Protoplast formation and regeneration in Lactobacillus delbrueckii |
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Authors: | Mamta Singhvi Dipti Joshi Shalaka Gaikaiwari Digambar V. Gokhale |
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Affiliation: | (1) Department of Biological Sciences, Faculty of Science, Shinshu University, Matsumoto Nagano, 390-8621, Japan;(2) Division of Science and Technology, Graduate School of Shinshu University, Matsumoto Nagano, 390-8621, Japan;(3) Present address: Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan; |
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Abstract: | Method for production and regeneration of Lactobacillus delbrueckii protoplasts are described. The protoplasts were obtained by treatment with a mixture of lysozyme and mutanolysin in protoplast buffer at pH 6.5 with different osmotic stabilizers. The protoplasts were regenerated on deMan, Rogosa and Sharpe (MRS) with various osmotic stabilizers. Maximum protoplast formation was obtained in protoplast buffer with sucrose as an osmotic stabilizer using a combination of lysozyme (1 mg/ml) and mutanolysin (10 μg/ml). Maximum protoplast regeneration was obtained on MRS medium with sucrose (0.5 M) as an osmotic stabilizer. The regeneration medium was also applicable to other species of lactobacilli as well. This is, to our knowledge, the first report on protoplast formation and efficient regeneration in case of L. delbrueckii. |
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Keywords: | Lactobacillus delbrueckii Protoplast formation Protoplast regeneration |
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