Nucleotide sequence of the Ruminococcus albus SY3 endoglucanase genes ce1A and ce1B (original) (raw)
Summary
The complete nucleotide sequences of Ruminococcus albus genes celA and celB coding for endoglucanase A (EGA) and endoglucanase B (EGB), respectively, have been determined. The celA structural gene consists of an open reading frame of 1095 bp. Confirmation of the nucleotide sequence was obtained by comparing the predicted amino acid sequence with that derived by N-terminal analysis of purified EGA. The celB structural gene consists of an open reading frame of 1227 bp; 7 by upstream of the translational start codnn of celB is a typical gram-positive Shine-Dalgarno sequence. The deduced N-terminal region of EGB conforms to the general pattern for the signal peptides of secreted prokaryotic proteins. The complete celB gene, cloned into pUC vectors, caused lethality in Escherichia coli. In contrast, celA cloned in pUC18, under the control of lacZp, directed high-level synthesis of EGA in E. coli JM83. EGA in cell-free extract, purified to near homogeneity by ionexchange chromatography, had a Mr of 44.5 kDa. Gene deletion and subcloning studies with celA revealed that EGA hydrolysed both CMC and xylan, and did not contain discrete functional domains. EGA and EGB showed considerable homology with each other, in addition to exhibiting similarity with Egl (R. albus), EGE (Clostridium thermocellum) and End (Butyrivibrio fibrisolvens).
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Abbreviations
CMC:
carboxymethylcellulose
CMCase:
carboxymethylcellulase
celA :
gene coding for EGA
EGA:
endoglucanase A
celB :
gene coding for EGB
EGB:
endoglucanase B
S-D:
Shine-Dalgarno
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Authors and Affiliations
- Department of Agricultural Biochemistry and Nutrition, University of Newcastle upon Tyne, NE1 7RU, Newcastle upon Tyne, UK
Debbie M. Poole & Harry J. Gilbert - Department of Biochemistry, AFRC Institute of Animal Physiology and Genetics Research, C132 4AT, Babraham, Cambridge, UK
Geoffrey P. Hazlewood, Judith I. Laurie & Patrick J. Barker
Authors
- Debbie M. Poole
- Geoffrey P. Hazlewood
- Judith I. Laurie
- Patrick J. Barker
- Harry J. Gilbert
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Communicated by H. Hennecke
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Poole, D.M., Hazlewood, G.P., Laurie, J.I. et al. Nucleotide sequence of the Ruminococcus albus SY3 endoglucanase genes ce1A and ce1B.Molec. Gen. Genet. 223, 217–223 (1990). https://doi.org/10.1007/BF00265057
- Received: 12 March 1990
- Issue date: September 1990
- DOI: https://doi.org/10.1007/BF00265057