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Bacillus licheniformis is recognised as an exceptional expression platform in biomanufacturing due to its ability to produce high-value products. Consequently, metabolic engineering of B. licheniformis is increasingly pursued to enhance its utility as a biomanufacturing vehicle. Effective B. licheniformis cell factories require promoters that enable regulated expression of target genes. This review discusses recent advancements in the characterisation, synthesis, and engineering of B. licheniformis promoters. We highlight the application of constitutive promoters, quorum sensing promoters, and inducible promoters in protein and chemical synthesis. Additionally, we summarise efforts to expand the promoter toolbox through hybrid promoter engineering, transcription factor-based inducible promoter engineering, and ribosome binding site (RBS) engineering. [ABSTRACT FROM AUTHOR]

Bacillus licheniformis is a significant industrial microorganism. Traditional gene editing techniques relying on homologous recombination often exhibit low efficiency due to their reliance on resistance genes. Additionally, the established CRISPR gene editing technology, utilizing Cas9 endonuclease, faces challenges in achieving simultaneous knockout of multiple genes. To address this limitation, the CRISPR-Cpf1 system has been developed, enabling multiplexed gene editing across various microorganisms. Key to the efficient gene editing capability of this system is the rigorous screening of highly effective expression elements to achieve conditional expression of protein Cpf1. In this study, we employed mCherry as a reporter gene and harnessed P mal for regulating the expression of Cpf1 to establish the CRISPR-Cpf1 gene editing system in Bacillus licheniformis . Our system achieved a 100 % knockout efficiency for the single gene vpr and up to 80 % for simultaneous knockout of the double genes epr and mpr . Furthermore, the culture of a series of protease-deficient strains revealed that the protease encoded by aprE contributed significantly to extracellular enzyme activity (approximately 80 %), whereas proteases encoded by vpr , epr , and mpr genes contributed to a smaller proportion of extracellular enzyme activity. These findings provide support for effective molecular modification and metabolic regulation in industrial organisms., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors.)

Background: Robot-assisted technology has been widely used in orthopedic surgery, which can provide surgeons with higher accuracy and reduce radiation exposure. In spinal surgery, robots are often used to assist pedicle screw implantation, while there are relatively few studies on robot-assisted percutaneous kyphoplasty (PKP) under local anesthesia., Methods: A total of 96 patients with single-segment OVCF who met the inclusion criteria were included in this study. Fifty-six patients underwent robot-assisted PKP and forty patients underwent conventional PKP by the same group of surgeons. Collect the relevant parameters., Results: The puncture time and fluoroscopy times during puncture in the robot group were significantly less than those in the manual group ( P  < 0.001). The success rate of first puncture in the robot group was 92.5%., Conclusions: PKP under local anesthesia assisted by the new spinal surgical robot effectively reduces the patient's intraoperative discomfort and has a low learning curve., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2024 Xue, Liu, Li, Xiao, Zhu, Wu and Zhang.)