Graphical Abstract and Lay Summary In our work, we have developed a simple approach to increase the expression level of recombinant proteins in E.coli. The developed method is based on the selection of the most optimal clone from a synthetic library of gene variants with synonymous substitutions in the first 11 codons. To select the clone with the highest expression level, the gene was fused with the sfGFP reporter protein, which allowed us to evaluate the expression level of the final gene::sfGFP constructs by the level of fluorescence of colonies on an agarized Petri dish. This method is sim ..read more

Graphical Abstract and Lay Summary CRISPR/Cas9 technology combined with somatic cell nuclear transfer (SCNT) provides the primary method for producing gene-edited pigs, with high-quality gene-edited nuclear donors closely associated with the forms of CRISPR/Cas9. The authors developed a platform based on fluorescence reporting signals and micropattern arrays to visually assess the efficiency of gene editing. It was efficiently determined that mRNA form targeting porcine cells had the highest editing efficiency. This platform also holds great potential for rapidly testing new gene editing tools ..read more

Graphical Abstract and Lay Summary Bone marrow mesenchymal stem cells (BMSCs) respond to external fluid shear stress (FSS) and hydrostatic pressure (HP) stimuli via the integrin-cytoskeleton and Piezo1 ion channels, respectively. This activation leads to downstream signaling involving transcription factors YAP1 and NFAT2. The activated YAP1 and NFAT2 proteins exhibit binding capabilities, potentially synergistically enhancing the osteogenic differentiation of BMSCs. This research contributes to a deeper comprehension of the crosstalk relationship between mechanical stimuli in cellular response ..read more

Graphical Abstract and Lay Summary Single-stranded DNA (ssDNA) is the foundation of synthetic biology, which promotes the third biotechnological revolution, while synthesizing ssDNA with high efficiency and low error rate remains major challenges. Six representative achievements and approaches were concluded and compared, including solid-phase oligonucleotide synthesis, terminal deoxynucleotidyl transferase-based ssDNA synthesis, reverse transcription, primer exchange reaction, asymmetric polymerase chain reaction, and rolling circle amplification. Envisioning comprehensive viewpoints on ssDNA ..read more

Graphical Abstract and Lay Summary Polyurethane is a polymer with a complex structure that contains urethane bonds as its repeating unit structure. The widespread use of PU products has led to a significant amount of plastic waste. Enzymatic depolymerization is a sustainable method for treating PU polymers, although there are few reported enzymes capable of hydrolyzing urethane bonds in PU. In this study, we introduced a combination of amidase GatA250 and cutinase LCC, which hydrolyze urethane bonds and ester bonds in PU, respectively. This approach promotes the complete depolymerization of PU ..read more

Graphical Abstract and Lay Summary Over the last years, utilization of enzymes in degradation of polymers has emerged as an environmentally friendly asset in tackling the pressing challenges of plastic pollution. In this study, the investigation of four phylogenetically distinct bacterial polyesterases took place. The enzymes, which were biochemically characterized and structurally analyzed, demonstrated degradation ability of various synthetic plastics. Our results point out the wide substrate spectrum of bacterial hydrolases and underscore the significant potential of PETase-like enzymes in ..read more

Graphical Abstract and Lay Summary Profiling the engineered strain with systems biology can help to understand the cellular metabolism and identify the potential bottleneck in optimizing the cell factory. In this study, combined metabolic analysis and proteomic analysis of targeted mevovanate (MVA) pathway showed that coordination of cytosolic and mitochondrial pathways had balanced metabolism, and genome integration of biosynthetic genes had less MVA enzymes and higher sabinene production. Comparative proteome analysis showed that compartmentalized mitochondria pathway had perturbations on ce ..read more

Graphical Abstract and Lay Summary Halogenase is an environmentally friendly method for the synthesis of halogenated products. A triple enzyme fusion of tryptophan-6-halogenase SpFDH was constructed to achieve a self-sufficient bi-coenzyme cycle system and applied to the continuous halogenation of tryptophan. This study provides a potential basis for the industrial application of flavin-dependent halogenase ABSTRACT The halogenase-based catalysis is one of the most environmentally friendly methods for the synthesis of halogenated products, among which flavin-dependent halogenases (FDHs) have ..read more

Biotechnology Journal, Volume 19, Issue 4, April 2024 ..read more

Graphical Abstract and Lay Summary This work demonstrated a novel concept of an artificial and stable two-component protein system able to sustain a good NADPH regeneration rate from hydrogen. The system proposed here is the first of its kind, based on a highly active and oxygen-resistant [FeFe]-hydrogenase and a reductase (BMR) as non-physiological partners. As a proof-of-concept, a cascade reaction involving the [FeFe]-hydrogenase, BMR and a mutant Baeyer-Villiger monooxygenase is presented, showing how the cascade can be exploited for indigo production in multiple reaction cycles. Abstract ..read more