User:Baileygentile/sandbox

Halobacterium noricense is a halophilic, rod-shaped microorganism that thrives in environments with salt levels near saturation. Despite the implication of the name, Halobacterium is actually a genus of archaea, not bacteria. H. noricense can be isolated from environments with high salinity such as the Dead Sea and the Great Salt Lake in Utah. Members of the Halobacterium genus are excellent model organisms for DNA replication and transcription due to the stability of their proteins and polymerases when exposed to high temperatures. To be classified in the Halobacterium genus, a microorganism must exhibit a membrane composition consisting of ether-linked phosphoglycerides and glycolipids.

Typical morphology of Halobacterium species

Halobacterium noricense
Scientific classification
Domain:	Archaea
Kingdom:	Euryarchaeota
Phylum:	Euryarchaeota
Class:	Halobacteria
Order:	Halobacteriales
Family:	Halobacteriaceae
Genus:	Halobacterium
Species:	H. noricense
Binomial name
Halobacterium noricense Gruber et al. 2005
Type Strain
DSM 15987T, ATCC BAA-852T, NCIMB 13967T, NRC-1, DL1, R1, CBA1132

Scientific classification

This organism is a member of the Halobacterium genus and its taxonomic classification is as follows: Archaea, Euryarchaeota, Euryarchaeota, Halobacteria, Halobacteriales, Halobacteriaceae, Halobacterium, Halobacterium noricense. There are currently 19 known halophilic archaeal genera and 57 known species within the Halobacterium genus.

Relatives

Three reported strains Halobacterium salinarium NRC-1, Halobacterium sp. DL1, and Halobacterium salinarium R1 were compared to Halobacterium noricense strain CBA1132. The phylogenetic trees based on Multi-Locus Sequence Typing (MLST) and Average Nucleotide Identity (ANI) indicated that strain CBA1132 and strain DL1 are closely related while strains NRC-1 and R1 are closely related. Multi-Locus Sequence Typing is a technique that uses genomic information to establish evolutionary relationships between bacterial taxa. Average Nucleotide Identity is a genetic method used to compare the similarity between nucleotides of two strains based off of the coding regions of their genomes, which has allowed scientists to veer away from traditional methods of classifying prokaryotes based on phenotypic similarities. The defining characteristic between strains CBA1132 and DL1 is that they both contain high GC content in their chromosomes, providing stability in a harsh environment. Other close relatives of H. noricense within the Halobacterium genus include Halobacterium denitrificans, Halobacterium halobium, and Halobacterium volcanii.

Morphology

Halobacterium noricense is known to be free living, and it typically appears as red or pink colonies due to the presence of carotenoids and bacterioruberin in their membranes. The carotenoids have the ability to absorb light between the wavelengths of 330-600 nm, as determined by light spectroscopy. Typical colony morphology is round with a diameter of 0.4 mm. Under the microscope, they can typically be measured at around 5 μm and appear gram-negative and rod-shaped. H. noricense does not contain the gas vesicles that are present in their close relative, Halobacterium salinarium, which often appear as floating cultures. Halobacterium noricense may occasionally appear as coccus-shaped when grown in liquid broth rather than on solid media.

Discovery

Etymology

Halobacterium noricense is named after Noricum, Austria, which is the location of the salt deposit in which the organism was isolated. The archaeon was discovered in 2004 by a group of scientists led by Claudia Gruber. The group isolated two strains of H. noricense, along with other Halobacterium species including H. salinarium.

Source

The first two strains (A1 and A2) of Halobacterium noricense were isolated from samples taken out of a salt deposit in Austria. The salt deposit was approximately 400 meters below the surface and is believed to have been formed during the Permian period. To obtain the samples, the researchers used a pre-existing mine to travel below the Earth’s surface. They used a core drill to remove cylindrical sections of the salt deposit, which were then taken for sequencing. The deposit retained high salt levels over approximately 250 million years due to the surrounding clay and limestone. These conditions do not allow the salt to escape, which formed an ideal environment for halophilic archaea.

Media

Halobacterium noricense was isolated on ATCC 2185 medium with 250.0 grams of NaCl, 20.0 grams of MgSO₄ 7H₂O, 2.0 grams of KCl, 3.0 grams of yeast extract, 5.0 grams of tryptone, and other compounds required for the isolate's growth. After an incubation period of approximately 2 weeks, red circular colonies appeared. This is the characteristic colony morphology of H. noricense.

Growth Conditions

Halobacterium noricense is known to be a mesophile, where optimum growth temperature is approximately 37°C with an incubation period of 18 days. It thrives in acidic conditions at pH 5.2-7.0. NaCl concentration between 15-17% has resulted in the highest growth rates in previous studies. It has been found that Halobacterium can survive in high metal concentrations because they are extremely halophilic. This can be achieved through metal resistance, which indicates that the H. noricense strain CBA1132 might also be able to survive in these high metal ion concentrations.

Genome

H. noricense strains A1 and A2 from Gruber et. al had 97.1% similarity to Halobacterium genus through their 16S rDNA sequences. H. noricense genome, strain CBA1132, comprised of four contigs containing 3,012,807 base pairs, approximately 3,084 gene coding sequences, and 2,536 genes. It has a GC content of approximately 65.95%, and 687 of the genes in the H. noricense genome have unknown functions. Metabolism and amino acid transport-related genes make up the largest group of known genes. This group contains 213 known genes. The Halobacterium genus is currently known as monophyletic because their 16S rRNA have less than 80% similarity with their closest relatives, the methanogens.

Sequencing

According to Joint Genome Institute, another complete genome analysis of Halobacterium (strain DL1) species was sequenced using 454 GS FLX, Illumina GAIIx. Halobacterium noricense (strain CBA1132) was recently isolated from solar salt and a complete genomic analysis was performed by researchers from Korea in 2016. The researchers extracted the DNA using a QuickGene DNA tissue kit, which uses a membrane with extremely fine pores to collect DNA and nucleic acids. They purified the DNA using the MG Genomic DNA purification kit. Once extracted and purified, the strategy for sequencing the genome was Whole Genome Sequencing by the method of a PacBio RS II system. Lastly, the genome was analyzed and performed by the Rapid Annotation using Subsystem Technology (RAST) server.

Metabolism

According to Gruber et al., Halobacterium noricense cannot ferment glucose, galactose, sucrose, xylose or maltose. It is resistant to many antibiotics, including Vancomycin and Tetracycline, but can be killed by Anisomycin. This organism does not produce the enzymes gelatinase or amylase, so it cannot break down starch or gelatin. H. noricense is a chemoorganotroph and uses aerobic respiration in most environments, except when exposed to L-arginine or nitrate. In these cases, it can function as a facultative anaerobe. It is catalase positive, meaning it has the ability to break down hydrogen peroxide into water and oxygen. The most abundant carbon source found in hypersaline environments is glycerol due to the contribution of the green algae, Dunaliella, to reduce its' surrounding osmotic pressure. H. noricense is able to metabolize glycerol through phosphorylation to glycerol 3- phosphate and eventually, into the formation of dihydroxyacetone 5- phosphate (DHAP). NMR spectroscopy, used to locate local magnetic fields around atomic nuclei, revealed during aerobic respiration that 90% of pyruvate that is converted to acetyl Co-A by pyruvate synthase enters the Citric acid cycle while the other 10% is converted to oxaloacetate by biotin carboxylases to later be used in fatty acid degradation.

Significance

Halobacterium noricense has many applications that can benefit humans and industries including drug delivery, UV protection, and the unique characteristic of bacteriorhodopsin to be able to be isolated outside of its environment. H. noricense produces a high concentration of menaquinones (fat soluble vitamin K2) that can be used as a micelle to deliver drugs to specific places in the body. According to Nimptsch K, the presence of menaquinones can also reduce the risk of malignant cancer. Fermented foods are also found to have high levels of menaquinones due to the presence of bacteria, especially in cheeses. H. noricense requires high salt concentrations and is currently being explored to enhance the process of fermentation. H. noricense is also catalase positive, meaning it can break down reactive oxygen species (ROS), like hydrogen peroxide into harmless substances such as water. Not only does it produce enzymes to protect itself against ROS, but it contains a pigment, bacterioruberin, that allows H. noricense to tolerate gamma and UV radiation. Further research into bacterioruberin can lead to bioactive compounds with anticancer characteristics. Lastly, bacteriorhodopsin (also protects cells from UV light), a light proton pump, has allowed scientists to apply it to electronics and optics. Its mechanism involves capturing light and creating a proton gradient to produce chemical energy. Some practical uses include motion detection, holographic storage, and nanotechnology.