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1.
Microb Genom ; 7(4)2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33900907

RESUMO

Lactobacillus helveticus is a well characterized lactobacillus for dairy fermentations that is also found in malt whisky fermentations. The two environments contain considerable differences related to microbial growth, including the presence of different growth inhibitors and nutrients. The present study characterized L. helveticus strains originating from dairy fermentations (called milk strains hereafter) and malt whisky fermentations (called whisky strains hereafter) by in vitro phenotypic tests and comparative genomics. The whisky strains can tolerate ethanol more than the milk strains, whereas the milk strains can tolerate lysozyme and lactoferrin more than the whisky strains. Several plant-origin carbohydrates, including cellobiose, maltose, sucrose, fructooligosaccharide and salicin, were generally metabolized only by the whisky strains, whereas milk-derived carbohydrates, i.e. lactose and galactose, were metabolized only by the milk strains. Milk fermentation properties also distinguished the two groups. The general genomic characteristics, including genomic size, number of coding sequences and average nucleotide identity values, differentiated the two groups. The observed differences in carbohydrate metabolic properties between the two groups correlated with the presence of intact specific enzymes in glycoside hydrolase (GH) families GH1, GH4, GH13, GH32 and GH65. Several GHs in the milk strains were inactive due to the presence of stop codon(s) in genes encoding the GHs, and the inactivation patterns of the genes encoding specific enzymes assigned to GH1 in the milk strains suggested a possible diversification manner of L. helveticus strains. The present study has demonstrated how L. helveticus strains have adapted to their habitats.


Assuntos
Lactobacillus helveticus/isolamento & purificação , Lactobacillus helveticus/fisiologia , Leite/microbiologia , Vinho/microbiologia , Adaptação Fisiológica , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Bovinos , Etanol/metabolismo , Fermentação , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/metabolismo , Lactobacillus helveticus/classificação , Lactobacillus helveticus/genética
2.
PLoS One ; 16(1): e0245903, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33493208

RESUMO

Lactobacillus helveticus carries many properties such as the ability to survive gastrointestinal transit, modulate the host immune response, accumulate biopeptides in milk, and adhere to the epithelial cells that could contribute to improving host health. In this study, the applicability as functional cultures of four L. helveticus strains isolated from Italian hard cheeses was investigated. A preliminary strain characterization showed that the ability to produce folate was generally low while antioxidant, proteolytic, peptidase, and ß-galactosidase activities resulted high, although very variable, between strains. When stimulated moDCs were incubated in the presence of live cells, a dose-dependent release of both the pro-inflammatory cytokine IL-12p70 and the anti-inflammatory cytokine IL-10, was shown for all the four strains. In the presence of cell-free culture supernatants (postbiotics), a dose-dependent, decrease of IL-12p70 and an increase of IL-10 was generally observed. The immunomodulatory effect took place also in Caciotta-like cheese made with strains SIM12 and SIS16 as bifunctional (i.e., immunomodulant and acidifying) starter cultures, thus confirming tests in culture media. Given that the growth of bacteria in the cheese was not necessary (they were killed by pasteurization), the results indicated that some constituents of non-viable bacteria had immunomodulatory properties. This study adds additional evidence for the positive role of L. helveticus on human health and suggests cheese as a suitable food for delivering candidate strains and modulating their anti-inflammatory properties.


Assuntos
Queijo/microbiologia , Lactobacillus helveticus/isolamento & purificação , Microbiologia de Alimentos , Humanos , Itália , Lactobacillus helveticus/genética , Lactobacillus helveticus/metabolismo , Leucócitos Mononucleares/metabolismo
3.
Food Microbiol ; 94: 103651, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33279076

RESUMO

Lactobacillus (L.) helveticus is widely used in food industry due to its high proteolytic activity. However, such activity varies greatly between isolates, and the determining factors regulating the strength of proteolytic activity in L. helveticus are unclear. This study sequenced the genomes of 60 fermented food-originated L. helveticus and systemically examined the proteolytic activity-determining factors. Our analyses found that the strength of proteolytic activity in L. helveticus was independent of the isolation source, geographic location, phylogenetic closeness between isolates, and distribution of cell envelope proteinases (CEPs). Genome-wide association study (GWAS) identified two genes, the acetate kinase (ackA) and a hypothetical protein, and 15 single nucleotide polymorphisms (SNPs) that were associated with the strength of the proteolytic activity. Further investigating the functions of these gene components revealed that ackA and two cysteine peptidases coding genes (pepC and srtA) rather than the highly heterogeneous and intraspecific CEPs were linked to the level of proteolytic activity. Moreover, the sequence type (ST) defined by SNP analysis revealed a total of ten STs, and significantly weaker proteolytic activity was observed among isolates of ST2. This study provides practical information for future selection of L. helveticus of strong proteolytic activity.


Assuntos
Acetato Quinase/metabolismo , Proteínas de Bactérias/metabolismo , Laticínios/microbiologia , Grão Comestível/microbiologia , Alimentos Fermentados/microbiologia , Lactobacillus helveticus/enzimologia , Peptídeo Hidrolases/metabolismo , Acetato Quinase/química , Acetato Quinase/genética , Animais , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Bovinos , Genoma Bacteriano , Estudo de Associação Genômica Ampla , Lactobacillus helveticus/genética , Lactobacillus helveticus/isolamento & purificação , Lactobacillus helveticus/metabolismo , Peptídeo Hidrolases/química , Peptídeo Hidrolases/genética , Filogenia , Proteólise
4.
Food Microbiol ; 89: 103457, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32139001

RESUMO

Monitoring L. helveticus strain dynamics in natural whey starters is of great interest at the industrial level due to the key role that this bacterial population plays in Grana Padano cheese production. In this study, we aimed to develop a PCR-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) assay based on the slpH locus, in parallel with performing culture-dependent analysis of whey samples using optimized media to maximize the number of isolated strains. We designed new primers targeting the slpH locus to amplify a gene region that would be suitable for PCR-DGGE analysis and discriminating strains. Our results confirmed that the developed PCR-DGGE method was rapid and reliable for monitoring the L. helveticus population in whey starter cultures. All sequences of bands detected in the PCR-DGGE profiles from whey samples showed high similarity to S-layer genes of L. helveticus, and perfectly matched with the slpH locus sequences of dominant strains. Overall, our findings indicated that the target region of the slpH locus was sufficiently heterologous to discriminate L. helveticus strains, and that our PCR-DGGE analysis provided a more accurate picture of the population composition of whey starters compared to culture-dependent techniques that often fail to isolate the most abundant strains.


Assuntos
Eletroforese em Gel de Gradiente Desnaturante/métodos , Lactobacillus helveticus/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Soro do Leite/microbiologia , Técnicas de Tipagem Bacteriana , Queijo , DNA Bacteriano/análise , Lactobacillus helveticus/classificação
5.
Biomed Res Int ; 2020: 8889198, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33426082

RESUMO

Probiotic bacteria are of utmost importance owing to their extensive utilisation in dairy products and in the prevention of various intestinal diseases. The objective of this study was to assess the probiotic properties of bacteriocin-producing isolates of Lactobacillus helveticus and Lactobacillus plantarum isolated from traditional Pakistani yoghurt. In this study, ten bacteriocin-producing isolates were selected to screen for the probiotic property. The isolates showed resistance to acidic pH (6-6.5), bile salt (0.01-1%), and 1-7% NaCl salt and showed good growth at acidic pH and antibacterial activity against ten different foodborne pathogens. Interestingly, these isolates were proved to be effective against Actinobacter baumannii but least effective against Klebsiella pneumoniae and Pseudomonas aeruginosa. A few isolates were found to be resistant to some antibiotics like vancomycim, gentamycin, erythromycin, streptomycin, and clindamycin. Our results provide strong evidence in favour of traditional Pakistani yoghurts as a potential source of bacteriocin-producing bacteria with an added benefit of the probiotic property. Specifically, LBh5 was considered a good probiotic isolate as compared to other isolates used in the study. Further extensive research should be done on isolation and characterisation of probiotic isolates from local fermented foods, and then, these isolates should be used in the development of probiotic enriched food supplements in Pakistan.


Assuntos
Lactobacillus helveticus , Lactobacillus plantarum , Probióticos , Iogurte/microbiologia , Antibacterianos/farmacologia , Bacteriocinas/metabolismo , Farmacorresistência Bacteriana , Lactobacillus helveticus/efeitos dos fármacos , Lactobacillus helveticus/isolamento & purificação , Lactobacillus helveticus/metabolismo , Lactobacillus helveticus/fisiologia , Lactobacillus plantarum/efeitos dos fármacos , Lactobacillus plantarum/isolamento & purificação , Lactobacillus plantarum/metabolismo , Lactobacillus plantarum/fisiologia , Paquistão , Tolerância ao Sal
6.
J Appl Microbiol ; 124(2): 572-584, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29236340

RESUMO

AIMS: Classical microbiology techniques are the gold standard for probiotic enumeration. However, these techniques are limited by parameters of time, specificity and incapacity to detect viable but nonculturable (VBNC) micro-organisms and nonviable cells. The aim of the study was to evaluate flow cytometry as a novel method for the specific quantification of viable and nonviable probiotics in multistrain products. METHODS AND RESULTS: Custom polyclonal antibodies were produced against five probiotic strains from different species (Bifidobacterium bifidum R0071, Bifidobacterium longum ssp. infantis R0033, Bifidobacterium longum ssp. longum R0175, Lactobacillus helveticus R0052 and Lactobacillus rhamnosus R0011). Evaluation of specificity confirmed that all antibodies were specific at least at the subspecies level. A flow cytometry method combining specific antibodies and viability assessment with SYTO® 24 and propidium iodide was applied to quantify these strains in three commercial products. Analyses were conducted on two flow cytometry instruments by two operators and compared with classical microbiology using selective media. Results indicated that flow cytometry provides higher cell counts than classical microbiology (P < 0·05) in 73% of cases highlighting the possible presence of VBNC. Equivalent performances (repeatability and reproducibility) were obtained for both methods. CONCLUSIONS: This study showed that flow cytometry methods can be applied to probiotic enumeration and viability assessment. Combination with polyclonal antibodies can achieve sufficient specificity to differentiate closely related strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Flow cytometry provides absolute and specific quantification of viable and nonviable probiotic strains in a very short time (<2 h) compared with classical techniques (>48 h), bringing efficient tools for research and development and quality control.


Assuntos
Bifidobacterium longum subspecies infantis/crescimento & desenvolvimento , Citometria de Fluxo/métodos , Lacticaseibacillus rhamnosus/crescimento & desenvolvimento , Lactobacillus helveticus/crescimento & desenvolvimento , Probióticos/química , Bifidobacterium longum subspecies infantis/química , Bifidobacterium longum subspecies infantis/isolamento & purificação , Lactobacillus helveticus/química , Lactobacillus helveticus/isolamento & purificação , Lacticaseibacillus rhamnosus/química , Lacticaseibacillus rhamnosus/isolamento & purificação , Viabilidade Microbiana , Reprodutibilidade dos Testes
7.
Appl Microbiol Biotechnol ; 101(20): 7621-7633, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28695230

RESUMO

In a screening for proteolytically active lactic acid bacteria, three strains, Lactobacillus delbrueckii ssp. lactis 92202, Lactobacillus helveticus 92201, and Lactobacillus delbrueckii ssp. bulgaricus 92059, showed the highest activities following growth in milk. All three strains degraded α- and ß-casein, but did not hydrolyse κ-casein. HPLC analysis of skim milk fermentation revealed increasing amounts of peptides after 5 and 10 h with Lb. d. ssp. bulgaricus 92059. Hydrolysates obtained with Lb. d. ssp. lactis 92202 and Lb. d. ssp. bulgaricus 92059 revealed the highest angiotensin-converting enzyme-inhibitory effect. The effect was dose dependent. Almost no effect (<10%) was seen for Lb. helveticus 92201. For Lb. d. ssp. bulgaricus 92059, maximal inhibition of approx. 65% was reached after 25 h of fermentation. In an in vitro assay measuring potential immunomodulation, hydrolysates of the three strains yielded anti-inflammatory activities in the presence of TNF-α. However, the effects were more pronounced at lower hydrolysate concentrations. In the absence of TNF-α, slight pro-inflammatory effects were observed. The hydrolysate of Lb. d. ssp. bulgaricus 92059, when purified by means of solid-phase extraction, exhibited pro-inflammatory activity. Sour whey containing Lb. d. ssp. bulgaricus 92059 cells showed pro-inflammatory activity while cell-free sour whey was clearly anti-inflammatory. In the purified hydrolysate, 20 different α- and ß-casein (CN)-derived peptides could be identified by LC-MS. Most peptides originated from the central and C-terminal regions of ß-casein. Peptide length was between 9 (ß-CN(f 59-67)) and 22 amino acids (ß-CN(f 117-138)).


Assuntos
Fatores Biológicos/metabolismo , Lactobacillus delbrueckii/isolamento & purificação , Lactobacillus helveticus/isolamento & purificação , Peptídeo Hidrolases/metabolismo , Peptídeos/metabolismo , Proteólise , Animais , Inibidores Enzimáticos/isolamento & purificação , Inibidores Enzimáticos/metabolismo , Fatores Imunológicos/isolamento & purificação , Fatores Imunológicos/metabolismo , Lactobacillus delbrueckii/enzimologia , Lactobacillus delbrueckii/crescimento & desenvolvimento , Lactobacillus delbrueckii/metabolismo , Lactobacillus helveticus/enzimologia , Lactobacillus helveticus/crescimento & desenvolvimento , Lactobacillus helveticus/metabolismo , Programas de Rastreamento , Leite/microbiologia , Peptidil Dipeptidase A/metabolismo , Fator de Necrose Tumoral alfa/antagonistas & inibidores
8.
Int J Food Microbiol ; 241: 173-180, 2017 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-27792948

RESUMO

Lactobacillus helveticus is widely used in dairy fermentations and produces a range of enzymes, which upon cell lysis can be released into the cheese matrix and impact degradation of proteins, peptides and lipids. In our study we set out to explore the potential of Lb. helveticus DSM 20075 for increased autolytic capacity triggered by conditions such as low pH and high salt concentrations encountered in cheese environments. Lb. helveticus DSM 20075 was subjected to varied incubation temperatures (ranging from 37 to 50°C). High-temperature incubation (in the range of 45 to 50°C) allowed us to obtain a collection of six variant strains (V45-V50), which in comparison to the wild-type strain, showed higher growth rates at elevated temperatures (42°C-45°C). Moreover, variant strain V50 showed a 4-fold higher, in comparison to wild type, autolytic capacity in cheese-like conditions. Next, strain V50 was used as an adjunct in lab-scale cheese making trials to measure its impact on aroma formation during ripening. Specifically, in cheeses made with strain V50, the relative abundance of benzaldehyde increased 3-fold compared to cheeses made with the wild-type strain. Analysis of the genome sequence of strain V50 revealed multiple mutations in comparison to the wild-type strain DSM 20075 including a mutation found in a gene coding for a metal ion transporter, which can potentially be linked to intracellular accumulation of Mn2+ and benzaldehyde formation. The approach of high-temperature incubation can be applied in dairy industry for the selection of (adjunct) cultures targeted at accelerated cheese ripening and aroma formation.


Assuntos
Bacteriólise , Queijo/microbiologia , Lactobacillus helveticus/citologia , Queijo/análise , Fermentação , Temperatura Alta , Lactobacillus helveticus/genética , Lactobacillus helveticus/isolamento & purificação , Lactobacillus helveticus/metabolismo , Proteínas do Leite/metabolismo
9.
J Biotechnol ; 224: 18-9, 2016 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-26953745

RESUMO

Here we report the complete genome sequence of Lactobacillus helveticus CAUH18, a new strain isolated from traditional fermented dairy product koumiss. Its genome has a circular 2.16Mb chromosome with no plasmid. The genome sequence indicated that this strain harbors a gene cluster involved in a novel exopolysaccharides (EPS) biosynthesis and a gene encoding cell-surface aggregation-promoting factors (APFs) to facilitate its colonization in gastrointestinal tract (GIT). This genome sequence provides a basis for further studies about its molecular genetics and probiotic functions.


Assuntos
Cromossomos Bacterianos/genética , Lactobacillus helveticus/isolamento & purificação , Análise de Sequência de DNA/métodos , Composição de Bases , Laticínios/microbiologia , Tamanho do Genoma , Lactobacillus helveticus/genética , Família Multigênica , Probióticos
10.
Benef Microbes ; 7(1): 3-10, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26503737

RESUMO

The aim of the studies was to determine the effects of calcium carbonate and calcium phosphate supplementation on faecal Lactobacillus spp., with and without a probiotic supplement, in healthy adults. Study 1 comprised of a randomised, double-blind, crossover design; participants (n=15) received 2 capsules/d of 250 mg elemental calcium as calcium carbonate (Ca1) and calcium phosphate (Ca2) each for 2-week periods, with 2-week baseline and washout periods. Study 2 was a randomised, double-blind, crossover design; participants (n=17) received 2 capsules/d of Lactobacillus helveticus R0052 and Lactobacillus rhamnosus R0011 (probiotic) alone, the probiotic with 2 capsules/d of Ca1, and probiotic with 2 capsules/d of Ca2 each for 2-week periods with 2-week baseline and washout periods. In both studies, stools were collected during the baseline, intervention and washout periods for Lactobacillus spp. quantification and qPCR analyses. Participants completed daily questionnaires of stool frequency and compliance. In Study 1, neither calcium supplement influenced viable counts of resident Lactobacillus spp., genome equivalents of lactic acid bacteria or stool frequency. In Study 2, faecal Lactobacillus spp. counts were significantly enhanced from baseline when the probiotic was administered with Ca2 (4.83±0.30, 5.79±0.31) (P=0.02), but not with Ca1 (4.98±0.31) or with the probiotic alone (5.36±0.31, 5.55±0.29) (not significant). Detection of L. helveticus R0052 and L. rhamnosus R0011 was significantly increased with all treatments, but did not differ among treatments. There were no changes in weekly stool frequency. Calcium phosphate co-administration may increase gastrointestinal survival of orally-administered Lactobacillus spp.


Assuntos
Fosfatos de Cálcio/farmacologia , Fezes/microbiologia , Lacticaseibacillus rhamnosus/efeitos dos fármacos , Lactobacillus helveticus/efeitos dos fármacos , Probióticos/farmacologia , Adolescente , Adulto , Estudos Cross-Over , Suplementos Nutricionais , Método Duplo-Cego , Feminino , Humanos , Lactobacillus helveticus/isolamento & purificação , Lacticaseibacillus rhamnosus/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Adulto Jovem
11.
Food Funct ; 7(2): 789-97, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26648272

RESUMO

Lactobacillus helveticus, an obligatory hetero-fermentative LAB, is Generally Recognized as Safe (GRAS) and is gaining popularity for application in dairy products. Lactic acid bacteria (LAB) play a remarkable role in inhibiting the growth of pathogenic bacteria in food products, without disturbing the sensory attributes of the food. In this study, the screening of the antimicrobial potential of Lactobacillus helveticus KLDS 1.8701 against four food-borne pathogens including Listeria monocytogenes ATCC 19115, Salmonella typhimurium ATCC 14028, Staphylococcus aureus ATCC 25923, and Escherichia coli O157:H7 ATCC 43889 in vitro was inspected using the Oxford cup method and mixed culture inhibition assays. The organic acid production and antimicrobial potential of the cell-free supernatants (CFS) have been evaluated via different treatments and analysis using high performance liquid chromatography (HPLC). The analysis results revealed that KLDS 1.8701 exhibited the highest antimicrobial potential compared to other antimicrobial strains. The antimicrobial activity of KLDS 1.8701 resulted from the organic acids in the culture and CFS. From the study, it was found that carbon sources, as well as organic acid production, accelerate the antimicrobial activity of KLDS 1.8701 and the fructooligosaccharides (FOS) were considered the best for improving the proliferation of KLDS 1.8701 and supporting its antimicrobial action. Results of the mixed culture inhibition assays showed that part of the antimicrobial activity resulted from the inhibitory action of the bacteria itself in culture, and this action required cellular contact between the food-borne pathogens and KLDS 1.8701. Conversely, the results of the antimicrobial spectrum assay revealed that some Lactobacilli remained unaffected by KLDS 1.8701. KLDS 1.8701 might also be favorable for use as a supplementary starter in fermented dairy productions. Furthermore, KLDS 1.8701 could survive well under GI tract conditions. Further studies on in vivo inhibition assays and the probiotic effects are recommended.


Assuntos
Anti-Infecciosos/farmacologia , Queijo/microbiologia , Doenças Transmitidas por Alimentos/microbiologia , Lactobacillus helveticus/metabolismo , Probióticos/farmacologia , Anti-Infecciosos/metabolismo , China , Escherichia coli O157/efeitos dos fármacos , Fermentação , Humanos , Lactobacillus helveticus/química , Lactobacillus helveticus/classificação , Lactobacillus helveticus/isolamento & purificação , Listeria monocytogenes/efeitos dos fármacos , Probióticos/metabolismo , Staphylococcus aureus/efeitos dos fármacos
12.
BMC Microbiol ; 15: 196, 2015 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-26428623

RESUMO

BACKGROUND: Koumiss is a traditionally fermented mare's milk described with health-promoting potentials for decades. However, only a few studies focused on the probiotic strains isolated from koumiss. In this study, we collected koumiss samples from Inner Mongolian pasturing area of China and selected a promising strain of Lactobacillus helveticus, isolate NS8, based on the survival abilities in gastrointestinal tract (GIT) and adhesion to intestinal endothelial cells in vitro. As the ability to positively modulate host immune response is a feature of increasing importance in measuring the probiotic potential of a bacterial strain, our study mainly focus on the immunomodulatory properties of L. helveticus NS8 by using in vivo and ex vivo analyses. RESULTS: L. helveticus NS8 was identified by molecular-typing methods, both at genus and species levels. As a typical food niche-specific bacteria, NS8 showed a moderate survival ability in GIT environment in vitro. However, an excellent binding capacity to the human intestinal epithelial cells, along with significant autoaggregation and cell-surface hydrophobicity was observed. Additionally, the presence of S-layer protein was responsible for the cell surface properties of this strain. NS8 was found to be rather protective against TNBS (2,4,6-trinitrobenzene sulfonic acid)-induced murine colitis. In the meantime, co-culture with NS8 induced an increased level of secretion of anti-inflammatory cytokine IL-10 in peripheral blood mono-nuclear cells (PBMCs). Furthermore, NS8 was also able to diminish the proinflammatory effects of lipopolysaccharide (LPS) in mouse macrophage cell line RAW264.7 by inducing higher levels of IL-10. Specially, adding of the purified S-layer protein didn't influence the production of IL-10. The specific ligand-host receptor interactions on the NS8 specific immune responses need to be learned further. CONCLUSION: In summary, L. helveticus NS8 exhibited good probiotic and particularly immunomodulatory properties, with a potential for development of functional food commercially or therapeutic adjuvant for inflammatory diseases.


Assuntos
Anti-Inflamatórios/uso terapêutico , Colite/prevenção & controle , Produtos Fermentados do Leite/microbiologia , Lactobacillus helveticus/isolamento & purificação , Probióticos/uso terapêutico , Animais , Anti-Inflamatórios/farmacologia , Aderência Bacteriana , Células CACO-2 , Células Cultivadas , Colite/induzido quimicamente , Modelos Animais de Doenças , Trato Gastrointestinal/microbiologia , Humanos , Interleucina-10/metabolismo , Lactobacillus helveticus/genética , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/microbiologia , Glicoproteínas de Membrana/metabolismo , Camundongos , Tipagem Molecular , Mongólia , Probióticos/farmacologia , Células RAW 264.7 , Ácido Trinitrobenzenossulfônico/efeitos adversos
13.
J Biotechnol ; 209: 14-5, 2015 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-26065338

RESUMO

Lactobacillus helveticus MB2-1 is a probiotic bacterium producing exopolysaccharides (EPS), which was isolated from traditional Sayram ropy fermented milk in southern Xinjiang, China. The genome consists of a circular 2,084,058bp chromosome with no plasmid. The genome sequence indicated that this strain includes a 15.20kb gene cluster involved in EPS biosynthesis. Genome sequencing information has provided the basis for understanding the potential molecular mechanism behind the EPS production.


Assuntos
Genoma Bacteriano , Lactobacillus helveticus/genética , Análise de Sequência de DNA/métodos , Produtos Fermentados do Leite/microbiologia , Tamanho do Genoma , Lactobacillus helveticus/isolamento & purificação , Lactobacillus helveticus/metabolismo , Família Multigênica , Polissacarídeos Bacterianos/metabolismo , Probióticos
14.
J Dairy Sci ; 98(5): 2962-72, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25726109

RESUMO

Lactobacillus helveticus is an economically important lactic acid bacterium used in industrial dairy fermentation. In the present study, the population structure of 245 isolates of L. helveticus from different naturally fermented dairy products in China and Mongolia were investigated using an multilocus sequence typing scheme with 11 housekeeping genes. A total of 108 sequence types were detected, which formed 8 clonal complexes and 27 singletons. Results from Structure, SplitsTree, and ClonalFrame software analyses demonstrated the presence of 3 subpopulations in the L. helveticus isolates used in our study, namely koumiss, kurut-tarag, and panmictic lineages. Most L. helveticus isolates from particular ecological origins had specific population structures.


Assuntos
Produtos Fermentados do Leite/microbiologia , Lactobacillus helveticus/genética , Tipagem de Sequências Multilocus , Animais , Bovinos , China , Fermentação , Microbiologia de Alimentos , Lactobacillus helveticus/classificação , Lactobacillus helveticus/isolamento & purificação , Mongólia
15.
Int J Food Microbiol ; 197: 65-71, 2015 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-25557185

RESUMO

Airag is a traditional fermented milk of Mongolia that is usually made from raw mare's milk. Lactobacillus helveticus is one of the lactic acid bacteria most frequently isolated from airag. In this study, we investigated the genetic and physiological characteristics of L. helveticus strains isolated from airag and clarified their significance in airag by comparing them with strains from different sources. Six strains of L. helveticus were isolated from five home-made airag samples collected from different regions of Mongolia. The optimal temperature for acidification in skim milk was 30 to 35°C for all the Mongolian strains, which is lower than those for the reference strains (JCM 1554 and JCM 1120(T)) isolated from European cheeses. All of the strains had a prtH1-like gene encoding a variant type of cell envelope proteinase (CEP). The CEP amino acid sequence in Snow Brand Typeculture (SBT) 11087 isolated from airag shared 71% identity with PrtH of L. helveticus CNRZ32 (AAD50643.1) but 98% identity with PrtH of Lactobacillus kefiranofaciens ZW3 (AEG40278.1) isolated from a traditional fermented milk in Tibet. The proteolytic activities of the CEP from SBT11087 on artificial substrate (N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide) and pure casein were measured using an intact-cell degradation assay. The activity of the CEP from SBT11087 was observed to be weak and exhibited a lower optimal temperature (40°C) than those from the reference strains (45-50°C). The specificity of the SBT11087 CEP for αS1-casein was typical of the CEPs previously reported in L. helveticus, as determined through the degradation profiles obtained through gel electrophoresis and mass spectrometry analyses. In contrast, the degradation profile of ß-casein revealed that the CEP of SBT11087 primarily hydrolyzes its C-terminal domain and hydrolyzed nine of the 16 cleavage sites shared among the CEPs of other L. helveticus strains. Thus, the CEP of SBT11087 is distinct from those from previously reported L. helveticus strains in terms of its optimal temperature and its degradation of ß-casein. Therefore, the Mongolian L. helveticus strains differ from other strains of the species in different collections and are specifically suited for the natural lactic acid bacterial population in airag.


Assuntos
Membrana Celular/metabolismo , Laticínios/microbiologia , Endopeptidases/metabolismo , Lactobacillus helveticus/enzimologia , Lactobacillus helveticus/isolamento & purificação , Leite/microbiologia , Sequência de Aminoácidos , Animais , Caseínas/metabolismo , Endopeptidases/química , Endopeptidases/genética , Fermentação , Cavalos , Mongólia , Proteólise , Homologia de Sequência de Aminoácidos
16.
J Dairy Sci ; 98(3): 1467-78, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25582584

RESUMO

Scamorza is a semi-hard, pasta filata cheese resembling low-moisture Mozzarella cheese, with a short ripening time (<30d). Scamorza has a bland flavor and, to provide diversification from similar cheeses, it was manufactured using 2 types of milk in the current study: 100% Italian Friesian milk (F) or 90% F and 10% Jersey cow milk (mixed, M), and 2 types of starter: Streptococcus thermophilus or S. thermophilus with peptidolytic Lactococcus lactis, Lactobacillus helveticus, and Lactobacillus paracasei strains as adjuncts). The cheeses were ripened for 30d. The adjunct did not significantly affect acid production or growth of the primary starter; 2 of the species used in the adjunct (Lb. paracasei and Lb. helveticus) rapidly colonized the cheese and persisted until the end of ripening, whereas the counts of nonstarter lactic acid bacteria in the control cheese were low until the end of ripening. The use of adjuncts affected pH, microbial composition (as assessed by both culture-dependent and culture-independent methods), total free amino acid content, and volatile profile (measured using an electronic nose), whereas milk type had only a minor effect. Although differences in primary proteolysis were found, they were probably indirect and related to the effects on pH and moisture. We conclude that, even with a short ripening time (30d), use of a peptidolytic adjunct may significantly affect important features of Scamorza and may be used to create a product that is measurably different from competing products.


Assuntos
Queijo/microbiologia , Lactobacillus/isolamento & purificação , Streptococcus thermophilus/isolamento & purificação , Animais , Bovinos , Contagem de Colônia Microbiana , DNA Bacteriano/isolamento & purificação , Análise de Alimentos , Manipulação de Alimentos , Microbiologia de Alimentos , Concentração de Íons de Hidrogênio , Lactobacillus helveticus/isolamento & purificação , Lactococcus lactis/isolamento & purificação , Paladar , Compostos Orgânicos Voláteis
17.
J Dairy Sci ; 98(3): 1479-91, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25465632

RESUMO

The present study aimed to evaluate the effect of a peptidolytic adjunct (Lactococcus lactis, Lactobacillus helveticus, and Lactobacillus paracasei), as a tool to accelerate ripening, on sensory properties and acceptability of Scamorza cheese obtained using 2 types of milk (Friesian and Friesian+Jersey) and Streptococcus thermophilus as primary starter. A 10-member panel was trained using a specific frame of references and used a specific vocabulary to assess cheese sensory properties through quantitative descriptive analysis (QDA), whereas 87 consumers were used to evaluate product acceptability. Analysis of variance showed that milk type did not markedly alter cheese sensory properties. Conversely, panelists perceived higher intensities of butter, saltiness, and sweetness flavors in cheese without adjunct culture (ST), whereas the addition of the adjunct culture (ST+A) induced higher and sourness flavors, oiliness and grainy textures, and lower adhesiveness, moisture, springiness, and tenderness. Principal component analysis showed positive relationships between pH and tenderness, sweetness and saltiness and a negative correlation between pH and grainy, oiliness, color and structure uniformity, sourness, and milk. Most of the differences observed in QDA and most of the relationships observed in the principal component analysis were linked to the higher microbial activity induced by the adjunct culture. Independently of milk and starter types, consumers perceived Scamorza cheese as characterized by a good eating quality (mean liking scores were all above the neutral point of the hedonic scale). Although ST cheeses showed higher values for overall liking, 2 homogeneous groups of consumers were identified using partial least squares regression analysis. One group preferred ST cheeses with higher levels of tenderness, adhesiveness, springiness, and moisture in terms of texture, butter in terms of flavor, and sweetness in terms of taste, whereas a second group preferred ST+A products characterized by specific attributes of texture (cohesiveness and oiliness), flavor (milk), taste (sourness), and appearance (structure and color uniformity). We conclude that further studies for the development of short-ripened products based on the use of adjunct cultures should be conducted to promote product differentiation and meet the sensory requirements of particular segments of consumers.


Assuntos
Queijo/análise , Queijo/microbiologia , Comportamento do Consumidor , Preferências Alimentares , Paladar , Adulto , Cor , Feminino , Análise de Alimentos , Microbiologia de Alimentos , Humanos , Concentração de Íons de Hidrogênio , Lactobacillus/isolamento & purificação , Lactobacillus helveticus/isolamento & purificação , Lactococcus lactis/isolamento & purificação , Modelos Lineares , Masculino , Análise Multivariada , Odorantes , Análise de Componente Principal
18.
J Appl Microbiol ; 114(2): 404-12, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23082800

RESUMO

AIMS: To assess the structure and stability of a dominant lactic acid bacteria (LAB) population during the propagation of rye sourdough in an industrial semi-fluid production over a period of 7 months. METHODS AND RESULTS: The sourdough was started from a 6-year-old freeze-dried sourdough originating from the same bakery. A unique microbial consortium consisting mainly of bacteria belonging to species Lactobacillus helveticus, Lactobacillus panis and Lactobacillus pontis was identified based on culture-dependent (Rep-PCR) and culture-independent [denaturing gradient gel electrophoresis (DGGE)] methods. Three of the isolated Lact. helveticus strains showed remarkable adaptation to the sourdough conditions. They differed from the type strain by the ability to ferment compounds specific to plant material, like salicin, cellobiose and sucrose, but did not ferment lactose. CONCLUSION: We showed remarkable stability of a LAB consortium in rye sourdough started from lyophilized sourdough and propagated in a large bakery for 7 months. Lactobacillus helveticus was detected as the dominant species in the consortium and was shown to be metabolically adapted to the sourdough environment. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of an established and adapted microbial consortium as a starter is a good alternative to commercial starter strains.


Assuntos
Pão/microbiologia , Lactobacillus helveticus/isolamento & purificação , Consórcios Microbianos , Secale , Pão/análise , Eletroforese em Gel de Gradiente Desnaturante , Microbiologia de Alimentos , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Lactobacillus helveticus/classificação , Lactobacillus helveticus/metabolismo , Reação em Cadeia da Polimerase
19.
J Bacteriol ; 193(16): 4282-3, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21705605

RESUMO

Lactobacillus helveticus MTCC 5463 was isolated from a vaginal swab from a healthy adult female. The strain exhibited potential probiotic properties, with their beneficial role in the gastrointestinal tract and their ability to reduce cholesterol and stimulate immunity. We sequenced the whole genome and compared it with the published genome sequence of Lactobacillus helveticus DPC4571.


Assuntos
Genoma Bacteriano , Lactobacillus helveticus/genética , Probióticos/isolamento & purificação , Adulto , Feminino , Humanos , Índia , Lactobacillus helveticus/isolamento & purificação , Dados de Sequência Molecular , Vagina/microbiologia
20.
J Bacteriol ; 193(10): 2666-7, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21398542

RESUMO

Lactobacillus helveticus strain H10 was isolated from traditional fermented milk in Tibet, China. We sequenced the whole genome of strain H10 and compared it to the published genome sequence of Lactobacillus helveticus DPC4571.


Assuntos
DNA Bacteriano/química , DNA Bacteriano/genética , Genoma Bacteriano , Lactobacillus helveticus/genética , Animais , Lactobacillus helveticus/isolamento & purificação , Leite/microbiologia , Dados de Sequência Molecular , Análise de Sequência de DNA , Tibet
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