Starch-binding domain shuffling in Aspergillus niger glucoamylase

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Starch-binding domain shuffling in Aspergillus niger glucoamylase.

Aspergillus niger glucoamylase (GA) consists mainly of two forms, GAI [from the N-terminus, catalytic domain + linker + starch-binding domain (SBD)] and GAII (catalytic domain + linker). These domains were shuffled to make RGAI (SBD + linker + catalytic domain), RGAIDeltaL (SBD + catalytic domain) and RGAII (linker + catalytic domain), with domains defined by function rather than by tertiary st...

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Solution structure of the granular starch binding domain of Aspergillus niger glucoamylase bound to beta-cyclodextrin.

BACKGROUND Carbohydrate-binding domains are usually small and physically separate from the catalytic domains of hydrolytic enzymes. Glucoamylase 1 (G1) from Aspergillus niger, an enzyme used widely in the food and brewing industries, contains a granular starch binding domain (SBD) which is separated from the catalytic domain by a semi-rigid linker. The aim of this study was to determine how the...

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Thermal unfolding of the starch binding domain of Aspergillus niger glucoamylase.

A fragment of the starch-binding domain (SBDF) of Aspergillus niger glucoamylase was prepared using recombinant DNA techniques, and its thermal unfolding was investigated by high-sensitivity differential scanning calorimetry (DSC). Thermal unfolding of SBDF was found to be reversible at pH 7 as expected from a DSC study of the whole enzyme molecule [Tanaka A. et al., J. Biochem., 117, 1024-1028...

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The predicted unfolding order of the b-strands in the starch binding domain from Aspergillus niger glucoamylase

The unfolding of the b-strands in the starch binding domain from Aspergillus niger glucoamylase was predicted to follow the order of b3 ! b2 ! b6 ! b5 ! b4 ! b1 ! b7 by 600 ps molecular dynamics simulations at 300, 400, and 600 K. The interior region around b-strands 2 and 3 acts as the initiation site for unfolding. b-Strands 1 and 7 are probably stabilized by the disulfide bond formed between...

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Thermodynamic effects of disulfide bond on thermal unfolding of the starch-binding domain of Aspergillus niger glucoamylase.

The thermodynamic effects of the disulfide bond of the fragment protein of the starch-binding domain of Aspergillus niger glucoamylase was investigated by measuring the thermal unfolding of the wild-type protein and its two mutant forms, Cys3Gly/Cys98Gly and Cys3Ser/Cys98Ser. The circular dichroism spectra and the thermodynamic parameters of binding with beta-cyclodextrin at 25 degrees C sugges...

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ژورنال

عنوان ژورنال: Protein Engineering Design and Selection

سال: 2003

ISSN: 1741-0126,1741-0134

DOI: 10.1093/protein/gzg066