Publication: Metabolic networks and bioenergetics of Aurantiochytrium sp. B-072 during storage lipid formation
Issued Date
2012-07-01
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ISSN
16784405
15178382
15178382
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2-s2.0-84870660066
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Mahidol University
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SCOPUS
Bibliographic Citation
Brazilian Journal of Microbiology. Vol.43, No.3 (2012), 1192-1205
Suggested Citation
Montri Chaisawang, Cornelis Verduyn, Somchai Chauvatcharin, Manop Suphantharika Metabolic networks and bioenergetics of Aurantiochytrium sp. B-072 during storage lipid formation. Brazilian Journal of Microbiology. Vol.43, No.3 (2012), 1192-1205. doi:10.1590/S1517-83822012000300047 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/14296
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Title
Metabolic networks and bioenergetics of Aurantiochytrium sp. B-072 during storage lipid formation
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Abstract
Baffled shake flask cultivation of Aurantiochytrium sp. B-072 was carried out at in a glucose-monosodium glutamate mineral medium at different C/N-ratios (30-165) with glucose fixed at 90 g/L. With increasing C/N-ratio, a modest increase in lipid content (60 to 73 % w/w) was observed whereas fat-free biomass decreased but overall biomass showed little variation. FA-profiles were not affected to a large extent by C/N-ratio and absolute docosahexaenoic (DHA)-levels fell in narrow range (5-6 g/L). However at C/N > 64 a rapid decrease in lipid synthetic rate and/or incomplete glucose utilization occurred. Glucose and FAfluxes based on fat-free biomass peaked at a C/N ratio of 56. This condition was chosen for calculation of the redox balance (NAD(P)H) and energy (ATP) requirement and to estimate the in vivo P/O ratio during the main period of fatty acid biosynthesis. Several models with different routes for NADPH, acetyl-CoA formation and re-oxidation of OAA formed via ATP-citrate lyase were considered as these influence the redox- and energy balance. As an example, using a commonly shown scheme whereby NADPH is supplied by a cytosolic "transhydrogenase cycle" (pyruvate-OAA-malate-pyruvate) and OAA formed by ATP-citrate lyase is recycled via import into the mitochondria as malate, the calculated NADPH-requirement amounted to 5.5 with an ATP-demand of 10.5 mmol/(g fat-free biomass x h) and an in vivo P/O-ratio (not including non-growth associated maintenance) of 1.6. The lowest ATP requirement is found when acetyl-CoA would be transported directly from the mitochondria to the cytosol by carnitine acetyltransferase. Assay of some enzymes critical for NADPH supply indicates that activity of glucose-6-phosphate dehydrogenase, the first enzyme in the HMP pathway, is far insufficient for the required NADPH-flux and malic enzyme must be a major source. Activity of the latter (ca. 300 mU/mg protein) far exceeds that in oleaginous fungi and yeast.