초록
<P><B>Background</B></P><P>Since volatile and rising cost factors such as energy, raw materials and market competitiveness have a significant impact on the economic efficiency of biotechnological bulk productions, industrial processes need to be steadily improved and optimized. Thereby the current production hosts can undergo various limitations. To overcome those limitations and in addition increase the diversity of available production hosts for future applications, we suggest a Production Strain Blueprinting (PSB) strategy to develop new production systems in a reduced time lapse in contrast to a development from scratch.</P><P>To demonstrate this approach, <I>Bacillus pumilus</I> has been developed as an alternative expression platform for the production of alkaline enzymes in reference to the established industrial production host <I>Bacillus licheniformis</I>.</P><P><B>Results</B></P><P>To develop the selected <I>B. pumilus</I> as an alternative production host the suggested PSB strategy was applied proceeding in the following steps (dedicated product titers are scaled to the protease titer of Henkel’s industrial production strain <I>B. licheniformis</I> at lab scale): Introduction of a protease production plasmid, adaptation of a protease production process (44%), process optimization (92%) and expression optimization (114%). To further evaluate the production capability of the developed <I>B. pumilus</I> platform, the target protease was substituted by an α-amylase. The expression performance was tested under the previously optimized protease process conditions and under subsequently adapted process conditions resulting in a maximum product titer of 65% in reference to <I>B. licheniformis</I> protease titer.</P><P><B>Conclusions</B></P><P>In this contribution the applied PSB strategy performed very well for the development of <I>B. pumilus</I> as an alternative production strain. Thereby the engineered <I>B. pumilus</I> expression platform even exceeded the protease titer of the industrial production host <I>B. licheniformis</I> by 14%. This result exhibits a remarkable potential of <I>B. pumilus</I> to be the basis for a next generation production host, since the strain has still a large potential for further genetic engineering. The final amylase titer of 65% in reference to <I>B. licheniformis</I> protease titer suggests that the developed <I>B. pumilus</I> expression platform is also suitable for an efficient production of non-proteolytic enzymes reaching a final titer of several grams per liter without complex process modifications.</P>