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KMID : 0903519990420010012
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
1999 Volume.42 No. 1 p.12 ~ p.19
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Cell fusion and fusants characterization of Bacillus strains producing biopolymer
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Abstract
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To improve biopolymer productivity and properties of Bacillus strains, protoplast fusion was performed between biopolymer producing Bacillus subtilis K-1 and lactose utilizing Bacillus coagulans. The results were as follows; Protoplasts mixed in fusion fluid containing 33% PEG 6000, 1% PVP and 10 mM CaCl, were reacted for 5 min at 37℃ and then centrifused protoplasts were directly overlaid on the selective media containing 100 ㎍/㎖ antibiotics and incubated for 3 days. At this conditions, the frequency of protoplast fusion was generally in the range of 4.6 × 10^(-5) to 1.8 × 10^(-7) in ratio. Segregation ratio was observed between 1 to 6% indicating genetic stability of all the fusants. Fusants growth were also observed on the media contained amino acid and antibiotics as required marked materials. DNA contents of the selected fusants were 1.6 to 2.7 times more than that of parent strains. With observation by TEM microscopy, spherical protoplasts were first released from the swollen parental cells and then contracted to fuse in the process of fusion. And fused cells were observed representative vesicle. Originally, the parental cells were observed as in the morphology of thick-walled and double membrane-surrounded rod shape with TEM microscopy.
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