Characterization of a mildly alkalophilic and thermostable recombinant <Emphasis Type="Italic">Thermus thermophilus</Emphasis> laccase with applications in decolourization of dyes

Objective

To examine the potential for applications of TthLAC, a monomeric (~ 53 kDa) laccase encoded by the genome of Thermus thermophilus (strain HB 27) which can be produced at low cost in Escherichia coli.

Result

Functional, thermostable and mildly alkalophilic TthLAC of high purity (> 90%) was produced through simple heating of suspended (TthLAC overexpressing) E.coli cells at 65 °C. For reactions of short duration (< 1 h) the temperature for optimal activity is ~ 90 °C. However, TthLAC undergoes slow partial unfolding and thermal inactivation above 65 °C, making it unsuitable for long incubations above this temperature. With different substrates, optimal function was observed from pH 6 to 8. With the substrate, ABTS, catalytic efficiency (K _m) and maximum velocity (V_max) at 60 °C and pH 6.0 were determined to be 2.4 × 10³ µM and 0.04 × 10³ µM/min respectively. Ultra-pure, affinity-purified TthLAC was used to confirm and characterize the enzyme’s ability to oxidize known (laccase) substrates such as ABTS, syringaldazine and 4-fluoro-2-methylphenol. TthLAC decoloured up to six different industrial dyes, with or without the use of redox mediators such as ABTS.

Conclusions

Unlike versatile laccases from most other sources, which tend to be thermolabile as well as acidophilic, TthLAC is a versatile, thermostable, mildly alkalophilic laccase which can be produced at low cost in E.coli for various redox applications.

     

Localization of an Ataxia-Telangiectasia Gene to an −500-kb Interval on Chromosome 11q23.1: Linkage Analysis of 176 Families by an International Consortium

We describe a 20-point linkage analysis map of chromosome 11q22-23 that is based on genotyping 249 families (59 CEPH and 190 A-T). Monte Carlo linkage analyses of 176 ataxia-telangiectasia (A-T) families localizes the major A-T locus to the region between S1819(A4) and S1818(A2). When seven nonlinking families were excluded from subsequent analyses, a 2-lod support interval of ~500 kb was identified between S1819(A4) and S1294. No recombinants were observed between A-T and markers S384, B7, S535, or S1294. Only 17 of the international consortium families have been assigned to complementation groups. The available evidence favors either a cluster of A-T genes on chromosome 11 or intragenic defects in a single gene.     

Bactericidal Potency and Extended Serum Life of Stereo-Chemically Engineered Peptides Against Mycobacterium

International Journal of Peptide Research and Therapeutics - Tuberculosis is one of the leading causes of death, with an annual mortality rate of 2&nbsp;million. The present treatment regimen...     

Modeling bioaffinity-based targeted delivery of antimicrobials to Escherichia coli biofilms using yeast microparticles. Part I: Model development and numerical simulation

Biofilms are potential reservoirs for pathogenic microbes leading to a significant challenge for food safety, ecosystems, and human health. Various micro-and nanoparticles have been experimentally evaluated to improve biofilm inactivation by targeted delivery of antimicrobials. However, the role of transport processes and reaction kinetics of these delivery systems are not well understood. In this study, a mechanistic modeling approach was developed to understand the targeted delivery of chlorine to an Escherichia coli biofilm using a novel bioaffinity-based yeast microparticle. Biofilm inactivation by this delivery system was numerically simulated as a combination of reaction kinetics and transport phenomena. Simulation results demonstrate that the targeted delivery system achieved 7 log reduction within 16.2 min, while the equivalent level of conventional free chlorine achieved only 3.6 log reduction for the same treatment time. These numerical results matched the experimental observations in our previous study. This study illustrates the potential of a mechanistic modeling approach to improve fundamental understanding and guide the design of targeted inactivation of biofilms using biobased particles.     

Structure of Membrane-Bound Huntingtin Exon 1 Reveals Membrane Interaction and Aggregation Mechanisms

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How chromosomal rearrangements shape adaptation and speciation: Case studies in Drosophila pseudoobscura and its sibling species Drosophila persimilis

The gene arrangements of Drosophila have played a prominent role in the history of evolutionary biology from the original quantification of genetic diversity to current studies of the mechanisms for the origin and establishment of new inversion mutations within populations and their subsequent fixation between species supporting reproductive barriers. This review examines the genetic causes and consequences of inversions as recombination suppressors and the role that recombination suppression plays in establishing inversions in populations as they are involved in adaptation within heterogeneous environments. This often results in the formation of clines of gene arrangement frequencies among populations. Recombination suppression leads to the differentiation of the gene arrangements which may accelerate the accumulation of fixed genetic differences among populations. If these fixed mutations cause incompatibilities, then inversions pose important reproductive barriers between species. This review uses the evolution of inversions in Drosophila pseudoobscura and D. persimilis as a case study for how inversions originate, establish and contribute to the evolution of reproductive isolation.