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Using Mortierella alpina as a Novel Platform for Lipid Synthesis Inhibitor Screening
Advances in Bioscience and Bioengineering
Volume 3, Issue 1, February 2015, Pages: 1-10
Received: Apr. 7, 2015; Accepted: Apr. 11, 2015; Published: Apr. 23, 2015
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Authors
Md. Shofiul Azam, State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, People’s Republic of China; Center of Excellence for Functional Food and Health, School of Food Science and Technology, Jiangnan University, Wuxi, People’s Republic of China; Department of Food Engineering & Tea Technology, School of Applied Sciences and Technology, Shahjalal University of Science & Technology, Sylhet, Bangladesh
Zhennan Gu, State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, People’s Republic of China; Center of Excellence for Functional Food and Health, School of Food Science and Technology, Jiangnan University, Wuxi, People’s Republic of China
Haiqin Chen, State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, People’s Republic of China; Center of Excellence for Functional Food and Health, School of Food Science and Technology, Jiangnan University, Wuxi, People’s Republic of China
Yong Q. Chen, State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, People’s Republic of China; Center of Excellence for Functional Food and Health, School of Food Science and Technology, Jiangnan University, Wuxi, People’s Republic of China
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Abstract
The screening method for discovering new lipid synthesis inhibitors has not been well developed. Lipid synthesis genes are responsible for the synthesis of fatty acids in normal and tumor cells. Mortierella alpina(M. alpina) fungus has been found to produce large amounts of fatty acids because it possesses a full complement of lipid synthesis genes. We therefore hypothesized that M. alpina could be a good screening tool to find new compounds that inhibit fatty acid synthesis. We developed a M. alpina liquid culture based method to analyze the inhibitory effects of lipid synthesis inhibitors. We applied a color indicator method to monitor the oil production using some well documented lipid inhibitors (C75, Cerulenin) to verify the system, and analyzing pictures using image analysis software. The experimental drug nocodazole inhibited the lipid production in M. alpina almost to the same extent as the control inhibitors. Compared with biomass and protein levels, there was a profound effect on fatty acid level. We report the development of a fast and effective method for screening lipid synthesis inhibitors which can be used against obesity and cancer. This method can further be used to screen additional lipid and fatty acid synthesis inhibitors from natural compound libraries.
Keywords
Fatty Acid Synthase, Image Analysis Software, Lipid Synthesis Inhibitor, Mortierella alpina, Nocodazole, Screening System
To cite this article
Md. Shofiul Azam, Zhennan Gu, Haiqin Chen, Yong Q. Chen, Using Mortierella alpina as a Novel Platform for Lipid Synthesis Inhibitor Screening, Advances in Bioscience and Bioengineering. Vol. 3, No. 1, 2015, pp. 1-10. doi: 10.11648/j.abb.20150301.11
References
[1]
S.Y. Ho, Y. Jiang, F. Chen, Polyunsaturated fatty acids (PUFAs) content of the fungus Mortierella alpina isolated from soil, J Agric Food Chem 55 (2007) 3960-3966.
[2]
H.D. Jang, Y.Y. Lin, S.S. Yang, Effect of culture media and conditions on polyunsaturated fatty acids production by Mortierella alpina, Bioresour Technol 96 (2005) 1633-1644.
[3]
C. Ratledge, Fatty acid biosynthesis in microorganisms being used for Single Cell Oil production, Biochimie 86 (2004) 807-815.
[4]
S. Shimizu, S. Jareonkitmongkol, Mortierella Species (Fungi): Production of C20 Polyunsaturated Fatty Acids, in: Y.P.S. Bajaj (Ed.), Medicinal and Aromatic Plants VIII, Springer Berlin Heidelberg, 1995, pp. 308-325.
[5]
N. Amano, Y. Shinmen, K. Akimoto, H. Kawashima, T. Amachi, S. Shimizu, H. Yamada, Chemotaxonomic significance of fatty-acid composition in the genus Mortierella (Zygomycetes, Mortierellaceae), Mycotaxon 44 (1992) 257-265.
[6]
E. Sakuradani, S. Shimizu, Single cell oil production by Mortierella alpina, J Biotechnol 144 (2009) 31-36.
[7]
E. Sakuradani, Advances in the Production of Various Polyunsaturated Fatty Acids through Oleaginous Fungus Mortierella alpina Breeding, Bioscience, Biotechnology, and Biochemistry 74 (2010) 908-917.
[8]
E. Sakuradani, A. Ando, J. Ogawa, S. Shimizu, Improved production of various polyunsaturated fatty acids through filamentous fungus Mortierella alpina breeding, Appl Microbiol Biotechnol 84 (2009) 1-10.
[9]
F.P. Kuhajda, Fatty-acid synthase and human cancer: new perspectives on its role in tumor biology, Nutrition 16 (2000) 202-208.
[10]
L. Weiss, G.E. Hoffmann, R. Schreiber, H. Andres, E. Fuchs, E. Korber, H.J. Kolb, Fatty-acid biosynthesis in man, a pathway of minor importance. Purification, optimal assay conditions, and organ distribution of fatty-acid synthase, Biol Chem Hoppe Seyler 367 (1986) 905-912.
[11]
M. Ookhtens, R. Kannan, I. Lyon, N. Baker, Liver and adipose tissue contributions to newly formed fatty acids in an ascites tumor, 1984.
[12]
F.P. Kuhajda, Fatty acid synthase and cancer: new application of an old pathway, Cancer Res 66 (2006) 5977-5980.
[13]
A.C. Price, K.H. Choi, R.J. Heath, Z. Li, S.W. White, C.O. Rock, Inhibition of beta-ketoacyl-acyl carrier protein synthases by thiolactomycin and cerulenin. Structure and mechanism, J Biol Chem 276 (2001) 6551-6559.
[14]
M. Leibundgut, T. Maier, S. Jenni, N. Ban, The multienzyme architecture of eukaryotic fatty acid synthases, Curr Opin Struct Biol 18 (2008) 714-725.
[15]
C. Goswami, L. Goswami, Filamentous microtubules in the neuronal spinous process and the role of microtubule regulatory drugs in neuropathic pain, Neurochem Int 57 (2010) 497-503.
[16]
M.J. Egan, M.A. McClintock, S.L. Reck-Peterson, Microtubule-based transport in filamentous fungi, Curr Opin Microbiol 15 (2012) 637-645.
[17]
S. Heino, S. Lusa, P. Somerharju, C. Ehnholm, V.M. Olkkonen, E. Ikonen, Dissecting the role of the golgi complex and lipid rafts in biosynthetic transport of cholesterol to the cell surface, Proc Natl Acad Sci U S A 97 (2000) 8375-8380.
[18]
A.K. Franz, M.A. Danielewicz, D.M. Wong, L.A. Anderson, J.R. Boothe, Phenotypic screening with oleaginous microalgae reveals modulators of lipid productivity, ACS Chem Biol 8 (2013) 1053-1062.
[19]
J.C. Yarrow, G. Totsukawa, G.T. Charras, T.J. Mitchison, Screening for cell migration inhibitors via automated microscopy reveals a Rho-kinase inhibitor, Chem Biol 12 (2005) 385-395.
[20]
S. Cooper, G. Iyer, M. Tarquini, P. Bissett, Nocodazole does not synchronize cells: implications for cell-cycle control and whole-culture synchronization, Cell Tissue Res 324 (2006) 237-242.
[21]
R. Ashraf, N.P. Shah, Selective and differential enumerations of Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium spp. in yoghurt--a review, Int J Food Microbiol 149 (2011) 194-208.
[22]
S.H. Moussa, A.A. Tayel, A.A. Al-Hassan, A. Farouk, Tetrazolium/Formazan Test as an Efficient Method to Determine Fungal Chitosan Antimicrobial Activity, Journal of Mycology 2013 (2013) 1-7.
[23]
M. Zhu, L.J. Yu, Z. Liu, H.B. Xu, Isolating Mortierella alpina strains of high yield of arachidonic acid, Lett Appl Microbiol 39 (2004) 332-335.
[24]
P. Phattanawasin, U. Sotanaphun, I. Kanchanaphibool, L. Sriphong, N. Piyapolrungroj, A Comparison of Image Analysis Software for Quantitative TLC, Silpakorn U Science & Tech J 5 (2011).
[25]
L. Wang, W. Chen, Y. Feng, Y. Ren, Z. Gu, H. Chen, H. Wang, M.J. Thomas, B. Zhang, I.M. Berquin, Y. Li, J. Wu, H. Zhang, Y. Song, X. Liu, J.S. Norris, S. Wang, P. Du, J. Shen, N. Wang, Y. Yang, W. Wang, L. Feng, C. Ratledge, H. Zhang, Y.Q. Chen, Genome characterization of the oleaginous fungus Mortierella alpina, PLoS One 6 (2011) e28319.
[26]
E.G.a. Bligh, W.J. Dyer, A rapid method of total lipid extraction and purification, Can.J.Biochem.Physiol. 37 (1959) 911-917.
[27]
L.D. Metcalfe, A.A. Schmitz, J.R. Pelka, Rapid Preparation of Fatty Acid Esters from Lipids for Gas Chromatographic Analysis, Analytical Chemistry 38 (1966) 514-515.
[28]
S.W. White, J. Zheng, Y.M. Zhang, Rock, The structural biology of type II fatty acid biosynthesis, Annu Rev Biochem 74 (2005) 791-831.
[29]
V. Bhadauria, Y.L. Peng, Optimization of a protein extraction technique for fungal proteomics, Indian J Microbiol 50 (2010) 127-131.
[30]
M.M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal Biochem 72 (1976) 248-254.
[31]
C.M. Stoscheck, Quantitation of protein, 182 (1990) 50-68.
[32]
J.P. Palta, J. Levitt, E.J. Stadelmann, Plant viability assay, Cryobiology 15 (1978) 249-255.
[33]
D.A. Musser, A.R. Oseroff, The use of tetrazolium salts to determine sites of damage to the mitochondrial electron transport chain in intact cells following in vitro photodynamic therapy with Photofrin II, Photochem Photobiol 59 (1994) 621-626.
[34]
P.R. Rich, L.A. Mischis, S. Purton, J.T. Wiskich, The sites of interaction of triphenyltetrazolium chloride with mitochondrial respiratory chains, FEMS Microbiology Letters 202 (2001) 181-187.
[35]
R.J. Vasquez, B. Howell, A.M. Yvon, P. Wadsworth, L. Cassimeris, Nanomolar concentrations of nocodazole alter microtubule dynamic instability in vivo and in vitro, Mol Biol Cell 8 (1997) 973-985.
[36]
S.S. Niini, M. Raudaskoski, Response of Ectomycorrhizal Fungi to Benomyl and Nocodazole - Growth-Inhibition and Microtubule Depolymerization, Mycorrhiza 3 (1993) 83-91.
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