Hartmannjorgensen0137

© King Abdulaziz City for Science and Technology 2020.l-isoleucine dioxygenase (IDO) is an Fe (II)/α-ketoglutarate (α-KG)-dependent dioxygenase that specifically converts l-isoleucine (l-Ile) to (2S, 3R, 4S)-4-hydroxyisoleucine (4-HIL). 4-HIL is an important drug for the treatment and prevention of type 1 and type 2 diabetes but the yields using current methods are low. In this study, the CRISPR-Cas9 gene editing system was used to knockout sucAB and aceAK gene in the TCA cycle pathway of Escherichia coli (E. coli). For single-gene knockout, the whole process took approximately 7 days. However, the manipulation time was reduced by 2 days for each round of gene modification for multigene editing. Using the genome-edited recombinant strain E. coli BL21(DE3) ΔsucABΔaceAK/pET-28a(+)-ido (2Δ-ido), the bioconversion ratio of L-Ile to 4-HIL was enhanced by about 15% compared to E. coli BL21(DE3)/pET-28a(+)-ido [BL21(DE3)-ido] strain. The CRISPR-Cas9 editing strategy has the potential in modifying multiple genes more rapidly and in optimizing strains for industrial production. © King Abdulaziz City for Science and Technology 2020.The natural estrogen 17β-estradiol (17β-E2) is a major endocrine disruptor. Accordingly, due to their frequent presence in global surface waters, prolonged exposure to estrogen-contaminated water may disrupt sexual development in animals. It has adverse effects on wildlife and humans. To date, the most effective strategy for estrogen removal from the environment is biodegradation using microorganisms. To this end, we isolated a strain of Lysinibacillus sphaericus, namely DH-B01, from a contraceptive factory in Beijing. The experimental results revealed that the bacterium has a high capacity to degrade estrogen, with a 17β-E2 degradation rate of about 97%, and produces the secondary metabolite estrone. In addition, a series of genes involved in steroid metabolism and stress response in L. sphaericus sp. DH-B01 were predicted, and several key genes with high similarity to those of other strains were subjected to sequence alignment to find their conserved regions. This is the first study of the ability of L. sphent L. sphaericus strains. The differences play an important role and further enrich the functionality and diversity of L. sphaericus strains. In subsequent studies, the specificity of L. sphaericus sp. DH-B01 can be applied to different environments for future environmental restoration. © King Abdulaziz City for Science and Technology 2020.Boerhavia diffusa (BD) Linn. (Nyctaginaceae) is one of the most commonly used herbs in the Indian traditional system of medicine for the urinary disorders. The aim of the current investigation was to carry out initiation, development, and maintenance of BD callus cultures and quantitative estimation of punarnavine in plant and callus extracts. Leaves and stem of BD were used as explant for the tissue culture studies using Murashige and Skoog (MS) basal medium. MS Media comprising 2,4-Dichlorophenoxy acetic acid (2,4-D) (1 ppm) and 2,4-D (1 ppm) + Indole-3-acetic acid (IAA) (1.0 ppm) were found to yield friable callus from leaf explant; similarly, 2,4-D (0.3 ppm) + IAA (0.75 ppm) + Kinetin (0.3 ppm) and 2,4-D (0.5 ppm) + Naphthalene acetic acid (NAA) (1.5 ppm) + Kinetin (0.3 ppm) were found to yield friable callus from the stem explant. High-performance thin-layer chromatography method was been developed for the quantitative estimation of punarnavine (R f = 0.73) using mobile phase containing toluene ethyl acetate formic acid in the ratio (7.02.50.7, v/v/v) at 262 nm. The validated method was found linear (r 2 = 0.9971) in a wide range (100-1000 ng spot-1), precise, accurate, and robust. The values of limit of detection, LOD = 30.3 ng spot-1, and limit of quantification, LOQ = 100.0 ng spot-1. The robustness of the method was proved by applying the Box-Behnken design (BBD). The developed method found appropriate for the quality control of medicinal plants containing punarnavine as a constituent. © King Abdulaziz City for Science and Technology 2020.Phosphate-solubilizing bacteria (PSB) have been widely used as biological fertilizer. However, its impact on the local microbial community has less been known. In this study, a mixture of PSB was inoculated into the tomato growth alone or combined with manure fertilizer. The growth parameter results showed that the combination use of PSB and compost could significantly increase the tomato growth and yield. The use of PSB could significantly increase pH, available phosphorus and several kinds of trace elements both in the rhizosphere and non-rhizosphere soil. The quantitative PCR and high-throughput sequencing results showed that the inoculated PSB did not become the dominant strains in the rhizosphere. However, the soil bacterial community structure was changed. The relative abundance of several indigenous bacteria, such as Pseudomonas, decreased, while the population of several bacteria, including Bacillus, Anaerolineaceae, Cytophagaceae, and Gemmationadaceae, increased. The redundancy analysis result showed that the soil properties had a great influence on the indigenous microbial community. In conclusion, the inoculated PSB could not colonize in the soil with a single inoculation. The PSB secreted small molecular organic acids to dissolve inorganic phosphorus and changed the soil properties, which changed the rhizosphere microbial community indirectly. BAY 1217389 inhibitor © King Abdulaziz City for Science and Technology 2020.Liposomes are very useful biocompatible tools used in diverse scientific disciplines, employed for the vehiculation and delivery of lipophilic, ampiphilic or hydrophilic compounds. Liposomes have gained the importance as drug carriers, as the drugs alone have limited targets, higher toxicity and develop resistance when used in higher doses. Conventional liposomes suffer from several drawbacks like encapsulation inefficiencies and partially controlled particle size. The surface chemistry of liposome technology started from simple conventional vesicles to second generation liposomes by modulating their lipid composition and surface with different ligands. Introduction of polyethylene glycol to lipid anchor was the first innovative strategy which increased circulation time, delayed clearance and opsonin resistance. PEGylated liposomes have been found to possess higher drug loading capacity up to 90% or more and some drugs like CPX-1 encapsuled in such liposomes have increased the disease control up to 73% patients suffering from colorectal cancer.