Laccase-immobilized on superparamagnetic iron oxide nanoparticles incorporated polymeric ultrafiltration membrane for the removal of toxic pentachlorophenol.

A biocatalytic membrane offers an ideal alternative to the conventional treatment process for the removal of toxic pentachlorophenol (PCP). The limelight of the study is to utilize superparamagnetic iron oxide nanoparticles (SPIONs) incorporated (poly (methyl vinyl ether-alt-maleic acid) (PMVEAMA) and poly (ether - ether) sulfone (PEES)) membrane for immobilization of laccase and its application towards the removal of PCP. In regard to immobilization of Tramates versicolor laccase onto membranes, 5 mM glutaraldehyde with 10 h cross-linking time was employed, yielding 76.92% and 77.96% activity recovery for PEES/PMVEAMA/La and PEES/PMVEAMA/SPIONs/Lac, respectively. In the context of kinetics and stability studies, the immobilized laccase on PEES/PMVEAMA/Lac membrane outperforms the free and PEES/PMVEAMA laccases. At pH 7.0, the free enzyme loses half of its activity, while the immobilized laccases maintained more than 87% of their initial activity even after 480 min. With regard to PCP removal, the removal efficiency of immobilized laccase on the membrane was more than free enzyme. With 100 ppm of PCP, immobilized laccase on PEES/PMVEAMA/SPIONs membrane at pH 4.0 and 50 °C had a removal efficacy of 61.65% in 24 h. Furthermore, to perk up the removal of PCP, the laccase-aided system with mediators was investigated. Amongst, veratryl alcohol displayed 71.04% of PCP removal using immobilized laccase. The reusability of the laccase heightened after immobilization on PEES/PMVEAMA/SPIONs portraying 62.44% of the residual activity with 39.4% of PCP removal even after five cycles. The current investigation reveals the efficacy of the mediator-aided PEES/PMVEAMA/lac membrane system towards removing PCP from the aqueous solution, which can also be proposed for a membrane bioreactor.

The hazardous threat of Bisphenol A: Toxicity, detection and remediation.

Bisphenol A (or BPA) is a toxic endocrine disrupting chemical that is released into the environment through modern manufacturing practices. BPA can disrupt the production, function and activity of endogenous hormones causing irregularity in the hypothalamus-pituitary-gonadal glands and also the pituitary-adrenal function. BPA has immuno-suppression activity and can downregulate T cells and antioxidant genes. The genotoxicity and cytotoxicity of BPA is paramount and therefore, there is an immediate need to properly detect and remediate its influence. In this review, we discuss the toxic effects of BPA on different metabolic systems in the human body, followed by its mechanism of action. Various novel detection techniques (LC-MS, GC-MS, capillary electrophoresis, immunoassay and sensors) involving a pretreatment step (liquid-liquid microextraction and molecularly imprinted solid-phase extraction) have also been detailed. Mechanisms of various remediation strategies, including biodegradation using native enzymes, membrane separation processes, photocatalytic oxidation, use of nanosorbents and thermal degradation has been detailed. An overview of the global regulations pertaining to BPA has been presented. More investigations are required on the efficiency of integrated remediation technologies rather than standalone methods for BPA removal. The effect of processing operations on BPA in food matrices is also warranted to restrict its transport into food products.

The trade-off of availability and growth inhibition through copper for the production of copper-dependent enzymes by Pichia pastoris.

BACKGROUND: Copper is an essential chemical element for life as it is a part of prosthetic groups of enzymes including super oxide dismutase and cytochrome c oxidase; however, it is also toxic at high concentrations. Here, we present the trade-off of copper availability and growth inhibition of a common host used for copper-dependent protein production, Pichia pastoris. RESULTS: At copper concentrations ranging from 0.1 mM (6.35 mg/L) to 2 mM (127 mg/L), growth rates of 0.25 h(-1) to 0.16 h(-1) were observed with copper uptake of as high as 20 mgcopper/gCDW. The intracellular copper content was estimated by subtracting the copper adsorbed on the cell wall from the total copper concentration in the biomass. Higher copper concentrations led to stronger cell growth retardation and, at 10 mM (635 mg/L) and above, to growth inhibition. To test the determined copper concentration range for optimal recombinant protein production, a laccase gene from Aspergillus clavatus [EMBL: EAW07265.1] was cloned under the control of the constitutive glyceraldehyde-3-phosphate (GAP) dehydrogenase promoter for expression in P. pastoris. Notably, in the presence of copper, laccase expression improved the specific growth rate of P. pastoris. Although copper concentrations of 0.1 mM and 0.2 mM augmented laccase expression 4 times up to 3 U/mL compared to the control (0.75 U/mL), while higher copper concentrations resulted in reduced laccase production. An intracellular copper content between 1 and 2 mgcopper/gCDW was sufficient for increased laccase activity. The physiology of the yeast could be excluded as a reason for the stop of laccase production at moderate copper concentrations as no flux redistribution could be observed by (13)C-metabolic flux analysis. CONCLUSION: Copper and its pivotal role to sustain cellular functions is noteworthy. However, knowledge on its cellular accumulation, availability and distribution for recombinant protein production is limited. This study attempts to address one such challenge, which revealed the fact that intracellular copper accumulation influenced laccase production and should be considered for high protein expression of copper-dependent enzymes when using P. pastoris. The results are discussed in the context of P. pastoris as a general host for copper -dependent enzyme production.

Modeling of process parameters for enhanced production of coenzyme Q10 from Rhodotorula glutinis.

Coenzyme Q10 (CoQ10) plays an indispensable role in ATP generation through oxidative phosphorylation and helps in scavenging superoxides generated during electron transfer reactions. It finds extensive applications specifically related to oxidative damage and metabolic dysfunctions. This article reports the use of a statistical approach to optimize the concentration of key variables for the enhanced production of CoQ10 by Rhodotorula glutinis in a lab-scale fermenter. The culture conditions that promote optimum growth and CoQ10 production were optimized and the interaction of significant variables para-hydroxybenzoic acid (PHB, 819.34 mg/L) and soybean oil (7.78% [v/v]) was studied using response surface methodology (RSM). CoQ10 production increased considerably from 10 mg/L (in control) to 39.2 mg/L in batch mode with RSM-optimized precursor concentration. In the fed-batch mode, PHB and soybean oil feeding strategy enhanced CoQ10 production to 78.2 mg/L.