Isolation, Biochemical Characterisation and Identification of Thermotolerant and Cellulolytic Paenibacillus lactis and Bacillus licheniformis.

RESEARCH BACKGROUND: Cellulose is an ingredient of waste materials that can be converted to other valuable substances. This is possible provided that the polymer molecule is degraded to smaller particles and used as a carbon source by microorganisms. Because of the frequently applied methods of pretreatment of lignocellulosic materials, the cellulases derived from thermophilic microorganisms are particularly desirable. EXPERIMENTAL APPROACH: We were looking for cellulolytic microorganisms able to grow at 50 °C and we described their morphological features and biochemical characteristics based on carboxymethyl cellulase (CMCase) activity and the API® ZYM system. The growth curves during incubation at 50 °C were examined using the BioLector® microbioreactor. RESULTS AND CONCLUSIONS: Forty bacterial strains were isolated from fermenting hay, geothermal karst spring, hot spring and geothermal pond at 50 °C. The vast majority of the bacteria were Gram-positive and rod-shaped with the maximum growth temperature of at least 50 °C. We also demonstrated a large diversity of biochemical characteristics among the microorganisms. The CMCase activity was confirmed in 27 strains. Hydrolysis capacities were significant in bacterial strains: BBLN1, BSO6, BSO10, BSO13 and BSO14, and reached 2.74, 1.62, 1.30, 1.38 and 8.02 respectively. Rapid and stable growth was observed, among others, for BBLN1, BSO10, BSO13 and BSO14. The strains fulfilled the selection conditions and were identified based on the 16S rDNA sequences. BBLN1, BSO10, BSO13 were classified as Bacillus licheniformis, whereas BSO14 as Paenibacillus lactis. NOVELTY AND SCIENTIFIC CONTRIBUTION: We described cellulolytic activity and biochemical characteristics of many bacteria isolated from hot environments. We are also the first to report the cellulolytic activity of thermotolerant P. lactis. Described strains can be a source of new thermostable cellulases, which are extremely desirable in various branches of circular bioeconomy.

Selection of Thermotolerant Corynebacterium glutamicum Strains for Organic Acid Biosynthesis.

In recent years, Corynebacterium glutamicum has been considered as producer of many valuable chemical compounds. Among them, organic acids such as l-lactic and succinic acids are the most important ones. It is known that the wild-type C. glutamicum grows well in the temperature range between 25 and 37 °C. Above 40 °C, the biomass growth usually abruptly stops; however, the bacteria remain metabolically active. High temperature affects the metabolic activity of C. glutamicum cells and can lead to changes in the composition and quantity of the fermentation products. Therefore, in a series of subsequent selection steps, we tried to obtain prototrophic strains capable of growing at 44 °C from the culture of homoserine auxotroph C. glutamicum ATCC 13287. During selection, we used complex and mineral media containing succinic and citric acids. As a result, we obtained 47 clones able to grow at elevated temperature. Moreover, the estimated optimal growth temperature for several of them was about 40 °C or higher. Under oxygen limitation conditions, C. glutamicum strains produce organic acids. Regardless of the tested clone, l-lactic acid was the main product. However, its concentration was the highest in the cultures performed at 44 °C. The elevated temperature also affected the biosynthesis of other organic acids. Compared to the parental strain, the concentration of acetic acid increased, and of succinic acid decreased in the cultures of thermotolerant strains. Strain RCG44.3 exhibited interesting properties; it was able to synthesise 27.1 g/L l-lactic acid, with production yield of 0.57 g/g, during 24 h of fermentation at 44 °C.

Designing laser-modified surface structures on titanium alloy custom medical implants using a hybrid manufacturing technology.

The hybrid technology combines an efficient material-removal process and implant surface treatment by the laser reducing time of manufacture process compared to currently used machining technologies. It also permits precise structuring of the implant material surface. Six structures of the Ti6Al4V ELI surface were designed and studied how the structure topography prepared with the hybrid technology affected the Escherichia coli adhesion to the surface and viability, as well as the growth, adhesion, and viability of human osteogenic Saos-2 cells cultured on the investigated surfaces. Results have confirmed that the microtopography of medical titanium alloy plays a beneficial role in bacterial adhesion and viability (number of bacteria found on reference surface: [5.9 ± 0.44] × 106 CFU/ml, sample no. 3: [8.8 ± 0.93] × 104 CFU/ml, and sample no. 5: [1.2 ± 0.23] × 107 CFU/ml; CFU - Colony Forming Unit). All tested structured surfaces enabled good cell attachment and proliferation of Saos-2 cells (viability of Saos-2 cells [% of control] for reference surface: 81.93%; sample no. 3: 75% and sample no. 5: 100%). Transcriptome analysis of genes commonly expressed in the process of osseointegration demonstrated that the use of hybrid technology allows designing structures that enhance osseointegration but it should be coupled with other methods of preventing bacterial growth, or with a different strategy to limit microbial colonization with the satisfactory osseointegration potential.

Advances in Chemical and Biological Methods to Identify Microorganisms-From Past to Present.

Fast detection and identification of microorganisms is a challenging and significant feature from industry to medicine. Standard approaches are known to be very time-consuming and labor-intensive (e.g., culture media and biochemical tests). Conversely, screening techniques demand a quick and low-cost grouping of bacterial/fungal isolates and current analysis call for broad reports of microorganisms, involving the application of molecular techniques (e.g., 16S ribosomal RNA gene sequencing based on polymerase chain reaction). The goal of this review is to present the past and the present methods of detection and identification of microorganisms, and to discuss their advantages and their limitations.