Production of a novel N‐terminal PEGylated crisantaspase

Torres-Obreque, Karin Mariana; Meneguetti, Giovanna Pastore; Custodio, Débora Fernandes; Monteiro, Gisele; Pessoa, Adalberto; Rangel-Yagui, Carlota de Oliveira

doi:10.1002/bab.1723

Cited by 12 publications

(8 citation statements)

References 22 publications

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“…There are three forms of treatment for ALL using Lasparaginase in clinical practice: the native enzyme derived from Escherichia coli, the pegylated enzyme, and an isolated ASNase from Erwinia chrysanthemi [6][7][8]. Treatment with bacterial L-asparaginase has produced some important side effects, such as neurological crisis, anaphylaxis, coagulation disorders, leukopenia, thrombosis, pancreatitis, hyperglycemia, and hepatotoxicity [2].…”

Section: Introductionmentioning

confidence: 99%

Optimization of aqueous two-phase micellar system for partial purification of L-asparaginase from Penicillium sp. grown in wheat bran as agro-industrial residue

et al. 2020

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L-asparaginase has been used in the remission of malignant neoplasms such as acute lymphoblastic leukemia. The search for new sources of this enzyme has become attractive for therapeutics. Traditional methods for biomolecule purification involve several steps. A two-phase system may be a good strategy to anticipate one of these stages. This study aimed to produce and purify a fungal L-asparaginase through an aqueous two-phase micellar system (ATPMS) using Triton X-114. The fungus Penicillium sp.-encoded 2DSST1 was isolated from Cerrado soil. Plackett-Burman design followed by a 2 4 full factorial design was used to determine the best conditions to produce L-asparaginase. The evaluated variables were L-asparagine, L-proline, wheat bran, potato dextrose broth, ammonium sulfate, yeast extract, sucrose and glucose concentrations, incubation temperature, incubation period, and initial pH of the culture medium. L-asparaginase quantification was valued by the formation of β-aspartyl hydroxamate. The significant positive variables, L-asparagine, L-proline, potato dextrose broth, and sucrose concentrations, were evaluated at 2 levels (+ 1 and − 1) with triplicate of the central point. After 34 runs, maximum activity (2.33 IU/mL) was achieved at the factorial design central point. A central composite design was performed in ATPMS at two levels (+ 1 and − 1) varying Triton X-114 concentration (w/v), separation phase temperature, and crude extract concentration (w/v). The Lasparaginase partition coefficient (K) was considered the experimental design response. Out of the 16 systems that were examined, the most promising presented a purification factor of 1.4 and a yield of 100%.

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Section: Introductionmentioning

confidence: 99%

Optimization of aqueous two-phase micellar system for partial purification of L-asparaginase from Penicillium sp. grown in wheat bran as agro-industrial residue

et al. 2020

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“…Tabela 2. Características dos principais biofármacos de L-asparaginase in vivo (OBREQUE, 2017). Por outro lado, a L-asparaginase nativa de E. coli é a opção que apresenta maior afinidade ao substrato, embora requeira maiores dosagens e atenções com efeitos alergênicos.…”

Section: L-asparaginase E Leucemia Linfoblástica Agudaunclassified

“…A etapa de extração consiste na retirada da L-asparaginase que, neste caso, é encontrada tanto no citoplasma como no espaço periplasmático, não sendo a enzima transportada para o meio extracelular (Torres-Obreque et al, 2019).…”

Section: Seção De Downstreamunclassified

“…Neste sentido, Torres-Obreque (2017) avaliou o processo de extração sob dois aspectos: Figura 14. Diagrama de blocos das principais operações envolvidas no processo de produção de L-asparaginase do tipo Erwinase purificada, baseado no processo desenvolvido por Torres-Obreque (2017).…”

Section: Seção De Downstreamunclassified

“…A Tabela 6 mostra os principais parâmetros de processo assumidos para a simulação do bioprocesso de Torres-Obreque (2017.…”

Section: Dimensionamento Da Planta E Do Processounclassified

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Análise Técnico-Econômica para a produção de biofármacos de L-asparaginase

Fuganholi¹

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Muito além deste projeto de dissertação e refletindo em todos que fizeram parte de minha trajetória, oferecendo e proporcionando apoio, privilégios e, principalmente, sempre depositando confiança em meus planos e decisões, agradeço imensamente à minha família por sempre estar ao meu lado, em especial: minha mãe Valéria, minhas irmãs e meu pai (em memória).Agradeço, também, aos meus amigos e às pessoas especiais que rodeiam a minha vida, por sempre estarem comigo, proporcionando todos os incentivos, apoios e ajudas para dar conta de tantas atividades que aconteceram nos últimos anos.Agradeço, também, a todos os professores que passaram por meu caminho e que com certeza contribuíram à minha formação nos mais variados aspectos.Ao nosso grupo de pesquisa, agradeço pelo acolhimento, pela paciência, pela disponibilidade e pela parceria quando necessário. Presto minha gratidão especialmente ao Professor Adalberto, por ter acolhido o projeto e me acompanhado durante este percurso, e também a importantes e brilhantes profissionais que cruzaram meu caminho, principalmente: Larissa, Rafael, Professor Adriano e Adamo.Ao Universo agradeço pela oportunidade de conviver e receber tantas pessoas especiais, que confiam, suportam e apostam em minhas decisões. RESUMO FUGANHOLI, N. S. Análise Técnico-Econômica para a produção de biofármacos de L-asparaginase. 2021. Dissertação (Mestrado) -Faculdade de Ciências Farmacêuticas,

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Recombinant production of a highly efficient photolyase from Thermus thermophilus

Torres‐Obreque,

Gonçalves,

Ferraro

et al. 2024

Biotechnology Journal

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Ultraviolet (UV) radiation from sunlight can damage DNA, inducing mutagenesis and eventually leading to skin cancer. Topical sunscreens are used to avoid the effect of UV irradiation, but the topical application of DNA repair enzymes, such as photolyase, can provide active photoprotection by DNA recovery. Here we produced a recombinant Thermus thermophilus photolyase expressed in Escherichia coli, evaluated the kinetic parameters of bacterial growth and the kinetics and stability of the enzyme. The maximum biomass (𝑋𝑚𝑎𝑥) of 2.0 g L−1 was reached after 5 h of cultivation, corresponding to 𝑃X = 0.4 g L−1 h. The µ𝑚𝑎𝑥 corresponded to 1.0 h−1. Photolyase was purified by affinity chromatography and high amounts of pure enzyme were obtained (3.25 mg L−1 of cultivation). Two different methods demonstrated the enzyme activity on DNA samples and very low enzyme concentrations, such as 15 µg mL−1, already resulted in 90% of CPD photodamage removal. We also determined photolyase kM of 9.5 nM, confirming the potential of the enzyme at very low concentrations, and demonstrated conservation of enzyme activity after freezing (‐20°C) and lyophilization. Therefore, we demonstrate T. thermophilus photolyase capacity of CPD damage repair and its potential as an active ingredient to be incorporated in dermatological products.

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Production of a novel N‐terminal PEGylated crisantaspase

Cited by 12 publications

References 22 publications

Optimization of aqueous two-phase micellar system for partial purification of L-asparaginase from Penicillium sp. grown in wheat bran as agro-industrial residue

Optimization of aqueous two-phase micellar system for partial purification of L-asparaginase from Penicillium sp. grown in wheat bran as agro-industrial residue

Análise Técnico-Econômica para a produção de biofármacos de L-asparaginase

Recombinant production of a highly efficient photolyase from Thermus thermophilus

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