scholarly journals Unleashing natural competence inLactococcus lactisby induction of the competence regulator ComX

2017 ◽  
Author(s):  
Joyce Mulder ◽  
Michiel Wels ◽  
Oscar P. Kuipers ◽  
Michiel Kleerebezem ◽  
Peter A. Bron
Keyword(s):  
Lactococcus Lactis ◽  
Genetic Manipulation ◽  
Bacterial Species ◽  
Streptococcus Thermophilus ◽  
Starter Cultures ◽  
Natural Competence ◽  
Competence Gene ◽  
Gene Sets ◽  
Novel Traits ◽  
Derivatives Of

AbstractIn biotechnological work horses likeStreptococcus thermophilusandBacillus subtilisnatural competence can be induced, which facilitates genetic manipulation of these microbes. However, in strains of the important dairy starterLactococcus lactisnatural competence has not been established to date. However,in silicoanalysis of complete genome sequences of 43L. lactisstrains revealed complete late-competence gene-sets in 2L. lactissubspeciescremorisstrains (KW2 and KW10) and 8L. lactissubspecieslactisstrains, including the model strain IL1403 and the plant-derived strain KF147. The remainder of the strains, including all dairy isolates, displayed genomic decay in one or more of the late competence genes. Nisin-controlled expression of the competence regulatorcomXinL. lactissubsp.lactisKF147 resulted in the induction of expression of the canonical competence regulon, and elicited a state of natural competence in this strain. By contrast,comXexpression inL. lactisNZ9000, predicted to encode an incomplete competence gene-set, failed to induce natural competence. Moreover, mutagenesis of thecomEA-ECoperon in strain KF147, abolished thecomXdriven natural competence, underpinning the involvement of the competence machinery. Finally, introduction of nisin-induciblecomXexpression intonisRK-harboring derivatives of strains IL1403 and KW2 allowed the induction of natural competence also in these strains, expanding this phenotype to otherL. lactisstrains of both subspecies.Significance statementSpecific bacterial species are able to enter a state of natural competence in which DNA is taken up from the environment, allowing the introduction of novel traits. Strains of the speciesLactococcus lactisare very important starter cultures for the fermentation of milk in the cheese production process, where these bacteria contribute to the flavor and texture of the end-product. The activation of natural competence in this industrially relevant organism can accelerate research aiming to understand industrially relevant traits of these bacteria, and can facilitate engineering strategies to harness the natural biodiversity of the species in optimized starter strains.

scholarly journals Unleashing Natural Competence in Lactococcus lactis by Induction of the Competence Regulator ComX

2017 ◽  
Vol 83 (20) ◽  
Author(s):  
Joyce Mulder ◽  
Michiel Wels ◽  
Oscar P. Kuipers ◽  
Michiel Kleerebezem ◽  
Peter A. Bron
Keyword(s):  
Lactococcus Lactis ◽  
Genetic Manipulation ◽  
Bacterial Species ◽  
Streptococcus Thermophilus ◽  
Starter Cultures ◽  
Natural Competence ◽  
Content Type ◽  
Competence Gene ◽  
Gene Sets ◽  
Novel Traits

ABSTRACT In biotechnological workhorses like Streptococcus thermophilus and Bacillus subtilis, natural competence can be induced, which facilitates genetic manipulation of these microbes. However, in strains of the important dairy starter Lactococcus lactis, natural competence has not been established to date. However, in silico analysis of the complete genome sequences of 43 L. lactis strains revealed complete late competence gene sets in 2 L. lactis subsp. cremoris strains (KW2 and KW10) and at least 10 L. lactis subsp. lactis strains, including the model strain IL1403 and the plant-derived strain KF147. The remainder of the strains, including all dairy isolates, displayed genomic decay in one or more of the late competence genes. Nisin-controlled expression of the competence regulator comX in L. lactis subsp. lactis KF147 resulted in the induction of expression of the canonical competence regulon and elicited a state of natural competence in this strain. In contrast, comX expression in L. lactis NZ9000, which was predicted to encode an incomplete competence gene set, failed to induce natural competence. Moreover, mutagenesis of the comEA-EC operon in strain KF147 abolished the comX-driven natural competence, underlining the involvement of the competence machinery. Finally, introduction of nisin-inducible comX expression into nisRK-harboring derivatives of strains IL1403 and KW2 allowed the induction of natural competence in these strains also, expanding this phenotype to other L. lactis strains of both subspecies. IMPORTANCE Specific bacterial species are able to enter a state of natural competence in which DNA is taken up from the environment, allowing the introduction of novel traits. Strains of the species Lactococcus lactis are very important starter cultures for the fermentation of milk in the cheese production process, where these bacteria contribute to the flavor and texture of the end product. The activation of natural competence in this industrially relevant organism can accelerate research aiming to understand industrially relevant traits of these bacteria and can facilitate engineering strategies to harness the natural biodiversity of the species in optimized starter strains.

scholarly journals A Cryptic Non-Inducible Prophage Confers Phage-Immunity on the Streptococcus thermophilus M17PTZA496

Viruses ◽  
2018 ◽  
Vol 11 (1) ◽  
pp. 7 ◽  
Author(s):  
Vinícius da Silva Duarte ◽  
Sabrina Giaretta ◽  
Stefano Campanaro ◽  
Laura Treu ◽  
Andrea Armani ◽  
...  
Keyword(s):  
Mitomycin C ◽  
Defense Mechanisms ◽  
Large Subunit ◽  
Bacterial Species ◽  
Dairy Industry ◽  
Model Organism ◽  
Streptococcus Thermophilus ◽  
Starter Cultures ◽  
Economic Losses ◽  
Important Species

Streptococcus thermophilus is considered one of the most important species for the dairy industry. Due to their diffusion in dairy environments, bacteriophages can represent a threat to this widely used bacterial species. Despite the presence of a CRISPR-Cas system in the S. thermophilus genome, some lysogenic strains harbor cryptic prophages that can increase the phage-host resistance defense. This characteristic was identified in the dairy strain S. thermophilus M17PTZA496, which contains two integrated prophages 51.8 and 28.3 Kb long, respectively. In the present study, defense mechanisms, such as a lipoprotein-encoding gene and Siphovirus Gp157, the last associated to the presence of a noncoding viral DNA element, were identified in the prophage M17PTZA496 genome. The ability to overexpress genes involved in these defense mechanisms under specific stressful conditions, such as phage attack, has been demonstrated. Despite the addition of increasing amounts of Mitomycin C, M17PTZA496 was found to be non-inducible. However, the transcriptional activity of the phage terminase large subunit was detected in the presence of the antagonist phage vB_SthS-VA460 and of Mitomycin C. The discovery of an additional immune mechanism, associated with bacteriophage-insensitive strains, is of utmost importance, for technological applications and industrial processes. To our knowledge, this is the first study reporting the capability of a prophage integrated into the S. thermophilus genome expressing different phage defense mechanisms. Bacteriophages are widespread entities that constantly threaten starter cultures in the dairy industry. In cheese and yogurt manufacturing, the lysis of Streptococcus thermophilus cultures by viral attacks can lead to huge economic losses. Nowadays S. thermophilus is considered a well-stablished model organism for the study of natural adaptive immunity (CRISPR-Cas) against phage and plasmids, however, the identification of novel bacteriophage-resistance mechanisms, in this species, is strongly desirable. Here, we demonstrated that the presence of a non-inducible prophage confers phage-immunity to an S. thermophilus strain, by the presence of ltp and a viral noncoding region. S. thermophilus M17PTZA496 arises as an unconventional model to study phage resistance and potentially represents an alternative starter strain for dairy productions.

scholarly journals Insertional Mutagenesis To Generate Lantibiotic Resistance in Lactococcus lactis

2007 ◽  
Vol 73 (14) ◽  
pp. 4677-4680 ◽  
Author(s):  
Caitriona M. Guinane ◽  
Paul D. Cotter ◽  
Elaine M. Lawton ◽  
Colin Hill ◽  
R. Paul Ross
Keyword(s):  
Lactococcus Lactis ◽  
Pathogenic Bacteria ◽  
Insertional Mutagenesis ◽  
Starter Cultures ◽  
Food Spoilage ◽  
Genetic Loci ◽  
The Creation ◽  
Derivatives Of

ABSTRACT While the potential emergence of food spoilage and pathogenic bacteria with resistance to lantibiotics is a concern, the creation of derivatives of starter cultures and adjuncts that can grow in the presence of these antimicrobials may have applications in food fermentations. Here a bank of Lactococcus lactis IL1403 mutants was created and screened, and a number of novel genetic loci involved in lantibiotic resistance were identified.

scholarly journals Importância das Bactérias Ácido Láticas e não Starter (NSLAB) na Tecnologia de Produção dos Derivados Lácteos

2020 ◽  
Vol 24 (4) ◽  
pp. 348-352
Author(s):  
Maria Tereza Pereira ◽  
Elsa Helena Walter de Santana ◽  
Joice Sifuentes dos Santos
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactococcus Lactis ◽  
Lactobacillus Casei ◽  
Streptococcus Thermophilus ◽  
Raw Milk ◽  
Fermented Milk ◽  
Starter Cultures ◽  
Bacillus Spp ◽  
Lactobacillus Spp

Produtos lácteos fermentados contêm bactérias ácido lácticas (BAL), naturalmente presentes ou adicionadas na matriz láctea como culturas iniciadoras (starters), contribuindo com aroma, textura, valor nutricional e segurança microbiológica. Lactobacillus spp., Streptococcus spp., Lactococcus spp. e Leuconostoc spp. são utilizados como culturas starters em laticínios. As BAL podem ser classificadas em mesofílicas (ex Lactococcus lactis) e termofílicas (ex Streptococcus thermophilus), e de acordo com seus metabólitos de fermentação em homofermentativas (ácido lático) e heterofermentativas (ácido lático, dióxido de carbono, diacetil e outros compostos flavorizantes). Entre as BAL há um grupo de bactérias lácticas que não fazem parte da cultura láctica (non starter lactic acid bacteria - NSLAB), que são oriundas do leite cru, do ambiente de ordenha ou da indústria formando biofilmes. As NSLAB são representadas por espécies heterofermentativas de lactobacilos mesofílicos como Lactobacillus casei spp., L. paracasei spp., L. rhamnosus spp. e L. plantarum spp., e ainda por Pediococcus spp., Leuconostoc spp. e Micrococcus spp. NSLAB termoduricas como Bacillus spp. também são relatadas. As NSLAB em queijos podem ajudar a desenvolver sabor e aroma, porém também são associadas aos defeitos em queijos e leites fermentados. Problemas como odores estranhos, sabor amargo ou muito ácido, perda de viscosidade, perda de coloração, estufamento e formação de gás são associados com a presença e contaminação por NSLAB. Assim, as BAL são importantes micro-organismos na indústria láctea, garantindo sabores e aromas aos derivados. Já a presença de NSLAB podem ser associados com defeitos em queijos e leites fermentados, sendo um problema na indústria beneficiadora.   Palavras-chave: Característica Sensorial. Leites Fermentados. Queijo. Textura.                       Abstract Fermented dairy products contain acid bacteria (BAL) naturally present or added to the dairy matrix as starter cultures (starters), contributing to aroma, texture, nutritional value and microbiological safety. Lactobacillus spp., Streptococcus spp., Lactococcus spp. and Leuconostoc spp. are used as starter dairy crops. As BAL it can be classified as mesophilic (ex: Lactococcus lactis) and thermophilic (ex: Streptococcus thermophilus), and agree with its fermentation metabolites in homofermentative (lactic acid) and heterofermentative (lactic acid, carbon dioxide, diacetyl and other flavorings). Among the BAL, there is a group of lactic bacteria that are not part of the dairy culture (non-initiating lactic acid bacteria - NSLAB) that originate from raw milk, the milking environment or the biofilm-forming industry. NSLAB is represented by heterofermentative species of mesophilic lactobacilli such as Lactobacillus casei spp., L. paracasei spp., L. rhamnosus spp. and L. plantarum spp., and also by Pediococcus spp., Leuconostoc spp. and Micrococcus spp. Termoduric NSLAB such as Bacillus spp. are also related. NSLAB in cheeses may help develop flavor and aroma, and they are also associated with defects in fermented cheeses and milks. Problems such as strange odors, bitter or very acidic taste, loss of viscosity, loss of color, establishment and gas training are associated with the presence and contamination by NSLAB. Thus,  BALs are important microorganisms in the dairy industry, contributing to the dairy flavors and aromas. The presence of NSLAB, on the other hand, can be associated with defects in fermented milk and cheese, being a problem in the processing industry.   Keywords: Cheese. Fermented Milk. Sensory Characteristic. Texture.

scholarly journals Functional strain redundancy and persistent phage infection in Swiss hard cheese starter cultures

2021 ◽  
Author(s):  
Vincent Somerville ◽  
Hélène Berthoud ◽  
Remo S. Schmidt ◽  
Hans-Peter Bachmann ◽  
Yi Hélène Meng ◽  
...  
Keyword(s):  
Microbial Communities ◽  
Evolutionary Dynamics ◽  
Temporal Dynamics ◽  
Negative Impact ◽  
Bacterial Species ◽  
Starter Culture ◽  
Streptococcus Thermophilus ◽  
Starter Cultures ◽  
Lactobacillus Delbrueckii ◽  
Shotgun Metagenomics

AbstractUndefined starter cultures are poorly characterized bacterial communities from environmental origin used in cheese making. They are phenotypically stable and have evolved through domestication by repeated propagation in closed and highly controlled environments over centuries. This makes them interesting for understanding eco-evolutionary dynamics governing microbial communities. While cheese starter cultures are known to be dominated by a few bacterial species, little is known about the composition, functional relevance, and temporal dynamics of strain-level diversity. Here, we applied shotgun metagenomics to an important Swiss cheese starter culture and analyzed historical and experimental samples reflecting 82 years of starter culture propagation. We found that the bacterial community is highly stable and dominated by only a few coexisting strains of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. lactis. Genome sequencing, metabolomics analysis, and co-culturing experiments of 43 isolates show that these strains are functionally redundant, but differ tremendously in their phage resistance potential. Moreover, we identified two highly abundant Streptococcus phages that seem to stably coexist in the community without any negative impact on bacterial growth or strain persistence, and despite the presence of a large and diverse repertoire of matching CRISPR spacers. Our findings show that functionally equivalent strains can coexist in domesticated microbial communities and highlight an important role of bacteria-phage interactions that are different from kill-the-winner dynamics.

scholarly journals Microencapsulation of Lactococcus lactis Gh1 with Gum Arabic and Synsepalum dulcificum via Spray Drying for Potential Inclusion in Functional Yogurt

Molecules ◽  
2019 ◽  
Vol 24 (7) ◽  
pp. 1422 ◽  
Author(s):  
Nurul Farhana Fazilah ◽  
Nurmelissa Hanani Hamidon ◽  
Arbakariya B. Ariff ◽  
Mohd Ezuan Khayat ◽  
Helmi Wasoh ◽  
...  
Keyword(s):  
Spray Drying ◽  
Lactococcus Lactis ◽  
Gum Arabic ◽  
Streptococcus Thermophilus ◽  
Viable Cell ◽  
Starter Cultures ◽  
Lactobacillus Delbrueckii ◽  
Bacterial Cells ◽  
Cell Counts

There has been an explosion of probiotic incorporated based product. However, many reports indicated that most of the probiotics have failed to survive in high quantity, which has limited their effectiveness in most functional foods. Thus, to overcome this problem, microencapsulation is considered to be a promising process. In this study, Lactococcus lactis Gh1 was encapsulated via spray-drying with gum Arabic together with Synsepalum dulcificum or commonly known as miracle fruit. It was observed that after spray-drying, high viability (~109 CFU/mL) powders containing L. lactis in combination with S. dulcificum were developed, which was then formulated into yogurt. The tolerance of encapsulated bacterial cells in simulated gastric juice at pH 1.5 was tested in an in-vitro model and the result showed that after 2 h, cell viability remained high at 1.11 × 106 CFU/mL. Incubation of encapsulated cells in the presence of 0.6% (w/v) bile salts showed it was able to survive (~104 CFU/mL) after 2 h. Microencapsulated L. lactis retained a higher viability, at ~107 CFU/mL, when incorporated into yogurt compared to non-microencapsulated cells ~105 CFU/mL. The fortification of microencapsulated and non-microencapsulated L. lactis in yogurts influenced the viable cell counts of yogurt starter cultures, Lactobacillus delbrueckii subs. bulgaricus and Streptococcus thermophilus.

scholarly journals In Silico Prediction of Horizontal Gene Transfer Events in Lactobacillus bulgaricus and Streptococcus thermophilus Reveals Protocooperation in Yogurt Manufacturing

2009 ◽  
Vol 75 (12) ◽  
pp. 4120-4129 ◽  
Author(s):  
Mengjin Liu ◽  
Roland J. Siezen ◽  
Arjen Nauta
Keyword(s):  
Gene Transfer ◽  
Horizontal Gene Transfer ◽  
Gene Cluster ◽  
In Silico ◽  
Bacterial Species ◽  
Streptococcus Thermophilus ◽  
Starter Cultures ◽  
Lactobacillus Helveticus ◽  
Lactobacillus Bulgaricus ◽  
Genetic Level

ABSTRACT Lactobacillus bulgaricus and Streptococcus thermophilus, used in yogurt starter cultures, are well known for their stability and protocooperation during their coexistence in milk. In this study, we show that a close interaction between the two species also takes place at the genetic level. We performed an in silico analysis, combining gene composition and gene transfer mechanism-associated features, and predicted horizontally transferred genes in both L. bulgaricus and S. thermophilus. Putative horizontal gene transfer (HGT) events that have occurred between the two bacterial species include the transfer of exopolysaccharide (EPS) biosynthesis genes, transferred from S. thermophilus to L. bulgaricus, and the gene cluster cbs-cblB(cglB)-cysE for the metabolism of sulfur-containing amino acids, transferred from L. bulgaricus or Lactobacillus helveticus to S. thermophilus. The HGT event for the cbs-cblB(cglB)-cysE gene cluster was analyzed in detail, with respect to both evolutionary and functional aspects. It can be concluded that during the coexistence of both yogurt starter species in a milk environment, agonistic coevolution at the genetic level has probably been involved in the optimization of their combined growth and interactions.

scholarly journals Novel Variants of Streptococcus thermophilus Bacteriophages Are Indicative of Genetic Recombination among Phages from Different Bacterial Species

2016 ◽  
Vol 83 (5) ◽  
Author(s):  
Paula Szymczak ◽  
Thomas Janzen ◽  
Ana Rute Neves ◽  
Witold Kot ◽  
Lars H. Hansen ◽  
...  
Keyword(s):  
Genetic Recombination ◽  
Genetic Relationships ◽  
Bacterial Species ◽  
Streptococcus Thermophilus ◽  
Starter Cultures ◽  
Sequencing Analysis ◽  
Content Type ◽  
Novel Variants ◽  
Positive Results

ABSTRACT Bacteriophages are the main cause of fermentation failures in dairy plants. The majority of Streptococcus thermophilus phages can be divided into either cos- or pac-type phages and are additionally characterized by examining the V2 region of their antireceptors. We screened a large number of S. thermophilus phages from the Chr. Hansen A/S collection, using PCR specific for the cos- or pac-type phages, as well as for the V2 antireceptor region. Three phages did not produce positive results with the assays. Analysis of phage morphologies indicated that two of these phages, CHPC577 and CHPC926, had shorter tails than the traditional S. thermophilus phages. The third phage, CHPC1151, had a tail size similar to those of the cos- or pac-type phages, but it displayed a different baseplate structure. Sequencing analysis revealed the genetic similarity of CHPC577 and CHPC926 with a subgroup of Lactococcus lactis P335 phages. Phage CHPC1151 was closely related to the atypical S. thermophilus phage 5093, homologous with a nondairy streptococcal prophage. By testing adsorption of the related streptococcal and lactococcal phages to the surface of S. thermophilus and L. lactis strains, we revealed the possibility of cross-interactions. Our data indicated that the use of S. thermophilus together with L. lactis, extensively applied for dairy fermentations, triggered the recombination between phages infecting different bacterial species. A notable diversity among S. thermophilus phage populations requires that a new classification of the group be proposed. IMPORTANCE Streptococcus thermophilus is a component of thermophilic starter cultures commonly used for cheese and yogurt production. Characterizing streptococcal phages, understanding their genetic relationships, and studying their interactions with various hosts are the necessary steps for preventing and controlling phage attacks that occur during dairy fermentations.

scholarly journals Cell Surface Polysaccharides Represent a Common Strategy for Adsorption among Phages Infecting Lactic Acid Bacteria: Lessons from Dairy Lactococci and Streptococci

mSystems ◽  
2021 ◽  
Vol 6 (4) ◽  
Author(s):  
Jennifer Mahony
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Cell Surface ◽  
Lactococcus Lactis ◽  
Streptococcus Thermophilus ◽  
Starter Cultures ◽  
Content Type ◽  
Common Strategy ◽  
Surface Polysaccharides

Food fermentations rely on the application of robust bacterial starter cultures, the majority of which are represented by members of the lactic acid bacteria including Lactococcus lactis and Streptococcus thermophilus . Bacteriophage (or phage) proliferation remains one of the most significant threats to the fermentation industry.

scholarly journals Functional strain redundancy and persistent phage infection in Swiss hard cheese starter cultures

2021 ◽  
Author(s):  
Vincent Somerville ◽  
Hélène Berthoud ◽  
Remo S. Schmidt ◽  
Hans- Peter Bachmann ◽  
Yi Hélène Meng ◽  
...  
Keyword(s):  
Microbial Communities ◽  
Evolutionary Dynamics ◽  
Temporal Dynamics ◽  
Negative Impact ◽  
Bacterial Species ◽  
Starter Culture ◽  
Streptococcus Thermophilus ◽  
Starter Cultures ◽  
Lactobacillus Delbrueckii ◽  
Shotgun Metagenomics

AbstractUndefined starter cultures are poorly characterized bacterial communities from environmental origin used in cheese making. They are phenotypically stable and have evolved through domestication by repeated propagation in closed and highly controlled environments over centuries. This makes them interesting for understanding eco-evolutionary dynamics governing microbial communities. While cheese starter cultures are known to be dominated by a few bacterial species, little is known about the composition, functional relevance, and temporal dynamics of strain-level diversity. Here, we applied shotgun metagenomics to an important Swiss cheese starter culture and analyzed historical and experimental samples reflecting 82 years of starter culture propagation. We found that the bacterial community is highly stable and dominated by only a few coexisting strains of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. lactis. Genome sequencing, metabolomics analysis, and co-culturing experiments of 43 isolates show that these strains are functionally redundant, but differ tremendously in their phage resistance potential. Moreover, we identified two highly abundant Streptococcus phages that seem to stably coexist in the community without any negative impact on bacterial growth or strain persistence, and despite the presence of a large and diverse repertoire of matching CRISPR spacers. Our findings show that functionally equivalent strains can coexist in domesticated microbial communities and highlight an important role of bacteria-phage interactions that are different from kill-the-winner dynamics.

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