The cellulase gene from Erwinia chrysanthemi coding for endoglucanase Z was subcloned into a broad-host-range plasmid pGSS33 in Escherichia coli. The recombinant pNB20 was transferred into Zymomonas mobilis ATCC 10988 by mobilization using the helper plasmid RP4. Plasmid pNB20 was stably maintained in E. coli and Z. mobilis hosts. The endoglucanase gene celZ was expressed efficiently and the level of expression was higher in Z. mobilis than in E. coli. The specific activity of the enzyme was comparable to that of the parent strain of Er. chrysanthemi. The proteins produced by Z. mobilis and Er. chrysanthemi presented identical immunological and electrophoretic properties. Biosynthesis of endoglucanase occurred during the exponential growth phase of Z. mobilis and about 35% of the enzyme was released into the medium in the absence of detectable cell lysis. The endoglucanase appeared to be located in the periplasmic space in Z. mobilis.
AfendraA.S.,
DrainasC.1987; Expression and stability of a recombinant plasmid in Zymomonas mobilis and Escherichia coli
. Journal of General Microbiology 133:127–134
BoyerH.W.,
Rouland-DussoixD.1969; A complementation analysis of the restriction and modification of DNA in Escherichia coli
. Journal of Molecular Biology 41:459–472
BoyerM.H.,
ChambostJ.P.,
MagnanM.,
CattaneoJ.1984a; Carboxymethyl-cellulase from Erwinia chrysanthemi. I. Production and regulation of extracellular carboxymethyl-cellulase. Journal of Biotechnology 1:229–239
BoyerM.H.,
ChambostJ.P.,
MagnanM.,
CattaneoJ.1984b; Carboxymethyl-cellulase from Erwinia chrysanthemi. II. Purification and partial characterization of an endo-β,4-glucanase. Journal of Biotechnology 1:241–252
BoyerM.H.,
CamiB.,
KotoujanskiA.,
ChambostJ.P.,
FrixonC.1987; Isolation of the gene encoding the major endoglucanase of Erwinia chrysanthemi. Homology between cel genes of two strains of Erwinia chrysanthemi
. FEMS Microbiology Letters 41:351–356
Brestic-GoachetN.,
GunasekaranP.,
CamiB.,
BarattiJ.1987; Transfer and expression of broad host range plasmids in Zymomonas mobilis
. Biotechnology Letters 9:13–18
ByunM.O.K.,
KaperJ.B.,
IngramL.O.1986; Construction of a new vector for the expression of foreign genes in Zymomonas mobilis
. Journal of Industrial Microbiology 1:9–15
CareyV.C.,
WaliaS.K.,
IngramL.O.1983; Expression of a lactose transposon (Tn957) in Zymomonas mobilis
. Applied and Environmental Microbiology 46:1163–1168
ConwayT.,
OsmanY.A.,
KonnanJ.I.,
HoffmanE.M.,
IngramL.O.1987b; Promoter and nucleotide sequences of the Zymomonas mobilis pyruvate decarboxylase. Journal of Bacteriology 169:949–954
DattaN.,
HedgesR.W.,
ShawE.J.,
SykesR.B.,
RichmondM.H.1971; Properties of an R factor from Pseudomonas aeruginosa
. Journal of Bacteriology 108:1244–1249
LejeuneA.,
EveleighD.,
ColsonC.1988; Expression of an endoglucanase gene of Pseudomonas fluorescens var.ellulosa in Zymomonas mobilis
. FEMS Microbiology Letters 49:363–366
NeuH.C.,
HeppelL.A.1965; The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. Journal of Biological Chemistry 240:3685–3692
NossalN.G.,
HeppelL.A.1966; The release of enzymes by osmotic shock from Escherichia coli in exponential phase. Journal of Biological Chemistry 241:3055–3062
SkotnickiM.L.,
LeeK.J.,
TribeD.E.,
RogersP.L.1981; Comparison of ethanol production by different Zymomonas strains. Applied and Environmental Microbiology 41:889–893
TeatherR.M.,
WoodP.J.1982; Use of Congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. Applied and Environmental Microbiology 43:777–780
TowbinH.,
StaehelinT.,
GordonJ.1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America 76:4350–4354
WillisR.C.,
CirakogluR.G.,
MorrisC.,
SchellenbergG.D.,
GerberN.H.,
FuriongC.E.1974; Preparation of the periplasmic binding proteins from Salmonella typhimurium and Escherichia coli
. Archives of Biochemistry and Biophysics 161:64–75
YanaseH.,
YasuiM.,
MiyazakiT.,
TonomuraK.1985; Fusion of spheroplasts and genetic recombination of Zymomonas mobilis
. Agricultural and Biological Chemistry 49:133–140