Cloning of Clostridium cellulovorans endo-1,4-β-glucanase genes

Oded Shoseyov; Tetsuo Hamamoto; Frances Foong; Roy H. Doi

doi:10.1016/0006-291X(90)90382-W

Cloning of Clostridium cellulovorans endo-1,4-β-glucanase genes

Oded Shoseyov^*
, Tetsuo Hamamoto
, Frances Foong
, Roy H. Doi

^*Corresponding author for this work

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

30 Scopus citations

Abstract

A Clostridium cellulovorans lambda gt1l gene bank was screened for endo-1,4-β-glucanase [EC 3.2.1.4, EGase, Carboxy Methyl Cellulase (CMCase)] genes using a chromogenic substrate. Three genes (engA, engB, and engC) were isolated. The engB expressed the most active CMCase. The engA encoded a bifunctional enzyme that displayed endo-1,4-β-glucanase and β-glucosidase activities. The three recombinant glucanases, when expressed in Escherichia coli, were partially degraded into multiform active enzymes as evidenced by their SDS-PAGE-CMC zymograms. None of the clones could degrade crystalline cellulose, thus supporting the hypothesis that the integrity of the C. cellulovorans cellulase complex was essential for its 'true cellulase' activity.

Original language	English
Pages (from-to)	667-672
Number of pages	6
Journal	Biochemical and Biophysical Research Communications
Volume	169
Issue number	2
DOIs	https://doi.org/10.1016/0006-291X(90)90382-W
State	Published - 15 Jun 1990
Externally published	Yes

Bibliographical note

Funding Information:
ACKNOWLEDGMENWTSe: thank Reinhold Bruckner for his suggestions on the cloning strategy. O.S. was supported by fellowship SI-0057-87 from The United State-Israel Binational Agricultural Research and Development Fund. The research was supported in part by Department of Energy grant #DE-FG03-87FR13705( RHD).

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title = "Cloning of Clostridium cellulovorans endo-1,4-β-glucanase genes",

abstract = "A Clostridium cellulovorans lambda gt1l gene bank was screened for endo-1,4-β-glucanase [EC 3.2.1.4, EGase, Carboxy Methyl Cellulase (CMCase)] genes using a chromogenic substrate. Three genes (engA, engB, and engC) were isolated. The engB expressed the most active CMCase. The engA encoded a bifunctional enzyme that displayed endo-1,4-β-glucanase and β-glucosidase activities. The three recombinant glucanases, when expressed in Escherichia coli, were partially degraded into multiform active enzymes as evidenced by their SDS-PAGE-CMC zymograms. None of the clones could degrade crystalline cellulose, thus supporting the hypothesis that the integrity of the C. cellulovorans cellulase complex was essential for its 'true cellulase' activity.",

author = "Oded Shoseyov and Tetsuo Hamamoto and Frances Foong and Doi, \{Roy H.\}",

note = "Funding Information: ACKNOWLEDGMENWTSe: thank Reinhold Bruckner for his suggestions on the cloning strategy. O.S. was supported by fellowship SI-0057-87 from The United State-Israel Binational Agricultural Research and Development Fund. The research was supported in part by Department of Energy grant \#DE-FG03-87FR13705( RHD).",

year = "1990",

month = jun,

day = "15",

doi = "10.1016/0006-291X(90)90382-W",

language = "אנגלית",

volume = "169",

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journal = "Biochemical and Biophysical Research Communications",

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AU - Shoseyov, Oded

AU - Hamamoto, Tetsuo

AU - Foong, Frances

AU - Doi, Roy H.

N1 - Funding Information: ACKNOWLEDGMENWTSe: thank Reinhold Bruckner for his suggestions on the cloning strategy. O.S. was supported by fellowship SI-0057-87 from The United State-Israel Binational Agricultural Research and Development Fund. The research was supported in part by Department of Energy grant #DE-FG03-87FR13705( RHD).

PY - 1990/6/15

Y1 - 1990/6/15

N2 - A Clostridium cellulovorans lambda gt1l gene bank was screened for endo-1,4-β-glucanase [EC 3.2.1.4, EGase, Carboxy Methyl Cellulase (CMCase)] genes using a chromogenic substrate. Three genes (engA, engB, and engC) were isolated. The engB expressed the most active CMCase. The engA encoded a bifunctional enzyme that displayed endo-1,4-β-glucanase and β-glucosidase activities. The three recombinant glucanases, when expressed in Escherichia coli, were partially degraded into multiform active enzymes as evidenced by their SDS-PAGE-CMC zymograms. None of the clones could degrade crystalline cellulose, thus supporting the hypothesis that the integrity of the C. cellulovorans cellulase complex was essential for its 'true cellulase' activity.

AB - A Clostridium cellulovorans lambda gt1l gene bank was screened for endo-1,4-β-glucanase [EC 3.2.1.4, EGase, Carboxy Methyl Cellulase (CMCase)] genes using a chromogenic substrate. Three genes (engA, engB, and engC) were isolated. The engB expressed the most active CMCase. The engA encoded a bifunctional enzyme that displayed endo-1,4-β-glucanase and β-glucosidase activities. The three recombinant glucanases, when expressed in Escherichia coli, were partially degraded into multiform active enzymes as evidenced by their SDS-PAGE-CMC zymograms. None of the clones could degrade crystalline cellulose, thus supporting the hypothesis that the integrity of the C. cellulovorans cellulase complex was essential for its 'true cellulase' activity.

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Cloning of Clostridium cellulovorans endo-1,4-β-glucanase genes

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Bibliographical note

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