High-throughput strategies for the discovery and engineering of enzymes for biocatalysis.

Innovations in novel enzyme discoveries impact upon a wide range of industries for which biocatalysis and biotransformations represent a great challenge, i.e., food industry, polymers and chemical industry. Key tools and technologies, such as bioinformatics tools to guide mutant library design, molecular biology tools to create mutants library, microfluidics/microplates, parallel miniscale bioreactors and mass spectrometry technologies to create high-throughput screening methods and experimental design tools for screening and optimization, allow to evolve the discovery, development and implementation of enzymes and whole cells in (bio)processes. These technological innovations are also accompanied by the development and implementation of clean and sustainable integrated processes to meet the growing needs of chemical, pharmaceutical, environmental and biorefinery industries. This review gives an overview of the benefits of high-throughput screening approach from the discovery and engineering of biocatalysts to cell culture for optimizing their production in integrated processes and their extraction/purification.

Feasibility study of date (Phoenix dactylifera L.) fruit syrup-based natural jelly using central composite design.

A feasibility study of natural fruit jelly from three Algerian raw materials, namely date (Phoenix dactylifera l.) fruit syrup and suspension of orange albedo powder (OAP) in lemon juice (LJ) was performed by response surface methodology (RSM) based on central composite design (CCD). The textural properties of the final jelly were investigated through two dependent variables: hardness and stickiness. The cooking temperature (X1), corresponding to that of thermo stated oil bath, and cooking time (X 2), taken for heating the initial fruit mixture in the oil bath (from ambient temperature without fixing however, the final temperature), were found to be the most influent factors, compared to °Brix of date syrup (X3) and temperature (X4) of the cooling stage following the cooking process. Results have also shown that the second-degree polynomial models correctly fit experimental data (R(2), adjusted R(2) (R(2) adj) and cross-validation (Q(2)) ≈ 1). Considering textural properties of commercial jellies as a reference, it was found that the cooking temperature of 155 °C for 10 min gave a jelly with suitable textural properties. On the other hand, FT-IR spectra revealed that the structure of such jelly was partially close to that of pectin molecules. Finally, the color analysis in the CIELab system of the fruit mixture over the cooking process showed that both lightness (L(*)) and a*/b* ratio were not affected by the experienced temperature range (80-155 °C).

Efficient size control of amphiphilic cyclodextrin nanoparticles through a statistical mixture design methodology.

PURPOSE: the aim of the study was to investigate size control of amphiphilic beta-cyclodextrin nanoparticles obtained by solvent displacement technique. METHODS: An experimental design methodology for mixture design was undertaken using D-optimal approach with the following technique variables: water fraction X1 (40-70% v/v), acetone fraction X2 (0-60% v/v) and ethanol fraction X3 (0-60% v/v). RESULTS: The resulting quadratic model obtained after logarithmic transformation of data and partial least-square regression was statistically validated and experimentally checked. Also, the morphology of the colloidal nanoparticles from selected experiments was observed by cryo-transmission electron microscopy. CONCLUSIONS: This experimental design approach allowed to produce interesting amphiphilic beta-cyclodextrin nanoparticles with a predicted mean size varying from 60 to 400 nm.