Protoplast formation and regeneration of a thermophilic Clostridium sp.

M Julia Pettinari

doi:10.1016/0378-1097(89)90048-7

Abstract

Protoplast formation and regeneration of a thermophilic Clostridium sp.

Highlights

Thermophilic Clostridium species capable of converting cellulose and hemicellulose to chemicals and fuels in a single step are currently of interest in biotechnology
Protoplast formation For protoplast formation, 5 ml of an early exponential phase culture, OD600 = 0.3, in C M 4 G were centrifuged and resuspended in one-tenth volume of either SMM buffer [9] or CM4 supplemented with 10% lactose (LPM)
Protoplast regeneration For protoplast regeneration the protoplast suspension was serially diluted in LPM containing 0.1 M N-acetylglucosamine and immediately plated on regeneration medium (RM)

Summary

Introduction

Thermophilic Clostridium species capable of converting cellulose and hemicellulose to chemicals and fuels in a single step are currently of interest in biotechnology. A procedure to prepare and regenerate protoplasts is essential for the genetic analysis of Correspondence to: Beatfiz S. Methods for the production and regeneration of protoplasts in Clostridium species have already been published for C. acetobutylicum [1,2,3], C. pasteurianum [4] and C. tertium [5]. Methods suitable for thermophilic Clostridium species are not available, stable L-phase variants were obtained from C. thermocellum [6].

Results

Conclusion