Despite the above evidence, the leaky plasma membrane hypothesis was discounted
by certain investigators in favor of an efflux system specific for glutamate and regulated by the energy state of the cell. The action of biotin was attributed to effects on intermediary metabolism, correlating with the activity of fatty acid syn38 1 History of Industrial Biotechnology thetases. Additional opinions discounting the permeability hypothesis attributed glutamate overproduction to a decrease in the activity of α - ketoglutarate dehydrogenase caused by biotin limitation or by addition of penicillin or surfactants. In 2001, however, the permeability modification hypothesis was supported. The various manipulations leading to glutamate overproduction were shown to cause increased permeability of the mycolic acid layer of the cell wall [70] . The glutamate - overproducing bacteria are characterized by a special cell envelope containing mycolic acids which surrounds the entire cell as a structured layer and is thought to be involved in permeation of solutes. The mycolic acids esterified with arabinogalactan and the non - covalently bound mycolic acid derivatives formed a second lipid layer, the cytoplasmic membrane being the first. As stated by these authors, “ The concepts of ‘ permeability of the cell wall ’ as originally used in the very fi rst work on L - glutamate production more than forty years ago now takes on a new meaning. ” Nampoothiri et al . [71] provided evidence that overexpression or inactivity of genes involved in lipid synthesis changed glutamate efflux dramatically, altered the chemical and physical properties of the cytoplasmic membrane, and that this was necessary to achieve efflux of L - glutamate. They state “ that altering the phosphoplipid content alone is sufficient to enable L - glutamate efflux.” Burkovski and Kraemer [72] further stated that “ There is no doubt that stimulation of glutamate excretion in C. glutamicum is directly or indirectly related to membrane and/or cell wall integrity. ”