Improved production of 3‐hydroxypropionaldehyde by complex formation with bisulfite during biotransformation of glycerol

3‐Hydroxypropionaldehyde (3HPA) is an important specialty chemical which can be produced from glycerol using resting cells of Lactobacillus reuteri. This biocatalytic route, however, suffers from substrate‐ and product‐mediated loss of enzyme activity within 2 h of biotransformation. In order to overcome the inhibitory effects of 3HPA, complex formation with sodium bisulfite was investigated, optimized and applied for in situ capture of the aldehyde during biotransformation of glycerol in a fed‐batch process. As a result, the activity of the cells was maintained for at least 18 h. The 3HPA produced per gram cell dry weight was increased 5.7 times compared to the batch production process, and 2.2 times compared to fed‐batch process without in situ complex formation. This approach may have potential for production and in situ removal of 3HPA after further process development. Biotechnol. Bioeng. 2013; 110: 1243–1248. © 2012 Wiley Periodicals, Inc.     

Thermoluminescence properties of gamma‐irradiated nano‐structure hydroxyapatite

The suitability of nano‐structured hydroxyapatite (HAP) for use as a thermoluminescence dosimeter was investigated. HAP samples were synthesized using a hydrolysis method. The formation of nanoparticles was confirmed by X‐ray diffraction and average particle size was estimated to be ~30 nm. The glow curve exhibited a peak centered at around 200 °C. The additive dose method was applied and this showed that the thermoluminescence (TL) glow curves follow first‐order kinetics due to the non‐shifting nature of T_m after different doses. The numbers of overlapping peaks and related kinetic parameters were identified from T_m–T_stop through computerized glow curve deconvolution methods. The dependence of the TL responses on radiation dose was studied and a linear dose response up to 1000 Gy was observed for the samples. Copyright © 2015 John Wiley & Sons, Ltd.     

Molecular Design,Expression and Evaluation of PASylated Human Recombinant Erythropoietin with Enhanced Functional Properties

Erythropoietin (EPO) is the principal hormone which, has somewhat short half-life involved in the differentiation and regulation of circulating red blood cells. The present study was carried out to evaluate the capability of a polyethylene glycol mimetic technology as a biological alternative to improve pharmaceutical properties of human recombinant EPO. In silico models of EPO fused to 200 amino acids of proline, alanine, and serine (PAS) were initially generated and assessed by molecular dynamic (MD) simulation. The fluctuations of the modeled structure reached a plateau after 6000 ps of MD simulation. The Phi and psi analysis showed >99.2% of residues were located in the allowed regions. An expression vector consisting of EPO cDNA tagged to PAS coding sequences was synthesized and expressed in CHO-K1 Cells. The produced PASylated molecule was purified and characterized by standard analytical methods. The molecular weight of fusion protein was expanded to 70 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis method. Analytical size exclusion chromatography revealed an approximately sevenfold increase in apparent size of produced protein. Although the in vitro potency of the fusion protein was significantly reduced (1.26?±?0.05 vs. 0.24?±?0.03 ng/ml) but, the in vivo activity was considerably increased up to 1.58?×?10⁵ IU/ml in normocythemic mice assay. Pharmacokinetic animal studies revealed strongly 15.6-fold plasma half-life extension for the PASylated EPO (83.16?±?13.28 h) in comparison to epoetin α (8.5?±?2.4 h) and darbepoetin α (25.3?±?2.2h).     

Effect of plant extracts on the mortality of root-knot nematodes’ J2, Meloidogyne javanica

Root-knot nematode, Meloidogyne javanica, is a major problem confronting greenhouse’s productions, field crops, vegetables, grapevines and almond rootstocks in Kermanshah province, Iran. Nematicides are not affordable to control this nematode. In the search for alternatives to chemicals control of nematodes, this research has dealt with nematicidal effects of crude herbal extracts on the root-knot nematodes. This study aimed to evaluate the effect of 21 endemic and exotic herbal extracts belong to 12 families of flowering plants in comparison with chicken manure and chemical nematicide (Temik) to control root-knot nematodes in in vitro conditions. The nematodes were pured and mass multiplied on tomato in the soil at greenhouse conditions. In order to study the effect of herbal extracts on mortality of second-stage juveniles (J₂), a 6?mL of each extract was poured in sterilised Petri dish and 54?±?4 juveniles were added. Distilled water was used as control and treatments replicated four times and incubated at ambient temperature. The LC₅₀ value of each extract was determined by assessing the mortality of juveniles (in the range of 5–95%) after 24, 48 and 72 h. Comparison between LC₅₀ value of the extracts indicated that Cinnamomum zeylanicum and Eugenia caryophillata are the most effective crude extracts on the mortality of juveniles and they were 15.4 and 17.9?mg?mL^?1, respectively. Meanwhile, the extract of tobacco, ferulago, garlic, eucalyptus, persan lilac, rattle, oliveria, licorice, russian knapweed, turnsole, sicilian sumac and chicken manure did not have any antinematode activity against fresh second-stage juveniles of the root-knot nematode.     

Modeling the time dependent distribution of a new 153Sm complex for targeted radiotherapy purpose

BackgroundFor radioimmunotherapy purposes, a chemical complex with high absorption in cancer tumor is required. New chemicals are to be examined for their concentration in tumor and healthy organs. These are labeled with β-emitting radioisotopes to irradiate the tumor while deposited inside it.AimTo study the capability of recently developed chemical complex in targeting cancer tumor and investigate the distribution of ¹⁵³Sm-TPTTC in rat organs as function of time.Materials and methodsThe chemical complex – [Tris(1,10-phenanthroline)Samarium(III)] trithiocyanate was prepared and labeled with ¹⁵³Sm radioisotope. The labeled complex was injected to a population of tumor bearing mice. In 2, 4, 24, 48, 96 h after injection the animals were sacrificed and the concentration of Samarium complex was measured in various organs such as blood, heart, intestine, colon, liver, spleen, kidney, sternum and bone.ResultsThe concentration of the radiopharmaceutical in various organs was measured at different times. The temporal behavior of biodistribution of ¹⁵³Sm-TPTTC was modeled and drawn as function of time.ConclusionIt is shown that ¹⁵³Sm-TPTTC is concentrated in tumor tissue and liver much more than in other organs. The variation of pharmaceutical concentration in all organs is described with summation of eight exponential terms and it approximates our experimental data with precision better than 2%.     

Membrane-active metabolites produced by soil actinomycetes using chromatic phospholipid/polydiacetylene vesicles

Increased resistance of pathogens toward existing antibiotics has compelled the research efforts to introduce new antimicrobial substances. Drugs with new and less resistant-prone targets to antimicrobial activity have a high priority for drug development activities. Cell membrane seems to be a potential target for new antibiotic agent development to overcome resistance. In this study, A total number of 67 actinomycetes were isolated from the soil samples collected from desert, farming and mineral parts of Iran. We used a chromatic sensor as a membrane model that was set up for the target of antimicrobial metabolites of actinomycetes isolated from the soil. The sensors particles were composed of phospholipid and polymerized polydiacetylene (PDA) lipids. These polymers exhibited color change following interaction with membrane-active metabolites. The color change was due to structural disorder in the lipids following their interaction with membrane-active metabolites. The resultant color change was recorded by fluorescent microscope and easily recognizable by naked eye as well. Sixteen strains were isolated which produced antimicrobial metabolites and were effective against test microorganisms (Escherichia coli, Candida albicans and Saccharomyces cerevisiae ). A total number of 3 out of 16 strains produced membrane-active metabolites. These 3 strains were identified using 16s rRNA as Streptomyces sp and submitted to GenBank (accession no. JN180853; JN180854; JN180855).     

Biotransformation of glycerol to 3-hydroxypropionaldehyde: Improved production by in situ complexation with bisulfite in a fed-batch mode and separation on anion exchanger

3-Hydroxypropionaldehyde (3HPA) is an important C3 chemical that can be produced from renewable glycerol by resting whole cells of Lactobacillus reuteri. However the process efficiency is limited due to substrate inhibition, product-mediated loss of enzyme activity and cell viability, and also formation of by-products. Complex formation of 3HPA with sodium bisulfite and subsequent binding to Amberlite IRA-400 was investigated as a means of in situ product recovery and for overcoming inhibition. The adsorption capacity and -isotherm of the resin were evaluated using the Langmuir model. The resin exhibited maximum capacity of 2.92 mmol complex/g when equilibrated with 45 mL solution containing an equilibrium mixture of 2.74 mmol 3HPA-bisulfite complex and 2.01 mmol free 3HPA. The dynamic binding capacity based on the breakthrough curve of 3HPA and its complex on passing a solution with 2.49 mmol complex and 1.65 mmol free 3HPA was 2.01 mmol/g resin. The bound 3HPA was desorbed from the resin using 0.20 M NaCl with a high purity as a mixture of complexed- and free 3HPA at a ratio of 0.77 mol/mol. Fed-batch biotransformation of glycerol (818.85 mmol) with in situ 3HPA complexation and separation on the bisulfite-functionalized resin resulted in an improved process with consumption of 481.36 mmol glycerol yielding 325.54 mmol 3HPA at a rate of 17.13 mmol/h and a yield of 68 mol%. Also, the cell activity was maintained for at least 28 h.     

Application of antimicrobial peptides in agriculture and food industry

Antimicrobial peptides have captured the attention of researchers in recent years because of their efficiency in fighting against pathogens. These peptides are found in nature and have been isolated from a wide range of organisms. Furthermore, analogs or synthetic derivatives have successfully been developed on the basis of natural peptide patterns. Long use of pesticides and antibiotics has led to development of resistance among pathogens and other pests as well as increase of environmental and health risks. Antimicrobial peptides are under consideration as new substitutes for conventional pesticides and antibiotics. Many plants and animals have been manipulated with antimicrobial peptide-encoding genes and several pesticides and drugs have been produced based on these peptides. Such strategies and products may still have a long way to go before being confirmed by regulatory bodies and others need to surmount technical problems before being accepted as applicable ones. In spite of these facts, several cases of successful use of antimicrobial peptides in agriculture and food industry indicate a promising future for extensive application of these peptides. In this review, we consider the developing field of antimicrobial peptide applications in various agricultural activities.     

Cultivation technology development of Rhodothermus marinus DSM 16675

Extremophiles - This work presents an evaluation of batch, fed-batch, and sequential batch cultivation techniques for production of R. marinus DSM 16675 and its exopolysaccharides (EPSs) and...     

Expression of a Novel Chimeric-Truncated tPA in Pichia pastoris with Improved Biochemical Properties

Thrombolytic therapy by plasminogen activators (PAs) has been a main goal in the treatment of acute myocardial infarction. Despite improved outcomes of currently available thrombolytic therapies, all these agents have different drawbacks that may result in less than optimal outcomes. In order to make tissue plasminogen activator (tPA) more potent, while being more resistant to plasminogen activator inhibitor-1 (PAI-1) and having a higher affinity to fibrin, a new chimeric-truncated form of tPA (CT tPA) was designed and expressed in Pichia pastoris. This novel variant consists of a finger domain of Desmoteplase, an epidermal growth factor (EGF) domain, a kringle 1 (K1) domain, a kringle 2 (K2) domain, in which the lysine binding site (LBS) was deleted, and a protease domain, where the four amino acids lysine 296, arginine 298, arginine 299, and arginine 304 were substituted by aspartic acid. The chimera CT tPA showed 14-fold increase in its activity in the presence of fibrin compared to the absence of fibrin. Furthermore, CT tPA showed about 10-fold more potency than commercially available full-length tPA (Actylase^®) and provided 1.2-fold greater affinity to fibrin. A residual activity of only 68 % was observed after incubation of Actylase^® with PAI-1, however, 91 % activity remained for CT tPA. These promising findings suggest that the novel CT tPA variant might be an acceptable PA with superior characteristics and properties.