Blocking sporulation by inhibiting SpoIIE

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing oxygen-containing organic compound

Reexamination Certificate

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C435S006120, C435S252300, C435S252700, C536S023100, C536S024100, C536S024500

Reexamination Certificate

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07432090

ABSTRACT:
We have shown that the control of solventogenesis and sporulation can be genetically uncoupled inC. acetobutylicum. In strain 824(pASspo), the absence of SpoIIE causes sporulation to be blocked at stage II. The cell remains in a vegetative state, and this allows solvent production to proceed for longer and for solvents to accumulate more rapidly and to a higher concentration. The characteristic drop in OD600 observed in wild type and control strains ofC. acetobutylicumafter 48-72 hours as the cells transition from the solventogenic phase to sporulation is notably absent in the fermentations of 824(pASspo). Mutant S (wild type background, spoIIE disrupted), Mutant BS (Mutant B background, spoIIE disrupted), Mutant HS (Mutant H background, spoIIE disrupted) and Mutant bukS (buk- background, spoIIE disrupted) were generated to create stable solvent producing bacteria with complete inactivation of the SpoIIE protein. Similarity between the SpoIIE protein ofC. acetobutylicum, B. subtilis, and otherClostridialspecies indicates that the techniques used inC. acetobutylicumcan be applied to other solvent producingClostridia.

REFERENCES:
patent: 2004/0248279 (2004-12-01), Sawada et al.
Desai et al. Applied and Environmental Microbiology vol. 65(3): 936-945, Mar. 1999.
Scothcer et al. Journal of Bacteriology vol. (6):1930-1936, Mar. 2005.

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