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The soil bacterium Corynebacterium glutamicum is the major organism for the production of the amino acid L-lysine, an important nutrient in animal feedstock. This study investigated new strategies for bioprocess and genetic engineering of C. glutamicum towards production of L-lysine. In aerobic cultivations, routinely performed in shake-flasks, the dissolved oxygen concentration is a critical often neglected parameter. Here, oxygen mass transfer was determined in disposable shake-flasks under different conditions. Based on the results, a mathematical correlation was developed allowing the prediction of the maximum possible cell concentration achievable without oxygen limitation. The developed tool is valuable for cultivations with costly carbon sources and nutrient additives. From a metabolic perspective, lysine production depends on a sufficient supply of precursors as well as an accurate regulation of metabolic fluxes. Thus, genetic tools were generated to modify selected genes. The strategies aimed to increase the availability of the essential cofactor NADPH and to manipulate gene expression levels using cofactor-engineering and a promoter library, respectively. In both cases modulation of individual target genes resulted in an increased lysine yield. The developed approaches seem valuable also for other fermentation processes.