RESUMEN
Vitamin B2 is an essential water-soluble vitamin. For most prokaryotes, a bifunctional enzyme called FAD synthase catalyzes the successive conversion of riboflavin to FMN and FAD. In this study, the plasmid pNEW-AZ containing six key genes for the riboflavin synthesis was transformed into strain R2 with the deleted FMN riboswitch, yielding strain R5. The R5 strain could produce 540.23 ± 5.40 mg/L riboflavin, which was 10.61 % higher than the R4 strain containing plasmids pET-AE and pAC-Z harboring six key genes. To further enhance the production of riboflavin, homology matching and molecular docking were performed to identify key amino acid residues of FAD synthase. Nine point mutation sites were identified. By comparing riboflavin kinase activity, mutations of T203D and N210D, which respectively decreased by 29.90 % and 89.32 % compared to wild-type FAD synthase, were selected for CRISPR/Cas9 gene editing of the genome, generating engineered strains R203 and R210. pNEW-AZ was transformed into R203, generating R6. R6 produced 657.38 ± 47.48 mg/L riboflavin, a 21.69 % increase compared to R5. This study contributes to the high production of riboflavin in recombinant E. coli BL21.
RESUMEN
The wide abuse of antibiotics has accelerated bacterial multiresistance, which means there is a need to develop tools for rapid detection and characterization of bacterial response to antibiotics in the management of infections. In the study, an electrochemical biosensor based on nanoporous alumina membrane and graphene quantum dots (GQDs) was developed for bacterial response to antibiotics detection. Anti-Salmonella antibody was conjugated with amino-modified GQDs by glutaraldehyde and immobilized on silanized nanoporous alumina membranes for Salmonella bacteria capture. The impedance signals across nanoporous membranes could monitor the capture of bacteria on nanoporous membranes as well as bacterial response to antibiotics. This nanoporous membrane and GQD-based electrochemical biosensor achieved rapid detection of bacterial response to antibiotics within 30 min, and the detection limit could reach the pM level. It was capable of investigating the response of bacteria exposed to antibiotics much more rapidly and conveniently than traditional tools. The capability of studying the dynamic effects of antibiotics on bacteria has potential applications in the field of monitoring disease therapy, detecting comprehensive food safety hazards and even life in hostile environment.
RESUMEN
We propose a general geometric definition of the V-parameter in photonic crystal fibers (PCF). We argue that the fundamental modal field in both square-lattice and triangular-lattice PCFs can be approached by certain proper assemblies of the fundamental mode in a one-dimensional planar waveguide. This observation suggests a geometric definition of the effective core radius and the single-mode regime of V≤π/2 in various PCFs. The numerical results imply that the suggestion is valid for all of the one-rod, seven-rod, 19-rod, and 37-rod core triangular-lattice PCFs and the one-rod, nine-rod, and 25-rod core square-lattice PCFs.
RESUMEN
A general model is proposed to describe thermal-induced mode distortion in the step-index fiber (SIF) high power lasers. Two normalized parameters in the model are able to determine the mode characteristic in the heated SIFs completely. Shrinking of the mode fields and excitation of the high-order modes by the thermal-optic effect are investigated. A simplified power amplification model is used to describe the output power redistribution under various guiding modes. The results suggest that fiber with large mode area is more sensitive on the thermally induced mode distortion and hence is disadvantaged in keeping the beam quality in high power operation. The model is further applied to improve the power scaling analysis of Yb-doped fiber lasers. Here the thermal effect is considered to couple with the optical damage and the stimulated Raman scattering dynamically, whereas direct constraint from the thermal lens is relaxed. The resulting maximal output power is from 67kW to 97kW, depending on power fraction of the fundamental mode.