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Jin-Lai Zhang, Yang-Zi Peng, Duo Liu, Hong Liu, Ying-Xiu Cao, Bing-Zhi Li, Chun Li, Ying-Jin Yuan
BACKGROUND: The oleaginous yeast Yarrowia lipolytica is a promising microbial cell factory due to their biochemical characteristics and native capacity to accumulate lipid-based chemicals. To create heterogenous biosynthesis pathway and manipulate metabolic flux in Y. lipolytica, numerous studies have been done for developing synthetic biology tools for gene regulation. CRISPR interference (CRISPRi), as an emerging technology, has been applied for specifically repressing genes of interest...
April 20, 2018: Microbial Cell Factories
Ryo Iwama, Mariho Hara, Aya Mizuike, Hiroyuki Horiuchi, Ryouichi Fukuda
In this study, we investigated the role of OSH6, which encodes a homolog of the oxysterol-binding protein, in the assimilation of n-alkanes in the yeast Yarrowia lipolytica. The deletion mutant of OSH6 showed growth defects on n-alkanes of 10-16 carbons. In the deletion mutant, production of the functional cytochrome P450 was not observed. However, transcription of ALK1, encoding a major P450 belonging to the CYP52 family that plays a critical role in n-alkane hydroxylation, and further translation of its transcript were noted in the deletion mutant as well as in the wild-type strain...
April 2, 2018: Biochemical and Biophysical Research Communications
Christophe Leplat, Jean-Marc Nicaud, Tristan Rossignol
Yarrowia lipolytica is a non-conventional oleaginous yeast that displays high lipid titers and yields; its production capacity holds significant promise for industrial biolipid applications. While its lipid metabolism has been widely studied, little is known about its transcriptional regulatory network. Deciphering the role of transcriptional regulators is crucial for understanding lipid accumulation, a complex phenomenon.To identify the transcription factors involved in lipid metabolism, we developed a systematic overexpression approach for 148 putative transcription factors...
March 29, 2018: FEMS Yeast Research
Lucille Garnier, Marcia Leyva Salas, Nicolas Pinon, Norman Wiernasz, Audrey Pawtowski, Emmanuel Coton, Jérôme Mounier, Florence Valence
In this study, we developed a high-throughput antifungal activity screening method using a cheese-mimicking matrix distributed in 24-well plates. This method allowed rapid screening of a large variety of antifungal agent candidates: bacterial fermented ingredients, bacterial isolates, and preservatives. Using the proposed method, we characterized the antifungal activity of 44 lactic acid bacteria (LAB) fermented milk-based ingredients and 23 LAB isolates used as protective cultures against 4 fungal targets (Mucor racemosus, Penicillium commune, Galactomyces geotrichum, and Yarrowia lipolytica)...
March 28, 2018: Journal of Dairy Science
Ashok Bankar, Smita Zinjarde, Manisha Shinde, Gita Gopalghare, Ameeta Ravikumar
Heavy metal tolerance of two marine strains of Yarrowia lipolytica was tested on solid yeast extract peptone dextrose agar plates. Based on minimum inhibitory concentration esteems, it is inferred that the two strains of Y. lipolytica were tolerant to heavy metals such as Pb(II), Cr(III), Zn(II), Cu(II), As(V), and Ni(II) ions. The impact of various heavy metal concentrations on the growth kinetics of Y. lipolytica was likewise assessed. With increased heavy metal concentration, the specific growth rate was reduced with delayed doubling time...
March 29, 2018: Extremophiles: Life Under Extreme Conditions
Mingjie Bian, Shan Li, Huanhuan Wei, Shiping Huang, Feng Zhou, Youming Zhu, Guoping Zhu
Yarrowia lipolytica, a nonpathogenic, nonconventional, aerobic and dimorphic yeast, is considered an oleaginous microorganism due to its excellent ability to accumulate large amounts of lipids. Glucose-6-phosphate dehydrogenase (G6PD) is one of two key enzymes involved in the lipid accumulation in this fungi, which catalyzes the oxidative dehydrogenation of glucose-6-phosphate to 6-phosphoglucono-δ-lactone with the reduction of NADP+ to NADPH. In this study, the full-length gene of G6PD from Y. lipolytica (YlG6PD) was cloned without intron and heterogeneously expressed in E...
March 23, 2018: Protein Expression and Purification
Cuijuan Gao, Yaqin Zheng
Succinic acid is a high value-added organic acid widely used in food, chemical and pesticide industries. As a new robust non-conventional yeast, Yarrowia lipolytica attracts more and more attention due to its potential for industrial applications. Previously, we obtained a succinic acid-producing strain through gene deletion of succinic acid dehydrogenase subunit encoding gene Ylsdh5, resulting in the strain of PGC01003. However, the recombinant strain produced large amount of acetic acid due to imbalance between glycolysis and TCA cycle which hindered the efficient production of succinic acid...
March 25, 2018: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
Weibing Zhang, Qiaoqiao Luo, Yan Zhu, Jiang Ma, Lei Cao, Min Yang, Pencheng Wen, Zhongmin Zhang, Xiaoling He
Douchi has been consumed as a flavoring ingredient for centuries. During production of douchi, numerous microorganisms play important roles in the hydrolysis and conversion of proteins and starch, which are related to the quality and flavor of the end product. Therefore, in the present study, the microbial diversity in two types of home-made traditional bacterial douchi from Gansu province in northwest China was studied by high-throughput sequencing, and a corresponding analysis of the bacterial and fungal communities were conducted...
2018: PloS One
Svetlana V Kamzolova, Roman V Shamin, Nadezda N Stepanova, Grigorii I Morgunov, Julia N Lunina, Ramil K Allayarov, Vladimir A Samoilenko, Igor G Morgunov
Isocitric acid exists in the form of four stereoisomers, of which only the threo -Ds-form (ICA) is a natural active compound, an intermediate of Krebs cycle, and suitable for nutritional and pharmaceutical use. In this paper, we propose a method for ICA production from ethanol by yeast Yarrowia lipolytica . The effects of temperature, pH of the medium, and aeration on the growth of the producer Y. lipolytica VKM Y-2373 and synthesis of ICA were studied. An optimal fermentation regime, which ensures a good growth of the producer and directed synthesis of the target product, was determined...
2018: BioMed Research International
In-Seung Jang, Byung Jo Yu, Ji Yeon Jang, Jonggeon Jegal, Ju Young Lee
Gene targeting is a challenge in Yarrowia lipolytica (Y. lipolytica) where non-homologous end-joining (NHEJ) is predominant over homologous recombination (HR). To improve the frequency and efficiency of HR in Y. lipolytica, the ku70 gene responsible for a double stand break (DSB) repair in the NHEJ pathway was disrupted, and the cell cycle was synchronized to the S-phase with hydroxyurea, respectively. Consequently, the HR frequency was over 46% with very short homology regions (50 bp): the pex10 gene was accurately deleted at a frequency of 60% and the β-carotene biosynthetic genes were integrated at the correct locus at an average frequency of 53%...
2018: PloS One
Nicholas J Morse, James M Wagner, Kevin Reed, Madan R Gopal, Lars Lauffer, Hal S Alper
Efficient guide RNA expression often limits CRISPR-Cas9 implementation in new hosts. To address this limitation in fungal systems, we demonstrate the utility of a T7 polymerase system to effectively express sgRNAs. Initially, we developed a methodology in Saccharomyces cerevisiae using a modified version of the T7 P266L mutant polymerase with an SV40 nuclear localization signal to allow guide RNA expression immediately downstream of a T7 promoter. To improve targeting efficiency, guide RNA design was found to be tolerant to three mismatches or up to three additional bases appended to the 5' end...
March 22, 2018: ACS Synthetic Biology
Pranjul Mishra, Na-Rae Lee, Meiyappan Lakshmanan, Minsuk Kim, Byung-Gee Kim, Dong-Yup Lee
BACKGROUND: Recently, there have been several attempts to produce long-chain dicarboxylic acids (DCAs) in various microbial hosts. Of these, Yarrowia lipolytica has great potential due to its oleaginous characteristics and unique ability to utilize hydrophobic substrates. However, Y. lipolytica should be further engineered to make it more competitive: the current approaches are mostly intuitive and cumbersome, thus limiting its industrial application. RESULTS: In this study, we proposed model-guided metabolic engineering strategies for enhanced production of DCAs in Y...
March 19, 2018: BMC Systems Biology
Kanchana Rueksomtawin Kildegaard, Belén Adiego-Pérez, David Doménech Belda, Jaspreet Kaur Khangura, Carina Holkenbrink, Irina Borodina
Astaxanthin is a red-colored carotenoid, used as food and feed additive. Astaxanthin is mainly produced by chemical synthesis, however, the process is expensive and synthetic astaxanthin is not approved for human consumption. In this study, we engineered the oleaginous yeast Yarrowia lipolytica for de novo production of astaxanthin by fermentation. First, we screened 12 different Y. lipolytica isolates for β-carotene production by introducing two genes for β-carotene biosynthesis: bi-functional phytoene synthase/lycopene cyclase ( crtYB ) and phytoene desaturase ( crtI ) from the red yeast Xanthophyllomyces dendrorhous ...
December 2017: Synthetic and Systems Biotechnology
Paul G Becher, Arne Hagman, Vasiliki Verschut, Amrita Chakraborty, Elżbieta Rozpędowska, Sébastien Lebreton, Marie Bengtsson, Gerhard Flick, Peter Witzgall, Jure Piškur
Yeast volatiles attract insects, which apparently is of mutual benefit, for both yeasts and insects. However, it is unknown whether biosynthesis of metabolites that attract insects is a basic and general trait, or if it is specific for yeasts that live in close association with insects. Our goal was to study chemical insect attractants produced by yeasts that span more than 250 million years of evolutionary history and vastly differ in their metabolism and lifestyle. We bioassayed attraction of the vinegar fly Drosophila melanogaster to odors of phylogenetically and ecologically distinct yeasts grown under controlled conditions...
March 2018: Ecology and Evolution
Asma Timoumi, Stéphane E Guillouet, Carole Molina-Jouve, Luc Fillaudeau, Nathalie Gorret
The yeast Yarrowia lipolytica is an industrially important microorganism with distinctive physiological and metabolic characteristics. A variety of external factors (e.g., pH, temperature, and nutrient availability) influences the behavior of the yeast and may act as stress conditions which the cells must withstand and adapt. In this mini review, the impacts of environmental factors on the morphology and metabolite production by Y. lipolytica are summarized. In this regard, detailed insights into the effectors involved in the dimorphic transition of Y...
March 9, 2018: Applied Microbiology and Biotechnology
Karen M Davies, Thorsten B Blum, Werner Kühlbrandt
We used electron cryo-tomography and subtomogram averaging to investigate the structure of complex I and its supramolecular assemblies in the inner mitochondrial membrane of mammals, fungi, and plants. Tomographic volumes containing complex I were averaged at ∼4 nm resolution. Principal component analysis indicated that ∼60% of complex I formed a supercomplex with dimeric complex III, while ∼40% were not associated with other respiratory chain complexes. The mutual arrangement of complex I and III2 was essentially conserved in all supercomplexes investigated...
March 8, 2018: Proceedings of the National Academy of Sciences of the United States of America
Yu Liu, Peng Zhang, Weilin Wang, Miren Dong, Min Wang, Changhao Gong, Zhihao Jia, Zhaoqun Liu, Anguo Zhang, Lingling Wang, Linsheng Song
DM9 is a novel protein domain with unknown function originally discovered in Drosophila melanogaster. Recently, a protein harboring DM9 repeats was identified as mannose-specific lectin (CgCGL1, renamed as CgDM9CP-1) from the Pacific oyster Crassostrea gigas. In the present study, another DM9 containing protein was identified from oyster C. gigas (designated as CgDM9CP-2). The open reading frame of CgDM9CP-2 gene was of 432 bp, encoding a polypeptide of 143 amino acids with two tandem DM9 repeats. The deduced amino acid sequence of CgDM9CP-2 shared 60...
March 5, 2018: Developmental and Comparative Immunology
P W Crous, M J Wingfield, T I Burgess, A J Carnegie, G E St J Hardy, D Smith, B A Summerell, J F Cano-Lira, J Guarro, J Houbraken, L Lombard, M P Martín, M Sandoval-Denis, A V Alexandrova, C W Barnes, I G Baseia, J D P Bezerra, V Guarnaccia, T W May, M Hernández-Restrepo, A M Stchigel, A N Miller, M E Ordoñez, V P Abreu, T Accioly, C Agnello, A Agustin Colmán, C C Albuquerque, D S Alfredo, P Alvarado, G R Araújo-Magalhães, S Arauzo, T Atkinson, A Barili, R W Barreto, J L Bezerra, T S Cabral, F Camello Rodríguez, R H S F Cruz, P P Daniëls, B D B da Silva, D A C de Almeida, A A de Carvalho Júnior, C A Decock, L Delgat, S Denman, R A Dimitrov, J Edwards, A G Fedosova, R J Ferreira, A L Firmino, J A Flores, D García, J Gené, A Giraldo, J S Góis, A A M Gomes, C M Gonçalves, D E Gouliamova, M Groenewald, B V Guéorguiev, M Guevara-Suarez, L F P Gusmão, K Hosaka, V Hubka, S M Huhndorf, M Jadan, Ž Jurjević, B Kraak, V Kučera, T K A Kumar, I Kušan, S R Lacerda, S Lamlertthon, W S Lisboa, M Loizides, J J Luangsa-Ard, P Lysková, W P Mac Cormack, D M Macedo, A R Machado, E F Malysheva, P Marinho, N Matočec, M Meijer, A Mešić, S Mongkolsamrit, K A Moreira, O V Morozova, K U Nair, N Nakamura, W Noisripoom, I Olariaga, R J V Oliveira, L M Paiva, P Pawar, O L Pereira, S W Peterson, M Prieto, E Rodríguez-Andrade, C Rojo De Blas, M Roy, E S Santos, R Sharma, G A Silva, C M Souza-Motta, Y Takeuchi-Kaneko, C Tanaka, A Thakur, M Th Smith, Z Tkalčec, N Valenzuela-Lopez, P van der Kleij, A Verbeken, M G Viana, X W Wang, J Z Groenewald
Novel species of fungi described in this study include those from various countries as follows: Antarctica : Cadophora antarctica from soil. Australia : Alfaria dandenongensis on Cyperaceae , Amphosoma persooniae on Persoonia sp., Anungitea nullicana on Eucalyptus sp . , Bagadiella eucalypti on Eucalyptus globulus , Castanediella eucalyptigena on Eucalyptus sp., Cercospora dianellicola on Dianella sp., Cladoriella kinglakensis on Eucalyptus regnans , Cladoriella xanthorrhoeae (incl. Cladoriellaceae fam. nov...
December 2017: Persoonia
Eun Ju Yun, James Lee, Do Hyoung Kim, Jungyeon Kim, Sooah Kim, Yong-Su Jin, Kyoung Heon Kim
Lipid production by oleaginous Yarrowia lipolytica depends highly on culture environments, such as carbon sources, carbon/nitrogen (C/N) ratios, types of media, and cellular growth phases. In this study, the effects of media and carbon sources on lipid and metabolite production were investigated by profiling fatty acids and intracellular metabolites of Y. lipolytica grown in various media. The highest total fatty acid yield 114.04 ± 6.23 mg/g dry cell weight was achieved by Y. lipolytica grown in minimal medium with glycerol (SCG) in the exponential phase...
February 27, 2018: Journal of Biotechnology
James M Wagner, Eden V Williams, Hal S Alper
Yarrowia lipolytica is a non-conventional yeast of interest to the biotechnology industry. However, the physiology, metabolism, and genetic regulation of Y. lipolytica diverge significantly from more well-studied and characterized yeasts such as Saccharomyces cerevisiae. To develop additional genetic tools for this industrially relevant host, we adapt the piggyBac transposon system to enable efficient generation of genome-wide insertional mutagenesis libraries and introduction of scarless, footprint-free genomic modifications in Y...
March 1, 2018: Biotechnology Journal
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