Improving AOX1 promoter efficiency by overexpression of Mit1 transcription factor.

BACKGROUND: Reprogramming in transcriptional regulation provides an effective tool for adjusting cellular metabolic activities. The strong methanol-inducible alcohol oxidase-1 promoter (pAOX1) is commonly used for heterologous gene expression in the yeast Pichia pastoris. Here, we present a novel Pichia pastoris strain engineered to co-express methanol-induced transcription factor 1 (Mit1) and the target protein. Mit1 upregulates pAOX1 in response to methanol. METHODS AND RESULTS: Two model proteins (VEGF and eGFP) have been used as the target proteins under the control of pAOX1. The sequence of Mit1 had obtained from the yeast genome and likewise cloned under the control of pAOX1. The results indicated a 1.9 and 2.2 fold increase in the detected VEGF and eGFP, respectively, when co-expressed with Mit1. Furthermore, the double-recombinant cells, containing Mit-1 and eGFP, produced 1.3 fold more eGFP when the methanol feeding concentration was doubled. The real-time PCR indicated a slight increase in the Mit1 expression, probably due to the negative regulatory feedback loop that exists for the intrinsic yeast Mit1. Overexpression of Mit1 also led to duplication of AOX1 enzyme activity, which may enhance the yeast cells' capacity for methanol detoxification. CONCLUSION: Overexpression of Mit1 could be considered a promising strategy for upregulation of target recombinant proteins in Pichia pastoris. Intracellular overexpression of Mit1 upregulates the heterologous target gene (eGFP) production, which is expressed under the control of pAOX1.

Niosomal formulation for antibacterial applications.

Infection is a disease that is mainly caused by different Gram-negative and Gram-positive bacteria. Treatment of infections requires a considerable amount of antibiotics, which can cause serious damage to the patient's body. Delivering the antibiotic only to the site of infection can prevent these destructive effects, such as the destruction of the normal intestinal flora. The drug delivery system through carriers will take antibiotics into a part of the body involved in the disease. Niosome nanoparticles, which have been made from non-ionic surfactants, have been emerging as ideal drug/antibiotics delivery vehicles. Recently, niosome formulations have been targeted to reduce toxicity and increase accumulation at the target site. Niosomes have performed well in the treatment of local infections, delivery of ocular drugs, and coating of orthopaedic bone/dental implants. This research aimed to highlight the molecular structure and physicochemical properties of niosomes and covered its manufacturing methodologies. Then we critically review the literature on niosomes for the mechanism of drug release, the carrier to deliver antibiotics, and its clinical effectiveness against bacterial infections.