Lactobacillus brevis
A Microbial Biorealm page on the genus Lactobacillus brevis
Classification
Higher order taxa
Bacteria; Firmicutes; Bacilli; Lactobacillales; Lactobacillaceae; Lactobacillus
Species
Lactobacillus brevis
Description and significance
Describe the appearance, habitat, etc. of the organism, and why it is important enough to have its genome sequenced. Describe how and where it was isolated. Include a picture or two (with sources) if you can find them.
As a general overview, lactic acid bacteria which include the species Lactobacillus, are all Gram-positive organisms whose main metabolic pathway involves fermenting hexose sugars to produce lactic acid.
The significance of all lactic acid bacteria throughout history and until today is their metabolism which is used for preservations of foods and beverages. When strains of this bacteria were domesticated, their metabolic simplicity resulted in the ability to carry out fermentations for many of our modern foods. In fact, this bacteria plays a prominent part in the worlds food supply and production of some beers and wine.
Lactobacillus brevis is rod-shaped strain of lactic acid bacteria that can be found mainly in the intestinal flora and in soil. In general Lactobacilli are gram-positive and in nonsporeforming rods. Along with other lactic acid-producing bacteria, L. brevis plays an integral role in the fermentation of certain foods such as sauerkraut and pickles and is likewise the most common cause of spoiled beer. L. brevis can be isolated from the food sources in inhabits, one example being kimchi.
Genome structure
Whole-genome sequencing was conducted at the U.S. Department of Energy for eight other species of Lactic acid bacteria the method used was shotgun sequencing, where different parts of a genome are sequenced and then the pieces were assembled. The completed sequenced genome of L. brevis shows that this bacteria is circular and has one chromosome with a length of 2.3 Mbp containing 2185 proteins and 82 RNAs. Two plasmids are also sequenced, much smaller in length, and have 11 and 12 proteins. (2)
The evolution of Lactobacillales reveals a lot about the heritage of species and the ancestrol gene losses, and may suggest where duplications and the addition of new and unique genes occurred. "Phylogenetic analyses, comparison of gene content across the group, and reconstruction of ancestral gene sets indicate a combination of extensive gene loss and key gene acquisitions via horizontal gene transfer during the coevolution of lactic acid bacteria with their habitats." (2) One notable difference in L. brevis from the other species in this order, includes the loss of genes that carry out the biosynthesis of arginine and aromatic amino acids.
Cell structure and metabolism
Lactobacillus brevis and the Lactobacilli in general, are Gram-positive, rod-shaped, nonsporeforming bacteria arranged in singles. This implies a thick cell wall and an inner membrane in its rod shape. Although the sequenced genome of L. brevis is relatively small, the lactic acid bacteria can encode a multitude of different transporters to fulfill the needs of both prototrophic and auxotrophic strains. (2) L. brevis bacteria are aerobe bacteria, they are motile, and probiotic. (3)
Fermentation is the one most common pathway in L. brevis and most other lactic acid bacteria. In this metabolic pathway, sugar is converted to lactic acid, and energy (ATP) is generated along the way by non-oxidative substrate level phoshorylation. The final product of fermentation, lactic acid contributes to production of such foods as cheese yogurt, fermented milks and others by inhibiting the growth of other organisms and lowering the pH of the products. Along the pathway, many metabolic reactions occur, one of them being the breakdown of milk products for cheese production. L. brevis specifically is used in industrial production to act as a starter culture for several types of beer and sourdough bread. (3)
Ecology
Describe any interactions with other organisms (included eukaryotes), contributions to the environment, effect on environment, etc.
One of the most interesting interaction is that with the human intestinal flora. Lactobacillus, when ingested, has probiotic effects as it improves the immune system. Most commonly however, this bacteria lives in the foods where it performs its metabolic pathways.
Pathology
How does this organism cause disease? Human, animal, plant hosts? Virulence factors, as well as patient symptoms.
L. brevis is not a pathogen and there are no cases where it has caused or contributed to disease in humans. In fact, it is the other way around. This bacteria is probiotic, and improves the human immune system.
Application to Biotechnology
Does this organism produce any useful compounds or enzymes? What are they and how are they used?
Current Research
1. "Proteomic approach for characterization of hop-inducible proteins in Lactobacillus brevis"
In order for lactic acid bacteria to cause beer spoilage during growth in beer, they must have a resistance to hops. A bacterial hop compound is also known as an ionophore which by definition is a small molecule whose main purpose is to transport ions through the lipid bilayer of a cell’s membrane. In the experiment, two strains of Lactobacillus brevis (TMW1.465 and variant TMW1.465A) were observed and the acid stress response and hop adaptation caused many detectable changes in the cells’ properties. To more clearly identify the cause of the changes in metabolism, membrane physiology, and cell wall composition, the scientists set up an experiment to identify related vital proteins despite the disadvantage of an incomplete genome sequence. Conclusively, hop resistance in the variant strain implied altered mechanisms that accounted for intracellular acidification, and changes in mechanisms that dealt with energy generation, genetic information, and enzyme functionality. Enzymes that are hop-regulated were also defined as being manganese or divalent cation dependent. Thus the ability to control the manganese level leads to the ability to tweak the metabolisms of applicable cells. One change found in metabolism in response to an environmental stress condition, was the shift into an energy-saving mode. Lastly this experiment demonstrated that bacteria in a hop stress environment were correlated with the depletion of proton motive force and with a limitation of the divalent cation, which as mentioned above is needed for hop-regulated enzymes. (4)
2. "Cloning, sequencing and expression of a novel glutamate decarboxylase gene from a newly isolated lactic acid bacterium"
A new strain of Lactobacillus brevis containing 84.292 mg/L/h of GABA (γ-aminobutyric acid) was isolated from a Korean fermented food, Kimchi. Lactobacillus brevis OPK-3 was found to have a core fragment of glutamate decarboxylase (GAD) DNA and this fragment was then isolated, cloned, and amplified (by method of PCR) to end up with a full-length piece of core GAD DNA and enough data for a nucleotide sequence analysis. The scientists found the open reading frame (ORF) of the DNA contained 1401 bases and had the ability to encode 467 amino acids for protein assembly. The molecular weight was found to be 53.4 kDa. When compared to the amino acid sequence of other Lactic Acid Bacteria GAD DNA, Lactobacillus plantarum GAD, Lactococcus lactis GAD, and Listeria monocytogenes, the LbGAD ORF was 83%, 71%, and 60% identical, respectively. Further in the experiment, the LbGAD gene was introduced into an Escherichia coli strain (UT481), and when the transformed E. coli was tested, a 53.4 kDa protein was apparent, and the bacteria also had heightened GAD activity. (5)
3. "Characterization of microflora in homemade semi-hard white Zlatar cheese"
In this experiment, researchers observed changes in lactic acid bacteria and in the chemical composition of Zlatar cheese, over the course of 60 days during which it was ripening. This cheese was produced from only nonpasteurized cow’s milk without any bacterial aid. The results were very interesting; it was found that the while the number of lactobacilli was slowly increasing over the 60 days, the percentage of cocci first decreased, and then began increasing again at the 30 day mark, eventually catching up with lactobacilli. Also, it was found that out of all bacteria, 57 isolates showed antimicrobial activity and were decreasing with cheese age, until no more was detected in samples of 60 day old cheese. In doing replication - PCR analysis, Lactobacillus brevis was found to be one of the main species of bacteria present in addition to Lactobacillus paracasei, Lactococcus lactis, and Enterococcuus faecium. (6)
References
1. NCBI Taxonomy, url: http://www.ncbi.nlm.nih.gov/Taxonomy/
4. "Lactobacillus brevis ATCC367", Integrated Microbial Genomes, Doe Joint Genome Institute.
8. David Gilbert, “Genomic comparison of lactic acid bacteria published”, Lawrence Livermore National Laboratory. Public Affairs : Newsroom : News Releases : 2006 News Release Archive :: NR-06-10-09.
9. Sakamoto, Kanta, “Beer spoilage bacteria and hop resistance in Lactobacillus brevis'”. University of Groningen, 2002. Chapter 1, p. 5-6.
10. Kagli DM, Vancanneyt M, Hill C, Vandamme P, Cogan TM, “Enterococcus and Lactobacillus contamination of raw milk in a farm dairy environment”. International Journal of Food Microbiology, 2007. V.114, Issue 2, p.243-251.
11. Hayashi N, Ito M, Horiike S, Taguchi H, “Molecular cloning of a putative divalent-cation transporter gene as a new genetic marker for the identification of Lactobacillus brevis strains capable of growing in beer”. Applied Microbiology Biotechnology, 2001. V. 55, Issue 5, p.596-603.
Edited by Ekaterina Magonova of Rachel Larsen and Kit Pogliano
