Strain improvement ofArthrobacter simplex by protoplast fusion |
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Authors: | Dr W-H Liu L-W Chow C-K Lo |
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Institution: | (1) Graduate Institute of Agricultural Chemistry, National Taiwan University, Taipei, Taiwan, Republic of China;(2) Present address: Department of Food Health, Chia-Nan College of Pharmacy, Tainan, Taiwan, Republic of China |
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Abstract: | Anl-tryptophan auxotroph and milky mutants were derived from an inducible cholesterol oxidase-producing bacterium,Arthrobacter simplex USA18, via UV-mutagenesis. Protoplasts of these mutants and a constitutive cholesterol oxidase producer, strain US3011, were prepared by growing cells in the presence of ampicillin (20 g ml–1) followed by digestion with lysozyme. Protoplast fusion between tested strains with complementary characteristics was achieved in the presence of 20–40% polyethylene glycol 6000. The fusion frequency was about 1.5–1.7×10–3. The cholesterol oxidase activity of four fusants in a cholesterol-containing medium was 20–60% higher than that of parental strains. This study demonstrated that protoplast fusion is applicable to strain improvement ofArthrobacter strains for enzyme production. |
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Keywords: | protoplast fusion protoplasting cholesterol oxidase Arthrobacter simplex |
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