Stability of microencapsulated lactic acid bacteria under acidic and bile juice conditions

The probiotic strains Lactobacillus brevis CCMA1284 and Lactobacillus plantarum CCMA0359 were microencapsulated by spray drying using different matrices – whey powder (W), whey powder with inulin (WI) and whey powder with maltodextrin (WM). Viability of the microencapsulated strains in acid and bile juices and during 90 days of storage (seven and 25 °C) was evaluated. The two strains exhibited high encapsulation efficiency (> 86%) by spray drying. The different matrices maintained L. plantarum viability above six log CFU g⁻¹ at 7 °C for 90 days, whereas similar results for L. brevis were observed only for W. The use of inulin as matrix of encapsulation did not enhance bacterial viability in the evaluated conditions. In general, the use of W and WM as matrices was effective for L. plantarum viability. However, only W was effective for L. brevis in the evaluated conditions. The spray drying technique was successfully adopted for the encapsulation of L. plantarum CCMA0359 and L. brevis CCMA1284 strains.     

Analysis of the essential oil of Mexican oregano (Lippia graveolens HBK)

The composition of the volatile oil of Mexican oregano (Lippia graveolens HBK) isolated by steam distillation was investigated by means of column chromatography, gas chromatography and mass spectrometry. A total of 33 components were identified including 22 hydrocarbons, 4 alcohols, 4 ethers, 2 phenols and 1 ketone. Of the 33 components observed in the present study 7 were identified in Mexican oregano for the first time.     

A Rapid and Eco-Friendly Method for Determination of Hydrolysable Tannins and Its Application to Honey Samples

This paper describes the development of a rapid and clean method for the determination of hydrolysable tannins in honey samples, focusing on an easy and green sample pretreatment and low generation of waste. The proposed method is based on the reaction between KIO₃ and hydrolysable tannin compounds in acetate buffer medium (pH 4.75), with formation of a colored product measured at 525 nm. The analytical conditions were optimized using experimental design (central composite design). The linear range obtained was 40.0–740 mg L^?1 (r?=?0.998), and the detection (LOD) and quantification (LOQ) limits were 12.2 and 40.8 mg L^?1, respectively. Acetate buffer was used to ensure formation of the colored product obtained by reaction between the chromogenic reagent and the analyte. The method showed good results when applied to honey samples, with recoveries in the range of 78.3–96.1 %, and the use of diffuse reflectance spectroscopy coupled with spot test analysis provided low reagent consumption and minimized waste generation. The results obtained with the proposed method were confirmed by a spectrophotometric method.     

A feasibility study of Lactobacillus plantarum in fruit powders after processing and storage

The aim of this study was to develop fruit powders (apple, banana and strawberry) enriched with a probiotic strain (Lactobacillus plantarum 299v). Two methodologies were proposed: (i) drying of the fruit with probiotic culture incorporated (by convection) or (ii) drying of fruit (by convection) and addition of spray‐dried probiotic culture. In the first methodology, processing caused a notable reduction in probiotic viable counts in apple, but this reduction was lower during drying of banana and strawberry. A large reduction in viable cells was also recorded during storage. In the second methodology, the survival of L. plantarum 299v was considerably higher during spray‐drying, and fruit powders with a microbial content suitable for a probiotic food (10⁸–10⁹ cfu g^?1) were obtained. The fruit powders incorporating L. plantarum 299v can be stored at 4 °C or at room temperature, for at least 3 months. This preliminary study demonstrated that fruit powders are good carriers of probiotic cultures, but the techniques used to produce them should be carefully considered.     

Identification of potential technologies for 1,4-Butanediol production using prospecting methodology

1,4-Butanediol (1,4-BDO) is a four-carbon diol used for industrial applications such as organic solvents, and the production of adhesives, fibers and polyurethanes. 1,4-BDO currently is produced through several petrochemical routes: hydrogenation of maleic anhydride, isomerization of propylene oxide, acetoxylation of butadiene, and the reaction between formaldehyde and acetylene. The current trends in 1,4-BDO production involve the utilization of renewable sources such as biomass. In this context, the present study aimed to identify promising technologies of 1,4-BDO production through prospecting methodology based on the analyses of patents and scientific article, describing the most relevant aspects of those emerging technologies. An increasing amount of 1,4-BDO production focused on biotechnological routes has been reported, with the US heavily involved in the development of new technologies. This study tracked three promising technologies which have potential for application in a biorefinery context because those processes involve (i) production of 1,4-BDO from sugars, classified herein as the biotechnological route; (ii) production of intermediates from sugar fermentation followed by catalytic conversion into 1,4-BDO, classified herein as the hybrid route, and (iii) furan/furfural conversion into 1,4-BDO. © 2020 Society of Chemical Industry     

Expression of genes encoding the E2 and E3 components of the Bacillus stearothermophilus pyruvate dehydrogenase complex and the stoichiometry of subunit interaction in assembly in vitro

Genes encoding the dihydrolipoyl acetyltransferase (E2) and dihydrolipoyl dehydrogenase (E3) components of the pyruvate dehydrogenase (PDH) multienzyme complex from Bacillus stearothermophilus were overexpressed in Escherichia coli. The E2 component was purified as a large soluble aggregate (molecular mass > 1 x 10(6) Da) with the characteristic 532 symmetry of an icosahedral (60-mer) structure, and the E3 as a homodimer with a molecular mass of 110 kDa. The recombinant E2 component in vitro was capable of binding either 60 E3(alpha2) dimers or 60 heterotetramers (alpha2beta2) of the pyruvate decarboxylase (E1) component (also the product of B. stearothermophilus genes overexpressed in E. coli). Assembling the E2 polypeptide chain into the icosahedral E2 core did not impose any restriction on the binding of E1 or E3 to the peripheral subunit-binding domain in each E2 chain. This has important consequences for the stoichiometry of the assembled complex in vivo. The lipoyl domain of the recombinant E2 protein was found to be unlipoylated, but it could be correctly post-translationally modified in vitro using a recombinant lipoate protein ligase from E. coli. The lipoylated E2 component was able to bind recombinant E1 and E3 components in vitro to generate a PDH complex with a catalytic activity comparable with that of the wild-type enzyme. Reversible unfolding of the recombinant E2 and E3 components in 6 M guanidine hydrochloride was possible in the absence of chaperonins, with recoveries of enzymic activities of 95% and 85%, respectively. However, only 26% of the E1 enzyme activity was recovered under the same conditions as a result of irreversible denaturation of both E1alpha and E1beta. This represents the first complete post-translational modification and assembly of a fully active PDH complex from recombinant proteins in vitro.     

Rat liver catechol-O-methyltransferase kinetics and assay methodology

In mammals, catechol-O-methyltransferase (COMT) is distributed throughout various organs, the highest activities being found in the liver and kidney. However, comparisons of the kinetic parameters are difficult to perform, since the experimental procedures in the enzyme assay vary quite considerably. The present work was aimed at studying the optimal liver COMT assay conditions for determining the kinetics of the enzyme. The COMT assay was performed with liver homogenates from 60 days old male Wistar rats with adrenaline (AD) as the substrate. Time course experiments using 100 microM S-adenosyl-L-methionine (SAMe) and 300 microM AD showed linearity of O-methylation reaction upto 10 min. Using 100 microM SAMe, Vmax (nmol mg protein-1 h-1) and Km (microM) values progressively decreased respectively from 22.1 and 104.8 at 5 min down to 5.8 and 24.62 at 60 min incubation periods. This decrease was not due to end-product inhibition. Using 2500 microM AD, Km values (microM) for the methyl donor SAMe increased progressively from 174 at 5 min upto 1192.5 at 60 min; upto 30 min of incubation Vmax values did not change. When a 5 min incubation period and 500 microM SAMe were used, Vmax and Km values for liver COMT were 63.4 nmol mg protein-1 h-1 and 261.1 microM, respectively. It is concluded that an incubation period of 5 min and a SAMe concentration of 500 microM provide optimal conditions for the liver homogenate COMT assay.     

Strongly Photosensitive and Fluorescent F8T2 Electrospun Fibers

Electrospun fibers of poly[(9,9‐dioctylfluorenyl‐2,7‐diyl)‐co‐bithiophene] (F8T2) with exceptional electro‐optical performance are obtained. The I/T characteristics measured in fibers with 7–15 µm diameter and 1 mm length show a semiconductor behavior; their thermal activation energy is 0.5 eV and the dark conductivity at RT is 5 × 10^?9 (Ω cm)^?1. Besides exhibiting a photosensitivity of about 60 under white light illumination with a light power intensity of 25 mW · cm^?2, the fibers also attain RT photoluminescence in the cyan, yellow, and red wavelength range under ultraviolet, blue, and green light excitation, respectively. Optical microscope images of F8T2 reveal homogeneous electrospun fibers, which are in good agreement with the uniformly radial fluorescence observed.