Sorele Batista Fiaux

Production of 6-pentyl-α-pyrone by trichoderma harzianum in solid-state fermentation

Muitas especies do genero Trichoderma sao capazes de produzir a substância 6-pentil-α-pirona (6-PP), uma lactona com aroma caracteristico de coco. No presente trabalho, varios parâmetros de cultura foram estudados para aumentar a producao de 6-PP por Trichoderma harzianum 4040 em fermentacao em estado solido. Po da casca de coco verde adicionado a uma solucao nutriente foi usado como material de suporte para a fermentacao. Um planejamento experimental de varredura segundo a tecnica de Plackett-Burman foi aplicado, seguido de um planejamento fatorial fracionario. No dominio experimental estudado, as melhores condicoes de cultura foram (100 g suporte)-1: sacarose, 3 g; NaNO3, 0,24 g; (NH4)2SO4, 0,18 g; KH2PO4, 0,1 g; producao do inoculo, 2,2 x 106 esporos; umidade, 55%. A temperatura estabelecida foi de 28oC. Esse estudo conduziu a concentracao de 6-PP seis vezes maior (5,0 mg/g de materia seca) do que a inicial (0,8 mg/g de materia seca) apos sete dias de cultivo.

Particle-size distribution (PSD) of pulverized hair: A quantitative approach of milling efficiency and its correlation with drug extraction efficiency

Different types of hair were submitted to different milling procedures and their resulting powders were analyzed by scanning electron microscopy (SEM) and laser diffraction (LD). SEM results were qualitative whereas LD results were quantitative and accurately characterized the hair powders through their particle size distribution (PSD). Different types of hair were submitted to an optimized milling conditions and their PSD was quite similar. A good correlation was obtained between PSD results and ketamine concentration in a hair sample analyzed by LC-MS/MS. Hair samples were frozen in liquid nitrogen for 5min and pulverized at 25Hz for 10min, resulting in 61% of particles <104μm and 39% from 104 to 1000μm. Doing so, a 359% increment on ketamine concentration was obtained for an authentic sample extracted after pulverization comparing with the same sample cut in 1mm fragments. When milling time was extended to 25min, >90% of particles were <60μm and an additional increment of 52.4% in ketamine content was obtained. PSD is a key feature on analysis of pulverized hair as it can affect the method recovery and reproducibility. In addition, PSD is an important issue on sample retesting and quality control procedures.

Asymmetric bioreduction of β-ketoesters derivatives by Kluyveromyces marxianus: influence of molecular structure on the conversion and enantiomeric excess

This study presents the bioreduction of six β-ketoesters by whole cells of Kluyveromyces marxianus and molecular investigation of a series of 13 β-ketoesters by hologram quantitative structure-activity relationship (HQSAR) in order to relate with conversion and enantiomeric excess of β-stereogenic-hydroxyesters obtained by the same methodology. Four of these were obtained as (R)-configuration and two (S)-configuration, among them four compounds exhibited >99% enantiomeric excess. The β-ketoesters series LUMO maps showed that the β-carbon of the ketoester scaffold are exposed to undergo nucleophilic attack, suggesting a more favorable β-carbon side to enzymatic reduction based on adopted molecular conformation at the reaction moment. The HQSAR method was performed on the β-ketoesters derivatives separating them into those provided predominantly (R)- or (S)-β-hydroxyesters. The HQSAR models for both (R)- and (S)-configuration showed high predictive capacity. The HQSAR contribution maps suggest the importance of β-ketoesters scaffold as well as the substituents attached therein to asymmetric reduction, showing a possible influence of the ester group carbonyl position on the molecular conformation in the enzyme catalytic site, exposing a β-carbon side to the bioconversion to (S)- and (R)-enantiomers.

Glicerol como solvente verde em eficiente obtenção de beta-hidroxiésteres

In this work, we report a new method for obtaining racemic β-hydroxyesters by reduction of β-ketoesters. The use of glycerol as a reactional medium in selective reduction of β-ketoesters into the corresponding alcohols was shown to be a viable and more efficient alternative compared with the conventional methodology, taking into account green chemistry prerogatives.

Solid-phase microextraction of 6-pentyl-α-pyrone produced by solid-state fermentation

The lactone 6-pentyl-a-pyrone (6-PP) is a coconut-like aroma compound approved by the American agency Food and Drug Administration (FDA) for food use. The aim of the present study was to improve the extraction conditions of headspace solid-phase microextraction coupled with gas chromatography to quantify the 6-PP produced by Trichoderma harzianum using green coir dust. To achieve this purpose, standard solutions of 6-PP were added to green coir dust. The compound was extracted by a polydimethylsiloxane fiber. Agitation and addition of a 25% NaCl solution (w/v) resulted in better reproducibility. A central composite design was applied to study three factors: extraction temperature, conditioning time, and extraction time. Good responses were obtained when the extraction temperature was 79 °C, and the extraction time was 29 minutes. The conditioning time effect was not significant and was established at 2 minutes. After the development of the analytical method, it was employed to determine the 6-PP production by Trichoderma harzianum in solid-state fermentation using green coir dust as the support material for the process.