Journal of Molecular Catalysis B: Enzymatic, ( ISI ), Volume (132), No (1), Year (2016-7) , Pages (75-83)

Title : ( Biochemical characterization and gene cloning of a novel alkaline endo-1-4-glucanase from Bacillus subtilis DR8806 )

Authors: somaye ramezani , Ahmad Asoodeh ,

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In the pre­sent study, an endo-1-4-glu­canase was iso­lated from Bacil­lus sub­tilis DR8806. The en­zyme was pu­ri­fied to ho­mo­gene­ity via salt pre­cip­i­ta­tion and ion-ex­change chro­matog­ra­phy. SDS-PAGE analy­sis re­vealed a mol­e­c­u­lar mass of 52 kDa. Op­ti­mum pH of en­zyme was 9.5 and the en­zyme was sta­ble at pH range of 8.5–10.5. The op­ti­mum tem­per­a­ture of en­zyme was found to be 55 °C and it showed a re­mark­able sta­bil­ity at tem­per­a­tures be­tween 40 and 60 °C. The en­zyme ac­tiv­ity was stim­u­lated by Co2+, K+, Mg2+, Ca2+ and Na+ ions while Hg2+, Mn2+, Pb2+ and Zn2+ ions were found to in­hibit the en­zyme ac­tiv­ity. The en­zyme ac­tiv­ity was de­creased by in­creas­ing in con­cen­tra­tion of β-mer­cap­toethanol, EDTA, SDS, PMSF and Tri­ton X-100. Or­ganic sol­vents such as hexane, 2-propanol, ace­tone and ethanol (20% v/​v) stim­u­lated en­zyme ac­tiv­ity by 110%, 114%, 119% and 128%, re­spec­tively. Im­i­da­zolium-based ionic liq­uids (ILs) had in­hibitory ef­fects on endo-1-4-glu­canase ac­tiv­ity. Us­ing car­boxymethyl cel­lu­lose as sub­strate, ki­netic pa­ra­me­ters of Km ap­par­ent and Vmax were cal­cu­lated to be 1.49% (W/​V) and 66.66 μM min−1 mg−1, re­spec­tively. Endo-1-4-glu­canase cod­ing gene of B. sub­tilis DR8806 was iden­ti­fied by T/​A cloning. The com­pu­ta­tional mod­el­ing of endo-1-4-glu­canase showed that two Glu residues are at ac­tive site. Our re­sults sug­gest that the en­zyme has great of po­ten­tial ap­pli­ca­tions in hy­drolyz­ing cel­lu­losic sub­strates

Keywords

, Endo-1-4-glu­canase, Bio­chem­i­cal char­ac­ter­i­za­tion, Gene
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@article{paperid:1057190,
author = {Ramezani, Somaye and Asoodeh, Ahmad},
title = {Biochemical characterization and gene cloning of a novel alkaline endo-1-4-glucanase from Bacillus subtilis DR8806},
journal = {Journal of Molecular Catalysis B: Enzymatic},
year = {2016},
volume = {132},
number = {1},
month = {July},
issn = {1381-1177},
pages = {75--83},
numpages = {8},
keywords = {Endo-1-4-glu­canase; Bio­chem­i­cal char­ac­ter­i­za­tion; Gene cloning},
}

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%0 Journal Article
%T Biochemical characterization and gene cloning of a novel alkaline endo-1-4-glucanase from Bacillus subtilis DR8806
%A Ramezani, Somaye
%A Asoodeh, Ahmad
%J Journal of Molecular Catalysis B: Enzymatic
%@ 1381-1177
%D 2016

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