Trimeric l‐N‐carbamoylase from newly isolated Brevibacillus reuszeriHSN1: A potential biocatalyst for production of l‐α‐amino acids

Abstract

l-N-carbamoylase was isolated from Brevibacillus reuszeri HSN1 and purified to homogeneity in three steps, which is a reasonably short protocol for native l-N-carbamoylase. The enzyme purification protocol resulted in ≈60-fold purification of l-N-carbamoylase with specific activity of 145 µmol/Min/mg. The subunit and native molecular mass were found to be 44.3 and 132 kDa, respectively. Temperature and pH optima were determined as 50°C and 8.5, respectively. The enzyme had retained ≈86% activity at 50°C when incubated for 60 Min and the half-life was determined as 180 Min at 50°C. N-carbamoyl-l-methionine (l-N-CMet) was found to be a preferred substrate with Km and Vmax values of ≈13.5 mM and ≈103 µmol/Min/mg, respectively. The broad substrate specificity with derivatives of N-carbamoyl amino acids is advantageous to be a better biocatalyst for production of corresponding l-α-amino acids. The enzyme activity was enhanced by 73% in the presence of 0.8 mM Mn(2+) ion during the biotransformation. In the batch experiment, ≈97% conversion of 5.0% l-N-CMet into enantiomerically pure l-methionine was achieved in 10 H when carried out at pH 8.0, 45°C, and 15% wet (w/v) cell loading, under controlled conditions. The overall merits of this enzyme show promise as a potential biocatalyst for l-α-amino acid production.

Structural and biochemical characterisation of the N‐carbamoyl‐β‐alanine amidohydrolase from Rhizobium radiobacterMDC 8606

“…The protein was analysed as purified and following elution from a desalting column to remove any loosely bound metals. Raw detected values for 55 Mn, 59 Co, 60 Ni, 63 Cu, 66 Zn and 111 Cd were compared with commercial standard curve preparations (Merck). 107 Ag and 115 In were used as internal standards.…”

Bacillales: From Taxonomy to Biotechnological and Industrial Perspectives

“…Many species of Brevibacillus have been considered biological resources for the production of various enzymes, especially cyclodextrin glycosyltransferase used in cosmetics, pharmaceutical, and food industries, serine proteases, biomaterials caused improvement of plants growth or nodulation of roots, antimicrobial peptides, insecticidal agents, extracellular polysaccharide substances (EPSs), polyhydroxyalkanoate copolymers, L-amino acids, etc. [188][189][190][191]. Additionally, Brevibacillus brevis strains can degrade many organic compounds such as triphenyltin, pyrene, triphenyl phosphate, or textile dyes, and their high resistance to heavy metals makes them the right choice to use in contaminated sites [192][193][194][195].…”

Novel chitinolytic bacteria from the shrimp shell processing waste

“…Several studies have reported the ability of B. reuszeri bacteria, including capable of producing extracellular metalloprotease/thermolysin inhibitors (Kobayashi et al 2004) to produce L-methionine-N-carbamoylase as potential biocatalyst for L-amino acid production (Nandanwar et al 2013), phosphate dilution, plant growthpromoting rhizobacteria (PGPR) (Pradesh et al 2019;Prasad et al 2019;Yildirim et al 2011), and antimicrobial bioactive compound production (Gholizadeh et al 2013;Yang and Yousef 2018).…”