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| {{STRUCTURE_1axk| PDB=1axk | SCENE= }} | | {{STRUCTURE_1axk| PDB=1axk | SCENE= }} |
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| '''ENGINEERED BACILLUS BIFUNCTIONAL ENZYME GLUXYN-1'''
| | ===ENGINEERED BACILLUS BIFUNCTIONAL ENZYME GLUXYN-1=== |
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| ==Overview==
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| The 1,3-1,4-beta-glucanase from Bacillus macerans (wtGLU) and the 1, 4-beta-xylanase from Bacillus subtilis (wtXYN) are both single-domain jellyroll proteins catalyzing similar enzymatic reactions. In the fusion protein GluXyn-1, the two proteins are joined by insertion of the entire XYN domain into a surface loop of cpMAC-57, a circularly permuted variant of wtGLU. GluXyn-1 was generated by protein engineering methods, produced in Escherichia coli and shown to fold spontaneously and have both enzymatic activities at wild-type level. The crystal structure of GluXyn-1 was determined at 2.1 A resolution and refined to R = 17.7% and R(free) = 22.4%. It shows nearly ideal, native-like folding of both protein domains and a small, but significant hinge bending between the domains. The active sites are independent and accessible explaining the observed enzymatic activity. Because in GluXyn-1 the complete XYN domain is inserted into the compact folding unit of GLU, the wild-type-like activity and tertiary structure of the latter proves that the folding process of GLU does not depend on intramolecular interactions that are short-ranged in the sequence. Insertion fusions of the GluXyn-1 type may prove to be an easy route toward more stable bifunctional proteins in which the two parts are more closely associated than in linear end-to-end protein fusions. | | The line below this paragraph, {{ABSTRACT_PUBMED_9618460}}, adds the Publication Abstract to the page |
| | (as it appears on PubMed at http://www.pubmed.gov), where 9618460 is the PubMed ID number. |
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| | {{ABSTRACT_PUBMED_9618460}} |
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| ==About this Structure== | | ==About this Structure== |
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| [[Category: Gluxyn-1]] | | [[Category: Gluxyn-1]] |
| [[Category: Hybrid enzyme]] | | [[Category: Hybrid enzyme]] |
| ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May 2 10:48:48 2008'' | | |
| | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jun 30 17:48:37 2008'' |
