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Title: Secretion of clostridium cellulase by E. coli

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.

Inventors:
 [1]
  1. (1885 California St., #62, Mountain View, CA 94041)
Publication Date:
Research Org.:
University of California
OSTI Identifier:
871590
Patent Number(s):
US 5759845
Assignee:
Yu, Ida Kuo (1885 California St., #62, Mountain View, CA 94041) OAK
DOE Contract Number:  
FG03-87ER13705
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
secretion; clostridium; cellulase; coli; encoding; endocellulase; newly; isolated; mesophilic; strain; iy-2; digest; bamboo; fibers; cellulose; rice; straw; sawdust; shotgun; cloning; expression; plasmid; plc2833; positive; clones; selected; based; ability; hydrolyze; milled; agar; plates; clone; contained; kb; dna; fragment; responsible; activity; western; blot; analyses; indicated; produced; secreted; mass; 58; 000; daltons; identical; size; subunit; major; cellulases; products; digestion; cloned; cellotetraose; soluble; polymers; signal; sequences; capable; directing; heterologous; proteins; host; describes; bioprocess; treatment; cellulosic; plant; materials; produce; cellular; growth; substrates; fermentation; suitable; production; liquid; fuels; solvents; acids; liquid fuel; liquid fuels; cloned dna; dna fragment; blot analyses; signal sequence; selected based; selected base; cellulase activity; /435/

Citation Formats

Yu, Ida Kuo. Secretion of clostridium cellulase by E. coli. United States: N. p., 1998. Web.
Yu, Ida Kuo. Secretion of clostridium cellulase by E. coli. United States.
Yu, Ida Kuo. Thu . "Secretion of clostridium cellulase by E. coli". United States. doi:. https://www.osti.gov/servlets/purl/871590.
@article{osti_871590,
title = {Secretion of clostridium cellulase by E. coli},
author = {Yu, Ida Kuo},
abstractNote = {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.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Thu Jan 01 00:00:00 EST 1998},
month = {Thu Jan 01 00:00:00 EST 1998}
}

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