Enhancing electricity production in microbial fuel cells using defined co-cultures

Microbial fuel cells (MFCs) hold great promise for the simultaneous treatment of wastewater and electricity production. However, the electricity recovery is currently poor, typically <10% of what is theoretically possible, and the extracellular electron transfer mechanisms are poorly understood. The influence of using cocultures as a way of improving substrate turnover rate and hence electricity produced was investigated using synthetic wastewater as a substrate. Cocultures used were (i) Shewanella oneidensis and Clostridium beijerinckii; (ii) combinations of Geobacter sulphurreducens, Clostridium beijerinckii and Saccharomyces cerevisiae. The relative abundances test showed mutualistic relationship within the cocultures and was determined using RT-PCR at the end of the investigation. The coculture of S.oneidensis and C.beijerinkii gave a maximum power density of 87mWm-2 compared to 60 mWm-2 for C.beijerinckii alone and 48 mWm-2 for S.oneidensis alone. In the second study the best coculture combination was a mixture of Geobacter sulphurreducens, Clostridium beijerinckii and Saccharomyces cerevisiae giving a maximum power density of 80 mWm-2. Another study investigated the contribution of direct electron transfer mechanism on electricity production by physically separating Shewanella oneidensis to/from the anode electrode using a dialysis membrane. The outcome of this study indicated a maximum power output of 114±6 mWm-2 when cells were restricted close to the anode, 3.5 times more than when the cells were restricted away from the anode. Without the membrane the maximum power output was 129±6 mWm-2. These results highlight the importance of cocultures and direct electron transfer mechanism in improving electricity recovery in microbial fuel cells. Further work will seek to heterologously express the proteins in Shewanella involved in direct electron transfer in E.coli

An Efficient Green Synthesis of 3-Amino-1H-chromenes Catalyzed by ZnO Nanoparticles Thin-film

A very simple and environmentally benign approach for the synthesis of 3-amino-1H-chromenes is described using ZnO nanoparticles thin-film as an efficient heterogeneous catalyst in green media, namely water. The mild reaction conditions, reusability of the catalyst, easy work-up and high yields of products make the present protocol sustainable and advantageous compared to conventional methods.KEYWORDS:  3-Amino-1H-chromene, aqueousmedium,hydrothermalsolution method, reusability of the catalyst,ZnOnanoparticles thin-film

Fungal Enzymes as Catalytic Tools for Polyethylene Terephthalate (PET) Degradation.

The ubiquitous persistence of plastic waste in diverse forms and different environmental matrices is one of the main challenges that modern societies are facing at present. The exponential utilization and recalcitrance of synthetic plastics, including polyethylene terephthalate (PET), results in their extensive accumulation, which is a significant threat to the ecosystem. The growing amount of plastic waste ending up in landfills and oceans is alarming due to its possible adverse effects on biota. Thus, there is an urgent need to mitigate plastic waste to tackle the environmental crisis of plastic pollution. With regards to PET, there is a plethora of literature on the transportation route, ingestion, environmental fate, amount, and the adverse ecological and human health effects. Several studies have described the deployment of various microbial enzymes with much focus on bacterial-enzyme mediated removal and remediation of PET. However, there is a lack of consolidated studies on the exploitation of fungal enzymes for PET degradation. Herein, an effort has been made to cover this literature gap by spotlighting the fungi and their unique enzymes, e.g., esterases, lipases, and cutinases. These fungal enzymes have emerged as candidates for the development of biocatalytic PET degradation processes. The first half of this review is focused on fungal biocatalysts involved in the degradation of PET. The latter half explains three main aspects: (1) catalytic mechanism of PET hydrolysis in the presence of cutinases as a model fungal enzyme, (2) limitations hindering enzymatic PET biodegradation, and (3) strategies for enhancement of enzymatic PET biodegradation

Genomic and molecular characterization of a novel quorum sensing molecule in Bacillus licheniformis

Quorum sensing molecules (QSMs) are involved in the regulation of complicated processes helping bacterial populations respond to changes in their cell-density. Although the QS gene cluster (comQXPA) has been identified in the genome sequence of some bacilli, the QS system B. licheniformis has not been investigated in detail, and its QSM (ComX pheromone) has not been identified. Given the importance of this antagonistic bacterium as an industrial workhorse, this study was aimed to elucidate B. licheniformis NCIMB-8874 QS. The results obtained from bioinformatics studies on the whole genome sequence of this strain confirmed the presence of essential quorum sensing-related genes. Although polymorphism was verified in three proteins of this cluster, ComQ, precursor-ComX and ComP, the transcription factor ComA was confirmed as the most conserved protein. The cell–cell communication of B. licheniformis NCIMB-8874 was investigated through further elucidation of the ComX pheromone as 13-amino acid peptide. The peptide sequence of the pheromone has been described through biochemical characterisation

A cell engineering strategy to enhance supercoiled plasmid DNA production for gene therapy

With the recent revival of the promise of plasmid DNA vectors in gene therapy, a novel synthetic biology approach was used to enhance the quantity, (yield), and quality of the plasmid DNA. Quality was measured by percentage supercoiling and supercoiling density, as well as improving segregational stability in fermentation. We examined the hypothesis that adding a Strong Gyrase binding Site (SGS) would increase DNA gyrase-mediated plasmid supercoiling. SGS from 3 different replicons, (the Mu bacteriophage and two plasmids, pSC101 and pBR322) were inserted into the plasmid, pUC57. Different sizes of these variants were transformed into E. coli DH5α, and their supercoiling properties and segregational stability measured. A 36% increase in supercoiling density was found in pUC57-SGS, but only when SGS was derived from the Mu phage and was the larger sized version of this fragment. These results were also confirmed at fermentation scale. Total % supercoiled monomer was maintained to 85-90%. A two-fold increase in plasmid yield was also observed for pUC57-SGS in comparison to pUC57. pUC57-SGS displayed greater segregational stability than pUC57-cer and pUC57, demonstrating a further potential advantage of the SGS site. These findings should augment the potential of plasmid DNA vectors in plasmid DNA manufacture. This article is protected by copyright. All rights reserved

Pichia pastoris (Komagataella phaffii) as a Cost-Effective Tool for Vaccine Production for Low- and Middle-Income Countries (LMICs)

Vaccination is of paramount importance to global health. With the advent of the more recent pandemics, the urgency to expand the range has become even more evident. However, the potential limited availability and affordability of vaccines to resource low‐ and middle‐income countries has created a need for solutions that will ensure cost‐effective vaccine production methods for these countries. Pichia pastoris (P. pastoris) (also known as Komagataella phaffii) is one of the most promising candidates for expression of heterologous proteins in vaccines development. It combines the speed and ease of highly efficient prokaryotic platforms with some key capabilities of mammalian systems, potentially reducing manufacturing costs. This review will examine the latest developments in P. pastoris from cell engineering and design to industrial production systems with focus on vaccine development and with reference to specific key case studies

Compact active duplexer based on CSRR and interdigital loaded microstrip coupled lines for LTE application

In this paper, a four-port compact active duplexer based on a complimentary split ring resonator (CSRR) and interdigital loaded microstrip coupled lines (CSRR-IL MCL) is presented. Interdigital capacitor is used on the top layer of the proposed structure and CSRR transmission lines are used on the bottom layer of the coupled lines in order to increase the coupling of the proposed circuit and create triple band resonances, respectively. The proposed active duplexer has one input port and three output ports operating in three distinct operation frequencies which are 1.4 GHz, 1.8 GHz, and 3.2 GHz. The active duplexer is designed to target LTE applications which are prevalent among the new technologies and devices. The input signal is split in terms of frequency into the three designed frequencies and is amplified by 13 dB gain of the amplifiers placed at the output ports. The fractional bandwidths of the proposed structure at 1.4 GHz, 1.8 GHz, and 3.2 GHz are 5.2%, 2.8%, and 9.4%, respectively. It is worth mentioning that the size of the proposed active duplexer is 0.29λ0 × 0.38λ0. The design guide of the proposed structure is presented, and it will be shown that the simulation as well as the measurement results of the proposed active duplexer have an acceptable agreement with each other. It should be noted that the VSWR of the proposed structure is less than 1.5 which means that the active duplexer has low return loss, and it is the plus point of it

DESIGNING A COMPETITIVE ADVANTAGE MODEL WITH TECHNOLOGY ORIENTED APPROACH USING FAHP TECHNIQUE: A CASE STUDY IN COIL INDUSTRY

One of the distinctive attributes of today’s successful companies is having at least one competitive advantage in one known area. Technological competency is an important advantage which helps improve the firm’s competitiveness. In fact, suitable use of new technologies can dramatically influence the innovation speed, decrease the time of product development cycle and also increase the rate of new product introduction. Firm-specific technological competencies help explain why a firm is different, how it changes over time, and whether it is capable of remaining competitive. In this study, technological competency factors (technology management, process technology, product technology) are prioritized according to the competitive advantage levels(customer satisfaction, brand reputation, new product introduction, market share) and competitive priorities (cost, price, quality, flexibility, time) using fuzzy Analytic hierarchy process (FAHP) with the aim of maximizing the nonfinancial performance at coil manufacture industry. The results indicate that within Iran coil industry, process technology is of greater importance than technology management and product technology

Caveolin-3 differentially orchestrates cholinergic and serotonergic constriction of murine airways

The mechanisms of controlling airway smooth muscle (ASM) tone are of utmost clinical importance as inappropriate constriction is a hallmark in asthma and chronic obstructive pulmonary disease. Receptors for acetylcholine and serotonin, two relevant mediators in this context, appear to be incorporated in specialized, cholesterol-rich domains of the plasma membrane, termed caveolae due to their invaginated shape. The structural protein caveolin-1 partly accounts for anchoring of these receptors. We here determined the role of the other major caveolar protein, caveolin-3 (cav-3), in orchestrating cholinergic and serotonergic ASM responses, utilizing newly generated cav-3 deficient mice. Cav-3 deficiency fully abrogated serotonin-induced constriction of extrapulmonary airways in organ baths while leaving intrapulmonary airways unaffected, as assessed in precision cut lung slices. The selective expression of cav-3 in tracheal, but not intrapulmonary bronchial epithelial cells, revealed by immunohistochemistry, might explain the differential effects of cav-3 deficiency on serotonergic ASM constriction. The cholinergic response of extrapulmonary airways was not altered, whereas a considerable increase was observed in cav-3â -/- intrapulmonary bronchi. Thus, cav-3 differentially organizes serotonergic and cholinergic signaling in ASM through mechanisms that are specific for airways of certain caliber and anatomical position. This may allow for selective and site-specific intervention in hyperreactive states

Influence of Pichia pastoris cellular material on polymerase chain reaction performance as a synthetic biology standard for genome monitoring

Advances in synthetic genomics are now well underway in yeasts due to the low cost of synthetic DNA. These new capabilities also bring greater need for quantitating the presence, loss and rearrangement of loci within synthetic yeast genomes. Methods for achieving this will ideally; i) be robust to industrial settings, ii) adhere to a global standard and iii) be sufficiently rapid to enable at-line monitoring during cell growth. The methylotrophic yeast Pichia pastoris (P. pastoris) is increasingly used for industrial production of biotherapeutic proteins so we sought to answer the following questions for this particular yeast species. Is time-consuming DNA purification necessary to obtain accurate end-point polymerase chain reaction (e-pPCR) and quantitative PCR (qPCR) data? Can the novel linear regression of efficiency qPCR method (LRE qPCR), which has properties desirable in a synthetic biology standard, match the accuracy of conventional qPCR? Does cell cultivation scale influence PCR performance? To answer these questions we performed e-pPCR and qPCR in the presence and absence of cellular material disrupted by a mild 30s sonication procedure. The e-pPCR limit of detection (LOD) for a genomic target locus was 50 pg (4.91 × 103 copies) of purified genomic DNA (gDNA) but the presence of cellular material reduced this sensitivity sixfold to 300 pg gDNA (2.95 × 104 copies). LRE qPCR matched the accuracy of a conventional standard curve qPCR method. The presence of material from bioreactor cultivation of up to OD600 = 80 did not significantly compromise the accuracy of LRE qPCR. We conclude that a simple and rapid cell disruption step is sufficient to render P. pastoris samples of up to OD600 = 80 amenable to analysis using LRE qPCR which we propose as a synthetic biology standard