N. Faria

Quantification of the morphology of sucrose crystals by image analysis

Automated image analysis procedures combined with discriminant factorial analysis (DFA) have been developed to classify agglomerated sucrose crystals according to their shape. The crystals are observed by optical microscopy. Agreement between manual and automated classifications is 90% in average. Each crystal is characterised by its own degree of agglomeration, calculated from the output of the classification. Mono-crystals are further classified into two types according to the habit of their projected silhouette. The use of these techniques is illustrated on commercial sucrose and batch-crystallised particles obtained in a lab-scale reactor in presence of impurities (dextran, raffinose, glucose, sodium carbonate) known to modify the sucrose crystal habit.

BacHBerry: BACterial Hosts for production of Bioactive phenolics from bERRY fruits

BACterial Hosts for production of Bioactive phenolics from bERRY fruits (BacHBerry) was a 3-year project funded by the Seventh Framework Programme (FP7) of the European Union that ran between November 2013 and October 2016. The overall aim of the project was to establish a sustainable and economically-feasible strategy for the production of novel high-value phenolic compounds isolated from berry fruits using bacterial platforms. The project aimed at covering all stages of the discovery and pre-commercialization process, including berry collection, screening and characterization of their bioactive components, identification and functional characterization of the corresponding biosynthetic pathways, and construction of Gram-positive bacterial cell factories producing phenolic compounds. Further activities included optimization of polyphenol extraction methods from bacterial cultures, scale-up of production by fermentation up to pilot scale, as well as societal and economic analyses of the processes. This review article summarizes some of the key findings obtained throughout the duration of the project.

Impact of the cultivation strategy on resveratrol production from glucose in engineered Corynebacterium glutamicum

The health benefits of polyphenols such as stilbenes and flavonoids for humans are increasingly attracting attention. Resveratrol is a well-characterized naturally-occurring stilbene and potent anti-oxidant, which is used as food supplement and cosmetic ingredient. Several microorganisms including Corynebacterium glutamicum were engineered for resveratrol production from glucose. Based on the cultivation of a resveratrol-producing C. glutamicum strain in shake flasks, different strategies for improving production under controlled conditions at bioreactor scale were tested. To this end, different cultivation parameters including substrate concentration and operation modes (batch and fed-batch) were evaluated. Whereas the highest biomass concentration was observed during fed-batch fermentation, the maximum resveratrol production was achieved in batch mode. The maximal titer obtained was 12mgL-1 of resveratrol without the addition of the fatty acid synthase inhibitor cerulenin, which was previously shown to be crucial for production with C. glutamicum. The specific growth rate during production seems to have a significant effect in resveratrol production and apparently low specific growth rates may redirect the metabolic bottleneck from p-coumaric acid formation to malonyl-CoA or ATP availability. We also show that high oxygen concentrations in the bioreactor negatively affected the obtained resveratrol titers with C. glutamicum, which is most likely due to the strong tendency of resveratrol to oxidize or oligomerize. Thus, up-scaling of the resveratrol production process is technically challenging and individual process parameters have to be optimized cautiously.

A dynamical model for the fermentative production of fructooligosaccharides

In this paper a detailed mathematical model is presented for the fermentative production of fructo-oligosaccharides with Aspergillus sp. The model accounts for hydrolysis and transfructolization reactions, as well as biomass formation and it contains 27 parameters that were determined from experimental data using a System Biology toolbox with the Simulated Annealing method for curve fitting. Several additional experiments were performed in bioreactors where the time variation of 7 state variables (Sucrose, Glucose, Fructose, 1-Kestose, Nystose, 1-fructosyl nystose and Biomass) was measured. Experimental data were compared with results from simulations using the estimated parameters and it was verified that the model can predict the FOS production profile. The good agreement between simulated and experimental data was verified by calculating the relative percentage deviation modulus, which was lower than 10% for all cases except one. The derived and validated model can be used for process optimization, for example for indicating which fed-batch strategy could be used to improve the production of FOS while minimizing glucose concentration.

Heterologous production of resveratrol in bacterial hosts: current status and perspectives

The polyphenol resveratrol (3,5,4′-trihydroxystilbene) is a well-known plant secondary metabolite, commonly used as a medical ingredient and a nutritional supplement. Due to its health-promoting properties, the demand for resveratrol is expected to continue growing. This stilbene can be found in different plants, including grapes, berries (blackberries, blueberries and raspberries), peanuts and their derived food products, such as wine and juice. The commercially available resveratrol is usually extracted from plants, however this procedure has several drawbacks such as low concentration of the product of interest, seasonal variation, risk of plant diseases and product stability. Alternative production processes are being developed to enable the biotechnological production of resveratrol by genetically engineering several microbial hosts, such as Escherichia coli, Corynebacterium glutamicum, Lactococcus lactis, among others. However, these bacterial species are not able to naturally synthetize resveratrol and therefore genetic modifications have been performed. The application of emerging metabolic engineering offers new possibilities for strain and process optimization. This mini-review will discuss the recent progress on resveratrol biosynthesis in engineered bacteria, with a special focus on the metabolic engineering modifications, as well as the optimization of the production process. These strategies offer new tools to overcome the limitations and challenges for microbial production of resveratrol in industry.