Awamori

Here we describe the purification of RHG, the cloning and characterization of the corresponding gene and (over)production of the enzyme in A. aculeatus and Aspergillus awamori. 

Figure 4. Western analysis of culture filtrate of A. aculeatus wild type (lanes 1-3) and A. aculeatus (lanes 4-6) and A. awamori multicopy transformants (lanes 7-9). Medium samples were applied undiluted (lanes 1,4,7), 10 times diluted (lanes 2,5,8) or 100 times diluted (lanes 3,6,9). Lane 10 contains purified rhamnogalacturonase.

Figure 6A-C. High-performance anion-exchange chromatography elution profile of isolated modified hairy regions without addition of enzymes (A), with Biopectinase OS (B) and with rhamnogalacturonase-containing culture filtrate from an A. awamori multicopy transformant (C). /xC ftCoulomb- Data taken from Ref. 6.

Hessing JGM, Rotterdam C van, Verbakel JMA, Roza M, Maat J, Gorcom REM van, Hondel CAMJJ van den (1994) Isolation and characterization of a 1,4-/3-endoxylanase gene of A. awamori. Curr Genet 26 228-232... 

Although proteins can be expressed in many heterologous production systems, including bacteria such as Proteus mirabilis [1], fungi such as Pichia pastoris [2, 3] and Aspergillus awamori [4] and insect cells [5, 6], the pharmaceutical industry has narrowed down process development to a small number of platform technologies ... 

Aleshin and coworkers (49) have reported the X-ray crystal structure at 2.2-A resolution of a G2-type variant produced by Aspergillus awamori. Meanwhile, an attempt was made to determine the amino acid residues that participate in the substrate binding and catalysis provided by G2 of A. niger (52). The results of the chemical approach indicated that the Asp-176, Glu-179, and Glu-180 form an acidic cluster crucial to the functioning of the enzyme. This conclusion was then tested by site-specific mutagenesis of these amino acid residues, which were replaced, one at a time, with Asn, Gin, and Gin, respectively (53). The substitution at Glu-179 provided an inactive protein. The other two substitutions affected the kinetic parameters but were not of crucial importance to the maintenance of activity. The crystal structure (49) supports the conclusion that Glu-179 functions as the catalytic acid but Asp-17 6 does not appear to be a good candidate for provision of catalytic base. Thus, there still exists considerable uncertainty as to how the disaccharide is accepted into the combining site for hydrolysis. Nevertheless, the kind of scheme presented by Svensson and coworkers (52) almost surely prevails. 

Agaricus bisporus Pleurotus ostreatus Trichoderma reesei Trichoderma spp. Schizophyllum commune Aspergillus awamori Aspergillus niger Tyromyces palustris Streptomyces flavogriseus Streptomyces olivochromogenes Streptomyces spp,... 

E. M. S. Harris, A. E. Aleshin, L. M. Firsov, and R. B. Honzatko, Refined structure of the complex of 1-deoxynojirimycin with glucoamylase from Aspergillus awamori var. X100 to 2.4-A resolution, Biochemistry, 32 (1993) 1618—1626. 

Gluco amylase Aspergillus awamori TOMAC/Revopal HV5/n-octanol/ Extraction [57,58]... 

Pavezzi, F. C., Gomes, E., da Silva, R. (2008). Production and characterization of glucoamylase from fungus Aspergillus awamori expressed in yeast Saccharomyces cerevisiae using different carbon sources. Braz. 

Ariff, A.B. and Webb, C., The influence of different fermenter configurations and modes of operation on glucoamylase production by Aspergillus awamori, Asia-Pacific J. Mol. Biol. Biotechnol. 1996 vol. 4, no. 3, pp. 183-195. 

Ariff and Webb studied production of glucoamylase using freely suspended cells of Aspergillus awamori in batch and continuous fermentations. Glucoamylase yields based on glucose consumed were 900 and 1080 U/g for batch and continuous fermentations, respectively. The immobilization of viable cells was achieved by adsorption to cubes of reticulated polyurethane foam. In comp uison with freely suspended cell fermentations, neither batch nor continuous fermentations of immobilized cells improved glucoamylase production significantly in tenns of yield or productivity. 

Human (Homo sapiens) Escherichia coli Aspergillus awamori, variant xlOO engrailed Homeodomain Drosophila melanogaster... 

Two forms of glucoamylase (often incorrectly called amyloglucosidase) have been observed in the culture supernatants of Aspergillus niger, A. awamori and... 

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