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Recent Developments in Enzyme Immobilization for Food Production

Recent Developments in Enzyme Immobilization for Food Production | iBB | Scoop.it

Greener, cost-effective methodologies for food processing and production have been actively looked after because of increased demand due to growing population. Enzymes, as green and highly efficient catalysts, have been a mainstay of food technology. However, the advantageous features of enzymes are negatively impacted by their long-term instability, harsh industrial operational conditions and challenges for enzyme recovery and reuse. Immobilized enzyme formulations have been developed to address these limitations. In a recently published book chapter, Filipe Carvalho and Pedro Fernandes (iBB-BERG) present the latest finding and trends on the design and application of immobilized enzyme for food production and processing. The chapter was published in the book Value Addition in Food Products and Processing using Enzyme Technology, published by Academic Press.

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Participation in the EU Project FuturEnzyme

Participation in the EU Project FuturEnzyme | iBB | Scoop.it

A team of researchers from iBB-BERG, including Carlos Rodrigues, Ricardo Pereira, Pedro Fernandes, Prof. Joaquim Sampaio Cabral and led by Carla de Carvalho, will contribute to the development of enzymes for a new generation of environmentally friendly consumer products in the EU project FuturEnzyme. The consortium of 16 leading academic and industrial partners, led by Dr. Manuel Ferrer from CSIC (Spain), will develop innovative solutions that will combine big biodata mining and bio-prospecting, disruptive machine learning and protein engineering, nano-biotechnology, fermentation and downstream-processing systems, and pre-industrial testing. Innovative solutions will be proposed to discover, design, optimize and formulate new enzymes to develop detergents, textiles and cosmetics that combine a high level of functionality with improved sustainability, thus responding to the demands from consumers and industries.

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Multi-Enzyme Systems in Flow Chemistry

Multi-Enzyme Systems in Flow Chemistry | iBB | Scoop.it

Recent years have witnessed a growing interest in the use of biocatalysts in flow reactors. This merging combines the high selectivity and mild operation conditions typical of biocatalysis with enhanced mass transfer and resource efficiency associated to flow chemistry. Additionally, it provides a sound environment to emulate Nature by mimicking metabolic pathways in living cells and to produce goods through the systematic organization of enzymes towards efficient cascade reactions. Moreover, by enabling the combination of enzymes from different hosts, this approach paves the way for novel pathways. In a recent paper published in the journal Processes, Pedro Fernandes and Carla de Carvalho (iBB-BERG) review the recent developments within the scope of flow chemistry involving multi-enzymatic cascade reactions. The paper was published in the Special Issue  "Biocatalysis, Enzyme and Process Engineering" edited by both authors.

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Eduardo Brás Defends PhD Thesis in Biotechnology and Biosciences

Eduardo Brás Defends PhD Thesis in Biotechnology and Biosciences | iBB | Scoop.it

Eduardo Brás defended his PhD thesis in Biotechnology and Biosciences on Tuesday the 15th september 2020 (14:30 H). During the last years, and under the supervision João Pedro Conde from INESC-MN and Pedro Fernandes from BERG-iBB, Eduardo explored how microfluidic technology and microfabrication could be coupled with enzymes with goal of developing biosensing in different fields of application. The title of the thesis is “Biocatalysis in Microfluidic Systems: Biosensing and Rapid Screening Applications”.

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Portable Platform for Point-of-Need Testing of Plant Health

Portable Platform for Point-of-Need Testing of Plant Health | iBB | Scoop.it

Microfluidic biosensing devices are becoming a key part in the development of point-of-care diagnosis systems for medical, animal and agricultural applications alike. However, to fully take advantage of the features presented by microfluidics technology, one has to be capable of providing the appropriate instrumentation for the actuation of the devices and transduction of the results produced by said devices. A versatile and portable platform for the optical transduction of microfluidic devices was recently the target of a publication in the IEEE Sensors Journal  In this work, thin-film, hydrogenated amorphous silicon photosensors are used to transduce the optical assay for the detection of azelaic acid in wine producing grapes infected with the fungus B. cinerea. The portable prototype provides all necessary electronics for liquid manipulation, light sources and electronic amplification for the assay read-out. The work was performed by Biotechnology PhD student Eduardo Brás (INESC-MN/IBB) and Biotechnology Alumni Dr. Rui Pinto under the supervision ofJoão Pedro Conde (INESC-MN/DBE) and Pedro Fernandes (BERG-iBB).

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Upgrading the Organic Fraction of Municipal Solid Waste to Biodegradable Bioplastics

Upgrading the Organic Fraction of Municipal Solid Waste to Biodegradable Bioplastics | iBB | Scoop.it

The organic fraction of municipal solid waste (OFMSW) accounts for approximately 30-40% of MSW in Europe. BERG-iBB researchers developed a process that uses this type of waste as a raw material or the production of the biodegradable biopolymer P(3HB). In a first step the complex carbohydrates in the waste are hydrolysed into simple monosaccharides. The hydrolysate is then used as a carbon source to feed and induce the bacterium Burkholderia sacchari to produce P(3HB). In order to overcome nutritional deficiencies and attain a significant polymer accumulation (58% g polymer/g CDW) the C/N ratio was adjusted and the hydrolysate was supplemented with minerals. This work demonstrates that an easily accessible waste can be transformed into valuable biodegradable bioplastics. The work was published in Bioresource Technology.

 

Photo details: municipal waste by OpenIDUser2, GFDL. 

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Eduardo Brás Delivers Poster Presentation at the 2019 Transducers-Eurosensors Joint Conference

Eduardo Brás Delivers Poster Presentation at the 2019 Transducers-Eurosensors Joint Conference | iBB | Scoop.it

Eduardo Brás delivered a poster presentation entitled "A Portable Microfluidic System for the Detection of Health Biomarkers in Grapes at the Point of Need", at the 2019 Transducers-Eurosensors joint conference, in Berlin, Germany. He demonstrated his work concerning the development of a portable device capable of fungal infection detection in grape trees through the detection of plant metabolites, through the use of a enzymatic reaction in a microfluidic device. This work was supervised by Prof. João Pedro Conde (INESC-MN) and Prof. Pedro Fernandes (IBB) and counted with the collaboration of Prof. Ana Margarida Fortes (FCUL).

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Special Issue "Biocatalysis, Enzyme, and Process Engineering"

Special Issue "Biocatalysis, Enzyme, and Process Engineering" | iBB | Scoop.it

Pedro Fernandes and Carla Carvalho from BERG-iBB are acting as guest editors of a special issue of the open-access journal "Processes" on “Biocatalysis, Enzyme, and Process Engineering”. The aim is to address novel advances in the development and application of enzymes in the production of goods of interest for large-scale production. Topics include but are not limited to the following: i) Enzyme engineering towards improved biocatalysts; ii) The development of novel process designs for biocatalysis; and iii) Multienzyme cascade reactions. Manuscripts can be submitted via the journal web page until November 30, 2019.

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Featured Photo: Cover of Biotechnol. Bioeng. Credits BERG-iBB Contribution

Featured Photo: Cover of Biotechnol. Bioeng. Credits BERG-iBB Contribution | iBB | Scoop.it

Description: Cover image of Biotechnology and Bioengineering, based on the article "Enzymatic synthesis of chiral amino-alcohols by coupling transketolase and transaminase-catalyzed reactions in a cascading continuous-flow microreactor system". Design Credit: Winter Mraz, Frozen Ink Designs., Copyright Wiley 2017.


Context: BERG-iBB researchers Pedro Fernandes and Filipe Carvalho, working in collaboration with UCL colleagues led by Nicolas Szita, have published a novel microreactor-based approach based on free enzymes for the synthesis of the chiral building block ABT. The work was performed in the context of iBB's Strategic Area 2: Advanced Bioprocess Engineering. (Click on title to learn more).

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Enzymatic Synthesis of Alkyl Esters in Miniemulsions

Enzymatic Synthesis of Alkyl Esters in Miniemulsions | iBB | Scoop.it

Oil-in-water miniemulsions display potential for application as a “green” system in the enzymatic alkyl ester synthesis. BERG-iBB researchers led by Luís Fonseca combined miniemulsion systems with medium engineering to increase the yields and produtivity of the cutinase-catalysed synthesis of medium-chain alkyl esters. Enzyme stability and reutilization were tuned by operating in a fed-batch mode, which allowed long-term utilization of the free enzyme. Click on title to learn more.


Photo details: 3D structure of cutinase from F. Solani pisi with catalytic triad in blue (PDB code: 1AGY).

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Enzymatic Synthesis of Chiral Amino-Alcohols in a Continuous-Flow Microreactor System

Enzymatic Synthesis of Chiral Amino-Alcohols in a Continuous-Flow Microreactor System | iBB | Scoop.it

Chiral amino-alcohols are key building blocks in the production of optically pure pharmaceuticals. While many of these compounds can be synthesized enzymatically, the rapid process development and intensification continues to be challenging. A recent contribution co-authored by BERG-iBB researchers Pedro Fernandes and Filipe Carvalho with UCL colleagues led by Nicolas Szita presents a novel microreactor-based approach based on free enzymes for the synthesis of the chiral building block ABT. The system couples a transketolase- and a transaminase-catalyzed reaction and is operated continuously. The presented approach illustrates how continuous-flow microreactors can be applied for the design and optimization of biocatalytic processes. Click on title to learn more about the work on Biotechnology and Bioengineering. Photo: 3D model of transketolase from Escherichia coli (pdb 1QGD).

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Sample Preparation With Aqueous Two-Phase Systems for On-site Immunodetection of Mycotoxins

Sample Preparation With Aqueous Two-Phase Systems for On-site Immunodetection of Mycotoxins | iBB | Scoop.it
Researchers at BERG-iBB and INESC-MN recently published a simple and versatile sample preparation methodology based on a green aqueous two-phase system for the simultaneous extraction and concentration of mycotoxins in multiple feeds, which are currently among the most relevant and widespread contaminants in the food and feed industry. This method was tailored for the subsequent rapid detection of ochratoxin A, aflatoxin B1 and deoxynivalenol at the relevant limits within the ng/g range, using a microfluidic multiplexed chemiluminescent immunoassay. Overall, sample preparation and detection were achieved in about 25 min using a “point-of-need” compatible process. Click on title to learn more.
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A Bespoke Capillary Wall-Coated Microreactor

A Bespoke Capillary Wall-Coated Microreactor | iBB | Scoop.it

The use of microscale set-ups for the fast development of bioprocesses has been gaining momentum. Within this, continuous flow operation for enzymatic reaction is particularly appealing, as it allows for steady state and ease of automation. In a paper recently published in the journal Catalysts, Pedro Fernandes and Filipe Carvalho from BERG-iBB, together with Marco Marques from UCL, presented a prototype of a home-made, easy to assemble capillary enzymatic microreactor. The assembly of the device only required materials commonly found in research labs, proved to be reproducible and reliable, and allowed for the detailed characterization of the model reaction system chosen, the production of invert syrup from sucrose. It is foreseen that the device can be easily adapted to other reaction systems and thus further disseminate the use of miniaturized set-ups for bioprocess development. Click on title to learn more.

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Special Issue "Bioprocess Engineering: Sustainable Manufacturing for a Green Society"

Special Issue "Bioprocess Engineering: Sustainable Manufacturing for a Green Society" | iBB | Scoop.it

The mild operational conditions typically required by biobased production processes, together with the selective nature of biocatalysts, microbial diversity and tunability, and the century-long record of fermentation, clearly place bioprocesses as the primary contenders, by far, in achieving the generalized implementation of efficient, green manufacturing strategies. Moreover, by overlapping with the biorefinery approach, the foundations needed for bioprocesses to embrace the circular economy concept are set. In this Special Issue on “Bioprocess Engineering: Sustainable Manufacturing for a Green Society", guest edited by Pedro Fernandes and Carla de Carvalho (iBB-BERG) in Processes, the papers will discuss new bioproducts and concomitant bioengineering approaches to achieve a sustainable and environmentally friendly economy.

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Special Issue "Biocatalysis, Enzyme and Process Engineering"

Special Issue "Biocatalysis, Enzyme and Process Engineering" | iBB | Scoop.it

Enzymes are acknowledged as effective catalysts, namely, on account of their selectivity, high turnover, and ability to operate under mild conditions. Despite these advantageous features, the industrial application of enzymes still lags behind their use on a laboratory scale. This pattern may be partly assigned to the intrinsic nature of enzymes: these biocatalysts have been optimized throughout the ages to be mostly active in aqueous media and to operate in environments where substrate and product concentrations are much lower than those required for industrial processes to be commercially feasible. In this Special Issue on “Biocatalysis, Enzyme, and Process Engineering”, guest edited by Pedro Fernandes and Carla de Carvalho (iBB-BERG) in Processes, several papers discuss the latest applications of enzymes.

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Mycobacterium vaccae adaptation to disinfectants and hand sanitisers

Mycobacterium vaccae adaptation to disinfectants and hand sanitisers | iBB | Scoop.it

In a year marked by the COVID-19 pandemic, the use of disinfectants and hand rubbing solutions became widespread. However, the use of such mixtures may influence bacteria that live/survive in human skin and surfaces, and increase their tolerance/resistance to e.g. antibiotics. In a study recently published in Antibiotics, Carla CCR de Carvalho, Raquel Teixeira and Pedro Fernandes (BERG-iBB), showed that Mycobacterium vaccae, which is being considered as an adjuvant to antituberculosis therapy, is being tested for the treatment of autoimmune diseases, and as an anti-depressive agent, adapted to disinfectants and hand rubbing solutions, and increased tolerance towards levofloxacin, thioridazine, and omeprazole. The use of disinfectants, especially those aimed at healthcare facilities, by the general public on a daily basis should probably be prevented.

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Microfluidic Packed-Bed Reactors

Microfluidic Packed-Bed Reactors | iBB | Scoop.it

Optimization of biocatalytic processes is crucial to drive down the price of pharmaceutical and other high-value molecules. L-DOPA and dopamine are two molecules with pharmaceutical relevance, often used in the treatment of neurological diseases such as Parkinson’s disease. In a paper published in the Journal of Biotechnology, Biotechnology PhD student Eduardo Brás and supervisors João Pedro Conde (INESC-MN/DBE) and Pedro Fernandes (IBB) used a microfluidic packed-bed reactor to optimize the enzymatic conversion of L-tyrosine to dopamine. The work culminated with the up-scale (780-fold) of the optimal conditions for L-DOPA synthesis, while maintaining the same output as obtained in the miniaturized system. This type of system can be used to expedite process development in the future.

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Eduardo Brás Presents Multiplexed Detection with Microfluidics at MicroTAS 2019

Eduardo Brás Presents Multiplexed Detection with Microfluidics at MicroTAS 2019 | iBB | Scoop.it

Biotechnology PhD student, Eduardo Brás, gave a poster presentation at the 23rd edition of the MicroTAS conference in Basel, Switzerland. The poster was about the multiplexed detection of plant health biomarkers in grapes, using a microfluidic platform. The system developed consists of a microfluidic chip containing sample preparation and detection modules for both Azelaic Acid and Salicylic Acid, coupled with a-Si:H for real time optical transduction. The work was carried out under the supervision of Prof. João Pedro Conde (INESC-MN) and Prof. Pedro Fernandes (iBB-BERG), with the collaboration of Prof. Ana Margarida Fortes (FCUL). Eduardo was also co-author of another poster presentation concerning the development of a microfluidic chip for targeted phage selection for personalized colorectal cancer treatment and received a travel grant to the conference.

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Point-of-need Pathogen Detection in Grapes

Point-of-need Pathogen Detection in Grapes | iBB | Scoop.it

Pathogenic infections in plant systems are on the rise, making their early detection in the field an important asset in order to avoid propagation amongst the rest of the crops. One biomarker of interest for this detection is Azelaic Acid (AzA). Based on this, a microfluidic device capable of detecting low concentrations of the acid was developed by Eduardo Brás under the supervision of Prof. João Pedro Conde (INESC-MN) and Prof. Pedro Fernandes (iBB), with the collaboration of Prof. Ana Margarida Fortes (FCUL). The results obtained using real samples as well as a description of the system, which exploits the inhibitory effect of AzA on the activity of the enzyme tyrosinase was published in the journal Analyst.

 

Photo details: Samir Budimčić - Own work, CC BY-SA 2.5, Wikimedia.

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The INMARE Legacy

The INMARE Legacy | iBB | Scoop.it

A video overview of the output of INMARE (‘Industrial Applications of Marine Enzymes’), a four-year European research project that set out to transform the enzyme biodiscovery process and to isolate new enzymes from the ocean, has been made available by the consortium. The goal of INMARE was to shorten & streamline the industrial enzyme pipeline by increasing the value of enzyme collections and to identify new lead products and prototypes during the projects lifetime. INMARE brought together several industrial and academic partners, including iBB-IST. The project received funding from the EU’s Horizon 2020 research and innovation programme under grant agreement No. 634486.

 

Photo details; sea shore by Cascais, Copyright Carla de Carvalho, 2017. 

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Eduardo Brás Presents Research on Point-of-care Devices for Plant Health at Eurosensors 2018

Eduardo Brás Presents Research on Point-of-care Devices for Plant Health at Eurosensors 2018 | iBB | Scoop.it

Eduardo Brás from BERG-iBB delivered an oral presentation at the 32nd edition of Eurosensors in Graz, Austria, concerning his work on the development of a Point-of-Care device for early plant health assessment. The system developed consists of a microfluidic chip containing sample preparation and detection modules, coupled with integrated photodetectors for real time optical transduction. The work is part of Eduardo´s PhD project on Biotechnology, which is supervised by João Pedro Conde from INESC-MN and Pedro Fernandes from BERG-iBB in collaboration with Margarida Fortes from (BioISI). 

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Microbial Enzymes for the Food Industry

Microbial Enzymes for the Food Industry | iBB | Scoop.it

The use of enzymes for food production and processing has a long tradition. This trend is related to their biocompatibility, selective nature and ability to operate under mild conditions. In a recent contribution to the book "Biotechnology of Microbial Enzymes" (Academic Press), BERG-researchers Pedro Fernandes and Filipe Carvalho provide an overview of the different application of enzymes in food production and processing, highlighting on the role of enzymes, their sources, and particular features and formulations required for targeted applications. The work provides the readers an updated and comprehensive perspective on the use of enzymes within the scope of the food industry. Click on title to access the book chapter. 


 Photo: Ralf Roletschek [GFDL 1.2 (http://www.gnu.org/licenses/old-licenses/fdl-1.2.html) or FAL], via Wikimedia Commons.

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Biocatalytic Process Intensification in Flow Bioreactors

Biocatalytic Process Intensification in Flow Bioreactors | iBB | Scoop.it

Biocatalysis has widened its scope and relevance with the availability of new molecular tools, e.g. improved expression systems, protein and metabolic engineering, and rational immobilization techniques. However, applications are still hampered by low productivity and scale-up difficulties. A practical step to improve biocatalyst (enzymes, whole-cells) performance is to use flow reactors. In a recent review published in Trends in Biotechnology, Pedro Fernandes from BERG-iBB together with colleagues from the Universities of Milan and Nottingham describe the state of the art on the design and use of biocatalysis in flow reactors. The successes of the technology, new opportunities, problems and advances are critically discussed. Click on title to learn more.

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A Microfluidic Toolbox for the Optimization of Aqueous Two Phase Antibody Extraction

A Microfluidic Toolbox for the Optimization of Aqueous Two Phase Antibody Extraction | iBB | Scoop.it
The use of aqueous two-phase extraction (ATPE) is a viable option for the purification of antibodies and other vluable proteins, but these systems are difficult to model and optimize. In a publication in the Journal of Chmoratography A, researchers at BERG-iBB and INESC-MN describe a methodology for the rapid screening of antibody extraction conditions that relies on a microfluidic channel-based toolbox. A first microfluidic structure allows a simple negative-pressure driven rapid screening of up to 8 extraction conditions simultaneously, using less than 20 μL of each phase-forming solution per experiment, while a second microfluidic structure allows the integration of multi-step extraction protocols based on the results obtained with the first device. Click on title to learn more.
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Finding Efficient Marine Biocatalysts

Finding Efficient Marine Biocatalysts | iBB | Scoop.it

Marine bacteria have evolved to survive in the marine environment by using unique physiological, biochemical and metabolic features and the ability to produce enzymes and compounds which may have commercial value. The Azores archipelago presents several ecosystems with strong volcanic activity where bacteria thrive under e.g. high temperatures. Carlos Rodrigues, Ricardo Pereira, Dr. Pedro Fernandes, Prof. Joaquim Cabral and Dr. Carla de Carvalho from BERG-iBB used a cultivation-based strategy to find efficient marine biocatalyst. Four Bacillus strains presenting high inulinase activity could be found and used in the development of bioprocesses for the hydrolysis of inulin. Click on title to learn more. 

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