Ahmad Ziad Sulaiman

Ultrasound mediated enzymatic hydrolysis of cellulose and carboxymethyl cellulose.

A recombinant Trichoderma reesei cellulase was used for the ultrasound‐mediated hydrolysis of soluble carboxymethyl cellulose (CMC) and insoluble cellulose of various particle sizes. The hydrolysis was carried out at low intensity sonication (2.4–11.8 W cm−2 sonication power at the tip of the sonotrode) using 10, 20, and 40% duty cycles. [A duty cycle of 10%, for example, was obtained by sonicating for 1 s followed by a rest period (no sonication) of 9 s.] The reaction pH and temperature were always 4.8 and 50°C, respectively. In all cases, sonication enhanced the rate of hydrolysis relative to nonsonicated controls. The hydrolysis of CMC was characterized by Michaelis‐Menten kinetics. The Michaelis‐Menten parameter of the maximum reaction rate Vmax was enhanced by sonication relative to controls, but the value of the saturation constant Km was reduced. The optimal sonication conditions were found to be a 10% duty cycle and a power intensity of 11.8 W cm−2. Under these conditions, the maximum rate of hydrolysis of soluble CMC was nearly double relative to control. In the hydrolysis of cellulose, an increasing particle size reduced the rate of hydrolysis. At any fixed particle size, sonication at a 10% duty cycle and 11.8 W cm−2 power intensity improved the rate of hydrolysis relative to control. Under the above mentioned optimal sonication conditions, the enzyme lost about 20% of its initial activity in 20 min. Sonication was useful in accelerating the enzyme catalyzed saccharification of cellulose.

Effect of flow rate, duty cycle, amplitude, and treatment Time of ultrasonic regimens towards Escherichia coli harbouring lipase

A full factorial design (FFD) approach was conducted to assess the effect of four factors, namely flow rate, duty cycle, amplitude, and treatment time of ultrasonic regimens towards Escherichia coli harbouring lipase. The 22 experiments were performed as the following values with six replicates of centre point: flow rate (0.1, 0.2, and 0.3 L/min), duty cycle (0, 20, and 40 ), amplitude (2, 6, and 10), and treatment time (10, 35, and 60 min). The FFD was employed as preliminary screening in shake flask cultivation to choose the significant factors (P< 0.05) for further optimisation process. In this study, zero duty cycle signified non-sonication of amplitude and no treatment time effect to the E. coli culture. Also, the designated flow rate and amplitude accordingly showed no effect towards the amount of dry cells weight (DCW). DCW1 was found significantly degraded after the exposure of high duty cycle and treatment time as other factors remained constant. Whereas for the lipase activity, no significant difference was observed in any main factors or interactions. Paired samples t-test confirms the result at a p-value of 0.625. This experimental study suggests the direct and continuous approach of sonication caused an adverse effect on the cells culture density.

Extractive fermentation for enhanced productivity of bio-ethanol by Zymomonas anaerobia

Plant materials. As initial explants were used shoot tips and mature seeds of Malus domestica Borkh. cv Golden Delicious, cv Starking. Disinfection of isolated buds. Isolated buds were treated with Bavistine 0.2% for 7 min, followed by a treatment with HgCl2 0.1% for 7 min and with ethanol 70% for a min. After every treatment on laminar flow, the buds were rinsed three times with sterile and distilled water. Nutrient medium and methods to avoid polyphenolic oxidation. For both types of explants was used MS nutrient medium supplemented with MS vitamins and combined with cytokinin BAP (1 mg l-1) and auxin IBA (0.1 mg l-1); sucrose 30 g l-1 and also agar 7%; the pH has been checked to be 5.8. Bud explants were treated by some methods in order to avoid the polyphenolic oxidation of the explants: Methods: i) MS + ascorbic acid 0.2%, 48 hours in the darkness and twice transfers in fresh medium, ii) MS + ascorbic acid 0.2% and 72 hours in the darkness, iii) MS + polyvinyl pyrrolidone (PVP) 0.1%, ascorbic acid 0.1%, citric acid 0.1%, 48 hours in darkness and some times of transfers in fresh medium, iv) MS + active charcoal 1g l-1 and 24 hours in darkness, v) MS + ascorbic acid 0.1%, citric acid 0.05%, 48 hours in darkness and twice transfers in fresh medium, vi) MS medium with ascorbic acid 0.1%, citric acid 0.05% and active charcoal 1g l-1, 2 days in darkness and onetime transfer to fresh medium. Conditions of in vitro culture chamber. The test tubes are placed in the culture chamber with controlled physical conditions (temperature: 23°C, photoperiod: 16 hours with light/ 24 hours). 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100% 5 10 15 Middle of April (Method I) End of April (Method II) End of April (Method III)T emergence of antibiotic resistant bacteria is one of the current greatest challenges of the healthcare system. Since multidrug-resistancy is rapidly spreading, development of novel antibiotics is not a feasible strategy. Therefore, new alternative antimicrobial agents are necessary. Using bacteriophages – the natural enemies of bacteria – is a promising alternative approach. Bacteriophages (phages) are viruses that are obligate intracellular parasites of bacteria, which could be applied to control also multidrug resistant bacteria. The consortium of Enviroinvest Co., University of Pecs and University of Szeged established a scientific center in order to develop phage therapeutic products against human-, animaland plant pathogenic bacteria. Staphylococcus aureus, is a Gram-positive bacterium which is responsible for numerous infections worldwide. The emergence methicillin-resistant (MRSA) and vancomycin-resistant (VRSA) cell lines made the treatment of S. aureus infections more difficult. The aim of this research was to find effective bacteriophages against S. aureus. Several potential candidates were isolated. Lysogenic bacteriophages are not suitable for therapeutic use, therefore our research focused on strictly lytic ones. Few candidates have been found which had strictly lytic phenotype. In order to use them in bacteriophage therapy, their morpoholgical characters, maximal titer, lytic spectra/host specificity and genomic sequence must be determined. Therefore, we have sequenced the viral genomes. In one lytic phage, several spontaneous mutations in the integrase gene could be recognized. These mutations could account for the strictly lytic phenotype of the bacteriophage. The complete genome sequence also allowed us to compare the new phage with previously sequenced ones.W pathway is a critical aspect in the complex network of signaling molecules implicated in cancer. In non-malignant cells, Wnt signal cascade is blocked by the Secreted Frizzled-related proteins (sFRPs), a family of glycoproteins. However, they are epigenetically silenced in cancer cells. In this study, we attempted to administer recombinant human sFRP1 purified from bacterial culture on to cervical cancer cells (HeLa) to determine its anti-cancer activity. Human sFRP1 gene was cloned into bacterial expression vector pGEX-4T2. Recombinant protein was over-expressed as inclusion bodies in E.coli BL21 (DE3). After optimization of the process of solubilization of protein using detergents, GST-sFRP1 was purified to homogeneity by glutathione agarose affinity chromatography. MALDI analysis verified peptide signature particular to GST-sFRP1 and structural integrity was characterized by circular dichroism spectra. Purified recombinant protein was found to exert significant anti-proliferative effect on HeLa cells at nanomolar concentrations. Western blotting with antibodies against beta-catenin and phosphorylated beta-catenin confirmed the implication of canonical Wnt pathway. Combination therapy exhibited that recombinant sFRP1 substantially chemo sensitized HeLa cells towards conventional chemotherapeutic drug cisplatin. Coating of the therapeutic protein over gold nano-cluster embedded novel composite nanoparticles facilitated sustained release of the protein and enhanced its functional activity. The remarkable fluorescence property of gold nano-clusters was exploited for tracking and imaging purposes. Overall, therapeutic sFRP1 based nano-system may prove to be revolutionary in the field of cancer theranostics.T human cannabinoid receptor CB2 belongs to the class A of heptahelical G protein-coupled receptors (GPCR) and is an attractive target for the development of drugs for management of pain, inflammation, osteoporosis and treatment of immunological disorders. High resolution structural studies are critical to obtain insights into the molecular mechanisms of ligand binding and activation of CB2.We developedmethods for expression in milligram quantities, purification, reconstitution in lipid bilayers and stabilization of the functional recombinant CB2 as well as efficient stable isotope labeling of CB2 by high density fermentation were developed enabling NMR studies. NMR analysis of labeled receptor reconstituted in proteoliposomes in agonist or inverse-agonist-bound form will be reported.Monoclonal antibodies wereraised against the purified CB2 and the affinity of interaction with CB2 was determined by surface plasmon resonance.Finally, we demonstrate the specific effects of lipids with negatively charged head group in activation of CB2 reconstituted in proteoliposomes as measured by an in vitro G protein activation which may have important physiological significance as manifested in natural membranes of various lipid compositions.S scrofa is considered an important animal for meat production. During the last decade pork quality is one of the main selection criteria for breeding in the swine industry and thus attracted much more research focus. Jeju Native Pig (JNP) is a typical pig breed in South Korea highly appreciated by Korean consumers; however the slower growth rate and lighter carcass weight than commercial pig breeds led the farmers move away from traditional pig farming to typically intensively muscular traits farming. One recent approach relies to identify the key gene and its SNPs as molecular markers and have been tried for their use in traceability systems in livestock. The present study focused on the association of polymorphisms in the Myosin binding protein H (MyBPH) with skeletal muscle development in the JNP and Berkshire pigs. Validation of expression of genes in the laboratory also supported the idea of differential expression of MyBPH conceived through the in silico analysis. Gene ontology analysis of MyBPH showed significant association with muscle contraction and muscle organ development. The concordant in silico analysis of MyBPH by using the various tools identified potential four nsSNPs (W93T, D218R, D230W and W315V) in JNP. Our nsSNPs analysis retrieved high deleterious effects at second coding region in JNP compared with Berkshire. This study is an in-depth pioneering in silico analysis of polymorphic MyBPH and will be a valuable resource for further targeted molecular diagnosis and population-based studies for improving the growth performance of the JNP.