scholarly journals Attenuated Lactococcus lactis and Surface Bacteria as Tools for Conditioning the Microbiota and Driving the Ripening of Semisoft Caciotta Cheese

2019 ◽  
Vol 86 (5) ◽  
Author(s):  
Maria Calasso ◽  
Fabio Minervini ◽  
Francesca De Filippis ◽  
Danilo Ercolini ◽  
Maria De Angelis ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactococcus Lactis ◽  
Lactobacillus Delbrueckii ◽  
Bacterial Strains ◽  
Multivariate Statistical ◽  
Content Type ◽  
Cheese Ripening ◽  
Brochothrix Thermosphacta ◽  
Staphylococcus Equorum ◽  
Metabolic Activities

ABSTRACT This study aimed at establishing the effects of attenuated starters and surface bacteria on various features of caciotta cheese. The cheese undergoes a ripening period during which the house microbiota contaminates the surface. Conventional cheese (the control cheese [CC]) is made using only primary starters. Primary starters and attenuated (i.e., unable to grow and synthesize lactic acid) Lactococcus lactis (Lc. lactis) subsp. lactis were used to produce caciotta cheese without (ATT cheese) or with an inoculum of surface bacteria: (i) Leuconostoc lactis (Le. lactis) (LL cheese), (ii) Vibrio casei (VC cheese), (iii) Staphylococcus equorum (SE cheese), (iv) Brochothrix thermosphacta (BX cheese), and (v) a mixture of all four (MIX cheese). Attenuated Lc. lactis increased microbial diversity during cheese ripening. At the core, attenuated starter mainly increased indigenous lactococci and Lactobacillus delbrueckii group bacteria. At the surface, the main effect was on Macrococcus caseolyticus. Autochthonous Le. lactis strains took advantage of the attenuated starter, becoming dominant. Adjunct Le. lactis positively affected Lactobacillus sakei group bacteria on the LL cheese surface. Adjunct V. casei, S. equorum, and B. thermosphacta did not become dominant. Surfaces of VC, SE, and BX cheeses mainly harbored Staphylococcus succinus. Peptidase activities were higher in cheeses made with attenuated starter than in CC, which had the lowest concentration of free amino acids. Based on the enzymatic activities of adjunct Le. lactis, LL and MIX cheeses exhibited the highest glutamate dehydrogenase, cystathionine-γ-lyase, and esterase activities. As shown by multivariate statistical analyses, LL and MIX cheeses showed the highest similarity for microbiological and biochemical features. LL and MIX cheeses received the highest scores for overall sensory acceptability. IMPORTANCE This study provides in-depth knowledge of the effects of attenuated starters and surface bacterial strains on the microbiota and related metabolic activities during cheese ripening. The use of attenuated Lc. lactis strongly impacted the microbiota assembly of caciotta cheese. This led to improved biochemical and sensory features compared to conventional cheese. Among surface bacterial strains, Le. lactis played a key role in the metabolic activities involved in cheese ripening. This resulted in an improvement of the sensory quality of caciotta cheese. The use of attenuated lactic acid bacteria and the surface adjunct Le. lactis could be a useful biotechnology to improve the flavor formation of caciotta cheese.

scholarly journals Isolation and Characterization of Lactic Acid Bacteria in Philippine Fermented Milkfish Chanos chanos-Rice Mixture (Burong Bangus)

2018 ◽  
Vol 6 (2) ◽  
pp. 500-508
Author(s):  
Julie Ann A. Arcales ◽  
Garner Algo L.Alolod
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Enzymatic Activity ◽  
Antimicrobial Property ◽  
Food Products ◽  
The Philippines ◽  
Lactobacillus Delbrueckii ◽  
Bacterial Strains ◽  
Isolation And Characterization

Isolation and characterization of bacteria in food products are important to determine and distinguish the beneficial or harmful effects of microbiota in certain samples. Lactic acid bacteria in food products had long been associated to good factors as food preservatives and with added fermentation metabolites. This study isolated and characterized lactic acid bacteria from burong bangus. The culture and purification process of bacteria isolation resulted to 4 strains of lactic acid bacteria namely Enterococcus faecalis, Tetragenococcus muriaticus, Lactobacillus delbrueckii subp. delbrueckii and Carnobacterium divergens. High enzymatic activity were observed with E. faecalis particularly on lipase and protease assay. While C. divergens have no enzymatic activity against lipase, protease, amylase and cellulase. The antimicrobial property of L. delbrueckii is only susceptible to amoxicillin unlike the other three bacteria isolates. No antagonistic activity were observed with the four bacterial strains against Bacillus subtilis, Staphylococcus aureus and Escherichia coli. The result of this study showed promising benefits to the industry especially in developing countries like the Philippines because population are not yet so aware of this organisms and the benefits that can be derived through their consumption.

scholarly journals Inulin Fermentation by Lactobacilli and Bifidobacteria from Dairy Calves

2020 ◽  
Vol 87 (1) ◽  
Author(s):  
Yuanting Zhu ◽  
Jinxin Liu ◽  
Julian M. Lopez ◽  
David A. Mills
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Gut Microbiome ◽  
Animal Health ◽  
Dairy Calves ◽  
Milk Replacer ◽  
Comparative Genomic ◽  
Bacterial Strains ◽  
Content Type ◽  
Long Chain

ABSTRACT Prebiotics are increasingly examined for their ability to modulate the neonate gut microbiota of livestock, and products such as inulin are commonly added to milk replacer used in calving. However, the ability of specific members of the bovine neonate microbiota to respond to inulin remains to be determined, particularly among indigenous lactobacilli and bifidobacteria, beneficial genera commonly enriched by inulin. Screening of Bifidobacterium and Lactobacillus isolates obtained from fresh feces of dairy calves revealed that lactobacilli had a higher prevalence of inulin fermentation capacity (58%) than bifidobacteria (17%). Several Ligilactobacillus agilis (synonym Lactobacillus agilis) isolates exhibited vigorous growth on, and complete degradation of, inulin; however, the phenotype was strain specific. The most vigorous inulin-fermenting strain, L. agilis YZ050, readily degraded long-chain inulin not consumed by bifidobacterial isolates. Comparative genomic analysis of both L. agilis fermenter and nonfermenter strains indicated that strain YZ050 encodes an inulinase homolog, previously linked to extracellular degradation of long-chain inulin in Lacticaseibacillus paracasei, that was strongly induced during growth on inulin. Inulin catabolism by YZ050 also generates extracellular fructose, which can cross-feed other non-inulin-fermenting lactic acid bacteria isolated from the same bovine feces. The presence of specific inulin-responsive bacterial strains within calf gut microbiome provides a mechanistic rationale for enrichment of specific lactobacilli and creates a foundation for future synbiotic applications in dairy calves aimed at improving health in early life. IMPORTANCE The gut microbiome plays an important role in animal health and is increasingly recognized as a target for diet-based manipulation. Inulin is a common prebiotic routinely added to animal feeds; however, the mechanism of inulin consumption by specific beneficial taxa in livestock is ill defined. In this study, we examined Lactobacillus and Bifidobacterium isolates from calves fed inulin-containing milk replacer and characterized specific strains that robustly consume long-chain inulin. In particular, novel Ligilactobacillus agilis strain YZ050 consumed inulin via an extracellular fructosidase, resulting in complete consumption of all long-chain inulin. Inulin catabolism resulted in temporal release of extracellular fructose, which can promote growth of other non-inulin-consuming strains of lactic acid bacteria. This work provides the mechanistic insight needed to purposely modulate the calf gut microbiome via the establishment of networks of beneficial microbes linked to specific prebiotics.

Influence of lactic cultures in denaturation of whey proteins during fermentation of milk

1988 ◽  
Vol 55 (3) ◽  
pp. 443-448 ◽  
Author(s):  
Nataraja Iyer Vaitheeswaran ◽  
Gajanan S. Bhat
Keyword(s):  
Lactic Acid ◽  
Whey Protein ◽  
Lactococcus Lactis ◽  
Whey Proteins ◽  
Skim Milk ◽  
Lactobacillus Delbrueckii ◽  
Streptococcus Salivarius ◽  
Dye Binding

SummaryUndenatured whey protein (UWP) content of skim milk acidified with lactic acid or cultured with lactic cultures was estimated by a dye-binding method. The UWP content decreased with increase in acidity and the denaturation was only partly reversible on neutralization to the original acidity. The decrease in UWP was higher in cultured milk than in the milk acidified to the same extent with lactic acid, indicating the effect of lactic cultures in denaturation of whey proteins during fermentation of milk. Among the lactic cultures the denaturation effect of Lactobacillus delbrueckii subsp. bulgaricus was highest, followed by Streptococcus salivarius subsp. thermophilus, Lactococcus lactis subsp. lactis and Lact. lactis biovar diacetylactis. Denaturation of whey proteins by lactic cultures was found to be partly irreversible.

Changes in Galactose and Lactic Acid Content of Sweet Whey during Storage

2004 ◽  
Vol 67 (2) ◽  
pp. 403-406 ◽  
Author(s):  
R. D. RAO ◽  
W. L. WENDORFF ◽  
K. SMITH
Keyword(s):  
Lactic Acid ◽  
Lactococcus Lactis ◽  
Lactic Acid Production ◽  
Starter Culture ◽  
Storage Conditions ◽  
Starter Cultures ◽  
Lactobacillus Helveticus ◽  
Lactobacillus Delbrueckii ◽  
The Other ◽  
Time Period

Whey is often stored or transported for a period of time prior to processing. During this time period, galactose and lactic acid concentrations may accumulate, reducing the quality of spray-dried whey powders in regard to stickiness and agglomeration. This study surveyed industry samples of Cheddar and mozzarella cheese whey streams to determine how galactose and lactic acid concentrations changed with storage at appropriate (4°C) and abuse (37.8°C) temperatures. Samples stored at 4°C did not exhibit significant increases in levels of lactic acid or galactose. Mozzarella whey accumulated the greatest amount of galactose and lactic acid with storage at 37.8°C. Whey samples derived from cheese made from single strains of starter culture were also evaluated to determine each culture's contribution to galactose and lactic acid production. Starter cultures evaluated included Streptococcus salivarius ssp. thermophilus, Lactobacillus helveticus, Lactobacillus delbrueckii ssp. bulgaricus, Lactococcus lactis ssp. cremoris, and Lactococcus lactis ssp. lactis. Whey derived from L. helveticus accumulated a significantly greater amount of lactic acid upon storage at 37.8°C as compared with the other cultures. Galactose accumulation was significantly decreased in whey from L. lactis ssp. lactis stored at 37.8°C in comparison with the other cultures. Results from this study indicate that proper storage conditions (4°C) for whey prevent accumulation of galactose and lactic acid while the extent of accumulation during storage at 37.8°C varies depending on the culture(s) used in cheese production.

scholarly journals Spatial Distribution of Lactococcus lactis Colonies Modulates the Production of Major Metabolites during the Ripening of a Model Cheese

2015 ◽  
Vol 82 (1) ◽  
pp. 202-210 ◽  
Author(s):  
Clémentine Le Boucher ◽  
Valérie Gagnaire ◽  
Valérie Briard-Bion ◽  
Julien Jardin ◽  
Marie-Bernadette Maillard ◽  
...  
Keyword(s):  
Spatial Distribution ◽  
Lactococcus Lactis ◽  
Time Course ◽  
Bacterial Population ◽  
Spatial Distributions ◽  
Content Type ◽  
Cheese Ripening ◽  
Volatile Metabolites ◽  
Small Colony ◽  
Exchange Surface

ABSTRACTIn cheese, lactic acid bacteria are immobilized at the coagulation step and grow as colonies. The spatial distribution of bacterial colonies is characterized by the size and number of colonies for a given bacterial population within cheese. Our objective was to demonstrate that different spatial distributions, which lead to differences in the exchange surface between the colonies and the cheese matrix, can influence the ripening process. The strategy was to generate cheeses with the same growth and acidification of aLactococcus lactisstrain with two different spatial distributions, big and small colonies, to monitor the production of the major ripening metabolites, including sugars, organic acids, peptides, free amino acids, and volatile metabolites, over 1 month of ripening. The monitored metabolites were qualitatively the same for both cheeses, but many of them were more abundant in the small-colony cheeses than in the big-colony cheeses over 1 month of ripening. Therefore, the results obtained showed that two different spatial distributions ofL. lactismodulated the ripening time course by generating moderate but significant differences in the rates of production or consumption for many of the metabolites commonly monitored throughout ripening. The present work further explores the immobilization of bacteria as colonies within cheese and highlights the consequences of this immobilization on cheese ripening.

scholarly journals Oxygen Affects Gut Bacterial Colonization and Metabolic Activities in a Gnotobiotic Cockroach Model

2015 ◽  
Vol 82 (4) ◽  
pp. 1080-1089 ◽  
Author(s):  
Dorothee Tegtmeier ◽  
Claire L. Thompson ◽  
Christine Schauer ◽  
Andreas Brune
Keyword(s):  
Bacterial Colonization ◽  
Anaerobic Bacteria ◽  
Close Relative ◽  
Bacterial Strains ◽  
Fermentation Products ◽  
Content Type ◽  
Facultatively Anaerobic ◽  
Pure Cultures ◽  
Metabolic Activities

ABSTRACTThe gut microbiota of termites and cockroaches represents complex metabolic networks of many diverse microbial populations. The distinct microenvironmental conditions within the gut and possible interactions among the microorganisms make it essential to investigate how far the metabolic properties of pure cultures reflect their activities in their natural environment. We established the cockroachShelfordella lateralisas a gnotobiotic model and inoculated germfree nymphs with two bacterial strains isolated from the guts of conventional cockroaches. Fluorescence microscopy revealed that both strains specifically colonized the germfree hindgut. In diassociated cockroaches, the facultatively anaerobic strain EbSL (a new species ofEnterobacteriaceae) always outnumbered the obligately anaerobic strain FuSL (a close relative ofFusobacterium varium), irrespective of the sequence of inoculation, which showed that precolonization by facultatively anaerobic bacteria does not necessarily favor colonization by obligate anaerobes. Comparison of the fermentation products of the cultures formedin vitrowith those accumulatedin situindicated that the gut environment strongly affected the metabolic activities of both strains. The pure cultures formed the typical products of mixed-acid or butyrate fermentation, whereas the guts of gnotobiotic cockroaches accumulated mostly lactate and acetate. Similar shifts toward more-oxidized products were observed when the pure cultures were exposed to oxygen, which corroborated the strong effects of oxygen on the metabolic fluxes previously observed in termite guts. Oxygen microsensor profiles of the guts of germfree, gnotobiotic, and conventional cockroaches indicated that both gut tissue and microbiota contribute to oxygen consumption and suggest that the oxygen status influences the colonization success.

scholarly journals The KupA and KupB Proteins of Lactococcus lactis IL1403 Are Novel c-di-AMP Receptor Proteins Responsible for Potassium Uptake

2019 ◽  
Vol 201 (10) ◽  
Author(s):  
Ingrid M. Quintana ◽  
Johannes Gibhardt ◽  
Asan Turdiev ◽  
Elke Hammer ◽  
Fabian M. Commichau ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacterium ◽  
Lactococcus Lactis ◽  
Second Messenger ◽  
Cell Factory ◽  
Content Type ◽  
Potassium Homeostasis ◽  
Rna Molecules ◽  
High Affinity ◽  
Potassium Transporters

ABSTRACT Cyclic di-AMP (c-di-AMP) is a second messenger involved in diverse metabolic processes, including osmolyte uptake, cell wall homeostasis, and antibiotic and heat resistance. In Lactococcus lactis, a lactic acid bacterium which is used in the dairy industry and as a cell factory in biotechnological processes, the only reported interaction partners of c-di-AMP are the pyruvate carboxylase and BusR, the transcription regulator of the busAB operon for glycine betaine uptake. However, recent studies uncovered a major role of c-di-AMP in the control of potassium homeostasis, and potassium is the signal that triggers c-di-AMP synthesis. In this study, we have identified KupA and KupB, which belong to the Kup/HAK/KT family, as novel c-di-AMP binding proteins. Both proteins are high-affinity potassium transporters, and their transport activities are inhibited by binding of c-di-AMP. Thus, in addition to the well-studied Ktr/Trk potassium channels, KupA and KupB represent a second class of potassium transporters that are subject to inhibition by c-di-AMP. IMPORTANCE Potassium is an essential ion in every living cell. Even though potassium is the most abundant cation in cells, its accumulation can be toxic. Therefore, the level of potassium has to be tightly controlled. In many Gram-positive bacteria, the second messenger cyclic di-AMP plays a key role in the control of potassium homeostasis by binding to potassium transporters and regulatory proteins and RNA molecules. In the lactic acid bacterium Lactococcus lactis, none of these conserved c-di-AMP-responsive molecules are present. In this study, we demonstrate that the KupA and KupB proteins of L. lactis IL1403 are high-affinity potassium transporters and that their transport activity is inhibited by the second messenger c-di-AMP.

scholarly journals Animal Rennets as Sources of Dairy Lactic Acid Bacteria

2014 ◽  
Vol 80 (7) ◽  
pp. 2050-2061 ◽  
Author(s):  
Margherita Cruciata ◽  
Ciro Sannino ◽  
Danilo Ercolini ◽  
Maria L. Scatassa ◽  
Francesca De Filippis ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Streptococcus Thermophilus ◽  
Species Level ◽  
Lactobacillus Delbrueckii ◽  
Rrna Gene ◽  
Microbial Composition ◽  
Content Type

ABSTRACTThe microbial composition of artisan and industrial animal rennet pastes was studied by using both culture-dependent and -independent approaches. Pyrosequencing targeting the 16S rRNA gene allowed to identify 361 operational taxonomic units (OTUs) to the genus/species level. Among lactic acid bacteria (LAB),Streptococcus thermophilusand some lactobacilli, mainlyLactobacillus crispatusandLactobacillus reuteri, were the most abundant species, with differences among the samples. Twelve groups of microorganisms were targeted by viable plate counts revealing a dominance of mesophilic cocci. All rennets were able to acidify ultrahigh-temperature-processed (UHT) milk as shown by pH and total titratable acidity (TTA). Presumptive LAB isolated at the highest dilutions of acidified milks were phenotypically characterized, grouped, differentiated at the strain level by randomly amplified polymorphic DNA (RAPD)-PCR analysis, and subjected to 16S rRNA gene sequencing. Only 18 strains were clearly identified at the species level, asEnterococcus casseliflavus,Enterococcus faecium,Enterococcus faecalis,Enterococcus lactis,Lactobacillus delbrueckii, andStreptococcus thermophilus, while the other strains, all belonging to the genusEnterococcus, could not be allotted into any previously described species. The phylogenetic analysis showed that these strains might represent different unknown species. All strains were evaluated for their dairy technological performances. All isolates produced diacetyl, and 10 of them produced a rapid pH drop in milk, but only 3 isolates were also autolytic. This work showed that animal rennet pastes can be sources of LAB, mainly enterococci, that might contribute to the microbial diversity associated with dairy productions.

scholarly journals Natural DNA Transformation Is Functional in Lactococcus lactis subsp. cremoris KW2

2017 ◽  
Vol 83 (16) ◽  
Author(s):  
Blandine David ◽  
Amandine Radziejwoski ◽  
Frédéric Toussaint ◽  
Laetitia Fontaine ◽  
Marie Henry de Frahan ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactococcus Lactis ◽  
Natural Transformation ◽  
Single Point ◽  
Adaptor Protein ◽  
Single Point Mutation ◽  
Dna Transformation ◽  
Dna Uptake ◽  
Genetic Traits ◽  
Content Type

ABSTRACT Lactococcus lactis is one of the most commonly used lactic acid bacteria in the dairy industry. Activation of competence for natural DNA transformation in this species would greatly improve the selection of novel strains with desired genetic traits. Here, we investigated the activation of natural transformation in L. lactis subsp. cremoris KW2, a strain of plant origin whose genome encodes the master competence regulator ComX and the complete set of proteins usually required for natural transformation. In the absence of knowledge about competence regulation in this species, we constitutively overproduced ComX in a reporter strain of late competence phase activation and showed, by transcriptomic analyses, a ComX-dependent induction of all key competence genes. We further demonstrated that natural DNA transformation is functional in this strain and requires the competence DNA uptake machinery. Since constitutive ComX overproduction is unstable, we alternatively expressed comX under the control of an endogenous xylose-inducible promoter. This regulated system was used to successfully inactivate the adaptor protein MecA and subunits of the Clp proteolytic complex, which were previously shown to be involved in ComX degradation in streptococci. In the presence of a small amount of ComX, the deletion of mecA, clpC, or clpP genes markedly increased the activation of the late competence phase and transformability. Altogether, our results report the functionality of natural DNA transformation in L. lactis and pave the way for the identification of signaling mechanisms that trigger the competence state in this species. IMPORTANCE Lactococcus lactis is a lactic acid bacterium of major importance, which is used as a starter species for milk fermentation, a host for heterologous protein production, and a delivery platform for therapeutic molecules. Here, we report the functionality of natural transformation in L. lactis subsp. cremoris KW2 by the overproduction of the master competence regulator ComX. The developed procedure enables a flexible approach to modify the chromosome with single point mutation, sequence insertion, or sequence replacement. These results represent an important step for the genetic engineering of L. lactis that will facilitate the design of strains optimized for industrial applications. This will also help to discover natural regulatory mechanisms controlling competence in the genus Lactococcus.

scholarly journals Major Role of NAD-Dependent Lactate Dehydrogenases in the Production of l-Lactic Acid with High Optical Purity by the Thermophile Bacillus coagulans

2014 ◽  
Vol 80 (23) ◽  
pp. 7134-7141 ◽  
Author(s):  
Limin Wang ◽  
Yumeng Cai ◽  
Lingfeng Zhu ◽  
Honglian Guo ◽  
Bo Yu
Keyword(s):  
Lactic Acid ◽  
Lactate Dehydrogenase ◽  
Acid Production ◽  
Lactic Acid Production ◽  
Optical Purity ◽  
Bacillus Coagulans ◽  
Lactobacillus Delbrueckii ◽  
Content Type ◽  
High Optical Purity ◽  
Optically Pure

ABSTRACTBacillus coagulans2-6 is an excellent producer of optically purel-lactic acid. However, little is known about the mechanism of synthesis of the highly optically purel-lactic acid produced by this strain. Three enzymes responsible for lactic acid production—NAD-dependentl-lactate dehydrogenase (l-nLDH; encoded byldhL), NAD-dependentd-lactate dehydrogenase (d-nLDH; encoded byldhD), and glycolate oxidase (GOX)—were systematically investigated in order to study the relationship between these enzymes and the optical purity of lactic acid.Lactobacillus delbrueckiisubsp.bulgaricusDSM 20081 (ad-lactic acid producer) andLactobacillus plantarumsubsp.plantarumDSM 20174 (adl-lactic acid producer) were also examined in this study as comparative strains, in addition toB. coagulans. The specific activities of key enzymes for lactic acid production in the three strains were characterizedin vivoandin vitro, and the levels of transcription of theldhL,ldhD, and GOX genes during fermentation were also analyzed. The catalytic activities ofl-nLDH andd-nLDH were different inl-,d-, anddl-lactic acid producers. Onlyl-nLDH activity was detected inB. coagulans2-6 under native conditions, and the level of transcription ofldhLinB. coagulans2-6 was much higher than that ofldhDor the GOX gene at all growth phases. However, for the twoLactobacillusstrains used in this study,ldhDtranscription levels were higher than those ofldhL. The high catalytic efficiency ofl-nLDH toward pyruvate and the high transcription ratios ofldhLtoldhDandldhLto the GOX gene provide the key explanations for the high optical purity ofl-lactic acid produced byB. coagulans2-6.

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