iBB

Pathologies such as psoriasis and rheumatoid arthritis have an inflammatory base triggered, in part, by an impairment of antioxidant defense systems which leads to overaccumulation of oxidative species (ROS). In a recently published paper, entitled “Up-Regulation of the Nrf2/HO-1 Antioxidant Pathway in Macrophages by an Extract from a New Halophilic Archaea Isolated in Odiel Saltworks”, researchers of the Universidad de Sevilla, Universidad de Huelva and Carla C. C. R. de Carvalho (iBB), show that an extract of Haloarcula sp. OS, containing bacterioruberin and significant amounts of fatty acids, exhibits high antioxidant activity. The study shows, for the first time, that the extract reduces LPS-induced oxidative stress and acute inflammation in THP-1 macrophages by attenuating both ROS levels and the production of the pro-inflammatory cytokines TNF-a and IL-6. The mechanism involves the activation of the Nrf-2/HO-1signaling pathway, and supports the use of the extract as a therapeutic agent in the treatment of oxidative stress-related inflammatory diseases.

Marine bacteria present enzymatic activities that result from extensive evolution necessary for cell survival under the harsh and continuously changing conditions found in the marine environment. Metagenomic techniques and cultivation of not-yet-cultured bacteria, combined with high-throughput screening techniques, have allowed the identification of new biocatalysts and the development of industrial processes. In a recently published paper, Carlos J.C. Rodrigues and Carla C.C.R. de Carvalho (iBB) review the latest studies related to growth of marine bacteria under laboratorial conditions, screening techniques, and bioprocess development.

Six researchers from @IBB have been ranked among the World’s Top 2% of most widely cited scientists in the 2022 List of outstanding researchers prepared by Elsevier BV, and Stanford University (USA).   

Carla Carvalho, Joaquim Cabral, Isabel Sá Correia,  Mário Berberan e Santos, Miguel Prazeres and Pedro Fonte were recognized for the impact throughout their careers, while Pedro Fernandes was recognized as one of the World’s Top 2%  influent scientist in his field.

The updated list is based on the Sep 01-2022 SCOPUS database, updated to the end of the citation year 2021 and analyses 22 scientific areas and 176 disciplines.

Overall, 93 researchers from Técnico were highlighted in the study.  Congratulations to all!

It is usually accepted that only 0.1-1% of marine bacteria may be cultivated under laboratory conditions. This resulted mainly from the observation that only a small fraction of the total bacteria determined by microscopic counts could be enumerated by culture plate techniques, and has been a justification for the widely implementation of cultivation-independent molecular techniques. In a recent published paper, Carlos J.C. Rodrigues and Carla C.C.R. de Carvalho (BERG-iBB) promoted the growth of bacteria from a marine rock pond by culture dependent techniques and compared the results with culture independent methods. The highest efficiency of cultivation was 45% attained in marine agar. The study shows that it is possible to grow more than the traditionally considered 1% of bacteria from a seawater sample using standard agar plate techniques and laboratorial conditions, and to find bacteria not detected by molecular approaches.

The Norwegian Minister of Fisheries and Ocean Policy Bjørnar Selnes Skjæran, the Minister of International Development Anne Beathe Tvinnereim, and the Embassador in Lisbon Tove Bruvik Westberg, visited the Instituto Superior de Agronomia as promotor of projects funded by EEA grants. Anabela Raymundo and Cristiana Nunes (LEAF-ISA), Carla de Carvalho (iBB-IST) and António Pagarete (Pagarete Microalgae Solutions) presented the collaborations they have had with Norwegian research groups including the project Yum Algae. This projects aims at finding enzymes to improve the sensory quality of microalgae ingredients in foods and during the visit several products, such as bread, cheese and ice cream containing microalgae, were tasted. The visit also included members of the “Unidade Nacional de Gestão” of EEA Grants and of “Direção Geral de Política do Mar”.

Pathogenic Vibrio species are responsible for human and animal illness and one of the main causes of human infection is related to the ingestion of undercooked seafood. Due to their filter-feeding habit, marine invertebrates, such as clams, are known to be a natural reservoir of specific microbial communities. In a recently published study carried out by researchers of ESTM, Polytechnic of Leiria, and Carla C.C.R. de Carvalho (BERG-iBB), the profile of antimicrobial resistance and the presence of virulence genes in the bacterial isolates from clams were evaluated. The presence of the mobilisable colistin resistance gene mcr-1 in three Vibrio spp. isolates highlights a potential threat to public health.

Developing viable biorefineries is urgent to support the transition to a sustainable society. Lignocellulosic biomass is a crucial renewable feedstock, but usually a combination of pretreatments is necessary to extract both sugars and lignin. In a recent paper published in the journal Sustainability, the M.Sc. student Manuel Dias and Carla C.C.R. de Carvalho (BERG-iBB) collaborated with researchers from Technische Universität Wien (Vienna, Austria) under supervision of Prof Anton Friedl, to study liquid-hot-water (LHW) as pretreatment for hemicellulose hydrolysis. Assessment of different LHW conditions allowed the determination of those leading to high sugar concentration and low concentration of degradation products. The lignin remaining in the solid phase could be extracted and hydrolysed. By including the analysis of lignin hydrolysis into the standard sugar and degradation products in LHW, it was possible to make an integrated decision to valorize the different lignocellulosic components of wheat straw.

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.

The production of recombinant proteins is gaining increasing importance as several applications request high quality proteins. However, several process parameters affect both the growth of cells and product yields. In a paper recently published in the journal Molecules, Ricardo FS Pereira and Carla CCR de Carvalho (iBB-BERG) used high throughput systems and statistical methods to assess the influence of fermentation conditions in lab-scale bioreactors. Using partial least squares, it was found that the height-to-diameter ratio of the bioreactor, aeration rate, and PID controller parameters contributed significantly to the final biomass and cytochrome concentrations. This information was used to fine-tune the process parameters, which increased cytochrome production and yield several-fold.

Carlos JC Rodrigues and Carla CCR de Carvalho (iBB-BERG) presented their work in EFB2021, the major scientific conference organised by the European Federation of Biotechnology. The two studies presented focused on the marine environment as a source of biocatalysts for suitable bioprocesses, and on using microbial heterogeneity to improve bioprocesses. The virtual conference took place during May 10th-14th.

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.

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.

Enzymes may be the right catalysts for efficient and clean processes, contributing to energy savings and negligible by-products and waste. Following the success of the Special Issue on “Biocatalysis, Enzyme and Process Engineering” in Processes (MDPI), the editors Pedro Fernandes and Carla C. C. R. de Carvalho wrote an editorial paper discussing the different papers published on this subject.

Microbes are known to contaminate money. In the project CleanCoin, funded by INCM, Carla C.C.R. de Carvalho (iBB), Telmo Santos (UNIDEMI, UNL) and Maria José Caramujo (cE3c, FCUL, UL), studied the bacterial population present at the surface of coins. In a recently published white paper, the research team and collaborators from INCM, present the results and compare the number of viable bacteria in coins with those found on other objects such as bank cards and mobile phones.

The increasing number of life-threatening infections observed in cancer patients has been ascribed to chemotherapy-induced neutropenia and to invasive medical procedures. In a recent published study, Carla C.C.R. de Carvalho (BERG-iBB) questioned if the infections could also be favored by an increased resistance of bacteria due to the adaptation to antineoplastic agents used in chemotherapy. After exposure to several antineoplastic agents, it was observed that cells of Staphylococcus aureus, Mycobacterium vaccae, Pseudomonas aeruginosa, and Escherichia coli changed the fatty acid profile of their cellular membranes, produced exopolymeric substances, and formed aggregates that adhered to surfaces. Additionally, when exposed to high concentrations of these compounds, a persister sub-population could be identified. After adaptation to antineoplastic agents, the minimum inhibitory concentration (MIC) of several antibiotics increased considerably in the tested strains. This should be a major concern during oncology treatments.

Helichrysum italicum is a halophyte shrub with bright yellow flowers with a strong curry-like aroma that have been used in the production of cosmetics and pharmaceuticals, for seasoning and flavoring of food, and as natural food preservatives. In a study  recently published in Food Research International, researchers of Universidade Nova de Lisboa, Polytechnic of Leiria, University of Coimbra, University of Lincoln, the Quadram Institute, and Carla C.C.R. de Carvalho (BERG-iBB), characterize the composition, bioactive compounds, and nutritive value of H. italicum flowers. The flowers were found to be a good source of minerals, omega-6 and omega-3 PUFAs, polyphenols, carotenoids, curcuminoids and other bioactive compounds with relevant antibacterial, antifungal and antioxidant properties.

Photo details: Helichrysum italicum, Bj.schoenmakers, CC0, via Wikimedia Commons.

Carla CCR de Carvalho (iBB-BERG) presented the work entitled “Marine bioprocesses with bacteria from the Portuguese coast” at the “One Sustainable Ocean”, a side event of the UN Ocean conference (UNOC) held in Lisbon. The UN Ocean Conference, co-hosted by the Governments of Kenya and Portugal, aimed at starting a new chapter of global ocean action.

The ocean is an excellent source for new biocatalysts due to the tremendous genetic diversity of marine microorganisms, and it may contribute to the development of sustainable industrial processes. In a recent paper published in Microorganisms, Carlos J.C. Rodrigues and Carla C.C.R. de Carvalho (iBB-BERG) used a marine bacterium for the conversion of benzaldehyde to benzyl alcohol, which is an important chemical employed as a precursor for producing esters for cosmetics and other industries. A stirred reactor, using a fed-batch approach, enabled a 1.5-fold increase in benzyl alcohol productivity when compared with batch mode. However, product accumulation in the reactor hindered the conversion. The use of a continuous flow reactor packed with immobilized cells enabled a 9.5-fold increase in productivity when compared with the fed-batch stirred reactor system.

Consumer acceptance of microalgae products is often hampered by their colour, “fishy” smell and flavor. An international project with research teams from ISA (PI Prof. Anabela Raymundo), IST (Carla de Carvalho, iBB-BERG), Norce (Norway) and the company Pagarete Microalgae solutions (Portugal) is aimed at the use of enzymes to improve the overall sensory quality of microalgal ingredients. The project, entitled “YUM ALGAE - enzYmes for improved sensory qUality of MicroALGAE ingredients in foods”, is funded in ca. 1 million euros by the Iceland, Liechtenstein, Norway grants (EEA grants, Blue Growth programme).

The cyclic anionic lipopeptide daptomycin (DAP) is used in the treatment of severe infections caused by Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus (MRSA). DAP resistance, though rare, often results in treatment failure. Paradoxically, in MRSA, DAP resistance is usually accompanied by a concomitant decrease in b-lactam resistance in what is known as the ‘see-saw effect’. In a recent publication in the Journal of Antimicrobial Chemotherapy, Carla CCR de Carvalho (iBB-BERG), Agustina Taglialegna and Adriana E. Rosato (currently at Riverside University Health System, USA), showed that the post-translocational protein PrsA has a major role in orchestrating the cell membrane lipid adaptation associated with the see-saw effect, in addition to its function required for the posttranscriptional maturation of PBP 2a. This study contributes greatly to the understanding of MRSA biology and of the impact of DAP/b-lactam on the cell membrane/cell wall machinery.

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.

Six iBB researchers are among the most cited top scientists at the top 2% of their areas. Portugal is represented in this list compiled by Stanford University with 385 scientists affiliated to national institutions. The rankings are based upon a researcher’s citations for both a single year (2019) and cumulative across their careers. Titled “Updated science-wide author databases of standardized citation indicators,” the list uses algorithms that quantify and systematically rank individuals into consistent scientific fields. The scores are provided  with and without self-citations to lessen the impact of self-citations or the use of citation farms (small clusters of researchers massively citing each other’s work). Ranked within the careerlong citation impact list are these iBB researchers: Joaquim Cabral, Isabel Sá-Correia, Mário Berberan Santos, Miguel Prazeres, Carla Carvalho, also along with Carina Crucho in the
2017 single-year citations list.