Author(s): Lu Y, Kong R, Hu L
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Abstract This paper describes an enzymatic method for yielding protoplasts from the microalga Chlorella protothecoides. Four kinds of commercially available enzymes were tested. The enzymatic digestion was optimal with 2\% cellulase R-10 and 1\% snailase prepared in 25 mM Tris buffer (pH 6.0) containing 0.6 M D-mannitol, and the protoplast density could reach the peak after treatment at 30°C for 16 h. Nearly all liberated protoplasts were green in the presence of 0.01\% phenosafranin, indicating their high viability. The regeneration rate was about 70\% when 0.6 M D-mannitol was used as an osmotic stabilizer in the regeneration medium. This protocol will find useful applications in genetic studies of this algal species.
This article was published in World J Microbiol Biotechnol
and referenced in Journal of Fundamentals of Renewable Energy and Applications