Specific tracking of xylan using fluorescent-tagged carbohydrate-binding module 15 as molecular probe

Additional file 5: Figure S3. Low-resolution XPS spectrum of UBKP surface. UBKP: unbleached kraft pulp. Unextracted pulp samples were analysed

Rapid and efficient colony-PCR for high throughput screening of genetically transformed chlamydomonas reinhardtii

Microalgae biotechnologies are rapidly developing into new commercial settings. Several high value products already exist on the market, and biotechnological development is focused on genetic engineering of microalgae to open up future economic opportunities for food, fuel and pharmacological production. Colony-polymerase chain reaction (colony-PCR or cPCR) is a critical method for screening genetically transformed microalgae cells. However, the ability to rapidly screen thousands of transformants using the current colony-PCR method, becomes a very laborious and time-consuming process. Herein, the non-homologous transformation of Chlamydomonas reinhardtii using the electroporation and glass beads methods generated more than seven thousand transformants. In order to manage this impressive number of clones efficiently, we developed a high-throughput screening (HTS) cPCR method to rapidly maximize the detection and selection of positively transformed clones. For this, we optimized the Chlamydomonas transformed cell layout on the culture media to improve genomic DNA extraction and cPCR in 96-well plate. The application of this optimized HTS cPCR method offers a rapid, less expensive and reliable method for the detection and selection of microalgae transformants. Our method, which saves up to 80% of the experimental time, holds promise for evaluating genetically transformed cells and selection for microalgae-based biotechnological applications such as synthetic biology and metabolic engineering

Red light variation an effective alternative to regulate biomass and lipid profiles in Phaeodactylum tricornutum

Abstract: Marine water diatom Phaeodactylum tricornutum is a photosynthetic organism that is known to respond to the changing light environment and adapt to different temperatures to prevent photoinhibition and maintain its metabolic functions. The objective of the present study was to test whether light shift variations in different growth phases impact the growth and lipid metabolism of P. tricornutum. Thus, we investigated R exposure in different growth phases to find the most effective light shift condition. The results showed that substituting white light (W) by red light (R) under autotrophic conditions, a condition called red shift (RS), increased biomass and lipid content compared to levels found under continuous W or R exposure alone. We observed an increase by 2-fold biomass and 2.3-fold lipid content in RS as compared to W. No significant change was observed in the morphology of lipid droplets, but the fatty acid (FA) composition was altered. Specifically, polyunsaturated FAs were increased, whereas monounsaturated FAs decreased in P. tricornutum grown in RS compared to W control. Therefore, we propose that a light shift during the beginning of the stationary phase is a low-cost cultivation strategy to boost the total biomass and lipids in P. tricornutum

Genome Editing by CRISPR-Cas: A Game Change in the Genetic Manipulation of Chlamydomonas

Microalgae are promising photosynthetic unicellular eukaryotes among the most abundant on the planet and are considered as alternative sustainable resources for various industrial applications. Chlamydomonas is an emerging model for microalgae to be manipulated by multiple biotechnological tools in order to produce high-value bioproducts such as biofuels, bioactive peptides, pigments, nutraceuticals, and medicines. Specifically, Chlamydomonas reinhardtii has become a subject of different genetic-editing techniques adapted to modulate the production of microalgal metabolites. The main nuclear genome-editing tools available today include zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases (TALENs), and more recently discovered the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein (Cas) nuclease system. The latter, shown to have an interesting editing capacity, has become an essential tool for genome editing. In this review, we highlight the available literature on the methods and the applications of CRISPR-Cas for C. reinhardtii genetic engineering, including recent transformation methods, most used bioinformatic tools, best strategies for the expression of Cas protein and sgRNA, the CRISPR-Cas mediated gene knock-in/knock-out strategies, and finally the literature related to CRISPR expression and modification approaches

MOESM1 of Specific tracking of xylan using fluorescent-tagged carbohydrate-binding module 15 as molecular probe

Additional file 1: Figure S1. SDS-PAGE analysis of the OC15 probe purified by affinity chromatography. The expected molecular weight of the OC15 fusion protein is 44.68 kDa. A 12 % polyacrylamide gel was used for SDS-PAGE analysis. Well M: Precision plus protein standards (5 µg). Well OC15: Purified OC15 probe (10 µg)

Legislative Documents

Also, variously referred to as: Senate bills; Senate documents; Senate legislative documents; legislative documents; and General Court documents

MOESM6 of Determination of optimal biomass pretreatment strategies for biofuel production: investigation of relationships between surface-exposed polysaccharides and their enzymatic conversion using carbohydrate-binding modules

Additional file 6. Standard curves for the conversion of fluorescence intensities into µg of probes. A) GC3a, B) CC17, C) OC15 and D) CC27 probes

MOESM4 of Determination of optimal biomass pretreatment strategies for biofuel production: investigation of relationships between surface-exposed polysaccharides and their enzymatic conversion using carbohydrate-binding modules

Additional file 4. Affinity gel electrophoresis (AGE) of the probes. A) CC17, B) OC15 and C) CC27 probes. Panel a: control (no polysaccharide); Panel b: CMC; Panel c: xylan; Panel d: galactomannan. In each panel the first well contained BSA as a negative control (10 µg) and the second well was loaded with an appropriate probe (10 µg). All soluble polysaccharides were used at final concentration of 0.5% (w/v) and a 12% polyacrylamide gel was used for affinity analysis

Effect of a low melting temperature mixture on the surface properties of lignocellulosic flax bast fibers

Bast flax fibers were treated, with or without ultrasound assistance, using a low melting mixture (LMM) composed of lactic acid, d-glucose and water. This LMM treatment affected both lignin and hemicelluloses contents and modified the fibers properties identified as crucial parameters in an industrial context, i.e. coloration, wettability, crystallinity, fibers diameter and chemical composition. Surface chemistry of the fibers were investigated through fluorescent tagged carbohydrates binding modules revealing macromolecular rearrangements responsible of both a fibers crystallinity enhancement and an unexpected hydrophobicity. It has been found that LMM treatments bleach fibers, which is considered a beneficial effect independent of the treatments