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The effect of nitrogen limitation on acetyl-CoA carboxylase expression and fatty acid content in Chromera velia and Isochrysis aff. galbana (TISO)
Authors:Roger Huerlimann  Eike J. Steinig  Heather Loxton  Kyall R. Zenger  Dean R. Jerry  Kirsten Heimann
Affiliation:1. School of Marine and Tropical Biology, James Cook University, Townsville, 4811, Australia;2. Centre for Sustainable Tropical Fisheries and Aquaculture, Townsville, 4811, Australia;3. James Cook University, Comparative Genomics Centre, Townsville, Queensland 4811, Australia;4. Centre for Biodiscovery and Molecular Development of Therapeutics, James Cook University, Townsville, Queensland 4811, Australia
Abstract:
Lipids from microalgae have become a valuable product with applications ranging from biofuels to human nutrition. While changes in fatty acid (FA) content and composition under nitrogen limitation are well documented, the involved molecular mechanisms are poorly understood. Acetyl-CoA carboxylase (ACCase) is a key enzyme in the FA synthesis and elongation pathway. Plastidial and cytosolic ACCases provide malonyl-CoA for de novo FA synthesis in the plastid and FA elongation in the endoplasmic reticulum, respectively. The present study aimed at investigating the expression of plastidial and cytosolic ACCase in Chromera velia and Isochrysis aff. galbana (TISO) and their impact on FA content and elongation level when grown under nitrogen-deplete conditions. In C. velia, plastidial ACCase was significantly upregulated during nitrogen starvation and with culture age, strongly correlating with increased FA content. Conversely, plastidial ACCase of I. aff. galbana was not differentially expressed in nitrogen-deplete cultures, but upregulated during the logarithmic phase of nitrogen-replete cultures. In contrast to plastidial ACCase, the cytosolic ACCase of C. velia was downregulated with culture age and nitrogen-starvation, strongly correlating with an increase in medium-chain FAs. In conclusion, the expression of plastidial and cytosolic ACCase changed with growth phase and nutrient status in a species-specific manner and nitrogen limitation did not always result in FA accumulation.
Keywords:α-CT, α-carboxyltransferase   ß  -CT, ß  -carboxyltransferase   aa, amino acid   ACC1, plastidial ACCase in algae   ACC2, cytosolic ACCase in algae   ACCase, acetyl-CoA carboxylase   AFDW, ash-free dry weight   ARA, arachidonic acid   BC, biotin carboxylase   BCCP, biotin carboxyl carrier protein   CoA, coenzyme A   DHA, docosahexaenoic acid   DNA, deoxyribonucleic acid   EPA, eicosapentaenoic acid   FA, fatty acid   FAME, fatty acid methyl ester   FAS, fatty acid synthase   GAPDH, glyceraldehyde 3-phosphate dehydrogenase   L, logarithmic growth phase   LA, linoleic acid   LL, late logarithmic growth phase   N-, cultures grown under nitrogen-deplete starting conditions    +, cultures grown under nitrogen-replete starting conditions   NCBI, National Center for Biotechnology Information   nt, nucleotide   PUFA, polyunsaturated fatty acid   qPCR, quantitative polymerase chain reaction   RNA, ribonucleic acid   S, stationary growth phase   MC-FA, medium-chain fatty acid   SE, standard error   TAG, triacylglyceride   TUA, alpha-tubulin   TUB, beta-tubulin   VLC-PUFA, very long-chain polyunsaturated fatty acid
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