Cytochrome P450 BM-3 is a soluble fatty acid hydroxylase composed of a heme domain and a reductase domain on a single polypeptide chain. We recently described a laboratoryevolved variant of the P450 BM-3 heme domain which functions as an H2O2-driven hydroxylase (TMperoxygenase∫) and does not require NADPH, O2, or the reductase. This variant, which we named 21B3, allows us to carry out cytochrom...

The gene encoding aspartate aminotransferase of a thermophilic Bacillus species, YM-2, has been cloned and expressed efficiently in Escherichia coli. The primary structure of the enzyme was deduced from nucleotide sequences of the gene and confirmed mostly by amino acid sequences of tryptic peptides. The gene consists of 1,176 base pairs encoding a protein of 392 amino acid residues; the molecu...

Abstract Protein dynamics are often invoked in explanations of enzyme catalysis, but their design has proven elusive. Here we track the role evolution, starting from evolvable and thermostable ancestral protein Anc HLD-RLuc which catalyses both dehalogenase luciferase reactions. Insertion-deletion (InDel) backbone mutagenesis challenged scaffold dynamics. Screening for activities reveals InDel ...

Most substrate for esterification has the inherent problem of low miscibility which requires addition of solvents into the reaction media. In this contribution, we would like to present an alternative and feasible option for an efficient solvent-free synthesis of menthyl butyrate using a novel thermostable crude T1 lipase. We investigated the effects of incubation time, temperature, enzyme load...

To meet the demand for the application of high activity and thermostable cellulases in the production of new-generation bioethanol from nongrain-cellulose sources, a hyperthermostable β-1,4-endoglucase Cel12B from Thermotoga maritima was selected for further modification by gene site-directed mutagenesis method in the present study, based on homology modeling and rational design. As a result, t...

Over the past few years considerable research attention has been assigned to extremophiles as sources of extremozymes due to their applicability in industrial processes, and the development of eco-friendly technologies. The establishment of efficient production strategies for heterologous proteins is an empirical process requiring broad background knowledge on available expression systems toget...

Raw starch degrading enzyme specially glucoamylase with binding domain (SBD) has great values in the processing industry because it digests particles below gelatinization temperature by releasing glucose from non-reducing ends sequentially. The purpose of study was to measure secretion levels recombinant Pichia pastoris, using α-mating factor signal peptide (α-MF) and native Aspergillus flavus ...

BACKGROUND Domain fusion is limited at enzyme one terminus. The issue was explored by swapping a mesophilic Aspergillus niger GH11 xylanase (Xyn) with a hyper-thermophilic Thermotoga maritima glucanase (Glu) to construct two chimeras, Xyn-Glu and Glu-Xyn, with an intention to create thermostable xylanase containing glucanase activity. RESULTS When expressed in E. coli BL21(DE3), the two chime...

Pectin methylesterase (PME) is an important enzyme that can alter the texture of potato products during processing and storage. To manage textural properties based products, low and/or high temperature blanching be used to promote or inactivate PME, respectively. However, many cultivars being for may contain PMEs with a different thermal stability. Therefore, inactivation PME extract from three...

L-Amino acid oxidase (LAAO) is a flavin adenine dinucleotide (FAD)-dependent enzyme active on most proteinogenic L-amino acids, catalysing their conversion to α-keto acids by oxidative deamination of the substrate. For this oxidation reaction, molecular oxygen used as electron acceptor, generating hydrogen peroxide. LAAO can be detect for production peroxide an agent or antimicrobial agent, and...