Synthesis of structured triacylglycerols enriched in n-3 fatty acids by immobilized microbial lipase

The search for new biocatalysts has aroused great interest due to the variety of micro-organisms and their role as enzyme producers. Native lipases from Aspergillus niger and Rhizopus javanicus were used to enrich the n-3 long-chain polyunsaturated fatty acids content in the triacylglycerols of soybean oil by acidolysis with free fatty acids from sardine oil in solvent-free media. For the immobilization process, the best lipase/support ratios were 1:3 (w/w) for Aspergillus niger lipase and 1:5 (w/w) for Rhizopus javanicus lipase using Amberlite MB-1. Both lipases maintained constant activity for 6 months at 4 °C. Reaction time, sardine-free fatty acids:soybean oil mole ratio and initial water content of the lipase were investigated to determine their effects on n-3 long-chain polyunsaturated fatty acids incorporation into soybean oil. Structured triacylglycerols with 11.7 and 7.2% of eicosapentaenoic acid + docosahexaenoic acid were obtained using Aspergillus niger lipase and Rhizopus javanicus lipase, decreasing the n-6/n-3 fatty acids ratio of soybean oil (11:1 to 3.5:1 and 4.7:1, respectively). The best reaction conditions were: initial water content of lipase of 0.86% (w/w), sardine-free faty acids:soybean oil mole ratio of 3:1 and reaction time of 36 h, at 40 °C. The significant factors for the acidolysis reaction were the sardine-free fatty acids:soybean oil mole ratio and reaction time. The characterization of structured triacylglycerols was obtained using easy ambient sonic-spray ionization mass spectrometry. The enzymatic reaction led to the formation of many structured triacylglycerols containing eicosapentaenoic acid, docosahexaenoic acid or both polyunsaturated fatty acids.47410061013CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPSem informaçãoSem informaçã

Enzyme-assisted modification of flavonoids from Matricaria chamomilla: antioxidant activity and inhibitory effect on digestive enzymes

Matricaria chamomilla L. contains antioxidant flavonoids that can have their bioactivity enhanced by enzymatic hydrolysis of specific glycosyl groups. This study implements an untargeted metabolomics approach based on ultra-performance liquid chromatography coupled with electrospray ionisation quadrupole time-of-flight mass spectrometry technique operating in MSE mode (UPLC-QTOF-MSE) and spectrophotometric analysis of chamomile aqueous infusions, before and after hydrolysis by hesperidinase and ?-galactosidase. Several phenolic compounds were altered in the enzymatically treated infusion, with the majority being flavonoid derivatives of apigenin, esculetin, and quercetin. Although enzymatically modifying the infusion only led to a small increase in antioxidant activity (DPPH? method), its inhibitory effect on pancreatic lipase was of particular interest. The enzymatically treated infusion exhibited a greater inhibitory effect (EC50 of 35.6??M) than unmodified infusion and kinetic analysis suggested mixed inhibition of pancreatic lipase. These results are of great relevance due to the potential of enzymatically treated functional foods in human health3514249CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informaçãosem informação2012/20393–

Enzymic enhancement of W3 polyunsaturated fatty acids content in brazilian sardine oil

Se ensayaron cuatro lipasas microbianas con la intención de aumentar el contenido de los ácidos grasos poliinsaturados de la serie W3 (AGPI W3) en la fracción de glicéridos no hidrolizados después de la hidrólisis del aceite de sardina brasileño. La lipasa de Candida cylindracea fue la más eficiente entre las enzimas ensayadas, aumentando el contenido de ácido decosahexaenoico (DHA) del 10,2% en el aceite original al 22,5% en relación a los ácidos grasos totales después de 16 h de reacción a 35 °C, lo que equivale a un incremento del 120% en relación a la cantidad presente antes de la hidrólisis. Las cuatro lipasas probadas no fueron eficientes para aumentar el contenido del ácido eicosapentenoico (EPA) en el aceite de sardina brasileño en las condiciones del ensayo.In an attempt to concentrate the content of W3 polyunsaturated fatty acids ( W3 PUFA) in the partially hydrolyzed glycerides, Brazilian sardine oil was hydrolyzed with four kinds of microbial lipases. The enzyme from Candida cylindracea was the most effective for the production of oil with high concentration of docosahexaenoic acid (DHA). Sardine oil was treated at 35 °C with this lipase for 16 h reaction and 60.0% hydrolysis resulted in an increase in the DHA content from 10.2% in the original oil to 22.5% (2.20- fold enrichment) in the unhydrolysed acylglycerol. None the four lipases tested could raise the EPA content significantly.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Enzymic enhancement of W3 polyunsaturated fatty acids content in brazilian sardine oil

Se ensayaron cuatro lipasas microbianas con la intención de aumentar el contenido de los ácidos grasos poliinsaturados de la serie W3 (AGPI W3) en la fracción de glicéridos no hidrolizados después de la hidrólisis del aceite de sardina brasileño. La lipasa de Candida cylindracea fue la más eficiente entre las enzimas ensayadas, aumentando el contenido de ácido decosahexaenoico (DHA) del 10,2% en el aceite original al 22,5% en relación a los ácidos grasos totales después de 16 h de reacción a 35 °C, lo que equivale a un incremento del 120% en relación a la cantidad presente antes de la hidrólisis. Las cuatro lipasas probadas no fueron eficientes para aumentar el contenido del ácido eicosapentenoico (EPA) en el aceite de sardina brasileño en las condiciones del ensayo.In an attempt to concentrate the content of W3 polyunsaturated fatty acids ( W3 PUFA) in the partially hydrolyzed glycerides, Brazilian sardine oil was hydrolyzed with four kinds of microbial lipases. The enzyme from Candida cylindracea was the most effective for the production of oil with high concentration of docosahexaenoic acid (DHA). Sardine oil was treated at 35 °C with this lipase for 16 h reaction and 60.0% hydrolysis resulted in an increase in the DHA content from 10.2% in the original oil to 22.5% (2.20- fold enrichment) in the unhydrolysed acylglycerol. None the four lipases tested could raise the EPA content significantly.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Aplicação de lipases microbianas na obtenção de concentrados de ácidos graxos poliinsaturados

Several polyunsaturated fatty acids (PUFA) belonging to the ômega 6 series, such as cis-6,9,12 gamma-linolenic acid, as well as those of the ômega 3 series, such as cis-5,8,11,14,17-eicosapentaenoic acid and cis-4,7,10,13,16,19-docosahexaenoic acid are of considerable interest due to their nutritional and therapeutic properties. Methods used for the concentration of PUFA from natural sources include urea adduct formation, solvent winterization, supercritical fluid extraction and lipase-catalyzed reaction. Lipases are known to have little reactivity on PUFA and these acids can be enriched by selective hydrolysis, direct esterification of glycerol with PUFA and interesterification. Since lipase reactions are advantageous with respect to fatty acid, positional specificities and mild incubation condition, these enzymes are considered to be suitable for the production of PUFA concentrates for medical purposes

Anticancer Asparaginases: Perspectives in Using Filamentous Fungi as Cell Factories

The enzyme L-asparaginase (L-asparagine amidohydrolase) catalyzes the breakdown of L-asparagine into aspartate and ammonia, which leads to an anti-neoplastic activity stemming from its capacity to deplete L-asparagine concentrations in the bloodstream, and it is therefore used in cases of acute lymphoblastic leukemia (ALL) to inhibit malignant cell growth. Nowadays, this anti-cancer enzyme, largely produced by Escherichia coli, is well established on the market. However, E. coli L-asparaginase therapy has side effects such as anaphylaxis, coagulation abnormality, low plasma half-life, hepatotoxicity, pancreatitis, protease action, hyperglycemia, and cerebral dysfunction. This review provides a perspective on the use of filamentous fungi as alternative cell factories for L-asparaginase production. Filamentous fungi, such as various Aspergillus species, have superior protein secretion capacity compared to yeast and bacteria and studies show their potential for the future production of proteins with humanized N-linked glycans. This article explores the past and present applications of this important enzyme and discusses the prospects for using filamentous fungi to produce safe eukaryotic asparaginases with high production yields

Enzymatic Hydrolysis of Rutin: Evaluation of Kinetic Parameters and Anti-Proliferative, Mutagenic and Anti-Mutagenic Effects

The bioavailability of glucoside flavonoids is influenced by the nature of the sugar, glucosides being absorbed faster than rhamnoglucosides, for example. One strategy to enhance the bioavailability is enzymatic hydrolysis. In this study, some kinetic parameters of hesperidinase-mediated hydrolysis of rutin were evaluated using an UHPLC/QTOF-MSE analysis of the products of a bioconversion reaction. The resulting hydrolyzed rutins (after 4, 8 and 12 h of reaction) were submitted to anti-proliferative and Cytokinesis-Block Micronucleus (CBMN) assays in CHO-K1 cells. In the hesperidinase-mediated hydrolysis, the final concentration of quercetin-3-O-glucoside (Q3G) was directly proportional to the rutin concentration and inversely proportional to the reaction time. At an anti-proliferative concentration (2.5 μg/mL), hydrolyzed rutin derivatives did not show a mutagenic effect, except for the sample with a higher content of Q3G (after 4 h of the enzymatic hydrolysis of rutin). Moreover, the higher Q3G content in hydrolyzed rutin protected the CHO-K1 cells 92% of the time against methyl methanesulfonate-induced mutagenic damage. These results suggested that the anti-mutagenic effect of hydrolyzed rutin might be related to antioxidant and cell death induction. Presenting a good lipophilicity/hydrophilicity ratio, together with antioxidant and anti-mutagenic activities, the hesperidinase-mediated hydrolyzed rutin seemed to be a promisor raw material for the development of food supplements

A Transcript Finishing Initiative for Closing Gaps in the Human Transcriptome

We report the results of a transcript finishing initiative, undertaken for the purpose of identifying and characterizing novel human transcripts, in which RT-PCR was used to bridge gaps between paired EST clusters, mapped against the genomic sequence. Each pair of EST clusters selected for experimental validation was designated a transcript finishing unit (TFU). A total of 489 TFUs were selected for validation, and an overall efficiency of 43.1% was achieved. We generated a total of 59,975 bp of transcribed sequences organized into 432 exons, contributing to the definition of the structure of 211 human transcripts. The structure of several transcripts reported here was confirmed during the course of this project, through the generation of their corresponding full-length cDNA sequences. Nevertheless, for 21% of the validated TFUs, a full-length cDNA sequence is not yet available in public databases, and the structure of 69.2% of these TFUs was not correctly predicted by computer programs. The TF strategy provides a significant contribution to the definition of the complete catalog of human genes and transcripts, because it appears to be particularly useful for identification of low abundance transcripts expressed in a restricted set of tissues as well as for the delineation of gene boundaries and alternatively spliced isoforms