ABSTRACT
The non-conventional yeast Kluyveromyces marxianus is an emerging industrial producer for many biotechnological processes. Here, we show the application of a biomass-linked stoichiometric model of central metabolism that is experimentally validated, and mass and charge balanced for assessing the carbon conversion efficiency of wild type and modified K. marxianus. Pairs of substrates (lactose, glucose, inulin, xylose) and products (ethanol, acetate, lactate, glycerol, ethyl acetate, succinate, glutamate, phenylethanol and phenylalanine) are examined by various modelling and optimisation methods. Our model reveals the organism's potential for industrial application and metabolic engineering. Modelling results imply that the aeration regime can be used as a tool to optimise product yield and flux distribution in K. marxianus. Also rebalancing NADH and NADPH utilisation can be used to improve the efficiency of substrate conversion. Xylose is identified as a biotechnologically promising substrate for K. marxianus.
Subject(s)
Industrial Microbiology , Kluyveromyces/metabolism , Acetates/metabolism , Biomass , Calibration , Culture Media/chemistry , Ethanol/metabolism , Glucose/chemistry , Glutamic Acid/metabolism , Glycerol/metabolism , Inulin/chemistry , Kluyveromyces/genetics , Lactates/metabolism , Lactose/chemistry , Metabolic Engineering , Models, Molecular , Oxygen Consumption , Phenylalanine/metabolism , Phenylethyl Alcohol/metabolism , Reproducibility of Results , Succinic Acid/metabolism , Xylose/chemistryABSTRACT
The ethanol and temperature effects on the ratio between Zymomonas mobilis 113S extracellular levansucrase activities were studied using fermentation broth supernatant, "levan-levansucrase" sediment precipitated by ethanol and highly purified enzyme. The fructooligosaccharide (FOS) production at different temperatures in the presence of ethanol was investigated. An ethanol increases FOS biosynthesis activity part of levansucrase. Especially, this effect was pronounced at lower temperatures (35-40 °C) and using purified levansucrase. The inverse relationship between temperature and ratio synthetic activity/total activity of levansucrase was found. The FOS composition containing mostly 1-kestose, 6-kestose, and neokestose obtained in the presence of different ethanol concentrations was found relative constant, while the changes in the sucrose concentration and temperature gave slight changes in the ratio between 1-kestose and 6-kestose.
