Fermentation Optimization of Mycobacterium smegmatis Using Experimental Design
Sebastian O. Wendel
Technische HochschuleMittelhessen, Department of Biotechnology/Biopharmaceutical Technology, Wiesenstrasse 14, 35390 Giessen, Germany. & Kansas State University, Department of Chemical Engineering, 1036 Durland Hall, Manhattan, KS 66506, United States.
Ayomi S. Perera
Zansas State University, Department of Chemistry, Manhattan,201 CBC Building, KS 66506-0401, United States
Peter H. Pfromm
Kansas State University, Department of Chemical Engineering, 1036 Durland Hall, Manhattan, KS 66506, United States.
Peter Czermak
Technische HochschuleMittelhessen, Department of Biotechnology/Biopharmaceutical Technology, Wiesenstrasse 14, 35390 Giessen, Germany. & Kansas State University, Department of Chemical Engineering, 1036 Durland Hall, Manhattan, KS 66506, United States.
Stefan H. Bossmann
Kansas State University, Department of Chemistry, Manhattan,201 CBC Building, KS 66506-0401, United States.
*Author to whom correspondence should be addressed.
Abstract
The exceptionally stable mycobacterial protein porin A (MspA) from Mycobacterium smegmatis has potential applications in protein-based solar cells, and as a biotemplate for nano-wires and nano-dots. These applications would be enabled by an efficient and cost effective method to grow the host organism at high cell mass yields, and recover purified MspA.
In this work, the cell mass yield was maximized and costs lowered by applying experimental design (varying nitrogen and iron contents according to a Doehlert matrix) based on a minimal fermentation medium that was reported earlier. Glucose use was minimized by adjusting glucose feed based on analyzing residual glucose after fermentation. The costs for extracted and purified MspA were lowered by 67% for the minimum medium and the optimized composition derived here compared to commercial medium (7H9 Middlebrook).
Keywords: Mycobacterium smegmatis, MspA, medium optimization, protein extraction