Acidithiobacillus strains SP5/1 and HV2/2 are rod-shaped, Gram-negative, γ-proteobacteria, oxidizing aerobic pyrite and acidophilic bacteria which grow optimally at 30°C and 45°C respectively (Klingl, Moissl-Eichinger et al. 2011). The symmetry similarity of the S-layer molecular structure in closely related species can be used as a taxonomic feature that distinguishes the phylogenetic position of the organism (Baumeister and Lembcke 1992, Klingl, Moissl-Eichinger et al. 2011). The S-layer protein coated the surface of the bacteria, with the potential to assemble. Say no to plagiarism. Get a tailor-made essay on “Why violent video games should not be banned”? Get the original essay The protein is contained in 2D pseudo-crystalline with symmetry variation (p1, p2, p3, p4 or p6) (Eichler 2003). Two mentioned strains have an S-layer protein with p2 symmetry (Klingl 2007). The surf layer is first identified in the cell wall of a Spirillum sp. (Houwink and Le Poole 1952, Houwink 1953), and as of today it is found in almost all taxonomies of bacteria and archaeal shells (Rachel, Pum et al. 1997, Sleytr 1997). S-layer subunits are connected to the peptidoglycan layer or pseudomurein in archaea and Gram-positive bacteria. The subunits are associated with lipopolysaccharide (LPS) components in Gram-negative bacteria (Sára and Sleytr 2000). The only cell wall component bound to the cytoplasmic membrane of archaea is thought to be Slayer (König, Rachel et al. 2007). It represents the simplest protein in archaeal envelopes and bacteria (Sára and Sleytr 2000); moreover, due to the possession of approximately 10% protein cells in bacteria and archaea, they introduce a fascinating prototype for cellular study during cell growth and division and evolutionary associations (Whitman 1998 , Sleytr, Huber et al. Extracellular polymeric substances (EPS) provide contact between the cell and the sulfur energy source, playing a key role in organic film formation and bacterial interactions (Savage and Fletcher 1985). , composed of polysaccharides, proteins, lipids and nucleic acids, the quantity and composition of which vary depending on various energy sources (Gehrke, Telegdi et al. 1998). A. ferrooxidans ATCC 23270T Greater growth of EPS has been reported in media containing pyrite or sulfur compared to ferrous iron (Gehrke, Telegdi et al. 1998). Extracellular polysaccharides (EPS) are linked to the complexation of iron(III) ions (Sand and Gehrke 2006, Klingl, Moissl-Eichinger et al. 2011). The amount of adhesion forces and attached cells is reduced in the absence of EPSin A. ferrooxydans ATCC 23270 (Li, Wang et al. 2016). Factors, such as pH and energy source, affect cell surface properties; therefore, different cultures exhibited varied adhesion strengths to the same substrate (Li, Wang et al. 2016). Iron binding does not occur randomly (Sand and Gehrke 2006), a correlation between the amount of iron(III) ions complexed in the EPS and FeS2 cell attachment was reported for R1 and SPIII strains /3 of A. ferrooxydans (Sand and Gehrke 2006). ).Different electron microscopy methods such as negative staining and the freeze etch method provide considerable information about the S-layer mesh structure (Messner, Pum et al. 1986, Rachel 1999). As a phylogenetic approach, the characteristicsCommon surface layers such as symmetry and dimension can be used to elucidate the taxonomic position of the organism by electron microscopy (König, Rachel et al. 2007, Klingl, Moissl-Eichinger et al. 2011). High-resolution freezing techniques as the best method to consider S-layer structure and flagella in fresh, unwashed intact bacteria (Sleytr, Schuster et al. 2014). Isolation of S-layer bacteria by hydrogen bond-breaking protein proteins leads to their recrystallization into suspended monomolecular or double-layer networks, whether in the form of flat sheets or with open-ended cylinders ( Sleytr, Huber et al. 2007), (Jarosch, Egelseer et al. 2001); Additionally, some native bacterial strains lose their ability to produce S-layers due to loss of plasmids during prolonged laboratory culture (Jarosch, Egelseer et al. 2001, Blecha, Zarschler et al. 2005). the proteins will be heterologous expressed in a eukaryotic system like [PIY3 (Ahmad, Hirz et al. 2014). The stability of the expressed proteins has been proven in yeast (Blecha, Zarschler et al. 2005). ObjectivesThe surface layer proteins of Acidithiobacillus strains SP5/1 and HV2/2 will be isolated and localized to:Keep in mind: this is just a sample.Get a personalized article from our expert editors now. Get a custom essay Clarify the phylogenetic position of these two strains Comparison of the mutual characters of the two strains with other Acidithiobacillus species, as described by (Kelly and Wood 2000) Acidithiobacillus ferrooxydans SP5/1 is a member of the Acidithiobacillus species caldus; however, growing in a medium with Fe2+ as the only energy substrate and oxidizing pyrite, which suggests classifying strain SP5/1 into an appropriate new species “Candidatus Acidithiobacillus striatothermus”. The existence of an S layer has been proven in the SP5/1 strain (Klingl, Moissl-Eichinger et al. 2011). Furthermore, we hypothesize that the Afe_2303 protein has putative similarities to a surface layer protein of A. ferrooxidans ATCC 23270 (Klingl, Moissl-Eichinger et al. 2011). By studying the structure of the S layer, the phylogenetic position of the two strains will be clarified, however, by studying the nature of the bands in these two strains, and by comparing the mutual characters with other same species, it would be possible to obtain more detailed information to classify Acidithiobacillus species. Research Design, Methods and ProceduresThe bacterial strain will be used in this study as follows: Acidithiobacillus ferrooxidans SP5/1 strains were isolated in 1984 from the Solfatara volcano, in the Pisciarelli region of Italy and the sampling of Acidithiobacillus sp HV2/2 came from Hveravellier, the island and was isolated in 1986 by Dr. med. Harald Huber from the University of Regensburg. Strains and culture conditions Acidithiobacillus strains SP5/1 and HV2/2 will be cultured in 9-K medium (Silverman and Lundgren 1959) containing 125 g of pyrite in Erlenmeyer flasks. The pH medium will be adjusted to 2.5 with 50% sulfuric acid and will be incubated on a rotating incubator at 150 rpm at optimal growth temperatures of 30°C for the SP5/1 strain and 45°C for the strain HV2/2 for 72 hours. A. ferrooxydans ATCC 23270, A. caldus DSM 8584 will be used in this study. A deficient laboratory strain of A. ferrooxydans SP5/1, for its part, will be used as a control. The growth and morphology of the two strains will be monitored by optical microscopy and counted with the Thoma cell counting chamber. Electron microscopy. The appendix of the samples will be studied by negative staining, method of.