Chitinase from Enterobacter sp. NRG4: Its purification, characterization and reaction pattern

Enterobacter sp. NRG4 was shown to excrete chitinase into the culture supernatant when cultivated in medium containing chitin. A 60 kDa extracellular chitinase was purified to homogeneity and characterized. The enzyme hydrolyzed swollen chitin, colloidal chitin, regenerated chitin and glycol chitin...

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Autores principales:	Dahiya,Neetu, Tewari,Rupinder, Tiwari,Ram P, Singh Hoondal,Gurinder
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2005
Materias:	chemical modification chitinase Enterobacter sp. NRG4 purification substrate binding
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582005000200003
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spelling	oai:scielo:S0717-345820050002000032005-10-13Chitinase from Enterobacter sp. NRG4: Its purification, characterization and reaction patternDahiya,NeetuTewari,RupinderTiwari,Ram PSingh Hoondal,Gurinder chemical modification chitinase Enterobacter sp. NRG4 purification substrate binding Enterobacter sp. NRG4 was shown to excrete chitinase into the culture supernatant when cultivated in medium containing chitin. A 60 kDa extracellular chitinase was purified to homogeneity and characterized. The enzyme hydrolyzed swollen chitin, colloidal chitin, regenerated chitin and glycol chitin but did not hydrolyze chitosan. The chitinase exhibited Km and Vmax values of 1.43 mg ml-1 and 83.33 µM µg-1 h-1 for swollen chitin, 1.41 mg ml-1 and 74.07 µM µg-1 h-1 for colloidal chitin, 1.8 mg ml-1 and 40 µM µg-1 h-1 for regenerated chitin and 2.0 mg ml-1 and 33.33 µM µg-1 h-1 for glycol chitin, respectively. The optimal temperature and pH for activity were 45ºC and pH 5.5, respectively. Mg2+, K+ and Ca2+ stimulated chitinase activity by 13, 16 and 18%, respectively whereas Cu2+, Co2+, Ag+ and Hg2+ inhibited chitinase activity by 9.7, 15, 22 and 72.2%, respectively at 1 mM concentration. N-bromosuccinamide (NBS) at 1 mM and iodoacetamide at 10 mM concentration completely inhibited the enzyme activity. Dithiobisnitrobenzoic acid (DTNB) at 10 mM concentration inhibited chitinase activity by 97.2%. Chitin was hydrolyzed to chitobiose and N-acetyl D-glucosamine when incubated with the purified enzyme. The hydrolysis pattern of the purified enzyme indicated that the chitinase was an endochitinaseinfo:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.8 n.2 20052005-08-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582005000200003en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	chemical modification chitinase Enterobacter sp. NRG4 purification substrate binding
spellingShingle	chemical modification chitinase Enterobacter sp. NRG4 purification substrate binding Dahiya,Neetu Tewari,Rupinder Tiwari,Ram P Singh Hoondal,Gurinder Chitinase from Enterobacter sp. NRG4: Its purification, characterization and reaction pattern
description	Enterobacter sp. NRG4 was shown to excrete chitinase into the culture supernatant when cultivated in medium containing chitin. A 60 kDa extracellular chitinase was purified to homogeneity and characterized. The enzyme hydrolyzed swollen chitin, colloidal chitin, regenerated chitin and glycol chitin but did not hydrolyze chitosan. The chitinase exhibited Km and Vmax values of 1.43 mg ml-1 and 83.33 µM µg-1 h-1 for swollen chitin, 1.41 mg ml-1 and 74.07 µM µg-1 h-1 for colloidal chitin, 1.8 mg ml-1 and 40 µM µg-1 h-1 for regenerated chitin and 2.0 mg ml-1 and 33.33 µM µg-1 h-1 for glycol chitin, respectively. The optimal temperature and pH for activity were 45ºC and pH 5.5, respectively. Mg2+, K+ and Ca2+ stimulated chitinase activity by 13, 16 and 18%, respectively whereas Cu2+, Co2+, Ag+ and Hg2+ inhibited chitinase activity by 9.7, 15, 22 and 72.2%, respectively at 1 mM concentration. N-bromosuccinamide (NBS) at 1 mM and iodoacetamide at 10 mM concentration completely inhibited the enzyme activity. Dithiobisnitrobenzoic acid (DTNB) at 10 mM concentration inhibited chitinase activity by 97.2%. Chitin was hydrolyzed to chitobiose and N-acetyl D-glucosamine when incubated with the purified enzyme. The hydrolysis pattern of the purified enzyme indicated that the chitinase was an endochitinase
author	Dahiya,Neetu Tewari,Rupinder Tiwari,Ram P Singh Hoondal,Gurinder
author_facet	Dahiya,Neetu Tewari,Rupinder Tiwari,Ram P Singh Hoondal,Gurinder
author_sort	Dahiya,Neetu
title	Chitinase from Enterobacter sp. NRG4: Its purification, characterization and reaction pattern
title_short	Chitinase from Enterobacter sp. NRG4: Its purification, characterization and reaction pattern
title_full	Chitinase from Enterobacter sp. NRG4: Its purification, characterization and reaction pattern
title_fullStr	Chitinase from Enterobacter sp. NRG4: Its purification, characterization and reaction pattern
title_full_unstemmed	Chitinase from Enterobacter sp. NRG4: Its purification, characterization and reaction pattern
title_sort	chitinase from enterobacter sp. nrg4: its purification, characterization and reaction pattern
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2005
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582005000200003
work_keys_str_mv	AT dahiyaneetu chitinasefromenterobacterspnrg4itspurificationcharacterizationandreactionpattern AT tewarirupinder chitinasefromenterobacterspnrg4itspurificationcharacterizationandreactionpattern AT tiwariramp chitinasefromenterobacterspnrg4itspurificationcharacterizationandreactionpattern AT singhhoondalgurinder chitinasefromenterobacterspnrg4itspurificationcharacterizationandreactionpattern
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Chitinase from Enterobacter sp. NRG4: Its purification, characterization and reaction pattern

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