Flexible Material Formulations for 3D Printing of Porous Beds with Applications in Bioprocess Engineering

Background: 3D printing is revolutionizing many industrial sectors and has the potential to enhance also the biotechnology and bioprocessing fields. Here, we propose a new flexible material formulation to 3D print support matrices with complex, perfectly ordered morphology and with tuneable properties to suit a range of applications in bioprocess engineering. Findings: Supports for packed-bed operations were fabricated using functional monomers as the key ingredients, enabling matrices with bespoke chemistry such as charged groups, chemical moieties for further functionalization, and hydrophobic/hydrophilic groups. Other ingredients, e.g. crosslinkers and porogens, provide the opportunity to further tune the mechanical properties of the supports and the morphology of their porous network. Through this approach, we fabricated and demonstrated the operation of Schoen gyroid columns with I) positive and negative charges for ion-exchange chromatography, II) enzyme bioreactors with immobilized trypsin to catalyse hydrolysis, and III) bacterial biofilms bioreactors for fuel desulfurization. Conclusions: This study demonstrates a simple, cost-effective and flexible fabrication of customized 3D printed supports for different biotechnology and bioengineering applications.

Liquid chromatography-mass spectrometry method for isomer separation and detection of sugars, phophorylated sugars and organic acids v1

This standard operating procedure is used to achieve effective separation of a wide range of polar metabolites found in central carbon metabolism via a hybrid liquid chromatographic method (ion-exchange chromatography and hydrophilic interaction liquid chromatography (HILIC)) using an Intrada Organic Acid column (Imtakt) coupled with triple quadrupole mass spectrometry. This method gives improved resolution while showing enhanced sensitivity for the detection of low abundance phosphorylated sugars compared with standard HILIC methods.

Polysaccharides from Basidiocarps of the Polypore Fungus Ganoderma resinaceum: Isolation and Structure

In this study, we focused on the isolation and structural characterization of polysaccharides from a basidiocarp of polypore fungus Ganoderma resinaceum. Polysaccharide fractions were obtained by successive extractions with cold water at room temperature (20 °C), hot water under reflux (100 °C), and a solution of 1 mol L−1 sodium hydroxide. The purity of all fractions was controlled mainly by Fourier transform infrared (FTIR) spectroscopy, and their composition and structure were characterized by organic elemental analysis; neutral sugar and methylation analyses by gas chromatography equipped with flame ionization detector (GC/FID) and mass spectrometry detector (GC/MS), respectively; and by correlation nuclear magnetic resonance (NMR) spectroscopy. The aqueous extracts contained two main polysaccharides identified as a branched O-2-β-d-mannosyl-(1→6)-α-d-galactan and a highly branched (1→3)(1→4)(1→6)-β-d-glucan. Mannogalactan predominated in the cold water extract, and β-d-glucan was the main product of the hot water extract. The hot water soluble fraction was further separated by preparative anion exchange chromatography into three sub-fractions; two of them were identified as branched β-d-glucans with a structure similar to the corresponding polysaccharide of the original fraction. The alkaline extract contained a linear (1→3)-α-d-glucan and a weakly branched (1→3)-β-d-glucan having terminal β-d-glucosyl residues attached to O-6 of the backbone. The insoluble part after all extractions was identified as a polysaccharide complex containing chitin and β-d-glucans.

Identification and partial purification of thermally stable peroxidase isoenzymes from seedlings of Vigna sp. (V) landrace Vn

Several soluble peroxidase isoenzymes are expressed in a landrace of Vigna sp. cultivated in the north of Cameroon (landrace called Vn in previous study) during seed germination. There are at least two cathodic peroxidases and eight major anodic peroxidases as shown by their electrophoretic migration at pH 7.4 under native conditions. These isoperoxidases are more expressed in roots than in shoots. They have different thermal stability, so that heat inactivation kinetics of crude peroxidase extracts from roots do not fit the first-order model. The slow and intermediate migrating groups of anodic isoperoxidases retains a substantial activity after ten minutes of incubation at 80°C and 85°C. An anodic isoperoxidase (named A6 in this study) shows in addition to this great thermal stability, a high activity in seedlings and is expressed both in roots and shoots. The combination of those characteristics makes this isoperoxidase a potential candidate for biotechnological applications. Three major anodic isoperoxidases, of which A6 and another thermostable isoperoxidase, were successfully separated from each other by ion exchange chromatography on DEAE-cellulose, after precipitation of total proteins by ice-cold acetone. This offers the prospect of being able to characterize these isoperoxidases individually in future studies.

Review on Microbial Sources, Production, Purification and Potential Industrial Applications of Laccases

Laccase (EC 1.10.3.2) is a multicopper blue oxidase which are involved in the oxidation of a broad range of organic substrates, including phenols, polyphenols, anilines, and even certain inorganic compounds by a one-electron transfer mechanism. Laccases are widely distributed in bacteria, fungai, insects and higher plants. There are mainly two production techniques for cultivation of laccase such as submersed fermentation and solid- state fermentation. This paper briefly discuss the effect of carbon source, effect of nitrogen source, effect of inducers, effects of surfactants, effect of agitator, influence of metal ions and use of agro-industrial waste in production medium. The paper also discussed the purification techniques such as ammonium sulphate precipitation for extraction purpose followed by dialysis and ion-exchange chromatography as well characterization techniques. Laccases are known to show application ranging from pharmaceutical industries to textile sector as well as in biosensor development.

Improvements of 210Po determination method in thermal water samples

Determination of naturally radionuclides have been known well as an important topic in environmental study in recently. One of the most toxic radioisotope in nature, a daughter product of 238U decay chain is 210Po (polonium). The improvement and optimizations methods for determination of this attractive isotope are still presenting so far. In this paper, a new improved method was elaborated for 210Po determination in thermal water sample. In the proposed method, analytical optimization of spontaneous/auto deposition does not use Teflon cup, magnetic stirring or any preparing equipment/item only normal glass and a side of square silver. In addition, the optimization was neglected with absent of purification of polonium (Liquid-liquid extraction methods/Ion exchange chromatography/Extraction chromatographic separations). The outcome of optimal procedure were simplify, less time consuming, great reduction of costs with chemical recovery >80% and could apply for any liquid environmental samples.

Caseinolytic Proteins (Clp) in the Genus Klebsiella: Special Focus on ClpK

Caseinolytic proteins (Clp), which are present in both prokaryotes and eukaryotes, play a major role in cell protein quality control and survival of bacteria in harsh environmental conditions. Recently, a member of this protein family, ClpK was identified in a pathogenic strain of Klebsiella pneumoniae which was responsible for nosocomial infections. ClpK is linked to the thermal stress survival of this pathogen. The genome wide analysis of Clp proteins in Klebsiella spp. indicates that ClpK is present in only 34% of the investigated strains. This suggests that the uptake of the clpk gene is selective and may only be taken up by a pathogen that needs to survive harsh environmental conditions. In silico analyses and molecular dynamic simulations show that ClpK is mainly α-helical and is highly dynamic. ClpK was successfully expressed and purified to homogeneity using affinity and anion exchange chromatography. Biophysical characterization of ClpK showed that it is predominantly alpha-helical, and this is in agreement with in silico analysis of the protein structure. Furthermore, the purified protein is biologically active and hydrolyses ATP in a concentration- dependent manner.

Nutritional Evaluation of Brillantaisia patula Leaves

Aim: To determine the nutritional and anti-nutritional compositions of Brillantaisia patula leaves using standard analytical methods. Place and Duration of Study: The proximate, mineral and anti-nutritional compositions were determined in the chemistry laboratory of Ekiti State University, Ado – Ekiti while the amino acid was determined at the Analytical Laboratory of Multi-Environmental Management Consultant, Lagos, Nigeria. Methodology: The proximate composition was carried out using the methods of Association of Official Chemists (AOAC) while mineral and anti-nutritional compositions were determined using standard analytical methods. Amino acid analysis was carried out through ion exchange chromatography (IEC) using the Technicon Sequential Multisample (TSM) Amino Acid Analyser. Results: The proximate composition ranged from 3.18% (crude fat) to 38.6% (carbohydrate). The major mineral constituents of the sample were: P (1061 mg/100g), K (874 mg/100g), Ca (799 mg/100g), Na (82.6 mg/100g) and Mg (24.3 mg/100g) while the minor mineral constituents were: Fe (26.9 mg/100g), Zn (7.7 mg/100g) and Mn (6.05 mg/100g). The evaluated anti-nutritional contents were: 6.71 mg/g oxalate, 5.37 % saponin, 1.0 mg/100g tannin and 4.72 mg/kg cyanide. Additional results showed that the leaves contained eighteen amino acids with values ranging from 0.504 g/100g cp (tryptophan) to 14.0 g/100g cp (glutamic acid). The value of the total essential amino acids (TEAA) with histidine was 45.6g/100g cp while the total non-essential amino acid (TNEAA) was evaluated to be 46.5 g/100g cp. Conclusion: Brillantaisia patula leaves could be utilized as a good source of essential amino acids and important mineral elements.

A Sulfuryl Group Transfer Strategy to Selectively Prepare Sulfated Steroids and Isotopically Labelled Derivatives

The treatment of common steroids: estrone, estradiol, cortisol, and pregnenolone with tributylsulfoammonium betaine (TBSAB) provides a convenient chemoselective conversion of the steroids alcohol/phenol moiety to the corresponding steroidal organosulfate. An important feature of the disclosed methodology is the millimolar scale of the reaction, and the isolation of the corresponding steroid sulfates as their biologically relevant sodium salts without the need for ion-exchange chromatography. The scope of the method was further explored in the estradiol and pregnanediol steroid systems with the bis-sulfated derivatives. Ultimately, a method to install an isotopic label, deuterium (2H) combined with estrone sulfation is a valuable tool for its mass-spectrometric quantification in biological studies.