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Two new thermostable alpha-L-rhamnosidases from a novel thermophilic bacterium

Birgisson, Hakon LU ; Hreggvidsson, G O; Fridjonsson, O H; Mort, A; Kristjansson, J K and Mattiasson, Bo LU (2004) In Enzyme and Microbial Technology 34(6). p.561-571
Abstract
Two new thermostable alpha-L-rhamnosidases with novel substrate hydrolysis pattern were cloned and expressed from a new thermophilic bacterium. Fragments of the two alpha-L-rhamnosidase genes, rhmA and rhmB were identified in a partially sequenced genome of the bacterium. Whole genes were recovered by amplifying flanking sequences with single specific primers and nonspecific walking primers. The recovered Genes were then cloned into Escherichia coli and their enzymes produced and purified. Both enzymes were dimers and the MW of the monomers. were 104 and 107 kDa for RhmA and RhmB, respectively. Both rhamnosidases had a temperature optimum at 70degreesC. RhmA had pH optimum at 7.9 and RhmB had a broad pH optimum of 5.0 to 6.9 and RhmA had... (More)
Two new thermostable alpha-L-rhamnosidases with novel substrate hydrolysis pattern were cloned and expressed from a new thermophilic bacterium. Fragments of the two alpha-L-rhamnosidase genes, rhmA and rhmB were identified in a partially sequenced genome of the bacterium. Whole genes were recovered by amplifying flanking sequences with single specific primers and nonspecific walking primers. The recovered Genes were then cloned into Escherichia coli and their enzymes produced and purified. Both enzymes were dimers and the MW of the monomers. were 104 and 107 kDa for RhmA and RhmB, respectively. Both rhamnosidases had a temperature optimum at 70degreesC. RhmA had pH optimum at 7.9 and RhmB had a broad pH optimum of 5.0 to 6.9 and RhmA had over 50% activity in the pH interval 5.0 to 8.7 and RhmB in the pH interval 4.0 to 7.9. Both enzymes had over 20% residual activity after 24-h incubation at 60degreesC. RhmA and RhmB had K values of 0.46 and 0.66 mM and V-max values of 134 and 352 U mg(-1) respectively, on p-nitrophenyl-alpha-L-rhamnopyrano side. Both rhamnosidases were active on both alpha-1,2- and alpha-1,6-linkages to beta-D-glucoside. (C) 2004 Elsevier Inc. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Cloning, α-Image-Rhamnosidases, Thermostable
in
Enzyme and Microbial Technology
volume
34
issue
6
pages
561 - 571
publisher
Elsevier
external identifiers
  • wos:000221037000005
  • scopus:1842610070
ISSN
0141-0229
DOI
10.1016/j.enzmictec.2003.12.012
language
English
LU publication?
yes
id
25a13e2d-007b-45f2-8855-e7f98e9ea434 (old id 140693)
date added to LUP
2007-07-02 08:14:26
date last changed
2017-11-19 03:25:38
@article{25a13e2d-007b-45f2-8855-e7f98e9ea434,
  abstract     = {Two new thermostable alpha-L-rhamnosidases with novel substrate hydrolysis pattern were cloned and expressed from a new thermophilic bacterium. Fragments of the two alpha-L-rhamnosidase genes, rhmA and rhmB were identified in a partially sequenced genome of the bacterium. Whole genes were recovered by amplifying flanking sequences with single specific primers and nonspecific walking primers. The recovered Genes were then cloned into Escherichia coli and their enzymes produced and purified. Both enzymes were dimers and the MW of the monomers. were 104 and 107 kDa for RhmA and RhmB, respectively. Both rhamnosidases had a temperature optimum at 70degreesC. RhmA had pH optimum at 7.9 and RhmB had a broad pH optimum of 5.0 to 6.9 and RhmA had over 50% activity in the pH interval 5.0 to 8.7 and RhmB in the pH interval 4.0 to 7.9. Both enzymes had over 20% residual activity after 24-h incubation at 60degreesC. RhmA and RhmB had K values of 0.46 and 0.66 mM and V-max values of 134 and 352 U mg(-1) respectively, on p-nitrophenyl-alpha-L-rhamnopyrano side. Both rhamnosidases were active on both alpha-1,2- and alpha-1,6-linkages to beta-D-glucoside. (C) 2004 Elsevier Inc. All rights reserved.},
  author       = {Birgisson, Hakon and Hreggvidsson, G O and Fridjonsson, O H and Mort, A and Kristjansson, J K and Mattiasson, Bo},
  issn         = {0141-0229},
  keyword      = {Cloning,α-Image-Rhamnosidases,Thermostable},
  language     = {eng},
  number       = {6},
  pages        = {561--571},
  publisher    = {Elsevier},
  series       = {Enzyme and Microbial Technology},
  title        = {Two new thermostable alpha-L-rhamnosidases from a novel thermophilic bacterium},
  url          = {http://dx.doi.org/10.1016/j.enzmictec.2003.12.012},
  volume       = {34},
  year         = {2004},
}