LB Bulgaricum PLC

Sofia, Bulgaria

LB Bulgaricum PLC

Sofia, Bulgaria
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Grant
Agency: European Commission | Branch: FP7 | Program: BSG-SME | Phase: SME-2013-1 | Award Amount: 1.29M | Year: 2013

Fermented food (e.g. cheese, yoghurt, bread, wine) constitute a large part of our diet. They are converted from a substrate (milk, fruits, cereals) by the action of micro-organisms (yeast or lactic acid bacteria). When such starter cultures also exert a health benefit (eg. prevention of diarrhea) they are known as probiotic strains. Throughout the EU starters and probiotic strains are mainly supplied by SMEs to end-users for application in food products. The SMEs compete with a few large companies providing such cultures. The competitive edge of SMEs is at risk in part due to stricter safety rules to be imposed by EFSA. EFSA will demand full genome characterisation to exclude presence of sequences posing a potential health risk (such as virulence factors, antibiotic resistance and toxins) and to allow traceability of distinct strains as proprietary strains are unique. Genome analyses and bio-informatics are not the current core of SMEs in this consortium as they have focus on primarily cost effectiveness. GENOBOX exploits the strength of four SME starter culture companies and probiotic producers in this sector combined with skills and expertise of two renowned RTD performers. The RTD performers will sequence and analyse 48 bacterial strains provided by SMEs and coming from culture collections. A genomics toolbox will be created, allowing every SME to exclusively study the detailed results on their own strains while benefitting from results of the collective. The SMEs will perform in-house experiments and demonstrations to validate the results of the project for optimised production yields. Also workshops will be organised by the RTD performers to develop data-exploitation strategies for the SMEs involved. During this project the SMEs will be able to abide to EFSA ruling, enable to patent their strains for specific purposes, exploit strain functionalities and health benefits and maximise their production yields by optimising strain survival during processing.


Ishlimova D.,LB Bulgaricum PLC | Urshev Z.,LB Bulgaricum PLC
Biotechnology and Biotechnological Equipment | Year: 2013

Streptococcus thermophilus LBB.A is a component of industrial starters for traditional Bulgarian yogurt. Eight genetically distinguishable cos-type phage cultures: φA1, φA2, φA5, φA6, φDmA, φSfA, φPtA, and φA7, were found to infect strain LBB.A. Bacteriophage-Resistant Mutants (BIMs) of strain LBB.A were isolated and their Clustered Regularly Interspaced Short Palindrome Repeats (CRISPR1 and CRISPR3) were amplified by PCR and compared to the parent strain. Challenging S. thermophilus LBB.A with φA1 resulted in the isolation of "first order" BIM AR3 A1, which was resistant to six of the phages, but remained sensitive to φPtA and φA7. In AR3 A1 the size of the CRISPR1 and CRISPR3 loci increased with approximately 66 bp each, compared to the parent strain, corresponding to the insertion of one additional repeat unit. Next, AR3 A1 was challenged with φA7 to obtain "second order" BIM AR3-3 A1A7, which was resistant to all of the eight phages. Strain AR3-3 A1A7 was found to have acquired only another 65 bp in its CRISPR1 locus with no change in CRISPR3. The newly acquired sequences in CRISPR1 and CRISPR3 of AR3 A1 and AR3-3 A1A7 showed similarity to sequences in already published S. thermophilus phage genomes. The phage-resistant mutant AR3-3 A1A7 acidified milk at a rate comparable to the parent strain LBB.A and, consequently, AR3-3 A1A7 was recommended as a potential replacement of the sensitive parent strain in yogurt production. The difference in the size of the amplified region of CRISPR1 and CRISPR3 can be used as a genetic marker to differentiate between BIMs and their phage sensitive parent strain.


Urshev Z.,LB Bulgaricum Plc | Ninova-Nikolova N.,LB Bulgaricum Plc | Ishlimova D.,LB Bulgaricum Plc | Pashova-Baltova K.,LB Bulgaricum Plc | And 2 more authors.
Biotechnology and Biotechnological Equipment | Year: 2014

Among Streptococcus thermophilus cultures, the principle component of yoghurt and cheese starters, a minority of strains forms the group of ‘H’-strains which show an unusually high acidification rate, grow faster and coagulate milk 3–5 hours earlier than the typical S. thermophilus cultures. A large-scale screening study was performed to select ‘H’-strains of S. thermophilus from more than 100 samples of home-made yoghurt, industrial yoghurt starters and single cultures, maintained in the LBB culture collection. Only four strains – LBB.TN1, LBB.M23, LBB.M34 and LBB.M60 – were isolated/selected due to their ability to form large yellowish colonies on milk agar, supplemented with beta-glycerophosphate and bromocresol purple. While in general S. thermophilus is described as a species with limited proteolytic capacity and in contrast to all other tested S. thermophilus cultures, the four selected strains invariably gave positive amplification product with the polymerase chain reaction when primers, specific for the membrane proteinase-coding gene prtS were used. The macrorestriction profiles of the genomic DNA of the four strains confirmed that they are non-isogenic and not related to each other. When grown in milk and compared to the control industrial strain LBB.A, the four strains showed a dramatically faster acidification, coagulating milk within four hours. The application of strain TN1 or M23 as adjunct culture to industrial yoghurt starter LBB.BY5-12 resulted in shortening the fermentation time with more than 30 min. © 2014 The Author(s).


Dimitrov Z.,LB Bulgaricum Plc.
Biotechnology and Biotechnological Equipment | Year: 2012

Strain-discriminative differentiation of Bifidobacteria strains by help of Amplified Fragment Length Polymorphism (AFLP) with satisfactory discriminative power and reproducibility was developed. AFLP genotyping is based on restriction cleavage of DNA with enzyme couple Xho I and Taq I, specially designed adapters, preselective and selective PCR primers. AFLP derived fragments were used successfully as a source of strain-specific markers for one probiotic Bifidobacterium longum strain, and subsequently, based on their specific sequences - for design of strain-specific probe. It’s specificity was confirmed upon 70 Bifidobacteria strains. © 2012 Taylor and Francis Group, LLC.


Zvancharova T.,LB Bulgaricum PLC | Baltova K.,LB Bulgaricum PLC | Urshev Z.,LB Bulgaricum PLC
Bulgarian Journal of Agricultural Science | Year: 2013

Originally, Bulgarian yoghurt was prepared from sheep's milk, but the homemade product had only a few days of shelf life. Today under industrial conditions, yoghurt with shelf life of 20 days is routinely produced. Nevertheless, producers and exporters are still interested in sheep's milk yoghurt with even longer storage period. In this study, we produced yoghurt with three selected starters and followed up its quality during three months of storage at 5°C. All three starters had low post acidification in the products. The freshly produced yoghurt samples had a pH of 4.17-4.39 which decreased gradually, but after 90 days it was still in the range 4.12-4.30 which was within the acceptable range. Viable cells of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus remained above the critical threshold of 108 and 106 cfu.ml-1 respectively for the whole period of 90 days. With the selected starters, the accumulation of products of proteolysis remained low with a slight increase only during the first month of storage. The limited proteolysis and low post acidification by the starters was achieved by the selection of weakly proteolytic strains of L. bulgaricus and/or a fermentation process with final viable cells of L. bulgaricus not exceeding 107 cfu.ml-1. Organoleptic tests did not reveal any adverse change of product taste and aroma of yoghurt samples at day 90. In conclusion, production of sheep's milk yoghurt with selected starters can give a product that will preserve its nutritional and biological value for as long as three months.


This invention relates to polybacterial probiotic preparation, which includes new strains of lactic acid bacteria Lactobacillus gasseri 7/12 NBIMCC No. 8720, Lactobacillus plantarum F12 NBIMCC No 8722 and Lactobacillus helveticus A1, NBIMCC No 8721 and has the following set of properties: antioxidant, hypocholesterolemic effect, anti-inflammatory immunomodulating effect, ability to release bioactive peptides that inhibit angiotensin-converting enzyme (ACE), which is responsible for the high blood pressure. The object of invention is also the described probiotic strains of lactic acid bacteria and the developed bacterial products with useful properties. The incorporated probiotic strain Lactobacillus gasseri 7/12 NBIMCC No. 8720, has a number of properties: strong adhesion to the epithelial layer of the colon, the ability for reduction of cholesterol and anti-inflammatory effects by reducing the levels of proinflammatory cytokine interleukin-8. The invention relates to the use of strains of lactic acid bacteria and bacterial agents in the production of starter cultures, food additives, food, functional products and pharmaceutical preparations intended to influence the health of humans and animals.


PubMed | LB Bulgaricum Plc.
Type: Journal Article | Journal: Biotechnology, biotechnological equipment | Year: 2015

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