Development of a Higee bioreactor (HBR) for production of polyhydroxyalkanoate: Hydrodynamics, gas–liquid mass transfer and fermentation studies

Abstract This study addresses the hydrodynamics and mass transfer characterisation of a Higee bioreactor (HBR) for application to polyhydroxyalkanoate (PHA) production from Pseudomonas putida KT2442 fermentation. The motivation for this work is to address the potential oxygen transfer limitations which can severely impede the progress of this aerobic fermentation process and reduce PHA productivity in conventional bioreactors. It is shown that a maximum of 2.5 transfer units can be achieved in an oxygen-stripping operation where the presence of packing, higher rotor speeds, higher air flowrates and lower liquid flowrates all have a positive influence on the number of transfer units (NTU). W…

Glycosidase enzymes stabilization through immobilization onto nanoparticulated bimodal organosilicas

Multigram Scale Enzymatic Synthesis of (R)-1-(4′-Hydroxyphenyl)ethanol Using Vanillyl Alcohol Oxidase

The enantioselective oxyfunctionalisation of C−H bonds is a highly interesting reaction, as it provides access to chiral alcohols that are important pharmaceutical building blocks. However, it is hard to achieve using traditional methods. One way in which it can be achieved is through the action of oxidative enzymes. Although many reports of the oxyfunctionalisation capabilities of enzymes at an analytical scale have been published, reports on the use of enzymes to achieve oxyfunctionalisation on a synthetically relevant scale are fewer. Here, we describe the scale-up of the conversion of 4-ethylphenol to (R)-1-(4′-hydroxyphenyl)ethanol using the flavin-dependent enzyme vanillyl alcohol oxi…

Enlarged pore size in nanoparticulated bimodal porous silicas: Improving accessibility

Abstract Mass-transfer kinetics seems to be highly favored in siliceous materials constructed from the aggregation of mesoporous nanoparticles. Besides intra-particle mesopores, over the course of the aggregation process an inter-particle (textural) large pore system is generated. Diffusion constrains through the resulting hierarchically structured pore systems mainly depend on the characteristics of the intra-particle mesopores. By using alkanes as swelling agents, we have been able to significantly increase the intra-particle mesopore size in previously well characterized UVM-7 materials. The Winsor-III-like behavior associated with the presence of alkanes in the hydro-alcoholic reaction …

MCR-ALS on metabolic networks: Obtaining more meaningful pathways

[EN] With the aim of understanding the flux distributions across a metabolic network, i.e. within living cells, Principal Component Analysis (PCA) has been proposed to obtain a set of orthogonal components (pathways) capturing most of the variance in the flux data. The problems with this method are (i) that no additional information can be included in the model, and (ii) that orthogonality imposes a hard constraint, not always reasonably. To overcome these drawbacks, here we propose to use a more flexible approach such as Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) to obtain this set of biological pathways through the network. By using this method, different constraint…

Metabolic flux understanding of Pichia pastoris grown on heterogenous culture media

[EN] Within the emergent field of Systems Biology, mathematical models obtained from physical chemical laws (the so-called first principles-based models) of microbial systems are employed to discern the principles that govern cellular behaviour and achieve a predictive understanding of cellular functions. The reliance on this biochemical knowledge has the drawback that some of the assumptions (specific kinetics of the reaction system, unknown dynamics and values of the model parameters) may not be valid for all the metabolic possible states of the network. In this uncertainty context, the combined use of fundamental knowledge and data measured in the fermentation that describe the behaviour…

Hierarchical bimodal porous silicas and organosilicas for enzyme immobilization

This work shows the ability of a hierarchical porous silica-based network with pore systems at two different length scales for enzyme immobilization. Two different enzymes have been selected, lysozyme, a relatively small globular enzyme, and α-L-arabinofuranosidase, a large enzyme of interest in the winemaking industry. The lysozyme immobilization on several silica supports (bimodal porous silicas denoted UVM-7 materials and conventional silica xerogels) has been studied and the loading amounts can be correlated to the open nature and accessibility of the internal surface area. Bimodal UVM-7 silicas present a very quick adsorption rate and high enzyme loading. α-L-Arabinofuranosidase has be…

Study on the effects of several operational variables on the enzymatic batch saccharification of orange solid waste

In this work, batch enzyme-aided extraction and enzymatic saccharification of blade-milled orange waste was studied. The operation variables for this process were thoroughly analysed. It was determined that batch runs with initial pH values of 5.0 and 5.2 controlled during the first hour, 50°C and 300-500r.p.m. agitation resulted in the best yields, with a limited total and partial first-order enzyme deactivation (for cellulases and polygalacturonidase, respectively). Orange peel waste (OPW) at 6.7% w/w dry solid, 0.22 filter paper units (FPU)/g DS and proportional activities of other enzymes led to over 40g/L free monosaccharides and global yields to glucose over 80%. When using 10.1% w/w …

In silico prospection of microorganisms to produce polyhydroxyalkanoate from whey: Caulobacter segnis DSM 29236 as a suitable industrial strain

15 p.-6 fig.-6 tab.

Estimation of recombinant protein production in Pichia pastoris base don a constraint-based model

[EN] A previously validated constraint based model and possibilistic MFA have been used to design a simple estimator of protein production rate in Pichia pastoris cultures. A structured model of the yeast P. pastoris metabolism is used to predict the balance of key energetic equivalents such as ATP from available measurements, mainly substrate consumption, gases exchange rates and biomass specific growth. It has been shown that ATP flux can be related to biomass growth and protein productivity specific rates by linear regression. Cross-validation has been applied for robust parameter fitting on the basis of chemostat, steady-state experimental conditions. In this way, protein estimation can…

Genome sequence of the butanol hyperproducer Clostridium saccharoperbutylacetonicum N1-4

2 p.

Heat-Treated Bifidobacterium longum CECT-7347: A Whole-Cell Postbiotic with Antioxidant, Anti-Inflammatory, and Gut-Barrier Protection Properties

Non-viable preparations of probiotics, as whole-cell postbiotics, attract increasing interest because of their intrinsic technological stability, and their functional properties, such as immune system modulation, gut barrier maintenance, and protection against pathogens. However, reports on Bifidobacteria-derived postbiotics remain scarce. This study aims to demonstrate the functional properties of a heat-treated (HT), non-viable, Bifidobacterium longum strain, CECT-7347, a strain previously selected for its anti-inflammatory phenotype and ability to improve biomarkers of intestinal integrity in clinical trials. The study used the nematode Caenorhabditis elegans and HT-29 cell cultures as e…

Novel biocatalysts for glycerol conversion into 2,3-butanediol

Abstract Bioconversion of biodiesel-derived waste glycerol into high-value products is proposed as a solution to improve economic viability of biorefineries. Several microorganisms are able to metabolize glycerol into 2,3-butanediol (2,3-BD), a promising bulk chemical with wide applications: solvent, fuel additive, and feedstock for synthetic rubber production, among them. In the present work, a wide screening of microorganisms present both into the waste water treatment system in a biodiesel industry and in culture collections was carried out in order to evaluate their potential as new 2,3-BD producer biocatalysts. Two microorganisms for 2,3-BD production from glycerol as sole carbon sourc…

New waves underneath the purple strain.

Summary Successful merging of chemical and biotechnological operations is essential to achieve cost‐efficient industrialization of bio‐based processes. The demonstration of the use of syngas, derived from microwave assisted pyrolysis of municipal solid waste, for the improved growth and poly‐3‐hydroxybutyrate production in Rhodospirillium rubrum, stands out as an example of the synergistic contribution of chemical engineering and applied microbiology to sustainable biomaterial manufacturing, paving the way to similar applications for other syngas derived bioproducts.

Genome Sequence of the Methanotrophic Poly-β-Hydroxybutyrate Producer Methylocystis parvus OBBP

-- PAGS 2 5709-5710