Immobilization of α-Amylase from Anoxybacillus sp. SK3-4 on ReliZyme and Immobead Supports.

α-Amylase from Anoxybacillus sp. SK3-4 (ASKA) is a thermostable enzyme that produces a high level of maltose from starches. A truncated ASKA (TASKA) variant with improved expression and purification efficiency was characterized in an earlier study. In this work, TASKA was purified and immobilized through covalent attachment on three epoxide (ReliZyme EP403/M, Immobead IB-150P, and Immobead IB-150A) and an amino-epoxide (ReliZyme HFA403/M) activated supports. Several parameters affecting immobilization were analyzed, including the pH, temperature, and quantity (mg) of enzyme added per gram of support. The influence of the carrier surface properties, pore sizes, and lengths of spacer arms (functional groups) on biocatalyst performances were studied. Free and immobilized TASKAs were stable at pH 6.0-9.0 and active at pH 8.0. The enzyme showed optimal activity and considerable stability at 60 °C. Immobilized TASKA retained 50% of its initial activity after 5-12 cycles of reuse. Upon degradation of starches and amylose, only immobilized TASKA on ReliZyme HFA403/M has comparable hydrolytic ability with the free enzyme. To the best of our knowledge, this is the first report of an immobilization study of an α-amylase from Anoxybacillus spp. and the first report of α-amylase immobilization using ReliZyme and Immobeads as supports.

A Comparative Analysis Assessing Growth Dynamics of Locally Isolated Chlorella sorokiniana and Chlorella vulgaris for Biomass and Lipid Production with Biodiesel Potential.

Prior research has emphasized the potential of microalgae in biodiesel production, driven by their ability to replace fossil fuels. However, the significant costs associated with microalgae cultivation present a major obstacle to scaling up production. This study aims to develop an eco-friendly microalgae cultivation system by integrating carbon dioxide from flue gas emissions with an affordable photobioreactor, providing a sustainable biomass production. The research evaluates the growth performance of Chlorella sorokiniana and Chlorella vulgaris across this integrated system for biomass and lipid production. Results indicate substantial biomass yields of 1.97 and 1.84 g/L, with lipid contents of 35 % and 41 % for C. sorokiniana and C. vulgaris, respectively. The macrobubble photobioreactor demonstrates high potential for microalgae biomass and lipid production, yielding quality fatty acid methyl esters such as palmitic, linoleic and stearic. This study presents an environmentally friendly system for efficient microalgae cultivation, generating lipid-rich biomass suitable for biodiesel production.

Epidermal and fibroblast growth factors incorporated polyvinyl alcohol electrospun nanofibers as biological dressing scaffold.

In this study, single, mix, multilayer Polyvinyl alcohol (PVA) electrospun nanofibers with epidermal growth factor (EGF) and fibroblast growth factor (FGF) were fabricated and characterized as a biological wound dressing scaffolds. The biological activities of the synthesized scaffolds have been verified by in vitro and in vivo studies. The chemical composition finding showed that the identified functional units within the produced nanofibers (O-H and N-H bonds) are attributed to both growth factors (GFs) in the PVA nanofiber membranes. Electrospun nanofibers' morphological features showed long protrusion and smooth morphology without beads and sprayed with an average range of 198-286 nm fiber diameter. The fiber diameters decrement and the improvement in wettability and surface roughness were recorded after GFs incorporated within the PVA Nanofibers, which indicated potential good adoption as biological dressing scaffolds due to the identified mechanical properties (Young's modulus) in between 18 and 20 MPa. The MTT assay indicated that the growth factor release from the PVA nanofibers has stimulated cell proliferation and promoted cell viability. In the cell attachment study, the GFs incorporated PVA nanofibers stimulated cell proliferation and adhered better than the PVA control sample and presented no cytotoxic effect. The in vivo studies showed that compared to the control and single PVA-GFs nanofiber, the mix and multilayer scaffolds gave a much more wound reduction at day 7 with better wound repair at day 14-21, which indicated to enhancing tissue regeneration, thus, could be a projected as a suitable burn wound dressing scaffold.

Catalyst-Free Crosslinking Modification of Nata-de-Coco-Based Bacterial Cellulose Nanofibres Using Citric Acid for Biomedical Applications.

Bacterial cellulose (BC) has gained attention among researchers in materials science and bio-medicine due to its fascinating properties. However, BC's fibre collapse phenomenon (i.e., its inability to reabsorb water after dehydration) is one of the drawbacks that limit its potential. To overcome this, a catalyst-free thermal crosslinking reaction was employed to modify BC using citric acid (CA) without compromising its biocompatibility. FTIR, XRD, SEM/EDX, TGA, and tensile analysis were carried out to evaluate the properties of the modified BC (MBC). The results confirm the fibre crosslinking phenomenon and the improvement of some properties that could be advantageous for various applications. The modified nanofibre displayed an improved crystallinity and thermal stability with increased water absorption/swelling and tensile modulus. The MBC reported here can be used for wound dressings and tissue scaffolding.

Current Status and Potential Applications of Underexplored Prokaryotes.

Thousands of prokaryotic genera have been published, but methodological bias in the study of prokaryotes is noted. Prokaryotes that are relatively easy to isolate have been well-studied from multiple aspects. Massive quantities of experimental findings and knowledge generated from the well-known prokaryotic strains are inundating scientific publications. However, researchers may neglect or pay little attention to the uncommon prokaryotes and hard-to-cultivate microorganisms. In this review, we provide a systematic update on the discovery of underexplored culturable and unculturable prokaryotes and discuss the insights accumulated from various research efforts. Examining these neglected prokaryotes may elucidate their novelties and functions and pave the way for their industrial applications. In addition, we hope that this review will prompt the scientific community to reconsider these untapped pragmatic resources.

Effects of single and co-immobilization on the product specificity of type I pullulanase from Anoxybacillus sp. SK3-4.

Type I pullulanase from Anoxybacillus sp. SK3-4 (PulASK) is an unusual debranching enzyme that specifically hydrolyzes starch α-1,6 linkages at long branches producing oligosaccharides (≥G8), but is nonreactive against short branches; thus, incapable of producing reducing sugars (G1-G7). We report on the effects of both single and co-immobilization of PulASK on product specificity. PulASK was purified and immobilized through covalent attachment to three epoxides (ReliZyme EP403/M, Immobead IB-150P, and Immobead IB-150A) and an amino-epoxide (ReliZyme HFA403/M) activated supports. Following immobilization, all PulASK derivatives were active on both short and long branches in starch producing reducing sugars (predominantly maltotriose) and oligosaccharides (≥G8), respectively, a feature that is absent in the free enzyme. This study also demonstrated that co-immobilization of PulASK and α-amylase from Anoxybacillus sp. SK3-4 (TASKA) on ReliZyme HFA403/M significantly changed the product specificity compared to the free enzymes alone or individually immobilized enzymes. In conclusion, individual or co-immobilization caused changes in the product specificity, presumably due to changes in the enzyme binding pocket caused by the influence of carrier surface properties (hydrophobic or hydrophilic) and the lengths of the spacer arms.