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Differences and similarities in enzymes from the neopullulanase subfamily isolated from thermophilic species

Nordberg Karlsson, Eva LU ; Labes, Antje; Turner, Pernilla LU ; Fridjonsson, Olafur H.; Wennerberg, Christina LU ; Pozzo, Tania; Hreggvidson, Gudmundur O.; Kristjansson, Jakob K. and Schoenheit, Peter (2008) 5th International Symposiium Biology and Taxonomy of Green Algae In Biologia 63(6). p.1006-1014
Abstract
Six glycoside hydrolase (GH) family 13 members, classified under the polyspecific neopullulanase subfamily GH13_20 (also termed cyclomaltodextrinase) were analysed. They originate from thermophilic bacterial strains (Anoxybacillus flavithermus, Laceyella sacchari, and Geobacillus thermoleovorans) or from environmental DNA, collected after in situ enrichments in Icelandic hot springs. The genes were isolated following the CODEHOP consensus primer strategy, utilizing the first two of the four conserved sequence regions in GH13. The typical domain structure of GH13 20, including an N-terminal domain (classified as CBM34), the catalytic module composed of the A- and B- domains, and a C- terminal domain, was found in five of the encoded enzymes... (More)
Six glycoside hydrolase (GH) family 13 members, classified under the polyspecific neopullulanase subfamily GH13_20 (also termed cyclomaltodextrinase) were analysed. They originate from thermophilic bacterial strains (Anoxybacillus flavithermus, Laceyella sacchari, and Geobacillus thermoleovorans) or from environmental DNA, collected after in situ enrichments in Icelandic hot springs. The genes were isolated following the CODEHOP consensus primer strategy, utilizing the first two of the four conserved sequence regions in GH13. The typical domain structure of GH13 20, including an N-terminal domain (classified as CBM34), the catalytic module composed of the A- and B- domains, and a C- terminal domain, was found in five of the encoded enzymes (abbreviated Amy1, 89, 92, 98 and 132). These five enzymes degraded cyclomaltodextrins (CDs) and starch, while only three, Amy92 (L. sacchari), Amy98 (A. flavithermus) and Amy132 (environmental DNA), also harboured neopullulanase activity. The L. sacchari enzyme was monomeric, but with CD as the preferred substrate, which is an unusual combination. The sixth enzyme (Amy29 from environmental DNA), was composed of the ABC-domains only. Preferred substrate for Amy29 was pullulan, which was degraded to panose, and the enzyme had no detectable activity on CDs. In addition to its different activity pro. le and domain composition, Amy29 also displayed a different conservation (LPKF) in the fifth conserved region (MPKL) proposed to identify the subfamily. All enzymes had apparent temperature optima in the range 50-65 degrees C, while thermostability varied, and was highest for Amy29 with a half-life of 480 min at 80 degrees C. Calcium dependent activity or stability was monitored in four enzymes, but could not be detected for Amy29 or 98. Tightly bound calcium can, however, not be ruled out, and putative calcium ligands were conserved in Amy98. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
keywords
starch, environmental DNA, GH13, cyclomaltodextrinase, calcium, neopullulanase, maltogenic amylase
in
Biologia
volume
63
issue
6
pages
1006 - 1014
publisher
Versita
conference name
5th International Symposiium Biology and Taxonomy of Green Algae
external identifiers
  • wos:000261401600036
  • scopus:57349146948
ISSN
0006-3088
DOI
10.2478/s11756-008-0171-3
language
English
LU publication?
yes
id
21a1b29e-2396-40cd-af32-c3336a7e2a5e (old id 1305758)
date added to LUP
2009-03-23 10:16:25
date last changed
2017-07-02 04:01:11
@inproceedings{21a1b29e-2396-40cd-af32-c3336a7e2a5e,
  abstract     = {Six glycoside hydrolase (GH) family 13 members, classified under the polyspecific neopullulanase subfamily GH13_20 (also termed cyclomaltodextrinase) were analysed. They originate from thermophilic bacterial strains (Anoxybacillus flavithermus, Laceyella sacchari, and Geobacillus thermoleovorans) or from environmental DNA, collected after in situ enrichments in Icelandic hot springs. The genes were isolated following the CODEHOP consensus primer strategy, utilizing the first two of the four conserved sequence regions in GH13. The typical domain structure of GH13 20, including an N-terminal domain (classified as CBM34), the catalytic module composed of the A- and B- domains, and a C- terminal domain, was found in five of the encoded enzymes (abbreviated Amy1, 89, 92, 98 and 132). These five enzymes degraded cyclomaltodextrins (CDs) and starch, while only three, Amy92 (L. sacchari), Amy98 (A. flavithermus) and Amy132 (environmental DNA), also harboured neopullulanase activity. The L. sacchari enzyme was monomeric, but with CD as the preferred substrate, which is an unusual combination. The sixth enzyme (Amy29 from environmental DNA), was composed of the ABC-domains only. Preferred substrate for Amy29 was pullulan, which was degraded to panose, and the enzyme had no detectable activity on CDs. In addition to its different activity pro. le and domain composition, Amy29 also displayed a different conservation (LPKF) in the fifth conserved region (MPKL) proposed to identify the subfamily. All enzymes had apparent temperature optima in the range 50-65 degrees C, while thermostability varied, and was highest for Amy29 with a half-life of 480 min at 80 degrees C. Calcium dependent activity or stability was monitored in four enzymes, but could not be detected for Amy29 or 98. Tightly bound calcium can, however, not be ruled out, and putative calcium ligands were conserved in Amy98.},
  author       = {Nordberg Karlsson, Eva and Labes, Antje and Turner, Pernilla and Fridjonsson, Olafur H. and Wennerberg, Christina and Pozzo, Tania and Hreggvidson, Gudmundur O. and Kristjansson, Jakob K. and Schoenheit, Peter},
  booktitle    = {Biologia},
  issn         = {0006-3088},
  keyword      = {starch,environmental DNA,GH13,cyclomaltodextrinase,calcium,neopullulanase,maltogenic amylase},
  language     = {eng},
  number       = {6},
  pages        = {1006--1014},
  publisher    = {Versita},
  title        = {Differences and similarities in enzymes from the neopullulanase subfamily isolated from thermophilic species},
  url          = {http://dx.doi.org/10.2478/s11756-008-0171-3},
  volume       = {63},
  year         = {2008},
}