Cloning, sequence analysis, and expression of genes encoding Xylan-degrading enzymes from the thermophile 'Caldocellum saccharolyticum'

E. Luthi; D. R. Love; J. McAnulty; C. Wallace; P. A. Caughey; D. Saul; P. L. Bergquist

Cloning, sequence analysis, and expression of genes encoding Xylan-degrading enzymes from the thermophile 'Caldocellum saccharolyticum'

E. Luthi, D. R. Love, J. McAnulty, C. Wallace, P. A. Caughey, D. Saul, P. L. Bergquist^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

111 Citations (Scopus)

Abstract

A λ recombinant bacteriophage coding for xylanase and β-xylosidase activity has been isolated from a genomic library of the extremely thermophilic anaerobe 'Caldocellum saccharolyticum'. Partial Sau3AI fragments of the λ recombinant DNA were ligated into pBR322. A recombinant plasmid with an insertion of ca. 7 kilobases of thermophilic DNA expressing both enzymatic activities was isolated. The location of the genes has been established by analyzing deletion derivatives, and the DNA sequence of 6.067 kilobases of the insert has been determined. Five open reading frames (ORFs) were found, one of which (ORF1; M(r) 40,455) appears to code for a xylanase (XynA) which also acts on o-nitrophenyl-β-D-xylopyranoside. Another, ORF5 (M(r) 56,365), codes for a β-xylosidase (XynB). The xynA gene product shows significant homology with the xylanases from the alkalophilic Bacillus sp. strain C125 and Clostridium thermocellum.

Original language	English
Pages (from-to)	1017-1024
Number of pages	8
Journal	Applied and Environmental Microbiology
Volume	56
Issue number	4
Publication status	Published - 1990
Externally published	Yes

Cite this

@article{32adf69300fb4573a9714228a37adbaf,

title = "Cloning, sequence analysis, and expression of genes encoding Xylan-degrading enzymes from the thermophile 'Caldocellum saccharolyticum'",

abstract = "A λ recombinant bacteriophage coding for xylanase and β-xylosidase activity has been isolated from a genomic library of the extremely thermophilic anaerobe 'Caldocellum saccharolyticum'. Partial Sau3AI fragments of the λ recombinant DNA were ligated into pBR322. A recombinant plasmid with an insertion of ca. 7 kilobases of thermophilic DNA expressing both enzymatic activities was isolated. The location of the genes has been established by analyzing deletion derivatives, and the DNA sequence of 6.067 kilobases of the insert has been determined. Five open reading frames (ORFs) were found, one of which (ORF1; M(r) 40,455) appears to code for a xylanase (XynA) which also acts on o-nitrophenyl-β-D-xylopyranoside. Another, ORF5 (M(r) 56,365), codes for a β-xylosidase (XynB). The xynA gene product shows significant homology with the xylanases from the alkalophilic Bacillus sp. strain C125 and Clostridium thermocellum.",

author = "E. Luthi and Love, {D. R.} and J. McAnulty and C. Wallace and Caughey, {P. A.} and D. Saul and Bergquist, {P. L.}",

year = "1990",

language = "English",

volume = "56",

pages = "1017--1024",

journal = "Applied and Environmental Microbiology",

issn = "0099-2240",

publisher = "American Society for Microbiology",

number = "4",

}

TY - JOUR

T1 - Cloning, sequence analysis, and expression of genes encoding Xylan-degrading enzymes from the thermophile 'Caldocellum saccharolyticum'

AU - Luthi, E.

AU - Love, D. R.

AU - McAnulty, J.

AU - Wallace, C.

AU - Caughey, P. A.

AU - Saul, D.

AU - Bergquist, P. L.

PY - 1990

Y1 - 1990

N2 - A λ recombinant bacteriophage coding for xylanase and β-xylosidase activity has been isolated from a genomic library of the extremely thermophilic anaerobe 'Caldocellum saccharolyticum'. Partial Sau3AI fragments of the λ recombinant DNA were ligated into pBR322. A recombinant plasmid with an insertion of ca. 7 kilobases of thermophilic DNA expressing both enzymatic activities was isolated. The location of the genes has been established by analyzing deletion derivatives, and the DNA sequence of 6.067 kilobases of the insert has been determined. Five open reading frames (ORFs) were found, one of which (ORF1; M(r) 40,455) appears to code for a xylanase (XynA) which also acts on o-nitrophenyl-β-D-xylopyranoside. Another, ORF5 (M(r) 56,365), codes for a β-xylosidase (XynB). The xynA gene product shows significant homology with the xylanases from the alkalophilic Bacillus sp. strain C125 and Clostridium thermocellum.

AB - A λ recombinant bacteriophage coding for xylanase and β-xylosidase activity has been isolated from a genomic library of the extremely thermophilic anaerobe 'Caldocellum saccharolyticum'. Partial Sau3AI fragments of the λ recombinant DNA were ligated into pBR322. A recombinant plasmid with an insertion of ca. 7 kilobases of thermophilic DNA expressing both enzymatic activities was isolated. The location of the genes has been established by analyzing deletion derivatives, and the DNA sequence of 6.067 kilobases of the insert has been determined. Five open reading frames (ORFs) were found, one of which (ORF1; M(r) 40,455) appears to code for a xylanase (XynA) which also acts on o-nitrophenyl-β-D-xylopyranoside. Another, ORF5 (M(r) 56,365), codes for a β-xylosidase (XynB). The xynA gene product shows significant homology with the xylanases from the alkalophilic Bacillus sp. strain C125 and Clostridium thermocellum.

UR - http://www.scopus.com/inward/record.url?scp=0025237055&partnerID=8YFLogxK

M3 - Article

C2 - 2111111

AN - SCOPUS:0025237055

SN - 0099-2240

VL - 56

SP - 1017

EP - 1024

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

IS - 4

ER -

Cloning, sequence analysis, and expression of genes encoding Xylan-degrading enzymes from the thermophile 'Caldocellum saccharolyticum'

Abstract

Other files and links

Fingerprint

Cite this