Bac 5

Bacilysin is a dipeptide antibiotic composed of L-alanine and L-anticapsin produced by certain strains of B. subtilis. Bacilysin is gaining increasing attention in industrial agriculture and pharmaceutical industries due to its potent antagonistic effects on various bacterial, fungal, and algal pathogens. However, its use in industrial applications is hindered by its low production in the native producer. The biosynthesis of bacilysin is mainly based on the bacABCDEF operon. Examination of the sequence surrounding the upstream of the bac operon did not reveal a clear, strong ribosome binding site (RBS). Therefore, in this study, we aimed to investigate the impact of RBS as a potential route to improve bacilysin production. For this, the 5' untranslated region (5'UTR) of the bac operon was edited using the CRISPR/Cas9 approach by introducing a strong ribosome binding sequence carrying the canonical Shine-Dalgarno sequence (TAAGGAGG) with an 8 nt spacing from the AUG start codon. Strong RBS substitution resulted in a 2.87-fold increase in bacilysin production without affecting growth. Strong RBS substitution also improved the mRNA stability of the bac operon. All these data revealed that extensive RBS engineering is a promising key option for enhancing bacilysin production in its native producers.

Ultrasonic treatment has been shown to have a favorable effect on the regeneration of spent biological activated carbon (BAC) from drinking water treatment plants. In this study, the use of ultrasound as a regeneration method had a significant effect on the recovery of spent BAC after 7.5 years of use; it effectively increased the iodine value from 300 mg/g to 600 mg/g and restored the specific surface area and pore volume of BAC. Ultrasound effectively changed the structure of the biofilm inside and on the surfaces of BAC particles, on the basis of confocal laser scanning microscopy (CLSM) images. The thickness of the surface biofilm attached to BAC reached an "active" level (about 100 μm) at the regeneration frequency of 40 kHz. The dehydrogenase activity significantly improved from 4.50 mg TF/g BAC to 9.13 mg TF/g BAC, and the content of adenosine-triphosphate (ATP) in regenerated BAC was maintained at a high level (2.501 × 10-6g ATP/g BAC), thus allowing the development of microbial growth. The production of soluble microbial products (SMPs) from regenerated BAC decreased during the reuse process. The removal efficiency of DOC, CODMn, NH4+ and NO3- control increased by approximately 78%, 71%, 50% and 20%, respectively.

Nectin-like molecule-5 (Necl-5) is an immunoglobulin-like molecule that was originally identified as a poliovirus receptor and is often upregulated in cancer cells. It has been said that Necl-5 plays a role in not only cell-cell adhesion, but also cell migration, proliferation, and metastasis. In this study, we used a bronchioloalveolar carcinoma (BAC) cell line and fibroblasts to assess the expression of Necl-5 in the development of cancer-stroma communication by using an easy-to-prepare double-layered collagen gel hemisphere (DL-CGH) system that enables visualization of cell migration during invasion. The expression of Necl-5 was higher in BAC cells than in fibroblasts. This tendency didn't change even when the BAC cells were mixed with fibroblasts. To assess the role of Necl-5 in the invasive activity of the BAC cells, we knocked down its expression using RNA interference (RNAi). The invasion assay with DL-CGH revealed that inhibition of Necl-5 expression in the BAC cells was associated with suppressed invasiveness. In addition, Necl-5 knockdown inhibited the movement and proliferation of the BAC cells. Necl-5 expression in lung cancer cells is crucial for their invasiveness in the cancer-stromal interaction, suggesting that Necl-5 could be a favorable molecular target for the suppression of invasiveness in lung adenocarcinoma.