Influence of the transposition of the thermostabilizing domain of Clostridium thermocellum xylanase (XynX) on xylan binding and thermostabilization
In: Applied and environmental microbiology (Print), Jg. 68 (2002), Heft 7, S. 3496-3501
Online
academicJournal
- print, 31 ref
Zugriff:
A xylanase gene, xynX, of Clostridium thermocellum had one thermostabilizing domain (TSD) between the signal peptide sequence and the catalytic domain (CD). The TSD of a truncated xylanase gene, xynX'TSD-CD, was transpositioned from the N terminus to the C terminus of the CD by overlapping PCRs, and a modified product, xynX'CD-TSD, was constructed. XynX'TSD-CD had a higher optimum temperature (70°C versus 65°C) and was more thermostable (residual activity of 68% versus 46% after a 20-min preincubation at 70°C) than the one without the TSD, XynX'CD. However, the domain-transpositioned enzyme, XynX'CD-TSD, showed a lower optimum temperature (30°C) and thermostability (20%) than XynX'CD. Both XynX'TSD-CD and XynX'CD-TSD showed significantly higher binding capacity toward xylan than XynX'CD, and the domain transposition did not cause any change in the binding ability. XynX'TSD-CD and XynX'CD-TSD also showed considerable binding to lichenan but not to carboxymethyl cellulose and laminarin. XynX'TSD-CD and XynX'CD-TSD had higher activities for insoluble xylan than XynX'CD, while XynX'CD was more active against soluble xylan than XynX'TSD-CD and XynX'CD-TSD. These results indicate that the TSD of XynX has dual functions, xylan binding and thermostabilization, and the domain should also be classified as a xylan-binding domain (XBD). The binding capacity of the XBD was not affected by domain transpositioning within the gene.
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Influence of the transposition of the thermostabilizing domain of Clostridium thermocellum xylanase (XynX) on xylan binding and thermostabilization
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Autor/in / Beteiligte Person: | SHIN, Eun-Sun ; YANG, Mi-Jeong ; KYUNG HWA, JUNG ; KWON, Eun-Ju ; JAE SUNG, JUNG ; SEUR KEE, PARK ; KIM, Jungho ; HAN DAE, YUN ; KIM, Hoon |
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Zeitschrift: | Applied and environmental microbiology (Print), Jg. 68 (2002), Heft 7, S. 3496-3501 |
Veröffentlichung: | Washington, DC: American Society for Microbiology, 2002 |
Medientyp: | academicJournal |
Umfang: | print, 31 ref |
ISSN: | 0099-2240 (print) |
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