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Purification and characterization of chitinases from the nematophagous fungi Verticillium chlamydosporium and v. suchlasporium

Tikhonov, VE; Lopez-Llorca, LV; Salinas, J and Jansson, Hans-Börje LU (2002) In Fungal Genetics and Biology 35(1). p.67-78
Abstract
Culture filtrates of the nematophagous fungi Verticillium chlamydosporium and V. suchlasporium growing on colloidal chitin showed increasing chitinolytic activity and production of two (32- and 43-kDa) main proteins. Maximum activity was found 18-20 days after inoculation, but V. suchlasporium always displayed higher activity. Zymography of such filtrates on carboxymethyl-chitin-Remazol brilliant violet 5R/acrylamide gels showed five bands of substrate degradation for V. suchlasporium and three for V. chlamydosporium. Filtrates with maximum activity were chromatographed on macroporous cross-linked chitin affinity matrix, showing a peak of main (50-60%) activity, which only contained a 43-kDa protein for both fungi. Zymography and colloidal... (More)
Culture filtrates of the nematophagous fungi Verticillium chlamydosporium and V. suchlasporium growing on colloidal chitin showed increasing chitinolytic activity and production of two (32- and 43-kDa) main proteins. Maximum activity was found 18-20 days after inoculation, but V. suchlasporium always displayed higher activity. Zymography of such filtrates on carboxymethyl-chitin-Remazol brilliant violet 5R/acrylamide gels showed five bands of substrate degradation for V. suchlasporium and three for V. chlamydosporium. Filtrates with maximum activity were chromatographed on macroporous cross-linked chitin affinity matrix, showing a peak of main (50-60%) activity, which only contained a 43-kDa protein for both fungi. Zymography and colloidal chitin degradation showed that it was a single endochitinase (CHl43) with optimum pH range of 5.2-5.7. The main isoforms had pis of 7.6 for V. suchlasporium and 7.9 for V. chlamydosporium. Eggs of the nematode Globodera pallida treated with CHl43 and the serine protease P32 from V. suchlasporium alone or in combination showed surface damage in comparison with controls when examined by scanning electron microscopy. (C) 2002 Elsevier Science (USA). (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
nematophagous fungi, eggshell degradation, chitinases, affinity purification, biocontrol, plant parasitic nematodes
in
Fungal Genetics and Biology
volume
35
issue
1
pages
67 - 78
publisher
Elsevier
external identifiers
  • wos:000176534100006
  • pmid:11860266
  • scopus:0036352560
ISSN
1087-1845
DOI
10.1006/fgbi.2001.1312
language
English
LU publication?
yes
id
6c377e7d-8d30-4bc0-b659-4ce9f86610a4 (old id 334178)
date added to LUP
2007-11-16 09:19:15
date last changed
2017-12-10 04:35:22
@article{6c377e7d-8d30-4bc0-b659-4ce9f86610a4,
  abstract     = {Culture filtrates of the nematophagous fungi Verticillium chlamydosporium and V. suchlasporium growing on colloidal chitin showed increasing chitinolytic activity and production of two (32- and 43-kDa) main proteins. Maximum activity was found 18-20 days after inoculation, but V. suchlasporium always displayed higher activity. Zymography of such filtrates on carboxymethyl-chitin-Remazol brilliant violet 5R/acrylamide gels showed five bands of substrate degradation for V. suchlasporium and three for V. chlamydosporium. Filtrates with maximum activity were chromatographed on macroporous cross-linked chitin affinity matrix, showing a peak of main (50-60%) activity, which only contained a 43-kDa protein for both fungi. Zymography and colloidal chitin degradation showed that it was a single endochitinase (CHl43) with optimum pH range of 5.2-5.7. The main isoforms had pis of 7.6 for V. suchlasporium and 7.9 for V. chlamydosporium. Eggs of the nematode Globodera pallida treated with CHl43 and the serine protease P32 from V. suchlasporium alone or in combination showed surface damage in comparison with controls when examined by scanning electron microscopy. (C) 2002 Elsevier Science (USA).},
  author       = {Tikhonov, VE and Lopez-Llorca, LV and Salinas, J and Jansson, Hans-Börje},
  issn         = {1087-1845},
  keyword      = {nematophagous fungi,eggshell degradation,chitinases,affinity purification,biocontrol,plant parasitic nematodes},
  language     = {eng},
  number       = {1},
  pages        = {67--78},
  publisher    = {Elsevier},
  series       = {Fungal Genetics and Biology},
  title        = {Purification and characterization of chitinases from the nematophagous fungi Verticillium chlamydosporium and v. suchlasporium},
  url          = {http://dx.doi.org/10.1006/fgbi.2001.1312},
  volume       = {35},
  year         = {2002},
}