PL221959B1 - Lactobacillus plantarum lactic bacteria strain - Google Patents
Lactobacillus plantarum lactic bacteria strainInfo
- Publication number
- PL221959B1 PL221959B1 PL401554A PL40155412A PL221959B1 PL 221959 B1 PL221959 B1 PL 221959B1 PL 401554 A PL401554 A PL 401554A PL 40155412 A PL40155412 A PL 40155412A PL 221959 B1 PL221959 B1 PL 221959B1
- Authority
- PL
- Poland
- Prior art keywords
- plantarum
- strain
- nbrc101977
- lactobacillus plantarum
- cfu
- Prior art date
Links
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
Opis wynalazkuDescription of the invention
Przedmiotem wynalazku jest szczep bakterii mlekowych Lactobacillus plantarum o właściwościach probiotycznych.The subject of the invention is a strain of lactic bacteria Lactobacillus plantarum with probiotic properties.
Specyficzna i kontrolowana pozytywna regulacja składu mikroflory jelitowej ludzi i zwierząt może być osiągnięta na drodze probiozy czyli spożywania preparatów lub produktów żywnościowych (lub paszowych dla zwierząt) zawierających żywe bakterie, głównie z rodzaju Lactobacillus. Dlatego też producenci żywności i pasz, a także przemysł farmaceutyczny zainteresowani są pozyskiwaniem nowych szczepów tych bakterii o udokumentowanych właściwościach probiotycznych.Specific and controlled positive regulation of the composition of the intestinal microflora of humans and animals can be achieved by probiosis, i.e. consumption of food (or animal feed) preparations or products containing live bacteria, mainly of the Lactobacillus genus. Therefore, food and feed producers as well as the pharmaceutical industry are interested in acquiring new strains of these bacteria with documented probiotic properties.
Dotychczas znane są szczepy bakterii Lactobacillus o właściwościach probiotycznych jak L. acidophilus NCFM, L. acidophilus DDS-1, L. acidophilus SBT-2062, L. acidophilus R0011, L. rhamnosus R0052, L. acidophilus LA-1, L. paracasei CRL-431, L. casei Shirota, L. casei DN-114001, L fermentum RC-14, L. rhamnosus GR-1, L. johnsonii La-1, L. plantarum 299V, L. rhamnosus 271, L. reuteri SD2112, L. rhamnosus GG, L. rhamnosus LB21, L. salivarius UCC118, L. acidophilus LB, L. paracasei F19, L. casei ŁOCK 0900, L. casei ŁOCK 0908, Ł. paracasei ŁOCK 0919, L. brevis ŁOCK 0944, L. paracesei ŁOCK 0920.So far, strains of Lactobacillus bacteria with probiotic properties are known, such as L. acidophilus NCFM, L. acidophilus DDS-1, L. acidophilus SBT-2062, L. acidophilus R0011, L. rhamnosus R0052, L. acidophilus LA-1, L. paracasei CRL -431, L. casei Shirota, L. casei DN-114001, L fermentum RC-14, L. rhamnosus GR-1, L. johnsonii La-1, L. plantarum 299V, L. rhamnosus 271, L. reuteri SD2112, L. rhamnosus GG, L. rhamnosus LB21, L. salivarius UCC118, L. acidophilus LB, L. paracasei F19, L. casei ŁOCK 0900, L. casei ŁOCK 0908, Ł. Paracasei ŁOCK 0919, L. brevis LOCK 0944, L . paracesei LOCK 0920.
Wynalazek dotyczy nowego szczepu bakterii mlekowych z rodzaju Lactobacillus o właściwościach probiotycznych, który oznaczono symbolem ŁOCK 0860 i który zdeponowano pod nr 11/04/2012 w Zakładzie Mikrobiologii Molekularnej Narodowego Instytutu Leków z siedzibą w Warszawie. Szczep stanowi Gram-dodatnia pałeczka, nieruchliwa, nie przetrwalnikująca. Szczep ten wykazuje 99,9% homologii do gatunku Lactobacillus plantarum, co stwierdzono na podstawie wyników testów fermentacyjnych API 50CHL. Sekwencję nukleotydową regionu DNA kodującego gen 16S rRNA nowego szczepu, która określa przynależność gatunkową szczepu, przedstawiono na końcu opisu.The invention concerns a new strain of lactic acid bacteria of the genus Lactobacillus with probiotic properties, marked with the symbol ŁOCK 0860 and deposited under No. 11/04/2012 at the Department of Molecular Microbiology of the National Medicines Institute with its seat in Warsaw. The strain is a gram-positive rod, immobile, non-sporulating. This strain shows 99.9% homology to the species Lactobacillus plantarum, which was found on the basis of the results of API 50CHL fermentation tests. The nucleotide sequence of the DNA region encoding the 16S rRNA gene of the new strain, which defines the species affiliation of the strain, is provided at the end of the description.
W celu określenia przynależności gatunkowej nowego szczepu Lactobacillus plantarum ŁOCK 0860 zsekwencjonowano region DNA kodujący gen 16S rRNA o długości 1339 pz. Otrzymaną sekwencję nukleotydową genu 16S rRNA porównano za pomocą programu BLASTN 2.2.27+ z sekwencjami dostępnymi w bazie National Center of Biotechnology Information (NCBI). Porównanie sekwencji nukleotydowej genu 16S rRNA nowego szczepu Lactobacillus plantarum 0860 z sekwencją szczepu referencyjnego Lactobacillus plantarum NBRC101977 przedstawiono na końcu opisu. Na podstawie tego porównania stwierdzono podobieństwo nowego szczepu bakterii do gatunku Lactobacillus plantarum z prawdopodobieństwem 99,3%.In order to determine the species affiliation of the new Lactobacillus plantarum LOCK 0860 strain, the DNA region encoding the 1339 bp 16S rRNA gene was sequenced. The obtained nucleotide sequence of the 16S rRNA gene was compared using the BLASTN 2.2.27+ program with the sequences available in the National Center of Biotechnology Information (NCBI) database. The comparison of the nucleotide sequence of the 16S rRNA gene of the new Lactobacillus plantarum 0860 strain with the sequence of the Lactobacillus plantarum NBRC101977 reference strain is presented at the end of the description. Based on this comparison, the similarity of the new bacterial strain to the species Lactobacillus plantarum was found with a probability of 99.3%.
Szczep ten wykazuje 99,9% homologii do gatunku Lactobacillus plantarum, co stwierdzono na podstawie wyników testów fermentacyjnych API 50CHL.This strain shows 99.9% homology to the species Lactobacillus plantarum, which was found on the basis of the results of API 50CHL fermentation tests.
Nowy szczep otrzymuje się w wyniku hodowli na podłożu Rogosa (Difco) o składzie w g/l: pepton kazeinowy - 10,0, ekstrakt drożdżowy - 5,0, D(+)glukoza -20,0, fosforan potasu - 6,0, cytrynian amonu - 2,0, Tween 80 - 1,0, octan sodu - 15,0 siarczan magnezu - 0,575, siarczan żelaza II - 0,034, siarczan manganu - 0,12, agar - 15,0, o pH = 5,5 w postaci kremowych, owalnych kolonii, o gładkiej powierzchni i regularnych brzegach, o średnicy około 1 mm. Natomiast w podłożu płynnym MRS (Merck) o składzie w g/l: ekstrakt drożdżowy - 4,0, ekstrakt mięsny 10,0 - pepton z kazeiny 10,0, D(+)glukoza - 20,0, Tween 80-1,0, wodorocytrynian amonu - 2,0, fosforan dwupotasowy - 2,0, octan sodu - 5,0, siarczan magnezu 7-wodny - 0,20, siarczan magnezu 4-wodny - 0,05, nowy szczep bakterii rośnie tworząc jednorodne zmętnienie pożywki. W obrazie mikroskopowym bakterie mają kształt krótkich, prostych, zgrubiałych pałeczek. Pałeczki nie wytwarzają przetrwalników. Szczep rośnie w temperaturach 15 - 45°C, natomiast optymalna temperatura wzrostu szczepu wynosi 37°C. Do wzrostu preferuje środowisko wzbogacone w 5% (v/v) CO2. Szczep nie wytwarza katalazy. Wybarwia się na Gram (+).The new strain is obtained by culturing in Rogosa (Difco) medium with the composition in g / l: casein peptone - 10.0, yeast extract - 5.0, D (+) glucose -20.0, potassium phosphate - 6.0, ammonium citrate - 2.0, Tween 80 - 1.0, sodium acetate - 15.0, magnesium sulfate - 0.575, iron II sulfate - 0.034, manganese sulfate - 0.12, agar - 15.0, pH = 5.5 in the form of cream, oval colonies, with a smooth surface and regular edges, approximately 1 mm in diameter. However, in the liquid medium MRS (Merck) with the composition in g / l: yeast extract - 4.0, meat extract 10.0 - peptone from casein 10.0, D (+) glucose - 20.0, Tween 80-1.0 , ammonium hydrogen citrate - 2.0, dipotassium phosphate - 2.0, sodium acetate - 5.0, magnesium sulphate 7-water - 0.20, magnesium sulphate 4-water - 0.05, a new strain of bacteria grows forming a homogeneous turbidity of the medium . In the microscopic image, the bacteria appear in the shape of short, straight, thickened sticks. The sticks do not produce spores. The strain grows at temperatures of 15 - 45 ° C, while the optimal growth temperature for the strain is 37 ° C. It prefers an environment enriched in 5% (v / v) CO2 for growth. The strain does not produce catalase. It stains Gram (+).
Cechy fizjologiczne i biochemiczne szczepu ŁOCK 0860Physiological and biochemical characteristics of the ŁOCK 0860 strain
Test API 50 CHL wykazał, że szczep jest zdolny do fermentacji następujących sacharydów i ich pochodnych: rybozy, galaktozy, glukozy, fruktozy, mannozy, ramnozy, mannitolu, N-acetyloglukozoaminy, amygdaliny, arbutyny, esculiny, salicyny, celobiozy, maltozy, laktozy, melibiozy, sacharozy, trehalozy, melecytozy, skrobi, gencjobiozy, D-tagatozy, glukonianu.The API 50 CHL test showed that the strain is capable of fermenting the following saccharides and their derivatives: ribose, galactose, glucose, fructose, mannose, rhamnose, mannitol, N-acetylglucosamine, amygdalin, arbutin, esculin, salicin, cellobiose, maltose, lactose, melibiosis, sucrose, trehalose, melecytose, starch, gentiobiose, D-tagatose, gluconate.
Wyniki testu API ZYM wskazują, że szczep wykazuje aktywność następujących enzymów: esterazy, β-galaktozydazy, a-glukozydazy, β-glukozydazy, N-acetylo^-glukozaminidazy, kwaśnej fosfatazy, fosfohydrolazy naftylo-AS-BI, arylamidazy leucyny, arylamidazy waliny, arylamidazy cystyny.The results of the ZYM API test indicate that the strain exhibits the activity of the following enzymes: esterase, β-galactosidase, α-glucosidase, β-glucosidase, N-acetylβ-glucosaminidase, acid phosphatase, naphthyl-AS-BI phosphohydrolase, leucine arylamidase, valine arylamidase, cystine arylamidase.
Ponadto stwierdzono, że szczep nie wytwarza siarkowodoru, ani amoniaku z argininy, nie wykazuje zdolności rozkładu H2O2.Moreover, it was found that the strain does not produce hydrogen sulphide or ammonia from arginine, does not show the ability to decompose H2O2.
PL 221 959 B1PL 221 959 B1
Cechy probiotyczne szczepu ŁOCK 0860Probiotic characteristics of the ŁOCK 0860 strain
Szczep spełnia kryteria stawiane bakteriom probiotycznym, co stwierdzono wyniku badań in vitro wykonanych według procedur zalecanych przez FAO/WHO. Nowy szczep wykazuje odporność na niskie pH (kwasowość soku żołądkowego) w zakresie pH od 2,5 do 3,5. W pH 3,5 przeżywalność po 60 minutach wynosi 2,0, x 108 jtk/ml, a po 180 minutach 1,0 x 108 jtk/ml, przy poziomie wyjściowyThe strain meets the criteria for probiotic bacteria, which was confirmed by in vitro tests carried out according to the procedures recommended by the FAO / WHO. The new strain is resistant to low pH (gastric acidity) in the pH range from 2.5 to 3.5. At pH 3.5, the survival after 60 minutes is 2.0 x 10 8 cfu / ml, and after 180 minutes, 1.0 x 10 8 cfu / ml, at the baseline
3,7 x 109 jtk/ml. Natomiast w pH 2,5 po 60 minutach przeżywa 7,2 x 107 jtk/ml, a po 180 minutach3.7 x 10 9 CFU / ml. However, at pH 2.5, after 60 minutes, it survives 7.2 x 10 7 cfu / ml, and after 180 minutes
1,1 x 104 jtk/ml, przy poziomie wyjściowym 3,7 x 109 jtk/ml.1.1 x 10 4 cfu / ml and the output level of 3.7 x 10 9 cfu / ml.
Wykazuje odporność na sole żółci w stężeniach 2%, 4%, 6% w czasie do 48 godzin. Po 48 go5 dzinach inkubacji w obecności 2% żółci przeżywalność szczepu wynosi 5,5 x 105 jtk/ml, przy stężeniu 53 żółci 4% wynosi 5,0 x 105 jtk/ml, natomiast przy stężeniu 6% przeżywa 3,9 x 103 jtk/ml, przy wyjściowym poziomie bakterii rzędu 3,7 x 109 jtk/ml.It shows resistance to bile salts in concentrations of 2%, 4%, 6% for up to 48 hours. 48 a 5 djinns incubated in the presence of 2% bile survival rate of the strain is 5.5 x 10 5 cfu / ml, at a concentration of 53% bile 4 is 5.0 x 10 5 cfu / ml, whereas at a concentration of 6% survival 3.9 x 10 3 cfu / ml, with the initial bacterial level of 3.7 x 10 9 cfu / ml.
Wykazuje oporność w stosunku do następujących antybiotyków i chemioterapeutyków o działaniu przeciwbakteryjnym: penicyliny, amoksycyliny, doksycykliny, oksytetracykliny, tetracykliny, erytromycyny.It is resistant to the following antibiotics and chemotherapeutic agents with antibacterial activity: penicillin, amoxicillin, doxycycline, oxytetracycline, tetracycline, erythromycin.
Charakteryzuję się słabym przyleganiem do linii komórkowych Caco-2 oznaczoną jako (+). Adherencję określano półilościowo przyjmując następujące kryteria (-) brak przylegania, (+) pojedyncze komórki bakterii w całym preparacie, (++) pojedyncze komórki bakterii w poszczególnych polach widzenia, liczne w preparacie, (+++) liczne bakterie w poszczególnych polach widzenia.It is characterized by poor adherence to Caco-2 cell lines marked as (+). Adherence was semi-quantified using the following criteria (-) no adherence, (+) single bacterial cells throughout the slide, (++) single bacterial cells in each visual field, numerous in the slide, (+++) numerous bacteria in each visual field.
Wykazuje aktywność antagonistyczną w stosunku do bakterii patogennych przenoszonych drogą pokarmową, jak Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis, Salmonella Enteritidis, Salmonella Typhimurium, Listeria monocytogenes, Campylobacter jejuni, Campylobacter coli.It shows antagonistic activity against pathogenic bacteria transmitted via the food route, such as Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecalis, Salmonella Enteritidis, Salmonella Typhimurium, Listeria monocytogenes, Campylobacter jejuni, Campylobacter coli.
Posiada niewielką aktywność fekalną. Spośród trzech enzymów: β-glukozydazy, β-glukuronidazy i ureazy aktywna jest tylko β-glukozydaza.Has little fecal activity. Of the three enzymes β-glucosidase, β-glucuronidase and urease, only β-glucosidase is active.
Cechy biotechnologiczne szczepu ŁOCK 0860Biotechnological features of the ŁOCK 0860 strain
Cechy biotechnologiczne szczepu określono w mleku. Szczep osiągnął fazę stacjonarną po 25 godzinach hodowli w mleku, a plon komórek wynosił 7,2 x 108 jtk/ml. Czas trwania fazy adaptacyjnej wynosił 2 godziny.The biotechnological characteristics of the strain were determined in milk. The strain reached stationary phase after 25 hours of culturing in milk, and the cell yield was 7.2 x 10 8 cfu / ml. The duration of the adaptation phase was 2 hours.
Maksymalna wartość właściwej szybkości wzrostu szczepu wynosiła 0,55 h-1, a produktywność hodowli 0,3 x 108.The maximum value of the specific growth rate of the strain was 0.55 h -1 , and the productivity of the culture was 0.3 x 10 8 .
Szczep w początkowym etapie fazy stacjonarnej osiągnął kwasowość wynoszącą 40° SH, gwarantującą czysto kwaśny smak produktu. Specyficzna aktywność kwasząca wyrażona w °SH na jednostkę tworzącą kolonie w fazie stacjonarnej wzrostu wynosiła dla szczepu 4,4° SH/108 jtk.The strain in the initial stage of the stationary phase reached an acidity of 40 ° SH, which guarantees a purely acidic taste of the product. The specific acidifying activity expressed in ° SH per colony forming unit in the stationary phase of growth for the strain was 4.4 ° SH / 10 8 cfu.
Szczep charakteryzuje się aktywnością β-galaktozydazy na poziomie 0,35 J, przy czym jednostka aktywności J jest definiowana jako ilość ąmoli ONP uwolnionych w czasie 1 minuty przez 1 mg suchej masy bakterii.The strain is characterized by a β-galactosidase activity of 0.35 J, the unit of J activity being defined as the number of µmoles of ONP released in 1 minute per 1 mg of bacteria dry weight.
Przedmiot wynalazku ilustrują poniższe przykłady.The following examples illustrate the subject matter of the invention.
P r z y k ł a d I.P r z x l a d I.
Szczep hodowano na podłożu płynnym MRS o składzie w g/l: ekstrakt drożdżowy - 4,0, ekstrakt mięsny - 10,0, pepton z kazeiny - 10,0, D(+)glukoza - 20,0, Tween 80 - 1,0, wodorocytrynian amonu - 2,0, fosforan dwupotasowy - 2,0, octan sodu - siarczan magnezu 7-wodny - 0,20, siarczan magnezu 4-wodny - 0,05, w temperaturze 37°C, w środowisku wzbogaconym w CO2 (5% v/v). Po 24 godzinach hodowli wyhodowane komórki oddzielono od podłoża na wstrząsarce uzyskując gęstość 8,5 x 108 komórek/ml. Tak otrzymaną zawiesinę bakterii umieszczono w środowisku o pH 3.0 i w temperaturze 37°C. Po 180 minutach przeżyło 93,1 komórek.The strain was grown on MRS liquid medium with the composition in g / l: yeast extract - 4.0, meat extract - 10.0, casein peptone - 10.0, D (+) glucose - 20.0, Tween 80 - 1.0 , ammonium hydrogen citrate - 2.0, dipotassium phosphate - 2.0, sodium acetate - magnesium sulphate 7-water - 0.20, magnesium sulphate 4-water - 0.05, at 37 ° C, in a CO2 enriched environment ( 5% v / v). After 24 hours of cultivation, the cultured cells were separated from the medium on a shaker to obtain a density of 8.5 x 10 8 cells / ml. The thus obtained bacterial suspension was placed in an environment with a pH of 3.0 and a temperature of 37 ° C. After 180 minutes, 93.1 cells had survived.
P r z y k ł a d II.P r z x l a d II.
Przygotowano liofilizowane mieszanki szczepu zawierające 10% mleko odtłuszczone, na nośniku węglowodanowym o składzie: hydrolizat skrobiowy MD 12,5 g/l, hydrolizat kazeiny 20 g/l, laktitol 0,4 g/l, sacharoza 2 g/l. Trwałość liofilizowanych preparatów oceniano w warunkach chłodniczych (4-5°C) oraz w temperaturze pokojowej (25°C) w trakcie 4 miesięcy przechowywania. Temperatura chłodnicza zapewnia większą stabilność i trwałość liofilizowanych preparatów niezależnie od czynnika ochronnego. Przechowywanie preparatu bakterii suszonego sublimacyjnie z mlekiem jako medium ochronnym przez 4 miesiące 5 spowodowało obniżenie liczby żywych komórek o 6,2% dla temperatury chłodniczej i o 13,0% dla temperatury pokojowej.Freeze-dried mixtures of the strain were prepared, containing 10% skim milk, on a carbohydrate carrier with the following composition: MD starch hydrolyzate 12.5 g / l, casein hydrolyzate 20 g / l, lactitol 0.4 g / l, sucrose 2 g / l. The stability of the freeze-dried preparations was assessed under refrigeration conditions (4-5 ° C) and at room temperature (25 ° C) during 4 months of storage. The cooling temperature ensures greater stability and durability of the freeze-dried preparations regardless of the protective factor. The storage of the freeze-dried bacterial preparation with milk as a protective medium for 4 months 5 resulted in a reduction of the number of viable cells by 6.2% for the cooling temperature and by 13.0% for the room temperature.
Wykaz sekwencji nukleotydowej regionu DNA kodującego gen 16S rRNA nowego szczepu Lactobacillus plantarum ŁOCK 0860, która określa przynależność gatunkową szczepu:List of the nucleotide sequence of the DNA region encoding the 16S rRNA gene of the new Lactobacillus plantarum ŁOCK 0860 strain, which defines the species affiliation of the strain:
PL 221 959 B1PL 221 959 B1
GCGGCGTGCC TAATACATGC AAGTCGAACG AACTCTGGTA TTGATTGGTG CTTGCATCAT GATTTACATT TGAGTGAGTG GGGAACTGGT GAGTAACACGGCGGCGTGCC TAATACATGC AAGTCGAACG AACTCTGGTA TTGATTGGTG CTTGCATCAT GATTTACATT TGAGTGAGTG GGGAACTGGT GAGTAACACG
101 TGGGAAACCT GCCCAGAAGC GGGGGATAAC ACCTGGAAAC AGATGCTAAT101 TGGGAAACCT GCCCAGAAGC GGGGGATAAC ACCTGGAAAC AGATGCTAAT
ACCGCATAAC AACTTGGACC GCATGGTCCG AGTTTGAAAG ATGGCTTCGGACCGCATAAC AACTTGGACC GCATGGTCCG AGTTTGAAAG ATGGCTTCGG
201 CTATCACTTT TGGATGGTCC CGCGGCGTAT TAGCTAGATG GTGGGGTAAC201 CTATCACTTT TGGATGGTCC CGCGGCGTAT TAGCTAGATG GTGGGGTAAC
GGCTCACCAT GGCAATGATA CGTAGCCGAC CTGAGAGGGT AATCGGCCACGGCTCACCAT GGCAATGATA CGTAGCCGAC CTGAGAGGGT AATCGGCCAC
301 ATTGGGACTG AGACACGGCC CAAACTCCTA CGGGAGGCAG CAGTAGGGAA301 ATTGGGACTG AGACACGGCC CAAACTCCTA CGGGAGGCAG CAGTAGGGAA
TCTTCCACAA TGGACGAAAG TCTGATGGAG CAACGCCGCG TGAGTGAAGATCTTCCACAA TGGACGAAAG TCTGATGGAG CAACGCCGCG TGAGTGAAGA
401 AGGGTTTCGG CTCGTAAAAC TCTGTTGTTA AAGAAGAACA TATCTGAAAA401 AGGGTTTCGG CTCGTAAAAC TCTGTTGTTA AAGAAGAACA TATCTGAAAA
TAACTGTTCA GGTATTGACG GTATTTAACC AGAAAGCCAC GGCTAACTACTAACTGTTCA GGTATTGACG GTATTTAACC AGAAAGCCAC GGCTAACTAC
501 GTGCCAGCAG CCGCGGTAAT ACGTAGGTGG CAAGCGTTGT CCGGATTTAT501 GTGCCAGCAG CCGCGGTAAT ACGTAGGTGG CAAGCGTTGT CCGGATTTAT
TGGGCGTAAA GCGAGCGCAG GCGGTTTTTT AAGTCTGATG TGAAAGCCTTTGGGCGTAAA GCGAGCGCAG GCGGTTTTTT AAGTCTGATG TGAAAGCCTT
601 CGGCTCAACC GAAGAAGTGC ATCGGAAACT GGGAAACTTG AGTGCAGAAG601 CGGCTCAACC GAAGAAGTGC ATCGGAAACT GGGAAACTTG AGTGCAGAAG
AGGACAGTGG AACTCCATGT GTTAGCGGTG AAATGCGTAG ATATATGGAAAGGACAGTGG AACTCCATGT GTTAGCGGTG AAATGCGTAG ATATATGGAA
701 GAACACCAGT GGCGAAGGCG GCTGTCTGGT CTGTAACTGA CGCTGAGGCT701 GAACACCAGT GGCGAAGGCG GCTGTCTGGT CTGTAACTGA CGCTGAGGCT
CGAAAGTATG GGTAGCAAAC AGGATTAGAT ACCCTGGTAG TCCATACCGTCGAAAGTATG GGTAGCAAAC AGGATTAGAT ACCCTGGTAG TCCATACCGT
801 AAACGATGAA TGCTAAGTGT TGGAGGGTTT CCGCCCTTCA GTGGCAGCTA801 AAACGATGAA TGCTAAGTGT TGGAGGGTTT CCGCCCTTCA GTGGCAGCTA
ACGCATTAAG CATTCCGCCT GGGGAGTACG GCCGCAAGGC TGAAACTCAAACGCATTAAG CATTCCGCCT GGGGAGTACG GCCGCAAGGC TGAAACTCAA
901 AGGAATTGAC GGGGGCCCGC ACAAGCGGTG GAGCATGTGG TTTAATTCGA901 AGGAATTGAC GGGGGCCCGC ACAAGCGGTG GAGCATGTGG TTTAATTCGA
AGCTACGCGA AGAACCTTAC CAGGTCTTGA CATACTATGC AAATCTAAGAAGCTACGCGA AGAACCTTAC CAGGTCTTGA CATACTATGC AAATCTAAGA
1001 GATTAGACGT TCCCTTCGGG GACATGGATA CAGGTGGTGC ATGGTTGTCG1001 GATTAGACGT TCCCTTCGGG GACATGGATA CAGGTGGTGC ATGGTTGTCG
TCAGCTCGTG TCGTGAGATG TTGGGTTAAG TCCCGCAAAG AGCGCACCCCTCAGCTCGTG TCGTGAGATG TTGGGTTAAG TCCCGCAAAG AGCGCACCCC
1101 TTATTATCAG TTGCCAGCAT TAAGTTGGGC ACTCTGGTGA GACTGCCGGT1101 TTATTATCAG TTGCCAGCAT TAAGTTGGGC ACTCTGGTGA GACTGCCGGT
GACAAACCGG AGGAAGGTGG GGATGACGTC AAATCATCAT GCCCCTTATGGACAAACCGG AGGAAGGTGG GGATGACGTC AAATCATCAT GCCCCTTATG
1201 ACCTGGGCTA CACACGTGCT ACAATGGATG GTACAACGAG TTGCGAACTC1201 ACCTGGGCTA CACACGTGCT ACAATGGATG GTACAACGAG TTGCGAACTC
GCGAGAGTAA GCTAATCTCT TAAAGCCATT CTCAGTTCGG ATTGTAGGCTGCGAGAGTAA GCTAATCTCT TAAAGCCATT CTCAGTTCGG ATTGTAGGCT
1301 GCAACTCGCC TACATGAAGT CGGAATCGCT AG7AATCGC1301 GCAACTCGCC TACATGAAGT CGGAATCGCT AG7AATCGC
W której: a - oznacza adeninę, g - guaninę, c - cytozynę, t -tyminę.In which: a - is adenine, g - guanine, c - cytosine, t-thymine.
Porównanie sekwencji nukleotydowej genu 16S rRNA nowego szczepu Lactobacillus plantarum NBRC101977:Comparison of the nucleotide sequence of the 16S rRNA gene of the new Lactobacillus plantarum NBRC101977 strain:
PL 221 959 B1PL 221 959 B1
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860No. plantarum NBRC101977 Lb. plantarum 0860
Lb. plantarum NBRC101977 Lb. plantarum 0860 (1) (1) (51) (40) (101) (90)No. plantarum NBRC101977 Lb. plantarum 0860 (1) (1) (51) (40) (101) (90)
1151) (140) (201) (190) (251) (240) (301) (290) (351) (340) (401) (390) (451) (440) (501) (490) (551) (540) (601) (590) (651) (640) (700) (690) (750) (740) (800) (790) (850) (840) (900) (888) (950) (938) (1000) (988) (1050) (1038) (1100) (1088)1151) (140) (201) (190) (251) (240) (301) (290) (351) (340) (401) (390) (451) (440) (501) (490) (551) (540) (601) (590) (651) (640) (700) (690) (750) (740) (800) (790) (850) (840) (900) (888) (950) (938 ) (1000) (988) (1050) (1038) (1100) (1088)
5050
GACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAACGAACTCTGGTGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAACGAACTCTGGT
-----------GCGGCGTGCCTAATACATGCAAGTCGAACGAACTCTGGT----------- GCGGCGTGCCTAATACATGCAAGTCGAACGAACTCTGGT
100 attgattggtgcttgcatcatgatttacatttgagtgagtggIgaactgg attgattggtgcttgcatcatgatttacatttgagtgagtgg|gaactgg 101 150100 attgattggtgcttgcatcatgatttacatttgagtgagtggIgaactgg attgattggtgcttgcatcatgatttacatttgagtgagtgg | gaactgg 101 150
TGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAA TGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAA 151 200 cagatgctaataccgcataacaacttggaccgcatggtccgagtttgaaa CAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGTTTGAAA 201 250TGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAA TGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAA 151 200 cagatgctaataccgcataacaacttggaccgACATTGGACGATAGTGGCTAATAGATTGGACGACAGATTGGACGATAGATTGCTACGACAGATTGGACGATAGATTGGACGATAGATTGCTACGATAGATTGGACGA
GATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGAT GATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGAT 251 300GATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGAT GATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGAT 251 300
GGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGG GGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGG 301 350GGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGG GGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGG 301 350
TAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCA TAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCA 351 400TAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCA TAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCA 351 400
GCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGC GCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGC 401 450GCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGC GCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGC 401 450
GTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAAC GTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAAC 451 500GTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAAC GTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAAC 451 500
ATArCTGA|A|TAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCA ATArCTGA|A|TAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCA 501 550ATArCTGA | A | TAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCA ATArCTGA | A | TAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCA 501 550
CGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTG CGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTG 551 600CGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTG CGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTG 551 600
TCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGAT TCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGAT 601 650TCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGAT TCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGAT 601 650
GTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTT GTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTT 651 700GTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTT GTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTT 651 700
GAGTGCAGAAGAGGACAGTGGAACTCCATGTGT|AGCGGTGAAATGCGTA gagtgcagaagaggacagtggaactccatgtgt|agcggtgaaatgcgta 701 750 gatatatggaagaacaccagtggcgaaggcggctgtctggtctgtaactg gatatatggaagaacaccagtggcgaaggcggctgtctggtctgtaactg 751 800GAGTGCAGAAGAGGACAGTGGAACTCCATGTGT | AGCGGTGAAATGCGTA gagtgcagaagaggacagtggaactccatgtgt | agcggtgaaatgcgta 701 750 gatatatggaagaacacTCCATGTGTgactaggtctggacgataggtctgacgactaggtctgactgactaggtgactgacgatggtgactgacgatgcgaaggagaacgactaggtctgactgactaggtgactgacgatgcgaaggagaacgactaggtctgactgacagtaggtctgagact
ACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTA acgctgaggctcgaaagtatgggtagcaaacaggattagataccctggta 801 850ACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTA acgctgaggctcgaaagtatgggtagcaaacaggattagataccctggta 801 850
GTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTC GTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTC 851 900 agti^Bgcagctaacgcattaagcattccgcctggggagtacggccgcaa agtgBgcagctaacgcattaagcattccgcctggggagtacggccgcaa 901 950 ggctgaaactcaaaggaattgacgggggcccgcacaagcggtggagcatg ggctgaaactcaaaggaattgacgggggcccgcacaagcggtggagcatg 951 1000GTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTC GTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTC 851 900 agti ^ Bgcagctaacgcattaagcattccgcctggggagtacggccgcaa agtgBgcagctaacgcattaagcattccgcctggggagtacggccgcaa ggctgaaactcaaaggaattgacgggggcccgcacaagcggtggagcatg ggctgaaactcaaaggaattgacgggggcccgcacaagcggtggagcatg 901 950 951 1000
TGGTTrAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTA TGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTA 1001 1050TGGTTrAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTA TGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTA 1001 1050
TGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGG TGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGG 1051 1100TGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGG TGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGG 1051 1100
TGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCA TGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCA 1101 1150 aBgagcgcaWcccttattatcagttgccagcattaagttgggcactctgg a|gagcgca|cccttattatcagttgccagcattaagttgggcactctggTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCA TGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCA 1101 1150 aBgagcgcaWcccttattatcagttgggagcctccctcattatcagttggggcagttactcagggagcctcagttgggagcctcctca
PL 221 959 B1PL 221 959 B1
11511151
12001200
TGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCAT tgagactgccggtgacaaaccggaggaaggtggggatgacgtcaaatcat 1201 1250TGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCAT tgagactgccggtgacaaaccggaggaaggtggggatgacgtcaaatcat 1201 1250
CATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAAC CATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAAC 1251 1300CATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAAC CATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAAC 1251 1300
GAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTT GAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTT 1301 1350GAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTT GAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTT 1301 1350
CGGATTGTAGGCTGCAACTCGCCTACATGAAGrCGGAATCGCTAGTAATC CGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATC 1351CGGATTGTAGGCTGCAACTCGCCTACATGAAGrCGGAATCGCTAGTAATC CGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATC 1351
GCGC
GCGC
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