Plasmid pBUL1 derived from a lactobacillus and derivatives there

Details Plasmid pBUL1 derived from a lactobacillus and derivatives there Plasmid pBUL1 derived from a lactobacillus and derivatives there

1992-10-22

1995-06-20

Schwartz, Richard A.

Chemistry: molecular biology and microbiology

Micro-organism, per se ; compositions thereof; proces of...

Bacteria or actinomycetales; media therefor

43525231, 4353201, 536 231, 536 241, C12N 1511, C12N 121, C12N 1574, C12N 1575

Patent

active

054260477

DESCRIPTION:

BRIEF SUMMARY
Field of the Invention

The present invention relates to a novel circular double-stranded DNA plasmid pBUL1 derived from Lactobacillus delbrueckii subsp. bulgaricus, the utility and safety of which as a yogurt-producing bacterium are well known, and to derivatives thereof, as well as to microorganisms transformed with the plasmids.


Prior Art And Problems To Be Solved By The Invention

Lactic acid bacteria are very useful microorganisms which have long been used in production of various fermented foods. If a recombinant DNA technology which has rapidly been developed in recent years could be applied to lactic acid bacteria, greater enhancement of the utility of the bacteria would be expected. In fact, host-vector systems with a fairly high efficiency have already been reported for lactic acid bacteria of some species of, for example, Lactococcus lactis (reference 1), Streptococcus salivarius subsp. thermophilus (reference 2), Lactobacillus plantarum (reference 3) and Lactobacillus casei (reference 4), and the current stage of the technical improvement is on an industrial scale use of host-vector systems on an industrial scale. However, despite of great efforts by many researchers, there has hitherto been no report of transformation of Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus delbrueckii subsp. lactis (hereinafter referred to as Lb. bulgaricus, and Lb. lactis, respectively), which are used extensively as bacteria for producing milk products such as yogurt. Application of broad-host-range plasmids, such as pNZ12 (reference 5), pGK12 (reference 6) and pIL253 (reference 7), with which transformants of lactic acid bacteria of some species have been obtained, to the above-mentioned two subspecies has been attempted, but transformation was not successful. However, establishment of host-vector systems of the abovementioned two subspecies has been earnestly desired. If a recombinant DNA technology is applied to microorganisms to be used in production of foods, the safety of vectors to be used for transformation of them must be established. As vectors for this purpose, those vectors are desirable which exist naturally in microorganisms that have been used to produce foods which have traditional been and the safety of which has been historically confirmed. On the other hand, fermented milk products such as yogurt are foods which have been eaten for a long period of time and their safety has been well confirmed. Therefore, the plasmids derived from microorganisms from fermented milk products, for example, those of the abovementioned two subspecies, are useful vectors in order to construct transformants of microorganims for food production. In addition they are also available vectors for transformation for the purpose of producing various physiologically active substances.


Means For Solving The Problems

For the purpose of developing host-vector systems for Lb. bulgaricus and Lb. lactis, the present inventors variously investigated and studied plasmids of these subspecies. As a result, they are the first who have succeeded in isolating a plasmid from Lb. bulgaricus M-878 strain (FERM BP-3758) possessed by Meiji Institute of Health Science of Meiji Milk Products Co., Ltd., which has a length of about 7 kbp, which has a restriction endonuclease map as shown in FIG. 1, and which does not have recognition sites for BamHI, EcoRI, KpnI, PstI and SalI, the base sequence (SEQ ID No. 1) of the SmaI fragment with 1344 bp being represented as the Sequence Number 1 of the Sequence Table in Table 1 below. They named the plasmid pBUL1.


TABLE 1 __________________________________________________________________________ Sequence Table: Sequence Number: 1 Length of Sequence: 1344 Type of Sequence: Nucleic Acid Number of Strand: Double-stranded Topology: Linear Kind of Sequence: Other Nucleic Acid Origin: Name of Microorganism: Lactobacillus delbrueckii subsp. bulgaricus Name of Strain: M-878 Sequence: CCCGGGGCGA AACGACATGG GGCGCTCAAA CCATTGCTGA GGCGATCAAT TACGTGCAAG 60 CCC

REFERENCES:
Ohtaguchi et al. (1987), J. Chem. Eng. Japan 20(6): 557-562.
Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.) pp. 1.12-1.20.
Somkuti, Steinberg, General method for plasmid DNA isolation from thermophilic lactic acid bacteria, Journal of Biotechnology, vol. 3, pp. 323-332, 1986.
Langella, Chopin, Conjugal transfer to plasmid pIP501 from Lactococcus lactis to Lactobacillus delbruckii subsp. bulgaricus and Lactobacillus heleveticus, FEMS Microbiology Letters, vol. 60, pp. 149-152, 1989.
Todd R. Klaenhammer, "A General Method for Plasmid Isolation in Lactobacilli", Current Microbiology, vol. 10, 1984, pp. 23-28.
M. Vescovo, et al., "Evidence of Plasmid Deoxyribonucleic Acid in Lactobacillus", Microbiologica (Bologna), vol. 4, No. 4, 1981, pp. 413-420.
V. Miteva, et al., "Isolation and Characterization of Plasmids from Different Strains of Lactobacillus-bulgaricus Lactobacillus-. . . ", Acta Microbiologica Bulgarica, vol. 27, No. 0, 1991, pp. 3-8.
Helge Holo, et al., "High-Frequency Transformation, by Electroporation, of Lactococcus lactis subsp. cremoris Grown with Glycine in . . . ", Applied and Enviromental Microbiology, vol./351 10 55, No. 12, Dec. 1989, pp. 3119-3123.
Annick Mercenier, "Molecular genetics of Streptococcus thermophilus", FEMS Microbiology Reviews, vol. 87, 1990, pp. 61-78.
Trees Scheirlinck, "Integration and Expression of .alpha.-Amylase and Endoglucanase Genes in the Lactobacillus plantarum Chromosome", Applied and Environmental Microbiology, vol. 55, No. 9, Sep. 1989, pp. 2130-2137.
Bruce Chassey et al., "Transformation of Lactobacillus casei by electroporation", FEMS Microbiology Letters vol. 44, 1987, pp. 173-177.
Willem De Vos, "Gene cloning and expression in lactic streptococci", FEMS Microbiology Reviews, vol. 46, 1987, pp. 281-295.
Jan Kok, "Construction of Plasmid Cloning Vectors for Lactic Streptococci Which Also Replicate in Bacillus subtilis and Escherichia coli", Applied and Eviromental Microbiology, vol. 48, No. 4, Oct. 1984, pp. 726-731.
Simon et al., "Construction of a vector plasmid family and its use for molecilar cloning in Streptococcus latis", Biochimie, vol. 70, 1988, pp. 559-566.
Efthymiou et al., "An Antigenic analysis of Lactobacillus Acidophilus", University of Maryland Department of Microbiology, Dec. 18, 1961, pp. 258-267.
Anderson et al., "Simple and Rapid Method for Isolating Large Plasmid DNA from Lactic Streptococci", Applied and Environnmental Microbiology, vol. 46, No. 3, Sep. 1983, pp. 549-552.
LeBlanc et al., "Physical and Genetic Analyses of Streptococcal Plasmid pAM.beta.1 and Cloning of Its Replication Region", Journal of Bacteriology, vol. 157, No. 2, Feb. 1984, pp. 445-453.
Chang et al., "High Frequency Transformation of Bacillus subtilis Protoplasts by Plasmid DNA", Molecular Genetics, vol. 168, 1979, pp. 111-115.
Yamane et al., "Changes in the Properties and Molecular Weights of Bacillus subtilis M-type and N-type .alpha.-Amylases Resulting from a Spontaneous Deletion", J. Biochem., vol. 96, 1984, pp. 1849-1858.
Ando et al., "Kosoh-kin (Bacillus subtilis)", Lectures on Fundamental Microbiology, 1987, pp. 168-215.

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