Comparison of nitrogen depletion and repletion on lipid production in yeast and fungal species

Yang, Shihui; Wang, Wei; Wei, Hui; Van Wychen, Stefanie; Pienkos, Philip T.; Zhang, Min; Himmel, Michael E.

doi:10.3390/en9090685

Title: Comparison of nitrogen depletion and repletion on lipid production in yeast and fungal species

Journal Article · Mon Aug 29 00:00:00 EDT 2016 · Energies

DOI:https://doi.org/10.3390/en9090685· OSTI ID:1319280

^[1]; Wang, Wei ^[2]; Wei, Hui ^[2]; Van Wychen, Stefanie ^[2]; Pienkos, Philip T. ^[2]; Zhang, Min ^[2]; Himmel, Michael E. ^[2]

National Renewable Energy Lab. (NREL), Golden, CO (United States); Hubei Univ., Wuhan (China)
National Renewable Energy Lab. (NREL), Golden, CO (United States)

Although it is well known that low nitrogen stimulates lipid accumulation, especially for algae and some oleaginous yeast, few studies have been conducted in fungal species, especially on the impact of different nitrogen deficiency strategies. In this study, we use two promising consolidated bioprocessing (CBP) candidates to examine the impact of two nitrogen deficiency strategies on lipid production, which are the extensively investigated oleaginous yeast Yarrowia lipolytica, and the commercial cellulase producer Trichoderma reesei. We first utilized bioinformatics approaches to reconstruct the fatty acid metabolic pathway and demonstrated the presence of a triacylglycerol (TAG) biosynthesis pathway in Trichoderma reesei. We then examined the lipid production of Trichoderma reesei and Y. lipomyces in different media using two nitrogen deficiency strategies of nitrogen natural repletion and nitrogen depletion through centrifugation. Our results demonstrated that nitrogen depletion was better than nitrogen repletion with about 30% lipid increase for Trichoderma reesei and Y. lipomyces, and could be an option to improve lipid production in both oleaginous yeast and filamentous fungal species. The resulting distinctive lipid composition profiles indicated that the impacts of nitrogen depletion on yeast were different from those for fungal species. Under three types of C/N ratio conditions, C16 and C18 fatty acids were the predominant forms of lipids for both Trichoderma reesei and Y. lipolytica. In addition, while the overall fatty acid methyl ester (FAME) profiles of Trichoderma reesei were similar, the overall FAME profiles of Y. lipolytica observed a shift. The fatty acid metabolic pathway reconstructed in this work supports previous reports of lipid production in T. reesei, and provides a pathway for future omics studies and metabolic engineering efforts. Further investigation to identify the genetic targets responsible for the effect of nitrogen depletion on lipid production improvement will facilitate strain engineering to boost lipid production under more optimal conditions for productivity than those required for nitrogen depletion.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office (BETO)

Grant/Contract Number:: AC36-08GO28308

OSTI ID:: 1319280

Report Number(s):: NREL/JA-5100-67021; ENERGA

Journal Information:: Energies, Vol. 9, Issue 9; ISSN 1996-1073

Publisher:: MDPI AGCopyright Statement

Country of Publication:: United States

Language:: English

Citation Metrics:

Cited by: 9 works

Citation information provided by
Web of Science

References (62)

Comparative Studies of Oleaginous Fungal Strains (Mucor circinelloides and Trichoderma reesei) for Effective Wastewater Treatment and Bio-Oil Production Bhanja, Anshuman; Minde, Gauri; Magdum, Sandip Biotechnology Research International, Vol. 2014 https://doi.org/10.1155/2014/479370	journal	November 2014
Effect of different C/N ratios on carotenoid and lipid production by Rhodotorula glutinis Braunwald, Teresa; Schwemmlein, Lisa; Graeff-Hönninger, Simone Applied Microbiology and Biotechnology, Vol. 97, Issue 14 https://doi.org/10.1007/s00253-013-5005-8	journal	June 2013
Snf1 Is a Regulator of Lipid Accumulation in Yarrowia lipolytica Seip, John; Jackson, Raymond; He, Hongxian Applied and Environmental Microbiology, Vol. 79, Issue 23 https://doi.org/10.1128/AEM.02079-13	journal	September 2013
Perspectives and new directions for the production of bioethanol using consolidated bioprocessing of lignocellulose Xu, Qi; Singh, Arjun; Himmel, Michael E. Current Opinion in Biotechnology, Vol. 20, Issue 3, p. 364-371 https://doi.org/10.1016/j.copbio.2009.05.006	journal	June 2009
l -Sorbose induces cellulase gene transcription in the cellulolytic fungus Trichoderma reesei Nogawa, Masahiro; Goto, Masahiro; Okada, Hirofumi Current Genetics, Vol. 38, Issue 6 https://doi.org/10.1007/s002940000165	journal	January 2001
ACEI of Trichoderma reesei Is a Repressor of Cellulase and Xylanase Expression Aro, N.; Ilmen, M.; Saloheimo, A. Applied and Environmental Microbiology, Vol. 69, Issue 1 https://doi.org/10.1128/AEM.69.1.56-65.2003	journal	January 2003
Enhancement of lipid production using biochemical, genetic and transcription factor engineering approaches Courchesne, Noémie Manuelle Dorval; Parisien, Albert; Wang, Bei Journal of Biotechnology, Vol. 141, Issue 1-2, p. 31-41 https://doi.org/10.1016/j.jbiotec.2009.02.018	journal	April 2009
Yarrowia lipolytica as a model for bio-oil production Beopoulos, Athanasios; Cescut, Julien; Haddouche, Ramdane Progress in Lipid Research, Vol. 48, Issue 6, p. 375-387 https://doi.org/10.1016/j.plipres.2009.08.005	journal	November 2009
Microbial production of advanced transportation fuels in non-natural hosts Connor, Michael R.; Liao, James C. Current Opinion in Biotechnology, Vol. 20, Issue 3, p. 307-315 https://doi.org/10.1016/j.copbio.2009.04.002	journal	June 2009
Vectors for gene expression and amplification in the yeastYarrowia lipolytica Juretzek, Thomas; Le Dall, Marie-Th�r�se; Mauersberger, Stephan Yeast, Vol. 18, Issue 2 https://doi.org/10.1002/1097-0061(20010130)18:2<97::AID-YEA652>3.0.CO;2-U	journal	January 2001
A review of enzymes and microbes for lignocellulosic biorefinery and the possibility of their application to consolidated bioprocessing technology Hasunuma, Tomohisa; Okazaki, Fumiyoshi; Okai, Naoko Bioresource Technology, Vol. 135 https://doi.org/10.1016/j.biortech.2012.10.047	journal	May 2013
One-step transformation of yeast in stationary phase Chen, Dz-Chi; Yang, Bei-Chang; Kuo, Tsong-Teh Current Genetics, Vol. 21, Issue 1 https://doi.org/10.1007/BF00318659	journal	January 1992
Heterologous Expression of Xylanase Enzymes in Lipogenic Yeast Yarrowia lipolytica Wang, Wei; Wei, Hui; Alahuhta, Markus PLoS ONE, Vol. 9, Issue 12 https://doi.org/10.1371/journal.pone.0111443	journal	December 2014
Acidophilic Green Alga Pseudochlorella sp. YKT1 Accumulates High Amount of Lipid Droplets under a Nitrogen-Depleted Condition at a Low-pH Hirooka, Shunsuke; Higuchi, Sumio; Uzuka, Akihiro PLoS ONE, Vol. 9, Issue 9 https://doi.org/10.1371/journal.pone.0107702	journal	September 2014
Identification and microbial production of a terpene-based advanced biofuel Peralta-Yahya, Pamela P.; Ouellet, Mario; Chan, Rossana Nature Communications, Vol. 2, Issue 1 https://doi.org/10.1038/ncomms1494	journal	September 2011
Consolidated bioprocessing of cellulosic biomass: an update Lynd, Lee R.; van Zyl, Willem H.; McBride, John E. Current Opinion in Biotechnology, Vol. 16, Issue 5, p. 577-583 https://doi.org/10.1016/j.copbio.2005.08.009	journal	October 2005
Integrative transformation of the yeast Yarrowia lipolytica Davidow, Lance S.; Apostolakos, Diane; O'Donnell, Michele M. Current Genetics, Vol. 10, Issue 1 https://doi.org/10.1007/BF00418492	journal	October 1985
Microbial oil production from rice straw hydrolysate by Trichosporon fermentans Huang, Chao; Zong, Min-hua; Wu, Hong Bioresource Technology, Vol. 100, Issue 19 https://doi.org/10.1016/j.biortech.2009.04.022	journal	October 2009
Regulation of Trichoderma cellulase formation: lessons in molecular biology from an industrial fungus Schmoll, Monika; Kubicek, C. P. Acta Microbiologica et Immunologica Hungarica, Vol. 50, Issue 2-3 https://doi.org/10.1556/AMicr.50.2003.2-3.3	journal	June 2003
Lipid accumulation inTrichodermaspecies Serrano-Carreon, Leobardo; Hathout, Yetrib; Bensoussan, Maurice FEMS Microbiology Letters, Vol. 93, Issue 2 https://doi.org/10.1111/j.1574-6968.1992.tb05087.x	journal	June 1992
Engineering the push and pull of lipid biosynthesis in oleaginous yeast Yarrowia lipolytica for biofuel production Tai, Mitchell; Stephanopoulos, Gregory Metabolic Engineering, Vol. 15, p. 1-9 https://doi.org/10.1016/j.ymben.2012.08.007	journal	January 2013
Re-annotation of the CAZy genes of Trichoderma reesei and transcription in the presence of lignocellulosic substrates Häkkinen, Mari; Arvas, Mikko; Oja, Merja Microbial Cell Factories, Vol. 11, Issue 1 https://doi.org/10.1186/1475-2859-11-134	journal	January 2012
The glucose repressor genecre1 ofTrichoderma: Isolation and expression of a full-length and a truncated mutant form Ilmén, M.; Thrane, C.; Penttilä, M. Molecular and General Genetics MGG, Vol. 251, Issue 4 https://doi.org/10.1007/BF02172374	journal	June 1996
Fat production byTrichoderma reesei Brown, D. E.; Hasan, M.; Thornton, A. J. Biotechnology Letters, Vol. 10, Issue 4 https://doi.org/10.1007/BF01024414	journal	April 1988
Studies on the Role of the are a Gene in the Regulation of Nitrogen Catabolism in Aspergillus nidulans Hynes, Mj Australian Journal of Biological Sciences, Vol. 28, Issue 3 https://doi.org/10.1071/BI9750301	journal	January 1975
ACEII, a Novel Transcriptional Activator Involved in Regulation of Cellulase and Xylanase Genes of Trichoderma reesei Aro, Nina; Saloheimo, Anu; Ilmén, Marja Journal of Biological Chemistry, Vol. 276, Issue 26, p. 24309-24314 https://doi.org/10.1074/jbc.M003624200	journal	April 2001
Enhancement of Lipid Productivity in Oleaginous Colletotrichum Fungus through Genetic Transformation Using the Yeast CtDGAT2b Gene under Model-Optimized Growth Condition Dey, Prabuddha; Mall, Nikunj; Chattopadhyay, Atrayee PLoS ONE, Vol. 9, Issue 11 https://doi.org/10.1371/journal.pone.0111253	journal	November 2014
Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels Atsumi, Shota; Hanai, Taizo; Liao, James C. Nature, Vol. 451, Issue 7174, p. 86-89 https://doi.org/10.1038/nature06450	journal	January 2008
Transcriptional profiling of cellulase and expansin-related genes in a hypercellulolytic Trichoderma reesei Verbeke, Jonathan; Coutinho, Pedro; Mathis, Hugues Biotechnology Letters, Vol. 31, Issue 9 https://doi.org/10.1007/s10529-009-0030-5	journal	May 2009
Effect of temperature and pH on lipid accumulation by Trichoderma reesei Brown, D. E.; Hasan, M.; Lepe-Casillas, M. Applied Microbiology and Biotechnology, Vol. 34, Issue 3 https://doi.org/10.1007/BF00170054	journal	December 1990
Metabolic engineering of Yarrowia lipolytica for industrial applications Zhu, Quinn; Jackson, Ethel N. Current Opinion in Biotechnology, Vol. 36 https://doi.org/10.1016/j.copbio.2015.08.010	journal	December 2015
Genome evolution in yeasts Dujon, Bernard; Sherman, David; Fischer, Gilles Nature, Vol. 430, Issue 6995 https://doi.org/10.1038/nature02579	journal	July 2004
Accurate and reliable quantification of total microalgal fuel potential as fatty acid methyl esters by in situ transesterification Laurens, Lieve M. L.; Quinn, Matthew; Van Wychen, Stefanie Analytical and Bioanalytical Chemistry, Vol. 403, Issue 1 https://doi.org/10.1007/s00216-012-5814-0	journal	February 2012
Effects of nitrogen sources on cell growth and lipid accumulation of green alga Neochloris oleoabundans Li, Yanqun; Horsman, Mark; Wang, Bei Applied Microbiology and Biotechnology, Vol. 81, Issue 4 https://doi.org/10.1007/s00253-008-1681-1	journal	September 2008
Xyr1 (Xylanase Regulator 1) Regulates both the Hydrolytic Enzyme System and d-Xylose Metabolism in Hypocrea jecorina Stricker, Astrid R.; Grosstessner-Hain, Karin; Würleitner, Elisabeth Eukaryotic Cell, Vol. 5, Issue 12 https://doi.org/10.1128/EC.00211-06	journal	October 2006
Engineering towards a complete heterologous cellulase secretome in Yarrowia lipolytica reveals its potential for consolidated bioprocessing Wei, Hui; Wang, Wei; Alahuhta, Markus Biotechnology for Biofuels, Vol. 7, Issue 1 https://doi.org/10.1186/s13068-014-0148-0	journal	October 2014
Proteomic analysis of Chlorella vulgaris: Potential targets for enhanced lipid accumulation Guarnieri, Michael T.; Nag, Ambarish; Yang, Shihui Journal of Proteomics, Vol. 93 https://doi.org/10.1016/j.jprot.2013.05.025	journal	November 2013
Manipulation of culture conditions alters lipid content and fatty acid profiles of a wide variety of known and new oleaginous yeast species Sitepu, Irnayuli R.; Sestric, Ryan; Ignatia, Laura Bioresource Technology, Vol. 144 https://doi.org/10.1016/j.biortech.2013.06.047	journal	September 2013
Transcriptomic Analyses during the Transition from Biomass Production to Lipid Accumulation in the Oleaginous Yeast Yarrowia lipolytica Morin, Nicolas; Cescut, Julien; Beopoulos, Athanasios PLoS ONE, Vol. 6, Issue 11, Article No. e27966 https://doi.org/10.1371/journal.pone.0027966	journal	November 2011
High efficiency transformation by electroporation of Yarrowia lipolytica Wang, Jia-Hung; Hung, Wenpin; Tsai, Shu-Hsien The Journal of Microbiology, Vol. 49, Issue 3 https://doi.org/10.1007/s12275-011-0433-6	journal	June 2011
Yarrowia lipolytica: recent achievements in heterologous protein expression and pathway engineering Madzak, Catherine Applied Microbiology and Biotechnology, Vol. 99, Issue 11 https://doi.org/10.1007/s00253-015-6624-z	journal	May 2015
The CRE1 carbon catabolite repressor of the fungus Trichoderma reesei: a master regulator of carbon assimilation Portnoy, Thomas; Margeot, Antoine; Linke, Rita BMC Genomics, Vol. 12, Issue 1 https://doi.org/10.1186/1471-2164-12-269	journal	May 2011
Physiology and genetics of the dimorphic fungus Yarrowia lipolytica Barth, Gerold; Gaillardin, Claude FEMS Microbiology Reviews, Vol. 19, Issue 4, p. 219-237 https://doi.org/10.1111/j.1574-6976.1997.tb00299.x	journal	April 1997
Development of cellobiose-degrading ability in Yarrowia lipolytica strain by overexpression of endogenous genes Guo, Zhongpeng; Duquesne, Sophie; Bozonnet, Sophie Biotechnology for Biofuels, Vol. 8, Issue 1 https://doi.org/10.1186/s13068-015-0289-9	journal	August 2015
Lipid production from Yarrowia lipolytica Po1g grown in sugarcane bagasse hydrolysate Tsigie, Yeshitila Asteraye; Wang, Chun-Yuan; Truong, Chi-Thanh Bioresource Technology, Vol. 102, Issue 19, p. 9216-9222 https://doi.org/10.1016/j.biortech.2011.06.047	journal	October 2011
Cloning of the creA gene from Aspergillus nidulans: a gene involved in carbon catabolite repression Dowzer, Celia E. A.; Kelly, Joan M. Current Genetics, Vol. 15, Issue 6 https://doi.org/10.1007/BF00376804	journal	June 1989
The Biochemistry and Molecular Biology of Lipid Accumulation in Oleaginous Microorganisms Ratledge, Colin; Wynn, James P. Advances in Applied Microbiology https://doi.org/10.1016/S0065-2164(02)51000-5	book	January 2002
Comparison of the Heterologous Expression of Trichoderma reesei Endoglucanase II and Cellobiohydrolase II in the Yeasts Pichia pastoris and Yarrowia lipolytica Boonvitthya, Nassapat; Bozonnet, Sophie; Burapatana, Vorakan Molecular Biotechnology, Vol. 54, Issue 2 https://doi.org/10.1007/s12033-012-9557-0	journal	May 2012
Genome sequencing and analysis of the biomass-degrading fungus Trichoderma reesei (syn. Hypocrea jecorina) Martinez, Diego; Berka, Randy M.; Henrissat, Bernard Nature Biotechnology, Vol. 26, Issue 5 https://doi.org/10.1038/nbt1403	journal	May 2008
Advancing oleaginous microorganisms to produce lipid via metabolic engineering technology Liang, Ming-Hua; Jiang, Jian-Guo Progress in Lipid Research, Vol. 52, Issue 4, p. 395-408 https://doi.org/10.1016/j.plipres.2013.05.002	journal	October 2013
Genolevures complete genomes provide data and tools for comparative genomics of hemiascomycetous yeasts Sherman, D. Nucleic Acids Research, Vol. 34, Issue 90001 https://doi.org/10.1093/nar/gkj160	journal	January 2006
Biology and biotechnology of Trichoderma Schuster, André; Schmoll, Monika Applied Microbiology and Biotechnology, Vol. 87, Issue 3 https://doi.org/10.1007/s00253-010-2632-1	journal	May 2010
A new Zn(II)2Cys6-type transcription factor BglR regulates β-glucosidase expression in Trichoderma reesei Nitta, Mikiko; Furukawa, Takanori; Shida, Yosuke Fungal Genetics and Biology, Vol. 49, Issue 5 https://doi.org/10.1016/j.fgb.2012.02.009	journal	May 2012
Advanced biofuel production in microbes Peralta-Yahya, Pamela P.; Keasling, Jay D. Biotechnology Journal, Vol. 5, Issue 2, p. 147-162 https://doi.org/10.1002/biot.200900220	journal	January 2010
Lipid production by Yarrowia lipolytica growing on industrial glycerol in a single-stage continuous culture Papanikolaou, Seraphim; Aggelis, George Bioresource Technology, Vol. 82, Issue 1, p. 43-49 https://doi.org/10.1016/S0960-8524(01)00149-3	journal	March 2002
Nitrogen and sulfur deprivation differentiate lipid accumulation targets of Chlamydomonas reinhardtii Cakmak, Turgay; Angun, Pinar; Ozkan, Alper D. Bioengineered, Vol. 3, Issue 6 https://doi.org/10.4161/bioe.21427	journal	November 2012
Optimization of lipid production with a genome-scale model of Yarrowia lipolytica Kavšček, Martin; Bhutada, Govindprasad; Madl, Tobias BMC Systems Biology, Vol. 9, Issue 1 https://doi.org/10.1186/s12918-015-0217-4	journal	October 2015
Improving lipid production from bagasse hydrolysate with Trichosporon fermentans by response surface methodology Huang, Chao; Wu, Hong; Li, Ri-feng New Biotechnology, Vol. 29, Issue 3 https://doi.org/10.1016/j.nbt.2011.03.008	journal	February 2012
Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production Blazeck, John; Hill, Andrew; Liu, Leqian Nature Communications, Vol. 5, Issue 1 https://doi.org/10.1038/ncomms4131	journal	January 2014
Optimization of lipids production by Cryptococcus laurentii 11 using cheese whey with molasses Castanha, Rodrigo Fernandes; Mariano, Adriano Pinto; Morais, Lilia Aparecida Salgado de Brazilian Journal of Microbiology, Vol. 45, Issue 2 https://doi.org/10.1590/S1517-83822014000200003	journal	June 2014
Multiple-copy integration in the yeast Yarrowia lipolytica Le Dall, Marie-Thérèse; Nicaud, Jean-Marc; Gaillardin, Claude Current Genetics, Vol. 26, Issue 1 https://doi.org/10.1007/BF00326302	journal	July 1994
Examination of Triacylglycerol Biosynthetic Pathways via De Novo Transcriptomic and Proteomic Analyses in an Unsequenced Microalga Guarnieri, Michael T.; Nag, Ambarish; Smolinski, Sharon L. PLoS ONE, Vol. 6, Issue 10 https://doi.org/10.1371/journal.pone.0025851	journal	October 2011

Cited By (1)

Ameliorating the Metabolic Burden of the Co-expression of Secreted Fungal Cellulases in a High Lipid-Accumulating Yarrowia lipolytica Strain by Medium C/N Ratio and a Chemical Chaperone Wei, Hui; Wang, Wei; Alper, Hal S. Frontiers in Microbiology, Vol. 9 https://doi.org/10.3389/fmicb.2018.03276	journal	January 2019

Similar Records

Expression and secretion of fungal endoglucanase II and chimeric cellobiohydrolase I in the oleaginous yeast Lipomyces starkeyi

Journal Article · Mon Jul 24 00:00:00 EDT 2017 · Microbial Cell Factories · OSTI ID:1319280

Xu, Qi; Knoshaug, Eric P.; Wang, Wei; +8 more

Fatty alcohol production in Lipomyces starkeyi and Yarrowia lipolytica

Journal Article · Mon Oct 24 00:00:00 EDT 2016 · Biotechnology for Biofuels · OSTI ID:1319280

Wang, Wei; Wei, Hui; Knoshaug, Eric; +4 more

Engineering towards a complete heterologous cellulase secretome in Yarrowia lipolytica reveals its potential for consolidated bioprocessing

Journal Article · Thu Oct 16 00:00:00 EDT 2014 · Biotechnology for Biofuels · OSTI ID:1319280

Wei, Hui; Wang, Wei; Alahuhta, Markus; +6 more

Related Subjects

09 BIOMASS FUELS
59 BASIC BIOLOGICAL SCIENCES
advanced biofuel
lipids
nitrogen depletion
nitrogen repletion
Trichoderma reesei
Yarrowia lipolytica

Title: Comparison of nitrogen depletion and repletion on lipid production in yeast and fungal species

Citation Formats

References (62)

Cited By (1)

Similar Records

Related Subjects