Leishmaniasis is an infectious disease in which different clinical manifestations are classified into three primary forms: visceral, cutaneous and mucocutaneous. These disease forms are associated with parasite species of the protozoan genus Leishmania. For instance, Leishmania infantum and Leishmania tropica are typically linked with visceral (VL) and cutaneous (CL) leishmaniasis, respectively; however, these two species can also cause other form to a lesser extent. What is more alarming is this characteristic, which threatens current medical diagnosis and treatment, is started to be acquired by other species. Our purpose was to address this issue; therefore, gel-based and gel-free proteomic analyses were carried out on the species L. infantum to determine the proteins differentiating between the parasites caused VL and CL. In addition, L. tropica parasites representing the typical cases for CL were included. According to our results, electrophoresis gels of parasites caused to VL were distinguishable regarding the repetitive down-regulation on some specific locations. In addition, a distinct spot of an antioxidant enzyme, superoxide dismutase, was shown up only on the gels of CL samples regardless of the species. In the gel-free approach, 37 proteins that were verified with a second database search using a different search engine, were recognized from the comparison between VL and CL samples. Among them, 31 proteins for the CL group and six proteins for the VL group were determined differentially abundant. Two proteins from the gel-based analysis, pyruvate kinase and succinyl-coA:3-ketoacid-coenzyme A transferase analysis were encountered in the protein list of the CL group.

A Chromatographic Approach for Partial Purification of Polygalacturonase Produced byAspergillus sojae ABSTRACTThe aim of this study was to produce polygalacturonase from A. sojae mutant and partially purify the crude extract by chromatographic methods. As a preliminary step for the confirmation of its peptides, matrix-assisted laser-desorptionionization-time-of-flight mass spectrometry (Maldi-TOF MS) analysis was performed on in-gel digested sodiumdodecyl-sulphate-polyacrylamide-gel-electrophoresis (SDS-PAGE) gels. Three different carbon sources were employed in submerged and solid-state fermentations for the production of polygalacturonase. Crude extract was first purified by ion-exchange chromatography (IEXC) and followed further by size exclusion chromatography. Crude extracts obtained from sub-merged [of bitter orange peel, sugar beet molasses and (NH4)2SO4] and solid-state [of wheat bran, sugar beet and HCl] fermentation exhibited high levels of polygalacturonase enzyme activity (95.22 and 50.27 U/mL, respectively). Size exclusion of IEXC pooled fraction (180, 200 and 220 mM salt fractions) revealed the highest yield (36%) and purification fold (2.00). The likely polygalacturonase bands from SDS-PAGE were in-gel digested and analyzed by Maldi-TOF MS in route for peptides confirmation.

Proteomik yaklaşımları 2000 li yılların başlarına kadar mikroorganizmalar ve hayvansal kaynaklı örneklerde ağırlıklı olarak kullanıldı. Bu dönemde bitki proteomik çalışmaları yok denecek kadar azdır. Bitkisel dokulardaki sert hücre çeperleri, karmaşık ve çok çeşitli sekonder metabolitlerin varlığı, fazla miktardaki pigmentler, proteazlar, polifenoller, polisakkaritler, nişasta ve lipitler total protein örneklerinin hazırlanması ve proteinlerin ayrımı sırasında pek çok soruna neden olmuştur. Ancak her bir sorunun üstesinden gelmek üzere sürdürülen çabalar sayesinde bitki dünyasında da proteomik yaklaşım kullanımı yaygınlaşmıştır. Bu derlemede, örnek hazırlığından protein tanımlamaya kadar tüm basamaklar yöntemsel gelişmeleri de kapsayacak şekilde ayrıntılı olarak ele alınmış ve konuyla ilgili araştırıcıların maksimum yararlanabileceği bir kaynak oluşturulmaya çalışılmıştır.