Characterization of a Novel Thermostable Enzyme from Thermus sp. 2.9 with Phospholipase and Acyltransferase Activities

2018 ◽  
Vol 28 (3) ◽  
pp. 99-106
Author(s):  
Laura Emilce Navas ◽  
Mónica Florin-Christensen ◽  
Graciela Beatriz Benintende ◽  
Rubén Oreste Zandomeni ◽  
Marcelo Facundo Berretta
Keyword(s):  
Egg Yolk ◽  
Thermostable Enzyme ◽  
Industrial Processes ◽  
Gene Encoding ◽  
High Thermostability ◽  
Egg Yolk Phosphatidylcholine ◽  
Wide Range ◽  
Potential Applications ◽  
Four Blocks ◽  
Differential Properties

Phospholipases are classified in different enzyme families according to the ester bond they cleave within phospholipids. The use of phospholipases in industrial processes has prompted the search for new enzymes with differential properties. A gene encoding a novel phospholipase (PLP_2.9) was identified in the genome of the thermophilic strain <i>Thermus</i> sp. 2.9. The analysis of the primary sequence unveiled a patatin-like domain. The alignment of the amino acid sequence of PLP_2.9 to other bacterial patatin-related proteins showed that the four blocks characteristic of this type of phospholipases and the amino acids representing the catalytic dyad are conserved in this protein. PLP_2.9 was overexpressed in <i>Escherichia coli</i> and the purified enzyme was characterized biochemically. PLP_2.9<i></i> hydrolyzed <i>p</i>-nitrophenyl palmitate at alkaline pH over a wide range of temperatures (55–80°C), showing high thermostability. PLP_2.9 displayed phospholipase A and acyltransferase activities on egg yolk phosphatidylcholine. Due to its high thermostability, PLP_2.9 has potential applications as a catalyst in several industrial processes.

scholarly journals Screening of phospholipase A activity and its production by new actinomycete strains cultivated by solid-state fermentation

PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e3524 ◽  
Author(s):  
Priscila Sutto-Ortiz ◽  
María de los Angeles Camacho-Ruiz ◽  
Manuel R. Kirchmayr ◽  
Rosa María Camacho-Ruiz ◽  
Juan Carlos Mateos-Díaz ◽  
...  
Keyword(s):  
Solid State ◽  
Solid State Fermentation ◽  
High Throughput Screening ◽  
Egg Yolk ◽  
Specific Substrate ◽  
Phospholipase Activity ◽  
High Accumulation ◽  
Egg Yolk Phosphatidylcholine ◽  
Potential Applications ◽  
Routine Assay

Novel microbial phospholipases A (PLAs) can be found in actinomycetes which have been poorly explored as producers of this activity. To investigate microbial PLA production, efficient methods are necessary such as high-throughput screening (HTS) assays for direct search of PLAs in microbial cultures and cultivation conditions to promote this activity. About 200 strains isolated with selected media for actinomycetes and mostly belonging toStreptomyces(73%) andMicromonospora(10%) genus were first screened on agar-plates containing the fluorophore rhodamine 6G and egg yolk phosphatidylcholine (PC) to detect strains producing phospholipase activity. Then, a colorimetric HTS assay for general PLA activity detection (cHTS-PLA) using enriched PC (≈60%) as substrate and cresol red as indicator was developed and applied; this cHTS-PLA assay was validated with known PLAs. For the first time, actinomycete strains were cultivated by solid-state fermentation (SSF) using PC as inductor and sugar-cane bagasse as support to produce high PLA activity (from 207 to 2,591 mU/g of support). Phospholipase activity of the enzymatic extracts from SSF was determined using the implemented cHTS-PLA assay and the PC hydrolysis products obtained, were analyzed by TLC showing the presence of lyso-PC. Three actinomycete strains of theStreptomycesgenus that stood out for high accumulation of lyso-PC, were selected and analyzed with the specific substrate 1,2-α-eleostearoyl-sn-glycero-3-phosphocholine (EEPC) in order to confirm the presence of PLA activity in their enzymatic extracts. Overall, the results obtained pave the way toward the HTS of PLA activity in crude microbial enzymatic extracts at a larger scale. The cHTS-PLA assay developed here can be also proposed as a routine assay for PLA activity determination during enzyme purification,directed evolution or mutagenesis approaches. In addition, the production of PLA activity by actinomycetes using SSF allow find and produce novel PLAs with potential applications in biotechnology.

scholarly journals Current Advances in Ejector Modeling, Experimentation and Applications for Refrigeration and Heat Pumps. Part 2: Two-Phase Ejectors

Inventions ◽  
2019 ◽  
Vol 4 (1) ◽  
pp. 16 ◽  
Author(s):  
Zine Aidoun ◽  
Khaled Ameur ◽  
Mehdi Falsafioon ◽  
Messaoud Badache
Keyword(s):  
Heat Pumps ◽  
Operating Conditions ◽  
Industrial Processes ◽  
Experimental Investigations ◽  
Mixing Enhancement ◽  
Data Generation ◽  
Two Phase ◽  
Wide Range ◽  
Potential Applications ◽  
And Performance

Two-phase ejectors play a major role as refrigerant expansion devices in vapor compression systems and can find potential applications in many other industrial processes. As a result, they have become a focus of attention for the last few decades from the scientific community, not only for the expansion work recovery in a wide range of refrigeration and heat pump cycles but also in industrial processes as entrainment and mixing enhancement agents. This review provides relevant findings and trends, characterizing the design, operation and performance of the two-phase ejector as a component. Effects of geometry, operating conditions and the main developments in terms of theoretical and experimental approaches, rating methods and applications are discussed in detail. Ejector expansion refrigeration cycles (EERC) as well as the related theoretical and experimental research are reported. New and other relevant cycle combinations proposed in the recent literature are organized under theoretical and experimental headings by refrigerant types and/or by chronology whenever appropriate and systematically commented. This review brings out the fact that theoretical ejector and cycle studies outnumber experimental investigations and data generation. More emerging numerical studies of two-phase ejectors are a positive step, which has to be further supported by more validation work.

scholarly journals Development and Optimization of Lipase-Catalyzed Synthesis of Phospholipids Containing 3,4-Dimethoxycinnamic Acid by Response Surface Methodology

Catalysts ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 588
Author(s):  
Magdalena Rychlicka ◽  
Natalia Niezgoda ◽  
Anna Gliszczyńska
Keyword(s):  
Response Surface Methodology ◽  
Food Additives ◽  
Reaction Medium ◽  
Egg Yolk ◽  
Molar Ratio ◽  
Molar Ratios ◽  
Egg Yolk Phosphatidylcholine ◽  
Potential Applications ◽  
Substrate Molar Ratio ◽  
Enzyme Load

The interesterification reaction of egg-yolk phosphatidylcholine (PC) with ethyl ester of 3,4-dimethoxycinnamic acid (E3,4DMCA) catalyzed by Novozym 435 in hexane as a reaction medium was shown to be an effective method for the synthesis of corresponding structured O-methylated phenophospholipids. The effects of substrate molar ratios, time of the reaction and enzyme load on the process of incorporation of 3,4DMCA into PC were evaluated by using the experimental factorial design of three factors and three levels. The results showed that a substrate molar ratio is a crucial variable for the maximization of the synthesis of 3,4-dimethoxycinnamoylated phospholipids. Under optimized parameters of 1/10 substrate molar ratio PC/E3,4DMCA, enzyme load 30% (w/w), hexane as a medium and incubation time of 3 days, the incorporation of aromatic acid into phospholipid fraction reached 21 mol%. The modified phosphatidylcholine (3,4DMCA-PC) and modified lysophosphatidylcholine (3,4DMCA-LPC) were obtained in isolated yields of 3.5% and 27.5% (w/w), respectively. The developed method of phosphatidylcholine interesterification is the first described in the literature dealing with 3,4DMCA and allows us to obtain new O-methylated phenophospholipids with potential applications as food additives or nutraceuticals with pro-health activity.

scholarly journals Sphingomyelin exhibits greatly enhanced protection compared with egg yolk phosphatidylcholine against detergent bile salts

2000 ◽  
Vol 41 (6) ◽  
pp. 916-924 ◽  
Author(s):  
Antonio Moschetta ◽  
Gerard P. vanBerge-Henegouwen ◽  
Piero Portincasa ◽  
Giuseppe Palasciano ◽  
Albert K. Groen ◽  
...  
Keyword(s):  
Bile Salts ◽  
Egg Yolk ◽  
Egg Yolk Phosphatidylcholine

scholarly journals A Slot-Die Technique for the Preparation of Continuous, High-Area, Chitosan-Based Thin Films

Polymers ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1566
Author(s):  
Oliver J. Pemble ◽  
Maria Bardosova ◽  
Ian M. Povey ◽  
Martyn E. Pemble
Keyword(s):  
Thin Films ◽  
Mechanical Properties ◽  
High Volume ◽  
Mechanical Anisotropy ◽  
Diverse Range ◽  
Direction Parallel ◽  
High Area ◽  
Wide Range ◽  
Slot Die ◽  
Potential Applications

Chitosan-based films have a diverse range of potential applications but are currently limited in terms of commercial use due to a lack of methods specifically designed to produce thin films in high volumes. To address this limitation directly, hydrogels prepared from chitosan, chitosan-tetraethoxy silane, also known as tetraethyl orthosilicate (TEOS) and chitosan-glutaraldehyde have been used to prepare continuous thin films using a slot-die technique which is described in detail. By way of preliminary analysis of the resulting films for comparison purposes with films made by other methods, the mechanical strength of the films produced was assessed. It was found that as expected, the hybrid films made with TEOS and glutaraldehyde both show a higher yield strength than the films made with chitosan alone. In all cases, the mechanical properties of the films were found to compare very favorably with similar measurements reported in the literature. In order to assess the possible influence of the direction in which the hydrogel passes through the slot-die on the mechanical properties of the films, testing was performed on plain chitosan samples cut in a direction parallel to the direction of travel and perpendicular to this direction. It was found that there was no evidence of any mechanical anisotropy induced by the slot die process. The examples presented here serve to illustrate how the slot-die approach may be used to create high-volume, high-area chitosan-based films cheaply and rapidly. It is suggested that an approach of the type described here may facilitate the use of chitosan-based films for a wide range of important applications.

scholarly journals Fungal Melanins and Applications in Healthcare, Bioremediation and Industry

2021 ◽  
Vol 7 (6) ◽  
pp. 488
Author(s):  
Ellie Rose Mattoon ◽  
Radames J. B. Cordero ◽  
Arturo Casadevall
Keyword(s):  
Chemical Structure ◽  
Industrial Processes ◽  
Metal Chelation ◽  
Biological Functions ◽  
Societal Benefit ◽  
Potential Applications ◽  
Fungal Melanin ◽  
Human Use ◽  
Fungal Melanins ◽  
Isolated Species

Melanin is a complex multifunctional pigment found in all kingdoms of life, including fungi. The complex chemical structure of fungal melanins, yet to be fully elucidated, lends them multiple unique functions ranging from radioprotection and antioxidant activity to heavy metal chelation and organic compound absorption. Given their many biological functions, fungal melanins present many possibilities as natural compounds that could be exploited for human use. This review summarizes the current discourse and attempts to apply fungal melanin to enhance human health, remove pollutants from ecosystems, and streamline industrial processes. While the potential applications of fungal melanins are often discussed in the scientific community, they are successfully executed less often. Some of the challenges in the applications of fungal melanin to technology include the knowledge gap about their detailed structure, difficulties in isolating melanotic fungi, challenges in extracting melanin from isolated species, and the pathogenicity concerns that accompany working with live melanotic fungi. With proper acknowledgment of these challenges, fungal melanin holds great potential for societal benefit in the coming years.

scholarly journals Health Benefits of Uses and Applications of Moringa oleifera in Bakery Products

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 318
Author(s):  
Paula García Milla ◽  
Rocío Peñalver ◽  
Gema Nieto
Keyword(s):  
Moringa Oleifera ◽  
Essential Amino Acids ◽  
Bakery Products ◽  
Unani Medicine ◽  
Functional Value ◽  
Organoleptic Characteristics ◽  
Wide Range ◽  
Potential Applications ◽  
Medicinal Properties ◽  
Excellent Source

Moringa oleifera belongs to the Moringaceae family and is the best known of the native Moringa oleifera genus. For centuries, it has been used as a system of Ayurvedic and Unani medicine and has a wide range of nutritional and bioactive compounds, including proteins, essential amino acids, carbohydrates, lipids, fibre, vitamins, minerals, phenolic compounds, phytosterols and others. These characteristics allow it to have pharmacological properties, including anti-diabetic, anti-inflammatory, anticarcinogenic, antioxidant, cardioprotective, antimicrobial and hepatoprotective properties. The entire Moringa oleifera plant is edible, including its flowers, however, it is not entirely safe, because of compounds that have been found mainly in the root and bark, so the leaf was identified as the safest. Moringa oleifera is recognised as an excellent source of phytochemicals, with potential applications in functional and medicinal food preparations due to its nutritional and medicinal properties; many authors have experimented with incorporating it mainly in biscuits, cakes, brownies, meats, juices and sandwiches. The results are fascinating, as the products increase their nutritional value; however, the concentrations cannot be high, as this affects the organoleptic characteristics of the supplemented products. The aim of this study is to review the application of Moringa oleifera in bakery products, which will allow the creation of new products that improve their nutritional and functional value.

scholarly journals Sporulation in solventogenic and acetogenic clostridia

2021 ◽  
Author(s):  
Mamou Diallo ◽  
Servé W. M. Kengen ◽  
Ana M. López-Contreras
Keyword(s):  
Current Knowledge ◽  
Carbon Sources ◽  
Cost Effective ◽  
Biotechnological Production ◽  
Key Points ◽  
Wide Range ◽  
Potential Applications ◽  
A Cell ◽  
Sporulation Initiation ◽  
Solventogenic Clostridia

AbstractThe Clostridium genus harbors compelling organisms for biotechnological production processes; while acetogenic clostridia can fix C1-compounds to produce acetate and ethanol, solventogenic clostridia can utilize a wide range of carbon sources to produce commercially valuable carboxylic acids, alcohols, and ketones by fermentation. Despite their potential, the conversion by these bacteria of carbohydrates or C1 compounds to alcohols is not cost-effective enough to result in economically viable processes. Engineering solventogenic clostridia by impairing sporulation is one of the investigated approaches to improve solvent productivity. Sporulation is a cell differentiation process triggered in bacteria in response to exposure to environmental stressors. The generated spores are metabolically inactive but resistant to harsh conditions (UV, chemicals, heat, oxygen). In Firmicutes, sporulation has been mainly studied in bacilli and pathogenic clostridia, and our knowledge of sporulation in solvent-producing or acetogenic clostridia is limited. Still, sporulation is an integral part of the cellular physiology of clostridia; thus, understanding the regulation of sporulation and its connection to solvent production may give clues to improve the performance of solventogenic clostridia. This review aims to provide an overview of the triggers, characteristics, and regulatory mechanism of sporulation in solventogenic clostridia. Those are further compared to the current knowledge on sporulation in the industrially relevant acetogenic clostridia. Finally, the potential applications of spores for process improvement are discussed.Key Points• The regulatory network governing sporulation initiation varies in solventogenic clostridia.• Media composition and cell density are the main triggers of sporulation.• Spores can be used to improve the fermentation process.

scholarly journals Click Chemistry Enabling Covalent and Non-Covalent Modifications of Graphene with (Poly)saccharides

Polymers ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 142
Author(s):  
Hu Li ◽  
Raffaello Papadakis
Keyword(s):  
Click Chemistry ◽  
Life Science ◽  
Environmental Applications ◽  
Energy Materials ◽  
Sensors And Actuators ◽  
Polar Solvents ◽  
Wide Range ◽  
Potential Applications ◽  
Covalent Modifications

Graphene is a material with outstanding properties and numerous potential applications in a wide range of research and technology areas, spanning from electronics, energy materials, sensors, and actuators to life-science and many more. However, the insolubility and poor dispersibility of graphene are two major problems hampering its use in certain applications. Tethering mono-, di-, or even poly-saccharides on graphene through click-chemistry is gaining more and more attention as a key modification approach leading to new graphene-based materials (GBM) with improved hydrophilicity and substantial dispersibility in polar solvents, e.g., water. The attachment of (poly)saccharides on graphene further renders the final GBMs biocompatible and could open new routes to novel biomedical and environmental applications. In this review, recent modifications of graphene and other carbon rich materials (CRMs) through click chemistry are reviewed.

scholarly journals State of the Science in Dried Blood Spots

2018 ◽  
Vol 64 (4) ◽  
pp. 656-679 ◽  
Author(s):  
Jeffrey D Freeman ◽  
Lori M Rosman ◽  
Jeremy D Ratcliff ◽  
Paul T Strickland ◽  
David R Graham ◽  
...  
Keyword(s):  
Systematic Approach ◽  
Full Range ◽  
Dried Blood Spots ◽  
Blood Spots ◽  
Highly Sensitive ◽  
Wide Range ◽  
Potential Applications ◽  
Review Of Reviews ◽  
Near Term ◽  
State Of The Science

Abstract BACKGROUND Advancements in the quality and availability of highly sensitive analytical instrumentation and methodologies have led to increased interest in the use of microsamples. Among microsamples, dried blood spots (DBS) are the most well-known. Although there have been a variety of review papers published on DBS, there has been no attempt at describing the full range of analytes measurable in DBS, or any systematic approach published for characterizing the strengths and weaknesses associated with adoption of DBS analyses. CONTENT A scoping review of reviews methodology was used for characterizing the state of the science in DBS. We identified 2018 analytes measured in DBS and found every common analytic method applied to traditional liquid samples had been applied to DBS samples. Analytes covered a broad range of biomarkers that included genes, transcripts, proteins, and metabolites. Strengths of DBS enable its application in most clinical and laboratory settings, and the removal of phlebotomy and the need for refrigeration have expanded biosampling to hard-to-reach and vulnerable populations. Weaknesses may limit adoption in the near term because DBS is a nontraditional sample often requiring conversion of measurements to plasma or serum values. Opportunities presented by novel methodologies may obviate many of the current limitations, but threats around the ethical use of residual samples must be considered by potential adopters. SUMMARY DBS provide a wide range of potential applications that extend beyond the reach of traditional samples. Current limitations are serious but not intractable. Technological advancements will likely continue to minimize constraints around DBS adoption.

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