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Volume 146, Issue 10

Cell-associated degradation affects the yield of secreted engineered and heterologous proteins in the expression system Free

Abstract

A series of chimeric α-amylase genes derived from , which encodes the liquefying α-amylase from , were constructed using gene splicing techniques. The gene constructs were cloned in , where their ability to direct the synthesis and secretion of active α-amylase was determined. Detectable α-amylase activity was observed for some, but not all, of the chimeric proteins. Studies on the secretion of wild-type AmyL and its chimeric derivatives revealed that, whilst these proteins were stable in the extracellular milieu, all were subject to some degree of degradation during secretion. The chimeric enzymes were degraded to a greater extent than the native enzyme. These findings suggest that cell-associated proteolysis is a significant problem affecting the use of as host bacterium for the production of heterologous proteins.

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Keyword(s): Bacillus subtilis , chimeric proteins , industrial enzymes , protein secretion , proteolysis and SOE, splicing by overlap extension
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2000-10-01
2024-04-25
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Cell-associated degradation affects the yield of secreted engineered and heterologous proteins in the Bacillus subtilis expression system
Microbiology 146, 2583 (2000); https://doi.org/10.1099/00221287-146-10-2583
/content/journal/micro/10.1099/00221287-146-10-2583
/content/journal/micro/10.1099/00221287-146-10-2583
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