Power of Portieria hornemannii: influence on zebrafish antioxidant system-inflammatory cascade by combatting copper-induced inflammation.

Portieria hornemannii, a marine red alga, has attracted considerable attention due to its possible therapeutic characteristics. NFκB, a crucial transcription factor involved in regulating immune and inflammatory responses, plays a central role in controlling the expression of various genes associated with inflammation, such as MMP 9. This study employed an aqueous extract of Portieria hornemannii (Ph) and subjected it to UV-Visible, FTIR spectroscopy, and GC-MS analysis to identify its phytochemical composition. The presence of Vanillin, tert-butyldimethylsilyl ether, Dodecane, 1-fluoro-, and Pentanoic acid, 5-hydroxy-2,4-di-t-butylphenyl esters in Ph indicates their potential anti-inflammatory properties, suggesting their potential application in inflammation treatment. In summary, Ph exhibited anti-inflammatory properties by modulating the expressions of NFκB-associated MMP 9 in zebrafish embryos, potentially reducing the risk of inflammatory diseases.

Rapid, high-titer biosynthesis of melanin using the marine bacterium Vibrio natriegens.

Melanin is one of the most abundant natural biomolecules on Earth. These macromolecular biopolymers display several unique physical and chemical properties and have garnered interest as biomaterials for various commercial and industrial applications. To this end, extensive research has gone into refining methods for the synthesis and extraction of melanin from natural and recombinant sources. In this study, we developed and refined a procedure using a recombinant microbial system for the biosynthesis of melanin using the tyrosinase enzyme Tyr1 and tyrosine as a substrate. Using the emergent microbial chassis organisms Vibrio natriegens, we achieved maximal yields of 7.57 g/L, and one of the highest reported volumetric productivities of 473 mg L-1 h-1 with 100% conversion rates in an optimized, minimally defined medium. Additionally, we identified and investigated the use of a native copper responsive promoter in V. natriegens for stringent regulation of heterologous protein expression as a cost effective alternative to traditional IPTG-based induction. This research represents a promising advancement towards a green, rapid, and economical alternative for the biomanufacture of melanin.

Copper induction and differential expression of laccase in Aspergillus flavus.

Aspergillus flavus was isolated from soil and exhibited laccase activity under both constitutive and copper induced conditions. Spiking the medium with 1 mM copper sulfate resulted in an increase in the activity which reached 51.84 U/mL, a distinctive protein band was detected at 60 kDa. The extracellular enzyme was purified 81 fold using gel filtration chromatography and resulted in two different laccase fractions L1 and L2, the latter had a higher enzymatic activity which reached 79.57 U/mL and specific activity of 64.17 U/µg protein. The analysis of the spectrum of the L2 fraction showed a shoulder at 330 nm which is characteristic for T2/T3 copper centers; both copper and zinc were detected suggesting that this is an unconventional white laccase. Primers of laccase gene were designed and synthesized to recover specific gene from A. flavus . Sequence analysis indicated putative laccase (Genbank ID: JF683612) at the amino acid level suggesting a close identity to laccases from other genera containing the copper binding site. Decolorization of textile waste water under different conditions showed possible application in bioremediation within a short period of time. The effect of copper on A. flavus was concentration dependent.

Copper induction and differential expression of laccase in Aspergillus flavus

Aspergillus flavus was isolated from soil and exhibited laccase activity under both constitutive and copper induced conditions. Spiking the medium with 1 mM copper sulfate resulted in an increase in the activity which reached 51.84 U/mL, a distinctive protein band was detected at 60 kDa. The extracellular enzyme was purified 81 fold using gel filtration chromatography and resulted in two different laccase fractions L1 and L2, the latter had a higher enzymatic activity which reached 79.57 U/mL and specific activity of 64.17 U/μg protein. The analysis of the spectrum of the L2 fraction showed a shoulder at 330 nm which is characteristic for T2/T3 copper centers; both copper and zinc were detected suggesting that this is an unconventional white laccase. Primers of laccase gene were designed and synthesized to recover specific gene from A. flavus. Sequence analysis indicated putative laccase (Genbank ID: JF683612) at the amino acid level suggesting a close identity to laccases from other genera containing the copper binding site. Decolorization of textile waste water under different conditions showed possible application in bioremediation within a short period of time. The effect of copper on A. flavus was concentration dependent.

Toward an understanding of the effects of agitation and aeration on growth and laccases production by Pleurotus ostreatus.

Mycelial growth and laccase production by Pleurotus ostreatus CP50 cultured in a 10-L mechanically agitated bioreactor were assessed through a 2(3) factorial experimental design. The main effects and interactions of three factors (agitation, aeration and copper induction) over five responses (µ, αLacc, ßLacc, maximal volumetric laccase activity and maximal biomass concentration) were analyzed. P. ostreatus growth was significantly improved when culturing was conducted with high agitation (5.9kW/m(3)s) and aeration flow (0.5vvm) rates. Under the experimental conditions evaluated, no evidence of hydrodynamic stress affecting fungal growth was observed. However, the high agitation and aeration conditions were detrimental for the growth-associated laccase production constant (αLacc), leading to a very complex optimization of the process. The maximal laccase volumetric activity (1.2 and 3.8U/ml for non-induced and copper-induced cultures, respectively) was observed when the culturing was performed at a low agitation rate (0.9kW/m(3)s) and a high aeration flow rate (0.5vvm). Laccase proteolysis may explain the complex interactions observed between agitation and aeration and the effects of these factors on the laccase volumetric activity observed in the cultures.

Copper inducing effect on laccase production of white rot fungi native from Misiones (Argentina).

Fungi may be selected as models for gene expression studies and further adaptation for biotechnological enzyme production. The aim of this work was to evaluate laccase production and to analyze the effect of Cu(2+) on selected fungi natives of Misiones, Ganoderma applanatum (strain F), Peniophora sp. (BAFC 633), Pycnoporus sanguineus (BAFC 2126) and Coriolus versicolor f. antarcticus (BAFC 266). Fungi secretion system of G. applanatum, Peniophora sp., P. sanguineus and C. versicolor f. antarcticus is sensitive to stimulation by copper. Biomass values of G. applanatum, Peniophora sp. and C. versicolor f. antarcticus did not show differences between treatments. P. sanguineus biomass underwent a dramatic growth inhibition with 1mM Cu(2+) and marked delay in growth with 0.5mM Cu(2+). Proteins were increased with copper in Peniophora sp., C. versicolor and G. applanatum. G. applanatum and Peniophora sp. reached the highest enzyme activity at 10th day equivalent to 49.2-fold and 19.7-fold higher than the control samples, respectively. Copper produced an increase of constitutive laccases in all fungi and an additional inducible isoenzyme in Peniophora sp., C. versicolor f. antarcticus and G. applanatum.