Publications by authors named "Maria Alice Zarur Coelho"

12 Publications

  • Page 1 of 1

Improved production of biocatalysts by Yarrowia lipolytica using natural sources of the biopolyesters cutin and suberin, and their application in hydrolysis of poly (ethylene terephthalate) (PET).

Bioprocess Biosyst Eng 2021 Jun 24. Epub 2021 Jun 24.

Biochemical Engineering Department, School of Chemistry, Federal University of Rio de Janeiro, Av. Athos da Silveira Ramos, 149. Ilha do Fundão, Rio de Janeiro, 21941-909, Brazil.

Since plastic pollution emerged as an urgent environmental problem, different biocatalysts have been tested for poly(ethylene terephthalate) (PET) hydrolysis. This work evaluated three different possible inducers for lipases and/or esterases, two natural sources of biopolymers (apple peels and commercial cork) and PET, as supplements in the solid-state fermentation of soybean bran by Yarrowia lipolytica. The obtained enzymatic extracts displaying different levels of lipase and esterase activities were then tested for PET depolymerization. Supplementation with 5 or 20 wt% of commercial cork led to an increase of 16% in lipase activity and to an increase of 131% in esterase activity, respectively. PET supplementation also led to an increase in the esterase activity of the enzymatic extracts (up to 69%). Enzymes produced in the screening step were able to act as biocatalysts in PET hydrolysis. Enzymatic extracts obtained in fermentation samples supplemented with 20 wt% PET and 20 wt% apple peels led to the highest terephthalic acid concentration (21.2 µmol L) in 7 days, whereas enzymes produced in commercial cork media were more efficient for bis(2-hydroxyethyl) terephthalate (BHET) hydrolysis, one of the key-PET hydrolysis intermediates. Results suggest a good potential of the biocatalysts produced by Y. lipolytica IMUFRJ 50,682 in a low-cost media for subsequent utilization in PET depolymerization reactions. This is one of the few reports on the use of a yeast for this application.
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http://dx.doi.org/10.1007/s00449-021-02603-wDOI Listing
June 2021

A critical view on the technology readiness level (TRL) of microbial plastics biodegradation.

World J Microbiol Biotechnol 2021 Jun 14;37(7):116. Epub 2021 Jun 14.

Biochemical Engineering Department, School of Chemistry, Federal University of Rio de Janeiro, Av. Athos da Silveira Ramos, 149. Ilha do Fundão Rio de Janeiro, 21941-909, Rio de Janeiro, Brazil.

Accumulation of plastic wastes and their effects on the ecosystem have triggered an alarm regarding environmental damage, which explains the massive investigations over the past few years, aiming technological alternatives for their proper destination and valorization. In this context, biological degradation emerges as a green route for plastic processing and recycling in a circular economy approach. Some of the main polymers produced worldwide are poly(ethylene terephthalate) (PET), polyethylene (PE) and polypropylene (PP), which are among the most recalcitrant materials in the environment. In comparison to other polymers, PET biodegradation has advanced dramatically in recent years concerning microbial and enzymatic mechanisms, being positioned in a higher technology readiness level (TRL). Even more challenging, polyolefins (PE and PP) biodegradation is hindered by their high recalcitrance, which is mainly related to stable carbon-carbon bonds. Potential microbial biocatalysts for this process have been evaluated, but the related mechanisms are still not fully elucidated. This review aims to discuss the latest developments on key microbial biocatalysts for degradation of these polymers, addressing biodegradation monitoring, intellectual property, and TRL analysis of the bioprocessing strategies using biodegradation performance, process time and scale as parameters for the evaluation.
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http://dx.doi.org/10.1007/s11274-021-03089-0DOI Listing
June 2021

Process strategies to improve biocatalytic depolymerization of post-consumer PET packages in bioreactors, and investigation on consumables cost reduction.

Bioprocess Biosyst Eng 2021 Mar 28;44(3):507-516. Epub 2020 Oct 28.

Biotechnology Division, Research and Development Center, PETROBRAS, Av. Horácio Macedo, 950, Ilha do Fundão, Rio de Janeiro, 21941‑915, Brazil.

Massive plastics production has raised concerns about low recycling rates and disposal of these materials in nature, causing environmental and economic impacts. Poly(ethylene terephthalate) (PET) is one of main polymers used for manufacture of plastic packaging (e.g. bottles, trays). Enzymatic recycling of PET has been a route of increasing study aiming at to recover its monomers (terephthalic acid and ethylene glycol), resulting in a circular production chain. In this study, investigation of pH control and fractionation of enzyme feeding were explored in post-consumed PET (PC-PET) hydrolysis reactions catalyzed by Humicola insolens cutinase (HiC) in stirred reactors. It was found that the unbuffered reaction provided of pH control by 0.5 M NaOH addition showed 2.39-fold improvement in the released monomers (to a total of 26.3 mM), comparatively to the Tris-HCl-buffered reaction. In addition, it was observed a possibility of reducing the enzyme loading used in the process by half, leading to an increase of 2.41-fold in the specific terephthalic acid concentration released per protein amount, whilst maintaining a high products concentration (97 mM). A simplified cost analysis of reaction consumables was performed, and the data reported here demonstrates that these alternative process strategies contribute to costs reduction on the enzymatic depolymerization reactions of PET.
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http://dx.doi.org/10.1007/s00449-020-02461-yDOI Listing
March 2021

Construction of wild-type Yarrowia lipolytica IMUFRJ 50682 auxotrophic mutants using dual CRISPR/Cas9 strategy for novel biotechnological approaches.

Enzyme Microb Technol 2020 Oct 12;140:109621. Epub 2020 Jun 12.

Micalis Institute, INRAE, AgroParisTech, Université Paris-Saclay, 78350 Jouy-en-Josas, France. Electronic address:

Yarrowia lipolytica IMUFRJ 50682 is a Brazilian wild-type strain with potential application in bioconversion processes which can be improved through synthetic biology. In this study, we focused on a combinatorial dual cleavage CRISPR/Cas9-mediated for construction of irreversible auxotrophic mutants IMUFRJ 50682, which genomic information is not available, thought paired sgRNAs targeting upstream and downstream sites of URA3 gene. The disruption efficiency ranged from 5 to 28 % for sgRNAs combinations closer to URA3's start and stop codon and the auxotrophic mutants lost about 970 bp containing all coding sequence, validating this method for genomic edition of wild-type strains. In addition, we introduced a fluorescent phenotype and achieved cloning rates varying from 80 to 100 %. The ura3Δ strains IMUFRJ 50682 were also engineered for β-carotene synthesis as proof of concept. Carotenoid-producing strains exhibited a similar growth profile compared to the wild-type strain and were able to synthesized 30.54-50.06 mg/L (up to 4.8 mg/g DCW) of β-carotene in YPD and YNB flask cultures, indicating a promisor future of the auxotrophic mutants IMUFRJ 50682 as a chassis for production of novel value-added chemicals.
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http://dx.doi.org/10.1016/j.enzmictec.2020.109621DOI Listing
October 2020

Investigation of mitochondrial protein expression profiles of Yarrowia lipolytica in response to citric acid production.

Bioprocess Biosyst Eng 2020 Sep 3;43(9):1703-1715. Epub 2020 May 3.

Department of Biochemical Engineering, School of Chemistry, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, 21941-909, Brazil.

Nitrogen-limiting condition is essential for citric acid production by Yarrowia lipolytica. Mitochondrial protein expression profiles of Y. lipolytica IMUFRJ 50,682 cells cultivated in biomass proliferation medium (YPG medium, yeast extract, peptone and glycerol) and citric acid production medium (CA medium) were analyzed to identify differences in expressed proteins in response to medium composition. The identification of 45 proteins in mitochondria of YPG medium cells and 48 proteins in mitochondria of CA medium cells were possible with proteomic analyses. Only 11 proteins were common to both conditions, showing a different expression pattern in relation to limiting and non-limiting nitrogen conditions. For both conditions, most proteins (52%-CA medium, 46%-YPG medium) were related to energy metabolism. CA medium cells expressed more carbohydrate metabolism proteins (six proteins) then YPG medium cells (three proteins) and the opposite was detected for translation proteins.
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http://dx.doi.org/10.1007/s00449-020-02363-zDOI Listing
September 2020

Growth Parameters and Survivability of for Probiotic Alcoholic Beverages Development.

Front Microbiol 2019 10;10:2092. Epub 2019 Sep 10.

Programa de Pós-Graduação em Ciência de Alimentos, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.

The aim of this research was to optimize the growth parameters (pH, ethanol tolerance, initial cell concentration and temperature) for and its tolerance to gastrointestinal conditions for probiotic alcoholic beverage development. Placket-Burman screening was used to select only statistically significant variables, and the polynomial mathematical model for yeast growth was obtained by central composite rotatable design. Confirmation experiments to determine the kinetic parameters for yeast growth were carried out by controlling the temperature and pH. Soon after, the survivability of yeast was tested under conditions mimicking the human upper gastrointestinal transit. had suitable resistance to alcohol and gastrointestinal conditions for probiotic alcoholic beverage development.
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http://dx.doi.org/10.3389/fmicb.2019.02092DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6747048PMC
September 2019

Characterization and Application of Yarrowia lipolytica Lipase Obtained by Solid-State Fermentation in the Synthesis of Different Esters Used in the Food Industry.

Appl Biochem Biotechnol 2019 Nov 1;189(3):933-959. Epub 2019 Jun 1.

School of Chemistry, Technology Center, Federal University of Rio de Janeiro, Av. Athos da Silveira Ramos, 149, Rio de Janeiro, CEP 21941-909, Brazil.

Yarrowia lipolytica lipase obtained by solid-state fermentation was characterized and applied in the synthesis of esters with commercial value in the food industry. The effect of different conditions on the hydrolysis activity of this biocatalyst was evaluated in the presence of metal ions, solvents, detergents, several pH and temperature parameters, and different substrates. Storage stability was also studied. The solid biocatalyst produced in soybean meal was used in synthesis reactions aiming to produce short-, medium-, and long-chain esters. Results showed that the best fermentation condition to produce the biocatalyst was using soybean oil (3% w/w), moisture content (55% w/v), and inoculum of 2.1 mgg at 28 °C for 14 h. High substrate conversion for ethyl octanoate, cetyl stearate, and stearyl palmitate synthesis was achieved in the presence of non-polar solvents in less than 6 h using a substrate molar ratio of 1:1 at 38 °C with 10-15% (w/v) of biocatalyst. This work showed the high potential of Y. lipolytica lipase to be used in the synthesis of different esters. Also, that it can be considered an attractive and economical process alternative to obtain high-added value products.
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http://dx.doi.org/10.1007/s12010-019-03047-5DOI Listing
November 2019

Biocatalytic esterification of fatty acids using a low-cost fermented solid from solid-state fermentation with .

3 Biotech 2019 Feb 7;9(2):38. Epub 2019 Jan 7.

1Chemistry Institute, Rio de Janeiro State University, Rio de Janeiro, Brazil.

This study aimed to evaluate the use of a lyophilized fermented solid (named solid enzymatic preparation, SEP), with lipase activity, as a low-cost biocatalyst for esterification reactions of fatty acids present in acid raw materials for biodiesel synthesis. The SEP was obtained by solid-state fermentation (SSF) of soybean bran using the strain of IMUFRJ 50682 and contains the lipases secreted by this yeast. The esterification reaction of ethanol and the predominant fatty acids present in different acid oil sources for biodiesel production (oleic, linoleic, stearic and palmitic acids) was investigated. Oleic acid conversion of above 85% was obtained after 24 h, using 30 wt% of SEP and ethanol/oleic acid molar ratio of 1, at 30 °C, in a reaction medium with and without solvent (n-hexane). Similar results were achieved with stearic (79%), palmitic (82%) and linoleic (90%) acids. The reusability of SEP was investigated over ten successive batches by washing it with different solvents (ethanol, water or n-hexane) between the cycles of ethyl oleate synthesis. Washing with water allowed the SEP to be reused for six cycles maintaining over 80% of the conversion reached in the first cycle. These results show the potential of this biocatalyst to reduce the content of free fatty acids in acid oils for biodiesel synthesis with a potential to be applied in a broad plethora of raw materials.
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http://dx.doi.org/10.1007/s13205-018-1550-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6323043PMC
February 2019

Enzyme-enhanced extraction of phenolic compounds and proteins from flaxseed meal.

ISRN Biotechnol 2013 23;2013:521067. Epub 2012 Oct 23.

School of Chemistry, Federal University of Rio de Janeiro, Ilha do Fundão, 21945-970, Rio de Janeiro, RJ, Brazil.

Flaxseed (Linum usitatissimum) meal, the main byproduct of the flaxseed oil extraction process, is composed mainly of proteins, mucilage, and phenolic compounds. The extraction methods of phenolics either commonly employed the use of mixed solvents (dioxane/ethanol, water/acetone, water/methanol, and water/ethanol) or are done with the aid of alkaline, acid, or enzymatic hydrolysis. This work aimed at the study of optimal conditions for a clean process, using renewable solvents and enzymes, for the extraction of phenolics and proteins from flaxseed meal. After a screening of the most promising commercial preparations based on different carbohydrases and proteases, a central composite rotatable design and a mixture design were applied, achieving as optimal results a solution containing 6.6 and 152 g kg(-1) meal of phenolics and proteins, respectively. The statistical approach used in the present study for the enzyme-enhanced extraction of phenolics and proteins from the major flaxseed byproduct was effective. By means of the sequential experimental design methodology, the extraction of such compounds was increased 10-fold and 14-fold, when compared to a conventional nonenzymatic extraction.
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http://dx.doi.org/10.5402/2013/521067DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4403571PMC
May 2015

Production and use of lipases in bioenergy: a review from the feedstocks to biodiesel production.

Enzyme Res 2011 7;2011:615803. Epub 2011 Jul 7.

School of Chemistry, Federal University of Rio de Janeiro, 21941-970 Rio de Janeiro, RJ, Brazil.

Lipases represent one of the most reported groups of enzymes for the production of biofuels. They are used for the processing of glycerides and fatty acids for biodiesel (fatty acid alkyl esters) production. This paper presents the main topics of the enzyme-based production of biodiesel, from the feedstocks to the production of enzymes and their application in esterification and transesterification reactions. Growing technologies, such as the use of whole cells as catalysts, are addressed, and as concluding remarks, the advantages, concerns, and future prospects of enzymatic biodiesel are presented.
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http://dx.doi.org/10.4061/2011/615803DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3137985PMC
November 2011

Factorial design to optimize biosurfactant production by Yarrowia lipolytica.

J Biomed Biotechnol 2010 23;2010:821306. Epub 2010 Mar 23.

Departamento de Engenharia Bioquímica, Escola de Química, UFRJ, 21949-900, Rio de Janeiro, RJ, Brazil.

In order to improve biosurfactant production by Yarrowia lipolytica IMUFRJ 50682, a factorial design was carried out. A 2(4) full factorial design was used to investigate the effects of nitrogen sources (urea, ammonium sulfate, yeast extract, and peptone) on maximum variation of surface tension (Delta ST) and emulsification index (EI). The best results (67.7% of EI and 20.9 mN m(-1) of Delta ST) were obtained in a medium composed of 10 g 1(-1) of ammonium sulfate and 0.5 g 1(-1) of yeast extract. Then, the effects of carbon sources (glycerol, hexadecane, olive oil, and glucose) were evaluated. The most favorable medium for biosurfactant production was composed of both glucose (4% w/v) and glycerol (2% w/v), which provided an EI of 81.3% and a Delta ST of 19.5 mN m(-1). The experimental design optimization enhanced Delta EI by 110.7% and Delta ST by 108.1% in relation to the standard process.
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http://dx.doi.org/10.1155/2010/821306DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2846354PMC
July 2010

Characterization of commercial amylases for the removal of filter cake on petroleum wells.

Appl Biochem Biotechnol 2010 May 6;161(1-8):171-80. Epub 2009 Oct 6.

Centro de Tecnologia, Escola de Química, Universidade Federal do Rio de Janeiro, Bloco E, Lab 103, Cidade Universitária, 21949-900, Rio de Janeiro, Rio de Janeiro, Brazil.

Drilling fluid has many functions, such as carry cuttings from the hole permitting their separation at the surface, cool and clean the bit, reduce friction between the drill pipe and wellbore, maintain the stability of the wellbore, and prevent the inflow of fluids from the wellbore and form a thin, low-permeable filter cake. Filter cake removal is an important step concerning both production and injection in wells, mainly concerning horizontal completion. The drilling fluids are typically comprised of starch, the most important component of the filter cake. A common approach to remove this filter cake is the use of acid solutions. However, these are non-specific reactants. A possible alternative is the use of enzymatic preparations, like amylases, that are able to hydrolyze starch. Wells usually operate in drastic conditions for enzymatic preparations, such as high temperature, high salt concentration, and high pressure. Thus, the main objective of this work was to characterize four enzymatic preparations for filter cake removal under open hole conditions. The results showed that high salt concentrations (204,000 ppm NaCl) in completion fluid decreased amylolytic activity. All enzymatic preparations were able to catalyze starch hydrolysis at all temperatures tested (30, 65, 80, and 95 degrees C). An increase of amylolytic activity was observed with the increase of pressure (100, 500 and 1,000 psi) for one commercial amylase.
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http://dx.doi.org/10.1007/s12010-009-8773-yDOI Listing
May 2010
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