Functional characterization of two diacylglycerol acyltransferase 1 genes in Mortierella alpina.
Mortierella alpina
Saccharomyces cerevisiae
diacylglycerol acyltransferase 1
oleaginous fungi
triacylglycerol
Journal
Letters in applied microbiology
ISSN: 1472-765X
Titre abrégé: Lett Appl Microbiol
Pays: England
ID NLM: 8510094
Informations de publication
Date de publication:
Feb 2022
Feb 2022
Historique:
revised:
02
11
2021
received:
17
09
2021
accepted:
05
11
2021
pubmed:
11
11
2021
medline:
19
1
2022
entrez:
10
11
2021
Statut:
ppublish
Résumé
Diacylglycerol acyltransferase (DGAT) is a crucial enzyme in the triacylglycerol (TAG) biosynthesis pathway. The oleaginous fungus Mortierella alpina can accumulate large amounts of arachidonic acid (ARA, C20:4) in the form of TAG. Therefore, it is important to study the functional characteristics of its DGAT. Two putative genes MaDGAT1A/1B encoding DGAT1 were identified in M. alpina ATCC 32222 genome by sequence alignment. Sequence alignment with identified DGAT1 homologs showed that MaDGAT1A/1B contain seven conserved motifs that are characteristic of the DGAT1 subfamily. Conserved domain analysis showed that both MaDGAT1A and MaDGAT1B belong to the Membrane-bound O-acyltransferases superfamily. The transforming with MaDGAT1A/1B genes could increase the accumulation of TAG in Saccharomyces cerevisiae to 4·47 and 7·48% of dry cell weight, which was 7·3-fold and 12·3-fold of the control group, respectively, but has no effect on the proportion of fatty acids in TAG. This study showed that MaDGAT1A/1B could effectively promote the accumulation of TAG and therefore may be used in metabolic engineering aimed to increase TAG production of oleaginous fungi.
Substances chimiques
Fatty Acids
0
Triglycerides
0
Diacylglycerol O-Acyltransferase
EC 2.3.1.20
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
194-203Subventions
Organisme : National Natural Science Foundation of China
ID : 32021005
Organisme : National Natural Science Foundation of China
ID : 31722041
Organisme : National Natural Science Foundation of China
ID : 32072186
Organisme : the Jiangsu Province "Collaborative Innovation Center for Food Safety and Quality Control"
Informations de copyright
© 2021 The Society for Applied Microbiology.
Références
Bligh, E.G. and Dyer, W.J. (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37, 911-917.
Cao, H. (2011) Structure-function analysis of diacylglycerol acyltransferase sequences from 70 organisms. BMC Res Notes 4, 249.
Cases, S., Smith, S.J., Zheng, Y.W., Myers, H.M., Lear, S.R., Sande, E., Novak, S., Collins, C. et al. (1998) Identification of a gene encoding an acyl CoA : diacylglycerol acyltransferase, a key enzyme in triacylglycerol synthesis. Proc Natl Acad Sci USA 95, 13018-13023.
Chen, H., Hao, G., Wang, L., Wang, H., Gu, Z., Liu, L., Zhang, H., Chen, W. et al. (2015) Identification of a critical determinant that enables efficient fatty acid synthesis in oleaginous fungi. Sci Rep 5, 11247.
Giniger, E., Varnum, S.M. and Ptashne, M. (1985) Specific DNA binding of GAL4, a positive regulatory protein of yeast. Cell 40, 767-774.
Gong, Y., Zhang, J., Guo, X., Wan, X., Liang, Z., Hu, C.J. and Jiang, M. (2013) Identification and characterization of PtDGAT2B, an acyltransferase of the DGAT2 acyl-Coenzyme A: diacylglycerol acyltransferase family in the diatom Phaeodactylum tricornutum. FEBS Lett 587, 481-487.
Greer, M.S., Truksa, M., Deng, W., Lung, S., Chen, G. and Weselake, R.J. (2015) Engineering increased triacylglycerol accumulation in Saccharomyces cerevisiae using a modified type 1 plant diacylglycerol acyltransferase. Appl Microbiol Biotechnol 99, 2243-2253.
Hao, G., Chen, H., Wang, L., Gu, Z., Song, Y., Zhang, H., Chen, W. and Chen, Y.Q. (2014) Role of malic enzyme during fatty acid synthesis in the oleaginous fungus Mortierella alpina. Appl Environ Microbiol 80, 2672-2678.
Hofmann, K. (2000) A superfamily of membrane-bound O-acyltransferases with implications for Wnt signaling. Trends Biochem Sci 25, 111-112.
Jeennor, S., Veerana, M., Anantayanon, J., Panchanawaporn, S., Chutrakul, C. and Laoteng, K. (2017) Diacylglycerol acyltransferase 2 of Mortierella alpina with specificity on long-chain polyunsaturated fatty acids: A potential tool for reconstituting lipids with nutritional value. J Biotechnol 263, 45-51.
Kennedy, E.P. (1957) Metabolism of lipides. Annu Rev Biochem 26, 119-148.
Liu, Q., Siloto, R.M.P., Snyder, C.L. and Weselake, R.J. (2011) Functional and topological analysis of yeast acyl-CoA: diacylglycerol acyltransferase 2, an endoplasmic reticulum enzyme essential for triacylglycerol biosynthesis. J Biol Chem 286, 13115-13126.
Lu, H., Chen, H., Tang, X., Yang, Q., Zhang, H., Chen, Y.Q. and Chen, W. (2019) Ultra performance liquid chromatography-Q Exactive Orbitrap/mass spectrometry-based lipidomics reveals the influence of nitrogen sources on lipid biosynthesis of Mortierella alpina. J Agr Food Chem 67, 10984-10993.
Luo, X., Zhu, Y., Liu, T., Wang, X., Zhou, P., Bao, Z. and Yu, L. (2017) Identification and characterization of a novel diacylglycerol acyltransferase gene from Mortierella alpina. Biotechnol Lett 39, 883-888.
Mao, X., Wu, T., Kou, Y., Shi, Y., Zhang, Y. and Liu, J. (2019) Characterization of type I and type II diacylglycerol acyltransferases from the emerging model alga Chlorella zofingiensis reveals their functional complementarity and engineering potential. Biotechnol Biofuels 12, 28.
Ratledge, C. (2004) Fatty acid biosynthesis in microorganisms being used for single cell oil production. Biochimie 86, 807-815.
Sakuradani, E. and Shimizu, S. (2009) Single cell oil production by Mortierella alpina. J Biotechnol 144, 31-36.
Shockey, J.M., Gidda, S.K., Chapital, D.C., Kuan, J., Dhanoa, P.K., Bland, J.M., Rothstein, S.J., Mullen, R.T. et al. (2006) Tung tree DGAT1 and DGAT2 have nonredundant functions in triacylglycerol biosynthesis and are localized to different subdomains of the endoplasmic reticulum. Plant Cell 18, 2294-2313.
Siloto, R.M.P., Madhavji, M., Wiehler, W.B., Burton, T.L., Boora, P.S., Laroche, A. and Weselake, R.J. (2008) An N-terminal fragment of mouse DGAT1 binds different acyl-CoAs with varying affinity. Biochem Biophys Res Commun 373, 350-354.
Stone, S.J., Levin, M.C. and Farese, R.V. (2006) Membrane topology and identification of key functional amino acid residues of murine acyl-CoA: diacylglycerol acyltransferase-2. J Biol Chem 281, 40273-40282.
Turchetto-Zolet, A.C., Maraschin, F.S., de Morais, G.L., Cagliari, A., Andrade, C.M.B., Margis-Pinheiro, M. and Margis, R. (2011) Evolutionary view of acyl-CoA diacylglycerol acyltransferase (DGAT), a key enzyme in neutral lipid biosynthesis. BMC Evol Biol 11, 263.
Wang, L., Chen, W., Feng, Y., Ren, Y., Gu, Z., Chen, H., Wang, H., Thomas, M.J. et al. (2011) Genome characterization of the oleaginous fungus Mortierella alpina. PLoS One 6, e28319.
Weselake, R.J., Madhavji, M., Szarka, S.J., Patterson, N.A., Wiehler, W.B., Nykiforuk, C.L., Burton, T.L., Boora, P.S. et al. (2006) Acyl-CoA-binding and self-associating properties of a recombinant 13.3 kDa N-terminal fragment of diacylglycerol acyltransferase-1 from oilseed rape. BMC Biochem 7, 24.
Wynn, J.P., Hamid, A.B.A. and Ratledge, C. (1999) The role of malic enzyme in the regulation of lipid accumulation in filamentous fungi. Microbiology 145, 1911-1917.
Xin, Y., Shen, C., She, Y., Chen, H., Wang, C., Wei, L., Yoon, K., Han, D. et al. (2019) Biosynthesis of triacylglycerol molecules with a tailored PUFA profile in industrial microalgae. Mol Plant 12, 474-488.
Xue, Z., Sharpe, P.L., Hong, S., Yadav, N.S., Xie, D., Short, D.R., Damude, H.G., Rupert, R.A. et al. (2013) Production of omega-3 eicosapentaenoic acid by metabolic engineering of Yarrowia lipolytica. Nat Biotechnol 31, 734.
Yang, J., Brown, M.S., Liang, G., Grishin, N.V. and Goldstein, J.L. (2008) Identification of the acyltransferase that octanoylates ghrelin, an appetite-stimulating peptide hormone. Cell 132, 387-396.
Yen, C.E., Stone, S.J., Koliwad, S., Harris, C. and Farese, R.V. (2008) Thematic review series: glycerolipids. DGAT enzymes and triacylglycerol biosynthesis. J Lipid Res 49, 2283-2301.
Zhai, L., Chaturvedi, D. and Cumberledge, S. (2004) Drosophila Wnt-1 undergoes a hydrophobic modification and is targeted to lipid rafts, a process that requires porcupine. J Biol Chem 279, 33220-33227.
Zhang, L., Zhang, H. and Song, Y. (2018) Identification and characterization of diacylglycerol acyltransferase from oleaginous fungus Mucor circinelloides. J Agr Food Chem 66, 674-681.