
issn: 1470-8744
official_url: https://iubmb.onlinelibrary.wiley.com/doi/abs/10.1002/bab.1082
citation:   Nandanwar, Hemraj S and Vohra, Rakesh M and Hoondal, Gurinder S  (2013) Enhanced stability of newly isolated trimeric l-methionine-N-carbamoylase from Brevibacillus reuszeri HSN1 by covalent immobilization.  Biotechnology and applied biochemistry, 60 (3).  pp. 305-15.  ISSN 1470-8744