WO1993005161A1 - Lactobacillus presentant une activite hydrolytique genetiquement manipulee des sels biliaires, leur production et leur utilisation - Google Patents

Lactobacillus presentant une activite hydrolytique genetiquement manipulee des sels biliaires, leur production et leur utilisation Download PDF

Info

Publication number
WO1993005161A1
WO1993005161A1 PCT/NL1992/000158 NL9200158W WO9305161A1 WO 1993005161 A1 WO1993005161 A1 WO 1993005161A1 NL 9200158 W NL9200158 W NL 9200158W WO 9305161 A1 WO9305161 A1 WO 9305161A1
Authority
WO
WIPO (PCT)
Prior art keywords
bile salt
salt hydrolase
microorganism
gene
genetic manipulation
Prior art date
Application number
PCT/NL1992/000158
Other languages
English (en)
Inventor
Rob Leer
Mark Posno
Harry Christiaens
Willy Verstraete
Original Assignee
Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno
Universiteit Gent
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno, Universiteit Gent filed Critical Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno
Publication of WO1993005161A1 publication Critical patent/WO1993005161A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • C12N9/80—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in linear amides (3.5.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
    • C12N15/746—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for lactic acid bacteria (Streptococcus; Lactococcus; Lactobacillus; Pediococcus; Enterococcus; Leuconostoc; Propionibacterium; Bifidobacterium; Sporolactobacillus)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the invention relates to the modified bacteria themselves, to methods by which they can be obtained and to recombinant nucleotides useful therefor.
  • Bile salts are synthesized in the liver, starting from cholesterol. Via the bile, they are secreted into the liver
  • Bile salts are biological detergents which make fats and fatty acids from the food water-soluble by forming
  • chylomicrons (micelles). At the basis of these detergent properties lies their amphiphilic structure: on the
  • hydrophobic steroid chain stands a hydrophilic side-chain, consisting of a carbonyl group which, via an amide-bond, is conjugated with taurine or glycine, referred to as
  • tauroconjugates or glycoconjugates respectively (bile salt hydrolysis; Fig. 1).
  • This conjugation promotes the secretion into the intestinal tract.
  • Secreted into the intestinal tract, bile salts can be deconjugated (Fig. 1). This conversion is purely bacterial and cannot be carried out by the host itself.
  • the deconjugative bacteria include the following genera:
  • Streptococcus Streptococcus, Peptostreptococcus and Lactobacillus.
  • BSH Bile Salt Hydrolase
  • Bile salt deconjugation is considered an important cause of growth depression in animals (Feighner and Dashkevicz, 1987, 1988).
  • the hydrophilic side-chain is split off from the hydrophobic nucleus and the bile salt loses part of its amphiphilic character (Roda et al., 1990).
  • the deconjugated metabolite moreover has a higher pKa value than the original substrate (Fini and Roda, 1987) so that upon acidification it precipitates easily and is thus withdrawn from the fat digestion process.
  • Deconjugated bile salts are also pricipitated more easily by Ca-phosphates (Van der Meer, 1989).
  • BSH activity Repopulation of the gastrointestinal tract with bacteria having a reduced capacity with respect to the BSH synthesis could contribute to an improvement of the growth achievements of the animals by increased fat absorption.
  • Bile salt hydrolysis combines the two principles
  • the invention utilizes the properties of BSH.
  • bacterial hydrolysis leads to deconjugated bile salts which pricipitate as Ca-complexes (Van der Meer, 1989) and are thus withdrawn from the enterohepatic cycle.
  • the bile salt stock constitutes an endogenous source of glycine and taurine which is released by hydrolysis and, once resorbed, can reduce the serum cholesterol
  • bacteria having increased BSH activity have a favourable influence on man's health in that the increased bile salt hydrolytic activity interferes with the uptake of fats as well as with the enterohepatic bile salt cycle.
  • the addition of such bacteria to the diet could thus contribute to a reduction of the serum cholesterol concentration in humans.
  • lactobacilli play an important role in the preparation of a variety of dairy products (cheese, yoghurt, buttermilk, and the like), in the fermentation of vegetables and meats and in wine-making, this use can be realized via a number of routes.
  • Bile salts are by nature biological detergents having a strong bactericidal activity. In order to be able to survive the bile salt stress of the small intestines, microorganisms must therefore be bile salt resistant.
  • a BSH-active probiotic can be considered a prophylactic preventing the colonization of pathogenic microorganisms.
  • lactobacilli play an important role in the preparation of a variety of dairy products (cheese, yoghurt, buttermilk and the like), in the fermentation of vegetables and meats and in wine-making, this application can be realized via a number of routes.
  • the cholesterol reducing effect will be enhanced by increased BSH activity. Since the colonization of bile salt sensitive microorganisms is prevented, the proportion of lactobacilli in the microflora of the (small) intestines will increase. As a result, the total BSH activity in the intestines will be additionally increased, which will lead to a further reduction of the serum cholesterol concentration.
  • the invention relates to methods for the construction, using recombinant DNA techniques, of two classes of
  • Lactobacillus strains as well as to the genetically modified strains themselves and to their uses.
  • the gene that codes for an enzyme that hydrolyses bile salts has been changed such that no active enzyme is produced.
  • the method described can be used for the construction of Lactobacillus strains which can be used to favourably influence the growth achievements of animal species which are economically relevant.
  • the present invention provides a microorganism of the genus Lactobacillus which, as a result of genetic manipulation, is capable of expressing a bile salt hydrolase which it does not naturally express, or expresses a naturally expressed bile salt hydrolase to a greater or lesser extent or no longer.
  • this concerns a microorganism which, as a result of genetic manipulation with a recombinant
  • polynucleotide contains a gene coding for an active bile salt hydrolase and is capable of expressing the active bile salt hydrolase, preferably a microorganism wherein the gene coding for an active bile salt hydrolase has been incorporated into the genome of the microorganism or is located on an
  • the microorganism preferably contains several copies of the gene coding for an active bile salt hydrolase.
  • the invention concerns a microorganism which, as a result of genetic manipulation with a recombinant polynucleotide, contains an inactivated bile salt hydrolase gene and is incapable of expressing an active bile salt hydrolase, preferably a microorganism wherein a naturally present bile salt hydrolase gene has been replaced with an inactivated bile salt hydrolase gene through gene exchange, while for the gene exchange preferably use has been made of a non-replicating plasmid or DNA fragment on which an
  • interrupted or deleted bgh gene is located.
  • the bile salt hydrolase gene which may or may not be inactivated, introduced by genetic manipulation, is derived from the bile salt hydrolase gene of a bile salt hydrolase-producing
  • Lactobacillus strain preferably a bile salt hydrolase- producing Lactobacillus plantarum strain.
  • the modified microorganism belongs to a
  • Lactobacillus species which is permissible in foodstuffs for humans or animals or occurs in the gastrointestinal tract of humans or animals.
  • the invention extends to foodstuffs containing a
  • the invention provides a method for producing a microorganism of the genus Lactobacillus which is capable of expressing a bile salt hydrolase which it does not naturally express, wherein the microorganism, by means of genetic manipulation with a recombinant polynucleotide, is provided with a gene coding for an active bile salt hydrolase, such that the microorganism is capable of expressing the active bile salt hydrolase; a method for producing a microorganism of the genus Lactobacillus which no longer expresses a naturally expressed bile salt hydrolase, wherein a bile salt hydrolase gene of the microorganism is replaced with an inactivated bile salt hydrolase gene by means of genetic manipulation with a recombinant polynucleotide, such that the microorganism is no longer capable of expressing an active bile salt hydrolase; and a method for producing a microorganism of the genus
  • Lactobacillus which expresses a naturally expressed bile salt hydrolase to a greater or lesser extent, wherein a bile salt hydrolase gene of the microorganism is replaced with a
  • the invention provides a recombinant
  • polynucleotide (preferably consisting of recombinant DNA) which comprises a bile salt hydrolase gene derived from a bile salt hydrolase-producing Lactobacillus strain, which gene either codes for an inactive bile salt hydrolase or has been inactivated; and a recombinant cloning or expression vector for cloning or expressing in a microorganism of the genus
  • Lactobacillus a bile salt hydrolase gene, which may or may not be inactivated, this recombinant vector comprising a plasmid or bacteriophage transformation vector provided with an insertion of a polynucleotide according to the invention.
  • recombinant vectors examples include the recombinant plasmids pLP3537-BSH and pEI2-BSH to be described hereinafter.
  • the invention further extends to the uses. These include a method for improving the growth achievements of a human or an animal, comprising a step wherein at least a part of the bacteria present in the gastrointestinal tract are replaced with a microorganism of the genus Lactobacillus which, as a result of genetic manipulation, expresses a naturally
  • a microorganism of the genus Lactobacillus is introduced into the gastrointestinal tract, which microorganism, as a result, of genetic manipulation, is capable of expressing a bile salt hydrolase which it does not naturally express, or expresses a naturally expressed bile salt hydrolase to a greater extent.
  • rDNA recombinant DNA
  • the invention relates to a method by which the synthesis of BSH in BSH-forming lactobacilli is prevented and to a method by which the synthesis of BSH in lactobacilli is increased.
  • the gene from Lactobacillus plantarum which codes for a bile salt hydrolase was cloned in Escherichia coli (Christiaens et al., 1990).
  • the gene product could be isolated from the periplasmatic space of E. coli , which suggests that the enzyme is secreted extra-cellularly by L. plantarum.
  • the gene product is active in particular with respect to glycine conjugates (Table 1).
  • the pH optimum is between 4 and 6, the temperature optimum is between 30 and 45°C.
  • the nucleotide sequence of the gene was determined (SEQ ID NO:1; Fig. 2).
  • the gene codes for a protein of 42kD.
  • the Vector pEI2-TH1-GAM the coding sequence of the bsh gene was interrupted by introducing a gene which codes for resistance against chloramphenicol (Fig. 3).
  • the plasmid pEI2-TH1-CAM was made by linearizing the plasmid pEl2-TH1 with Clal and ligating thereto a Clal-TaqI fragment on which the cjal gene is located, interruption of the BSH-coding gene prevents the possibility of an enzymatically active product, i.e. BSH, being formed from this gene.
  • Lactobacillus bacteria in principle offers the possibility of targeted integration of the vector at a specific site of the Lactobacillus genome. Such a method had previously been described for other microorganisms (Chopin et al., 1989), but is not known for Lactobacillus.
  • the provision of parts of the BSH-coding gene in the vector both to the left and to the right of the cml gene creates the possibility that the entire vector (via a so-called single cross-over), or the cml gene and flanking sequences (via a so-called double cross-over) are integrated at the site of the bsh gene.
  • L. plantarum LP80 is very suitable for this purpose.
  • Lactobacillus strains which can be used as probiotics in animals having one stomach, for instance pigs, it is required that Lactobacillus strains are isolated from this organism, which satisfy the following criteria:
  • the BSH activity of the strain constitutes an important part of the total BSH activity in the intestines;
  • the strain is capable of maintaining itself in the intestines for a long time
  • the strain must be relatively simple to culture, in large amounts;
  • the BSH strains constructed in accordance with the invention can be used for the improvement of the growth achievements of different animal species.
  • the cloned gene was also incorporated into a shuttle vector (pLP3537); Posno et al., 1991) which is capable of autonomous replication in a large number of Lactobacillus strains, resulting in the vector pLP3537BSH (Fig. 5).
  • Transformation using pLP3537BSH of E. coli leads to the formation of enzymatically active bile salt hydrolase.
  • Transformation of L. plantarum results in a strong increase of BSH activity.
  • the activity can be detected by the formation of a precipitate of bile salts (halo) around the bacteria colony when the bacteria are plated on an agar culture medium to which taurine
  • Fig. 1 shows the general structure of bile salts and the process of bile salt deconjugation, where R represents the group CH 2 COOH in the case of glycoconjugates and the group CH 2 CH 2 SO 3 H in the case of tauroconjugates.
  • Fig. 2 shows the nucleotide sequence of the bsil gene of Lactobacillus plantarum LP80.
  • Fig. 3 shows the structure of the plasmid pEI2-TH1-CAM.
  • Fig. 4 schematically shows the exchange of the bsh gene of Lactobacillus plantarum LP80 for a bsll gene in which the chloramphenicol-resistance gene (cml) has been incorporated.
  • Fig. 5 shows the structure of the plasmid pLP3537BSH.
  • Fig. 6 is a reproduction of a photograph showing the formation of a precipitate of taurodeoxycholic acid of four Lactobacillus plantarum LP80 transformants containing the plasmid pLP3537BSH. In the center of the picture a non- transformed Lactobacillus plantarum LP80 strain is shown. References
  • GCT GAT AAA GTT AAT ATC ACA CCA TTT GAA TTA ATT CCT TGG TTA 315 Ala Asp Lys Val Asn lle Thr Pro Phe Glu Leu lle Pro Trp Leu

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Microorganisme du genre Lactobacillus pouvant, grâce à une manipulation génétique, exprimer une hydrolase de sel biliaire qu'il n'exprimerait pas de manière naturelle, ou exprimer dans une mesure plus ou moins grande ou plus du tout une hydrolase de sel biliaire exprimée de manière naturelle. On a également prévu un procédé de production d'un tel microorganisme et l'utilisation de celui-ci.
PCT/NL1992/000158 1991-09-11 1992-09-11 Lactobacillus presentant une activite hydrolytique genetiquement manipulee des sels biliaires, leur production et leur utilisation WO1993005161A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
NL9101537 1991-09-11
NL9101537A NL9101537A (nl) 1991-09-11 1991-09-11 Lactobacilli met genetisch gemodificeerde galzouthydrolytische activiteit, produktie en toepassing daarvan.

Publications (1)

Publication Number Publication Date
WO1993005161A1 true WO1993005161A1 (fr) 1993-03-18

Family

ID=19859689

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/NL1992/000158 WO1993005161A1 (fr) 1991-09-11 1992-09-11 Lactobacillus presentant une activite hydrolytique genetiquement manipulee des sels biliaires, leur production et leur utilisation

Country Status (3)

Country Link
AU (1) AU2646192A (fr)
NL (1) NL9101537A (fr)
WO (1) WO1993005161A1 (fr)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0671468A1 (fr) * 1994-03-11 1995-09-13 The Calpis Food Industry Co., Ltd. Bactéries lactiques du genre lactobacillus
EP0792586A3 (fr) * 1996-02-28 1999-04-07 Unilever N.V. Produits alimentaires contenant des bactéries ayant une activité hypocholestérolémique
WO2004076657A2 (fr) 2003-02-28 2004-09-10 Mcgill University Compositions cellulaires et enzymatiques destinees a la modulation d'acides biliaires, de cholesterols et de triglycerides
CN101948857A (zh) * 2010-08-16 2011-01-19 中国农业大学 重组干酪乳杆菌工程菌株及其制备方法
EP2419114B1 (fr) 2009-05-01 2016-04-20 UAS Laboratories LLC Compositions bactériennes destinées au traitement et à la prophylaxie de maladies dégénératives
CN105524907A (zh) * 2016-02-01 2016-04-27 江南大学 一种底物专一性提高的胆盐水解酶突变体
CN103992991B (zh) * 2013-11-11 2016-06-29 江南大学 一种胆盐水解酶突变体及其应用
WO2017123592A1 (fr) * 2016-01-11 2017-07-20 Synlogic, Inc. Bactérie manipulée pour traiter des troubles associés aux sels biliaires
US11291693B2 (en) 2015-06-25 2022-04-05 Synlogic Operating Company, Inc. Bacteria engineered to treat metabolic diseases

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
APPL. ENVIRON. MICROBIOL vol. 55, no. 7, July 1989, pages 1848 - 1851 G. TANNOCK ET AL 'Lactobacilli and bile salt hydrolase in the murine intestinal tract' *
APPL. ENVIRON. MICROBIOL. vol. 55, no. 7, July 1989, pages 1769 - 1774 M-C. CHOPIN ET AL 'Insertion and amplification of foreign genes in the Lactococcus subsp. lactis chromosome' cited in the application *
APPL. ENVIRONM. MICROBIOL. vol. 57, no. 6, June 1991, pages 1822 - 1828 M. POSNO ET AL 'Incompatability of Lactobacillus plasmids and segregational instability of the introduced vectors' *
DISSERTATION ABSTRACTS INT. B vol. 51, no. 7, 1991, page 3207 D WALKER 'The importance of bile salt hydrolase by Lactobacillus acidophilus and its genetic location' *
H. CHRISTIAENS ET AL. In: G. VENEMA ET AL: Proceedings of the Third Symposium on Lactic Acid Bacteria, 17-21-September 1990 Wageningen, The Netherlands, page 115 cited in the application *
J. BACTERIOLOGY vol. 172, no. 8, August 1990, pages 4171 - 4177 S.LUNDEEN EN D. SAVAGE 'Characterization and purification of bile salt hydrolase from Lactobacillus sp. strain 100-100' cited in the application *
JOURNAL OF DAIRY SCIENCE vol. 67, 1984, CHAPAIGN, ILLINOIS US pages 3045 - 3051 S. GILLILAND ET AL 'Importance of bile salt tolerance of Lactobacillus acidophilus used as a dietary adjunct' *

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0671468A1 (fr) * 1994-03-11 1995-09-13 The Calpis Food Industry Co., Ltd. Bactéries lactiques du genre lactobacillus
EP0792586A3 (fr) * 1996-02-28 1999-04-07 Unilever N.V. Produits alimentaires contenant des bactéries ayant une activité hypocholestérolémique
US9637729B2 (en) 2003-02-28 2017-05-02 Uas Laboratories Llc Cell and enzyme compositions for modulating bile acids, cholesterol and triglycerides
WO2004076657A2 (fr) 2003-02-28 2004-09-10 Mcgill University Compositions cellulaires et enzymatiques destinees a la modulation d'acides biliaires, de cholesterols et de triglycerides
WO2004076657A3 (fr) * 2003-02-28 2004-12-29 Univ Mcgill Compositions cellulaires et enzymatiques destinees a la modulation d'acides biliaires, de cholesterols et de triglycerides
US7939061B2 (en) 2003-02-28 2011-05-10 Micropharma Limited Cell and enzyme compositions for modulating bile acids, cholesterol and triglycerides
US8932578B2 (en) 2003-02-28 2015-01-13 Uas Laboratories Llc Cell and enzyme compositions for modulating ale acids, cholesterol and triglycerides
EP2419114B1 (fr) 2009-05-01 2016-04-20 UAS Laboratories LLC Compositions bactériennes destinées au traitement et à la prophylaxie de maladies dégénératives
US10660857B2 (en) 2009-05-01 2020-05-26 Uas Laboratories Llc Bacterial compositions for prophylaxis and treatment of degenerative disease
EP2419114B2 (fr) † 2009-05-01 2019-02-27 UAS Laboratories LLC Compositions bactériennes destinées au traitement et à la prophylaxie de maladies dégénératives
CN101948857A (zh) * 2010-08-16 2011-01-19 中国农业大学 重组干酪乳杆菌工程菌株及其制备方法
CN103992991B (zh) * 2013-11-11 2016-06-29 江南大学 一种胆盐水解酶突变体及其应用
US11291693B2 (en) 2015-06-25 2022-04-05 Synlogic Operating Company, Inc. Bacteria engineered to treat metabolic diseases
US11896627B2 (en) 2015-06-25 2024-02-13 Synlogic Operating Company, Inc. Bacteria engineered to treat metabolic diseases
WO2017123592A1 (fr) * 2016-01-11 2017-07-20 Synlogic, Inc. Bactérie manipulée pour traiter des troubles associés aux sels biliaires
CN105524907A (zh) * 2016-02-01 2016-04-27 江南大学 一种底物专一性提高的胆盐水解酶突变体

Also Published As

Publication number Publication date
AU2646192A (en) 1993-04-05
NL9101537A (nl) 1993-04-01

Similar Documents

Publication Publication Date Title
EP1002051B1 (fr) Souches probiotiques provenant de lactobacillus salivarius et agents antimicrobiens obtenus a partir de celles-ci
AU2004241895B2 (en) A probiotic composition comprising at least two lactic acid bacterial strains which are able to colonise the gastrointestinal tracts in combination with having intestinal survival property, intestinal binding property, an infection protection property and a fiber fermenting property
JP5875975B2 (ja) ロバの乳から分離されたプロバイオティクス微生物
Bengoa et al. Physicochemical, immunomodulatory and safety aspects of milks fermented with Lactobacillus paracasei isolated from kefir
US20140227789A1 (en) Stress tolerant bifidobacteria
Daly et al. Technological and health benefits of dairy starter cultures
Mollet Genetically improved starter strains: opportunities for the dairy industry
WO1993005161A1 (fr) Lactobacillus presentant une activite hydrolytique genetiquement manipulee des sels biliaires, leur production et leur utilisation
Burns et al. Bile-resistant derivatives obtained from non-intestinal dairy lactobacilli
AU753558B2 (en) Production of L(+)-Lactate
Molin et al. Effect of fermented oatmeal soup on the cholesterol level and the Lactobacillus colonization of rat intestinal mucosa
Sen et al. Molecular characterization and in vitro analyses of a sporogenous bacterium with potential probiotic properties
Banina et al. Characterization of natural isolate Lactobacillus acidophilus BGRA43 useful for acidophilus milk production
Gawai et al. Safety aspects of fermented and probiotic foods
JP3954711B2 (ja) 液体発酵によるビスパン菌の生産方法
WO2001088150A1 (fr) Bacteries d'acide lactique probiotiques incapables d'utiliser le lactose
US20040014221A1 (en) Plasmid originated from bifidobacterium, recombinant expression vector using the plasmid and transformation method
KR100621657B1 (ko) 내산성과 내담즙성이 좋고 효소의 생산이 우수한 신규바실러스 서브틸리스 유비티-엠02 균주 , 이 신규 균주를이용 함유하는 사료첨가제 및 이 사료첨가제를 함유하는가축 사료
O'Sullivan Primary sources of probiotic cultures
Ebrahimi et al. Native chicken-derived Lactobacillus spp. strains with high probiotic, cholesterol-assimilation and aflatoxin-degradation capabilities
Lee et al. Potential probiotic properties of Lactococcus lactis NK34 isolated from jeotgal
WO2005001068A1 (fr) Souche de bifidobacterium breve lmc520 contenant un plasmide pbc520, procede de preparation d'acides gras conjugues et de laits fermentes contenant de tels acides gras au moyen de ladite souche, utilisation d'un plasmide pbc520
JP4473570B2 (ja) 生体内での微生物の生残性を向上させる方法及び生残性が向上した微生物
Temirova Antibiotic resistance and probiotic properties of lactic acid bacteria isolated from camel milk and shubat
KR100804193B1 (ko) 락타아제를 생산하는 신균주 락토바실러스 아시도필러스ｌａ－１２

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
NENP Non-entry into the national phase

Ref country code: CA

122 Ep: pct application non-entry in european phase