Biotransformation of terpenes in flavor and functional compounds

Orientador: Glaucia Maria PastoreTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de AlimentosResumo: Este trabalho teve por objetivos o estudo da biotransformação de terpenos e avaliação de suas propriedades biológicas. A biotransformação de limoneno foi realizada por uma linhagem de Fusarium oxysporum cultivada em manipueira e transferida para meio mineral, sendo o óleo essencial de laranja, um resíduo da indústria do suco de laranja, a fonte do limoneno. Rendimentos da ordem de 450 mg/L de a-terpineol, o principal produto obtido, foram alcançados. Da mesma forma, a biotransformação do citronelol foi conduzida por uma linhagem de Penicillium sp. cultivada em manipueira e transferida para meio mineral. O principal produto obtido foi o cis-óxido de rosa em concentrações da ordem de 70 mg/L. A seleção de microrganismos biotransformadores de limoneno (cuja fonte foi óleo essencial de laranja) a- e b-pinenos (cuja fonte foi terebentina, resíduo industrial da indústria de papel) foi realizada por microextração em fase sólida (MEFS). A técnica mostrou-se eficaz para a recuperação de voláteis presentes no 'heapspace¿ de culturas esporuladas de superfície para a biotransformação. Os microrganismos selecionados por MEFS foram submetidos à biotransformação em cultura líquida e produção da ordem de 50 mg/L de verbenol por Mucor sp. 2276 e 70 mg/L de verbenona resultaram da biotransformação de a- e b-pinenos. Aspergillus sp. 2357 e Penicillium sp. 2360 produziram aproximadamente 90 e 10 mg/L de a-terpineol e álcool perílico respectivamente. A biotransformação de a-farneseno por linhagens de Aspergillus niger gerou compostos nunca relatados anteriormente na literatura. Quatro produtos principais foram obtidos. Apenas um composto pôde ser identificado por meio de CG-EM como 6-OH-farneseno. Análises de CG-olfatometria descreveram 6-OH-farneseno como aroma cítrico impactante. Estudos 'in vitro¿ e 'in vivo¿ com extrato da biotransformação de limoneno por Fusarium oxysporum e com padrões de monoterpenos revelaram o potencial desses compostos em atuarem como antioxidantes, gerando uma possibilidade para esses compostos serem utilizados industrialmente como aromas funcionaisAbstract: The biotransformation of terpenes and their functional properties were investigated in this study. The biotransformation of limonene was done by a Fusarium oxysporum strain grown in cassava waste water and transferred into a mineral medium for biotransformation. The limonene source was an orange essential oil from a orange juice industry. The main biotransformation product was a-terpineol, reaching around 450 mg/L. Similarly, biotransformation of citronellol was conducted by a Penicillium sp. strain grown also in cassava waste water and transferred into a mineral medium. The main product was cis-rose oxide, reaching concentrations higher than 70 mg/L. The screening of microrganisms for biotransformation of limonene (from orange essential oil) and a-, b-pinenes (from turpentine oil, residue from pulp industry) was done by solid phase microextraction (SPME). The technique was effective for the recovery of the volatiles from the headspace of sporulated surface biotransformation cultures. Liquid culture biotransformation experiments performed with the SPME screened strains resulted in the production of 50 mg/L of verbenol by Mucor sp. 2276 and 70 mg/L of verbenone form a-, b-pinenes and 90 mg/L of a-terpineol by Aspergillus sp. 2357 and 10 mg/L of perillyl alcohol by Penicillium sp. 2360. The biotransformation of a-farnesene by Aspergillus niger strains resulted in compounds never described in the literature before. Four main new compounds were obtained. Only one of them could be identified by GC-MS as 6-OH-farnesene. CG-O experiments revealed the impactant citrus aroma of 6-OH-farnesene. 'In vitro¿ and 'in vivo¿ experiments with the limonene biotransformation extract by Fusarium oxysporum and with the monoterpene standards present in the extract revealed the antioxidant potential of these compounds, which open a new perspective for the utilization of these compounds as functional aroma compoundsDoutoradoDoutor em Engenharia de Alimento

Carotenoids: properties, applications and biotransformation in flavor compounds

Carotenoids are widely distributed in nature, providing yellow, orange or red color in a great number of vegetables, microorganisms and in some animals. Carotenoids act as biological antioxidants and seem to play an important role in human health by protecting cells and tissues from the damaging effects of free radicals and singlet oxygen. Several authors describe the oxidative cleavage of carotenoids in flavor compounds as occuring through chemical or photochemical degradations or through biotechnological processes. Biotransformation of carotenoids seems to be a reasonable alternative to produce flavor compounds since these compounds are considered 'natural' ingredients. In this work we describe the properties of some carotenoids, as well as biotechnological approaches to obtain its oxyfunctionalized derivatives.61662

The Use of Endophytes to Obtain Bioactive Compounds and Their Application in Biotransformation Process

Endophytes are microorganisms that reside asymptomatically in the tissues of higher plants and are a promising source of novel organic natural metabolites exhibiting a variety of biological activities. The laboratory of Bioaromas (Unicamp, Brazil) develops research in biotransformation processes and functional evaluation of natural products. With the intent to provide subsidies for studies on endophytic microbes related to areas cited before, this paper focuses particularly on the role of endophytes on the production of anticancer, antimicrobial, and antioxidant compounds and includes examples that illustrate their potential for human use. It also describes biotransformation as an auspicious method to obtain novel bioactive compounds from microbes. Biotransformation allows the production of regio- and stereoselective compounds under mild conditions that can be labeled as “natural,” as discussed in this paper

Suplementação com ácido linoleico conjugado: conteúdo de lípides e histologia hepática de ratos Wistar saudáveis

This work aimed to evaluate the effects of the consumption of two commercial conjugated linoleic acid (CLA) mixtures on lipid content and liver histology of healthy rats. The investigation was carried out using thirty rats divided into three groups: C (control), AE (AdvantEdge®CLA), and CO (CLA One®). The concentration of CLA was 2% of feed consumption, and the animals were supplemented daily for 42 days. The total lipid content of the liver was determined, and the histology of the organ was examined by Transmission Electronic Microscopy. The results of total liver lipid contents did not exhibit significant differences between the groups. With regard to hepatic histology, it was observed that although fat globules were visibly present in higher numbers and bigger size in the CLA groups, the organ histology was considered normal since both cytoplasm and organelles showed integrity. It was concluded that even though liver microscopic images indicated the presence of fat globules in the liver, from a statistical point of view, the supplementation for 42 days did not bring about lipid accumulation, nor did it alter hepatic histology.O objetivo do trabalho foi avaliar o efeito da suplementação com duas misturas comerciais de ácido linoleico conjugado (CLA) sobre o conteúdo de lípides e a histologia hepática de ratos saudáveis. Trinta ratos foram divididos em três grupos: C (controle), AE (AdvantEdge®CLA) e CO (CLA One®). A concentração de CLA foi de 2% em relação ao consumo diário de dieta, e os animais foram suplementados diariamente durante 42 dias. O conteúdo de lípides totais foi determinado e a histologia hepática foi examinada por meio de Microscopia Eletrônica de Transmissão. Não houve diferença significativa nos resultados de conteúdo de lípides hepáticos entre os grupos. Quanto à histologia hepática, embora tenha sido observada a presença de glóbulos de gordura em maior número e tamanho nos grupos suplementados com CLA, a histologia do órgão apresentou-se normal, visto que citoplasma e organelas estavam íntegros. Concluiu-se que não houve acúmulo de lípides nem alteração da histologia hepática após 42 dias de suplementação com CLA.141146Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Comparação de volatilidade e compostos polifenólicos na própolis verde brasileira e sua origem botânica Baccharis dracunculifolia

Ethanolic extracts and essential oils from Green Propolis from southeastern Brazil and leaf buds from its botanical origin Baccharis dracunculifolia were analyzed by Reversed Phase High Performance Liquid Chromatography (RP-HPLC), Reversed Phase High Performance Thin Layer Chromatography (RP-HPTLC) and Gas Chromatography - Mass Spectrometry (GC-MS). The essential oils were obtained by hydro-distillation. Both ethanolic extracts and essential oils showed similar chromatographic profiles. Thirteen flavonoids were identified by RP-HPLC and RP-HPTLC analyses in both samples. Twenty-three volatile compounds were identified by GC-MS analyses. Seventeen were present in both essential oils. The major flavonoid compound in both extracts was artepillin C. The major volatile compound in both essential oils was nerolidol. The major compounds identified in this work could be used as chemical markers in order to classify and identify botanical origins of propolis.Extratos etanólicos e óleos essenciais de própolis verde do sudeste brasileiro e gemas de sua origem botânica (Baccharis dracunculifolia) foram analisados por CLAE-FR (Cromatografia Líquida de Alta Eficiência em Fase Reversa), CCDAE (Cromatografia em Camada Delgada de Alta Eficiência) e CG-EM (Cromatografia Gasosa acoplada a Espectrometria de Massas). Os óleos essenciais foram obtidos por hidro destilação. Extratos etanólicos e óleos essenciais de Baccharis dracunculifolia e de própolis mostraram perfis cromatográficos similares entre si. Treze flavonóides foram identificados por CLAE-FR e CCDAE em ambas as amostras. Vinte e três compostos voláteis foram identificados por CG EM, sendo dezessete deles presentes em ambos os óleos essenciais. Artepillin C foi o flavonóide encontrado em maiores concentrações em ambas as amostras, enquanto nerolidol foi o volátil majoritário em ambos os óleos essenciais. Os compostos majoritários identificados neste trabalho podem ser utilizados como marcadores químicos para classificar de forma prática e identificar origens botânicas de própolis.178181Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Carotenóides: propriedades, aplicações e biotransformação para formação de compostos de aroma

Carotenoids are widely distributed in nature, providing yellow, orange or red color in a great number of vegetables, microorganisms and in some animals. Carotenoids act as biological antioxidants and seem to play an important role in human health by protecting cells and tissues from the damaging effects of free radicals and singlet oxygen. Several authors describe the oxidative cleavage of carotenoids in flavor compounds as occuring through chemical or photochemical degradations or through biotechnological processes. Biotransformation of carotenoids seems to be a reasonable alternative to produce flavor compounds since these compounds are considered 'natural' ingredients. In this work we describe the properties of some carotenoids, as well as biotechnological approaches to obtain its oxyfunctionalized derivatives

Conjugated linoleic acid supplementation: lipid content and hepatic histology in healthy Wistar rats

This work aimed to evaluate the effects of the consumption of two commercial conjugated linoleic acid (CLA) mixtures on lipid content and liver histology of healthy rats. The investigation was carried out using thirty rats divided into three groups: C (control), AE (AdvantEdge®CLA), and CO (CLA One®). The concentration of CLA was 2% of feed consumption, and the animals were supplemented daily for 42 days. The total lipid content of the liver was determined, and the histology of the organ was examined by Transmission Electronic Microscopy. The results of total liver lipid contents did not exhibit significant differences between the groups. With regard to hepatic histology, it was observed that although fat globules were visibly present in higher numbers and bigger size in the CLA groups, the organ histology was considered normal since both cytoplasm and organelles showed integrity. It was concluded that even though liver microscopic images indicated the presence of fat globules in the liver, from a statistical point of view, the supplementation for 42 days did not bring about lipid accumulation, nor did it alter hepatic histology