Secretion of Clostridium cellulase by E. coli

Details Secretion of Clostridium cellulase by E. coli Secretion of Clostridium cellulase by E. coli

1993-08-11

1996-03-05

Wax, Robert A.

Chemistry: molecular biology and microbiology

Micro-organism, per se ; compositions thereof; proces of...

Bacteria or actinomycetales; media therefor

435209, 4352402, 43525233, 43525411, 4353201, C12N 115, C12N 121, C12N 510, C12N 942

Patent

active

054967252

ABSTRACT:
A gene, encoding an endocellulase from a newly isolated mesophilic Clostridium strain IY-2 which can digest bamboo fibers, cellulose, rice straw, and sawdust, was isolated by shotgun cloning in an E. coli expression plasmid pLC2833. E. coli positive clones were selected based on their ability to hydrolyze milled bamboo fibers and cellulose present in agar plates. One clone contained a 2.8 kb DNA fragment that was responsible for cellulase activity. Western blot analyses indicated that the positive clone produced a secreted cellulase with a mass of about 58,000 daltons that was identical in size to the subunit of one of the three major Clostridium cellulases. The products of cellulose digestion by this cloned cellulase were cellotetraose and soluble higher polymers. The cloned DNA contained signal sequences capable of directing the secretion of heterologous proteins from an E. coli host. The invention describes a bioprocess for the treatment of cellulosic plant materials to produce cellular growth substrates and fermentation end products suitable for production of liquid fuels, solvents, and acids.

REFERENCES:
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Ohmiya et al., Appl. Env. Microbiol. 55:2399-2402 (1989).
Fujino et al., J. Bacteriol. 171:4076-4079 (1989).
Yu, I. K. et al.; Abstr. Annu. Meet. Am. Soc. Microbiol., 87th Meeting, p. 272 (1987).
Shima, S.; Biol Abstr. 88(8): 84088 (1989).
Langsford, M. L. et al.; J. Gen. Microbiol. 130:1367-1376 (1984).

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