Abstract
Yarrowia lipolytica was usually transformed by heat shock, but linearized integrative vectors always resulted in a low transformation efficiency when electroporation was used. To develop a high efficiency integrative transformation method by electroporation of Y. lipolytica, we report here that pretreatment of Y. lipolytica with 150 mM LiAc for 1 h before electroporation will approximately 30-fold of increase transformation efficiency. A cell concentration of 1010/ml and instrument settings of 1.5 kV will generate the highest transformation efficiencies. We have developed a procedure to transform Y. lipolytica that will be able to yield an efficiency of 2.1×104 transformants/μg for integrative linear DNA. With our modifications, the electroporation procedures became a very efficient and reliable tool for Y. lipolytica transformation.
Citations
Citations to this article as recorded by
- Advances in synthetic biology tools paving the way for the biomanufacturing of unusual fatty acids using the Yarrowia lipolytica chassis
Kaifeng Wang, Tian-Qiong Shi, Lu Lin, Ping Wei, Rodrigo Ledesma-Amaro, Xiao-Jun Ji, He Huang
Biotechnology Advances.2022; 59: 107984. CrossRef - Orotic acid production by Yarrowia lipolytica under conditions of limited pyrimidine
Paul Swietalski, Frank Hetzel, Iris Klaiber, Eva Pross, Ines Seitl, Lutz Fischer
Yeast.2022; 39(3): 230. CrossRef - Recent advances in genetic technology development of oleaginous yeasts
Zhiqiang Wen, Naief H. Al Makishah
Applied Microbiology and Biotechnology.2022; 106(17): 5385. CrossRef - Identification and Characterization of the Mitochondrial Replication Origin for Stable and Episomal Expression in Yarrowia lipolytica
Zhiyong Cui, Huihui Zheng, Zhennan Jiang, Zhaoxuan Wang, Jin Hou, Qian Wang, Quanfeng Liang, Qingsheng Qi
ACS Synthetic Biology.2021; 10(4): 826. CrossRef - Recent advances in lipid metabolic engineering of oleaginous yeasts
Atrayee Chattopadhyay, Mohor Mitra, Mrinal K. Maiti
Biotechnology Advances.2021; 53: 107722. CrossRef - Yarrowia lipolytica Strains and Their Biotechnological Applications: How Natural Biodiversity and Metabolic Engineering Could Contribute to Cell Factories Improvement
Catherine Madzak
Journal of Fungi.2021; 7(7): 548. CrossRef - Revisiting the unique structure of autonomously replicating sequences in Yarrowia lipolytica and its role in pathway engineering
Carmen Lopez, Mingfeng Cao, Zhanyi Yao, Zengyi Shao
Applied Microbiology and Biotechnology.2021; 105(14-15): 5959. CrossRef - Optimization of electrotransformation (ETF) conditions in lactic acid bacteria (LAB)
Chao Wang, Yanhua Cui, Xiaojun Qu
Journal of Microbiological Methods.2020; 174: 105944. CrossRef - Understanding and Eliminating the Detrimental Effect of Thiamine Deficiency on the Oleaginous Yeast Yarrowia lipolytica
Caleb Walker, Seunghyun Ryu, Richard J. Giannone, Sergio Garcia, Cong T. Trinh, Haruyuki Atomi
Applied and Environmental Microbiology.2020;[Epub] CrossRef - Sustainable production of FAEE biodiesel using the oleaginous yeast Yarrowia lipolytica
Aiqun Yu, Yu Zhao, Jian Li, Shenglong Li, Yaru Pang, Yakun Zhao, Cuiying Zhang, Dongguang Xiao
MicrobiologyOpen.2020;[Epub] CrossRef - Secretion of a low and high molecular weight β-glycosidase by Yarrowia lipolytica
Paul Swietalski, Frank Hetzel, Ines Seitl, Lutz Fischer
Microbial Cell Factories.2020;[Epub] CrossRef - A novel electroporation procedure for highly efficient transformation of Lipomyces starkeyi
Hiroaki Takaku, Atsumi Miyajima, Haruka Kazama, Rikako Sato, Satoshi Ara, Tomohiko Matsuzawa, Katsuro Yaoi, Hideo Araki, Yosuke Shida, Wataru Ogasawara, Harutake Yamazaki
Journal of Microbiological Methods.2020; 169: 105816. CrossRef - Homology‐independent genome integration enables rapid library construction for enzyme expression and pathway optimization in Yarrowia lipolytica
Zhiyong Cui, Xin Jiang, Huihui Zheng, Qingsheng Qi, Jin Hou
Biotechnology and Bioengineering.2019; 116(2): 354. CrossRef - Electroporation of germinated conidia and young mycelium as an efficient transformation system for Acremonium chrysogenum
Jessica Cruz-Ramón, Francisco J. Fernández, Armando Mejía, Francisco Fierro
Folia Microbiologica.2019; 64(1): 33. CrossRef - Exceptional solvent tolerance in Yarrowia lipolytica is enhanced by sterols
Caleb Walker, Seunghyun Ryu, Cong T. Trinh
Metabolic Engineering.2019; 54: 83. CrossRef - Understanding Functional Roles of Native Pentose-Specific Transporters for Activating Dormant Pentose Metabolism in Yarrowia lipolytica
Seunghyun Ryu, Cong T. Trinh, Marie A. Elliot
Applied and Environmental Microbiology.2018;[Epub] CrossRef - Metabolic engineering in the host Yarrowia lipolytica
Ahmad M. Abdel-Mawgoud, Kelly A. Markham, Claire M. Palmer, Nian Liu, Gregory Stephanopoulos, Hal S. Alper
Metabolic Engineering.2018; 50: 192. CrossRef - Optimization of chemico-physical transformation methods for various bacterial species using diverse chemical compounds and nanomaterials
Jun Ren, Dokyun Na, Seung Min Yoo
Journal of Biotechnology.2018; 288: 55. CrossRef - Recent Advances in Metabolic Engineering of Yarrowia lipolytica for Lipid Overproduction
Si‐Yu Zeng, Hu‐Hu Liu, Tian‐Qiong Shi, Ping Song, Lu‐Jing Ren, He Huang, Xiao‐Jun Ji
European Journal of Lipid Science and Technology.2018;[Epub] CrossRef - High-efficiency transformation of Yarrowia lipolytica using electroporation
Kelly A Markham, Sofia Vazquez, Hal S Alper
FEMS Yeast Research.2018;[Epub] CrossRef - Synthesis and assembly of human vault particles in yeast
Meng Wang, Valerie A. Kickhoefer, Leonard H. Rome, Oliver K. Foellmer, Shaily Mahendra
Biotechnology and Bioengineering.2018; 115(12): 2941. CrossRef - Fast and efficient genetic transformation of oleaginous yeast Rhodosporidium toruloides by using electroporation
Hongdi Liu, Xiang Jiao, Yanan Wang, Xiaobing Yang, Wenyi Sun, Jihui Wang, Sufang Zhang, Zongbao Kent Zhao
FEMS Yeast Research.2017;[Epub] CrossRef - New recombinant strains of the yeast Yarrowia lipolytica with overexpression of the aconitate hydratase gene for the obtainment of isocitric acid from rapeseed oil
I. A. Laptev, N. A. Filimonova, R. K. Allayarov, S. V. Kamzolova, V. A. Samoilenko, S. P. Sineoky, I. G. Morgunov
Applied Biochemistry and Microbiology.2016; 52(7): 699. CrossRef - Overexpression of Δ12-Fatty Acid Desaturase in the Oleaginous Yeast Rhodosporidium toruloides for Production of Linoleic Acid-Rich Lipids
Yanan Wang, Sufang Zhang, Markus Pötter, Wenyi Sun, Li Li, Xiaobing Yang, Xiang Jiao, Zongbao K. Zhao
Applied Biochemistry and Biotechnology.2016; 180(8): 1497. CrossRef - Recent advances in bioengineering of the oleaginous yeast Yarrowia lipolytica
Murtaza Shabbir Hussain, Gabriel M Rodriguez, Difeng Gao, Michael Spagnuolo, Lauren Gambill, Mark Blenner
AIMS Bioengineering.2016; 3(4): 493. CrossRef - Metabolic Flux Analysis of Lipid Biosynthesis in the Yeast Yarrowia lipolytica Using 13C-Labled Glucose and Gas Chromatography-Mass Spectrometry
Huaiyuan Zhang, Chao Wu, Qingyu Wu, Junbiao Dai, Yuanda Song, Wei Ning Chen
PLOS ONE.2016; 11(7): e0159187. CrossRef - Comparison of Nitrogen Depletion and Repletion on Lipid Production in Yeast and Fungal Species
Shihui Yang, Wei Wang, Hui Wei, Stefanie Van Wychen, Philip Pienkos, Min Zhang, Michael Himmel
Energies.2016; 9(9): 685. CrossRef - Engineering Rhodosporidium toruloides for increased lipid production
Shuyan Zhang, Jeffrey M. Skerker, Charles D. Rutter, Matthew J. Maurer, Adam P. Arkin, Christopher V. Rao
Biotechnology and Bioengineering.2016; 113(5): 1056. CrossRef - Activating and Elucidating Metabolism of Complex Sugars in Yarrowia lipolytica
Seunghyun Ryu, Julie Hipp, Cong T. Trinh, D. Cullen
Applied and Environmental Microbiology.2016; 82(4): 1334. CrossRef - Simultaneous saccharification and fermentation of cellulose in ionic liquid for efficient production of α-ketoglutaric acid by Yarrowia lipolytica
Seunghyun Ryu, Nicole Labbé, Cong T. Trinh
Applied Microbiology and Biotechnology.2015; 99(10): 4237. CrossRef - Electroporation on microchips: the harmful effects of pH changes and scaling down
Yang Li, Mengxi Wu, Deyao Zhao, Zewen Wei, Wenfeng Zhong, Xiaoxia Wang, Zicai Liang, Zhihong Li
Scientific Reports.2015;[Epub] CrossRef - Improved electroporation procedure for genetic transformation ofDekkera/Brettanomyces bruxellensis
Marina Miklenić, Bojan Žunar, Anamarija Štafa, Ivan-Krešimir Svetec, Jens Nielsen
FEMS Yeast Research.2015; 15(8): fov096. CrossRef - Biotechnological applications of Yarrowia lipolytica: Past, present and future
Hu-Hu Liu, Xiao-Jun Ji, He Huang
Biotechnology Advances.2015; 33(8): 1522. CrossRef - The Strictly Aerobic Yeast Yarrowia lipolytica Tolerates Loss of a Mitochondrial DNA-Packaging Protein
Jana Bakkaiova, Kosuke Arata, Miki Matsunobu, Bungo Ono, Tomoyo Aoki, Dana Lajdova, Martina Nebohacova, Jozef Nosek, Isamu Miyakawa, Lubomir Tomaska
Eukaryotic Cell.2014; 13(9): 1143. CrossRef - Yarrowia lipolytica as an Oleaginous Cell Factory Platform for Production of Fatty Acid-Based Biofuel and Bioproducts
Ali Abghari, Shulin Chen
Frontiers in Energy Research.2014;[Epub] CrossRef - Regulatory properties of malic enzyme in the oleaginous yeast, Yarrowia lipolytica, and its non-involvement in lipid accumulation
Huaiyuan Zhang, Luning Zhang, Haiqin Chen, Yong Q. Chen, Colin Ratledge, Yuanda Song, Wei Chen
Biotechnology Letters.2013; 35(12): 2091. CrossRef
MSK
Cite this Article
