Characterization of aNeocallimastixpatriciarumxylanase gene and its product

1999 ◽  
Vol 45 (11) ◽  
pp. 970-974 ◽  
Author(s):  
Jin-Hao Liu ◽  
Brent L Selinger ◽  
Cheng-Fang Tsai ◽  
Kuo-Jaon Cheng
Keyword(s):  
Catalytic Domain ◽  
Xylanase Activity ◽  
Binding Domain ◽  
Glycosyl Hydrolases ◽  
Cellulose Binding Domain ◽  
Xylanase Gene ◽  
Neocallimastix Patriciarum ◽  
Cellulose Binding ◽  
Linker Domain

A xylanase gene (xynC) isolated from the anaerobic ruminal fungus Neocallimastix patriciarum was characterized. The gene consists of an N-terminal catalytic domain that exhibited homology to family 11 of glycosyl hydrolases, a C-terminal cellulose binding domain (CBD) and a putative dockerin domain in between. Each domain was linked by a short linker domain rich in proline and alanine. Deletion analysis demonstrated that the CBD was essential for optimal xylanase activity of the enzyme, while the putative dockerin domain may not be required for enzyme function.Key words: xylanase, cellulose binding domain, Neocallimastix patriciarum.

The cellulose-binding domain (CBDCex) of an exoglucanase fromCellulomonas fimi: Production inEscherichia coli and characterization of the polypeptide

1993 ◽  
Vol 42 (4) ◽  
pp. 401-409 ◽  
Author(s):  
Edgar Ong ◽  
Neil R. Gilkes ◽  
Robert C. Miller ◽  
R. Antony J. Warren ◽  
Douglas G. Kilburn
Keyword(s):  
Binding Domain ◽  
Inescherichia Coli ◽  
Cellulose Binding Domain ◽  
Cellulose Binding

scholarly journals Characterization of the cellulose-binding domain of the Clostridium cellulovorans cellulose-binding protein A.

1993 ◽  
Vol 175 (18) ◽  
pp. 5762-5768 ◽  
Author(s):  
M A Goldstein ◽  
M Takagi ◽  
S Hashida ◽  
O Shoseyov ◽  
R H Doi ◽  
...  
Keyword(s):  
Binding Protein ◽  
Protein A ◽  
Binding Domain ◽  
Cellulose Binding Domain ◽  
Clostridium Cellulovorans ◽  
Cellulose Binding

scholarly journals Characterization of a Cellobiohydrolase (MoCel6A) Produced by Magnaporthe oryzae

2010 ◽  
Vol 76 (19) ◽  
pp. 6583-6590 ◽  
Author(s):  
Machiko Takahashi ◽  
Hideyuki Takahashi ◽  
Yuki Nakano ◽  
Teruko Konishi ◽  
Ryohei Terauchi ◽  
...  
Keyword(s):  
Magnaporthe Oryzae ◽  
Magnaporthe Grisea ◽  
Catalytic Domain ◽  
Cellulose Binding Domain ◽  
Enhanced Activity ◽  
Hydrolytic Activities ◽  
Cellulose Binding ◽  
Hydrolysis Of ◽  
Increased Activity

ABSTRACT Three GH-6 family cellobiohydrolases are expected in the genome of Magnaporthe grisea based on the complete genome sequence. Here, we demonstrate the properties, kinetics, and substrate specificities of a Magnaporthe oryzae GH-6 family cellobiohydrolase (MoCel6A). In addition, the effect of cellobiose on MoCel6A activity was also investigated. MoCel6A contiguously fused to a histidine tag was overexpressed in M. oryzae and purified by affinity chromatography. MoCel6A showed higher hydrolytic activities on phosphoric acid-swollen cellulose (PSC), β-glucan, and cellooligosaccharide derivatives than on cellulose, of which the best substrates were cellooligosaccharides. A tandemly aligned cellulose binding domain (CBD) at the N terminus caused increased activity on cellulose and PSC, whereas deletion of the CBD (catalytic domain only) showed decreased activity on cellulose. MoCel6A hydrolysis of cellooligosaccharides and sulforhodamine-conjugated cellooligosaccharides was not inhibited by exogenously adding cellobiose up to 438 mM, which, rather, enhanced activity, whereas a GH-7 family cellobiohydrolase from M. oryzae (MoCel7A) was severely inhibited by more than 29 mM cellobiose. Furthermore, we assessed the effects of cellobiose on hydrolytic activities using MoCel6A and Trichoderma reesei cellobiohydrolase (TrCel6A), which were prepared in Aspergillus oryzae. MoCel6A showed increased hydrolysis of cellopentaose used as a substrate in the presence of 292 mM cellobiose at pH 4.5 and pH 6.0, and enhanced activity disappeared at pH 9.0. In contrast, TrCel6A exhibited slightly increased hydrolysis at pH 4.5, and hydrolysis was severely inhibited at pH 9.0. These results suggest that enhancement or inhibition of hydrolytic activities by cellobiose is dependent on the reaction mixture pH.

scholarly journals A modular esterase from Pseudomonas fluorescens subsp. cellulosa contains a non-catalytic cellulose-binding domain

1993 ◽  
Vol 294 (2) ◽  
pp. 349-355 ◽  
Author(s):  
L M Ferreira ◽  
T M Wood ◽  
G Williamson ◽  
C Faulds ◽  
G P Hazlewood ◽  
...  
Keyword(s):  
Pseudomonas Fluorescens ◽  
Catalytic Domain ◽  
Genomic Library ◽  
Binding Domain ◽  
Cellulose Binding Domain ◽  
Reading Frame ◽  
Conserved Sequence ◽  
Methoxycinnamic Acid ◽  
Cellulose Binding ◽  
Derivatives Of

The 5′ regions of genes xynB and xynC, coding for a xylanase and arabinofuranosidase respectively, are identical and are reiterated four times within the Pseudomonas fluorescens subsp. cellulosa genome. To isolate further copies of the reiterated xynB/C 5′ region, a genomic library of Ps. fluorescens subsp. cellulosa DNA was screened with a probe constructed from the conserved region of xynB. DNA from one phage which hybridized to the probe, but not to sequences upstream or downstream of the reiterated xynB/C locus, was subcloned into pMTL22p to construct pFG1. The recombinant plasmid expressed a protein in Escherichia coli, designated esterase XYLD, of M(r) 58,500 which bound to cellulose but not to xylan. XYLD hydrolysed aryl esters, released acetate groups from acetylxylan and liberated 4-hydroxy-3-methoxycinnamic acid from destarched wheat bran. The nucleotide sequence of the XYLD-encoding gene, xynD, revealed an open reading frame of 1752 bp which directed the synthesis of a protein of M(r) 60,589. The 5′ 817 bp of xynD and the amino acid sequence between residues 37 and 311 of XYLD were almost identical with the corresponding regions of xynB and xynC and their encoded proteins XYLB and XYLC. Truncated derivatives of XYLD lacking the N-terminal conserved sequence retained the capacity to hydrolyse ester linkages, but did not bind cellulose. Expression of truncated derivatives of xynD, comprising the 5′ 817 bp sequence, encoded a non-catalytic polypeptide that bound cellulose. These data indicate that XYLD has a modular structure comprising of a N-terminal cellulose-binding domain and a C-terminal catalytic domain.

scholarly journals Two Cellobiohydrolase-Encoding Genes from Aspergillus niger Require d-Xylose and the Xylanolytic Transcriptional Activator XlnR for Their Expression

1999 ◽  
Vol 65 (10) ◽  
pp. 4340-4345 ◽  
Author(s):  
Marco M. C. Gielkens ◽  
Ester Dekkers ◽  
Jaap Visser ◽  
Leo H. de Graaff
Keyword(s):  
Amino Acid ◽  
Aspergillus Niger ◽  
Northern Blot Analysis ◽  
Catalytic Domain ◽  
Transcriptional Activator ◽  
Glycosyl Hydrolases ◽  
Cellulose Binding Domain ◽  
Linker Peptide ◽  
Cellulose Binding ◽  
Encoding Genes

ABSTRACT Two cellobiohydrolase-encoding genes, cbhA andcbhB, have been isolated from the filamentous fungusAspergillus niger. The deduced amino acid sequence shows that CbhB has a modular structure consisting of a fungus-type cellulose-binding domain (CBD) and a catalytic domain separated by a Pro/Ser/Thr-rich linker peptide. CbhA consists only of a catalytic domain and lacks a CBD and linker peptide. Both proteins are homologous to fungal cellobiohydrolases in family 7 of the glycosyl hydrolases. Northern blot analysis showed that the transcription of thecbhA and cbhB genes is induced byd-xylose but not by sophorose and, in addition, requires the xylanolytic transcriptional activator XlnR.

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