Affiliation:
1. Laboratory of Microbiology and Immunology, National Institute of Dental Research, Bethesda, Maryland 20205
Abstract
Methods for the production and regeneration of
Lactobacillus casei
protoplasts are described. Protoplasts of
L. casei
strains were obtained by treatment with mutanolysin or with mutanolysin and lysozyme together in a protoplast formation buffer containing 0.02 M HEPES (
N
-2-hydroxyethylpiperazine-
N′
-2-ethanesulfonic acid) (pH 7.0), 1 mM MgCl
2
, 0.5% gelatin, and 0.3 M raffinose. Cells were regenerated on a complex medium supplemented with bovine serum albumin, MgCl
2
, CaCl
2
, gelatin, and raffinose. Lengthy digestion with lytic enzymes inhibited the capacity of protoplasts to regenerate. The optimum conditions of protoplast formation varied from strain to strain. Using predetermined optimal conditions it was possible to prepare protoplasts of several
L. casei
strains and regenerate them with 10 to 40% efficiency. The methods were applicable to other species of lactobacilli as well.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Reference39 articles.
1. Studies on regeneration media for Bacillus subtilus protoplasts;Akamatsu T.;Agric. Biol. Chem.,1981
2. Clostridium acetobutylicum protoplast formation and regeneration;Allcock E. R.;Appl. Environ. Microbiol.,1982
3. Protoplast fusion in Streptomyces: conditions for efficient genetic recombination and cell regeneration;Baltz R. H.;J. Gen. Microbiol.,1981
4. Transformation of plasmid DNA into Streptomyces at high frequency;Bibb M. J.;Nature (London),1978
5. Development of competence in the Bacillus subtilis transformation system;Bott K. F.;J. Bacteriol.,1967
Cited by
51 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献