Abstract
In this study, some of the ATP-citrate lyase genes (ACL1) were deleted and the copy number of the iso-citrate lyase gene (ICL1) was increased in the marine-derived yeast Yarrowia lipolytica SWJ-1b displaying the recombinant inulinase. It was found that lipid content and iso-citric acid in the transformant 30 obtained were greatly reduced and citric acid production was greatly enhanced. It was also found that the ACL1 gene expression and ATP-citrate lyase activity in the transformant 30 were declined and the ICL1 gene expression and iso-citrate lyase activity were promoted. During the 2-l fermentation, 84.0 g/l of citric acid and 1.8 g/l of iso-citric acid in the fermented medium were attained from 10.0 % of inulin by the transformant 30 within 214 h. The results showed that only 0.36 % of the residual reducing sugar and 1.0 % of the residual total sugar were left in the fermented medium, suggesting that 89.6 % of the total sugar was used for citric acid production and cell growth by the transformant 30.
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References
Alvarez-Vasquez F, González-Alcón C, Torres NV (2000) Metabolism of citric acid production by Aspergillus niger: model definition, steady-state analysis and constrained optimization of citric acid production rate. Biotechnol Bioeng 70:82–108
Antonucci S, Bravi M, Bubbico R, Michele AD, Verdone N (2001) Selectivity in citric acid production by Yarrowia lipolytica. Enzy Microb Technol 28:189–195
Arzumanov TE, Shishkanova NV, Finogenova TF (2000) Biosynthesis of citric acid by Yarrowia lipolytica repeat-batch culture on ethanol. Appl Microbiol Biotechnol 53:525–529
Camp BJ, Farmer L (1967) A rapid spectrophotometric method for the determination of citric acid in blood. Clin Chem 13:501–505
Chi ZM, Liu J, Zhang W (2001) Trehalose accumulation from soluble starch by Saccharomycopsis fibuligera sdu. Enzy Microb Technol 28:240–245
Chi ZM, Zhang T, Cao TS, Liu XY, Cui W, Zhao CH (2011) Biotechnological potential of inulin for bioprocesses. Biores Technol 102:4295–4303
Crolla A, Kennedy KJ (2001) Optimization of citric acid production from Candida lipolytica Y-1095 using n-paraffin. J Biotechnol 89:27–40
Darvishi F, Nahvi I, Zarkesh-Esfahani H, Momenbeik F (2009) Effect of plant oils upon lipase and citric acid production in Yarrowia lipolytica yeast. J Biomed Biotechnol 56:29–43
de Jongh WA, Nielsen J (2008) Enhanced citrate production through gene insertion in Aspergillus niger. Metab Eng 10:87–96
Folch J, Lees M, Slane-Stanley JA (1957) A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem 226:497–509
Förster A, Aurich A, Mauersberger S, Barth G (2007) Citric acid production from sucrose using a recombinant strain of the yeast Yarrowia lipolytica. Appl Microbiol Biotechnol 75:1409–1417
Gong F, Sheng J, Chi ZM, Li J (2007) Inulinase production by a marine yeast Pichia guilliermondii and inulin hydrolysis by the crude inulinase. J Ind Microbiol Biotechnol 34:179–185
Imandi SB, Bandaru VVR, Somalanka SR, Garapati HR (2007) Optimization of medium constituents for the production of citric acid from byproduct glycerol using Doehlert experimental design. Enzyme Microb Technol 40:1367–13
Jolivalt C, Madzak C, Brault A, Caminade E, Malosse C, Mougin C (2005) Expression of laccase IIIb from the white-rot fungus Trametes versicolor in the yeast Yarrowia lipolytica for environmental applications. Appl Microbiol Biotechnol 66:450–456
Kamzolova SV, Finogenova TV, Morgunov IG (2008) Microbiological production of citric and isocitric acids from sunflower oil. Food Technol Biotechnol 46:51–59
Lazar Z, Walczak E, Robak M (2011) Simultaneous production of citric acid and invertase by Yarrowia lipolytica SUC + transformants. Biores Technol 102:6982–6989
Lipmann F, Tuttle LC (1945) A specific micromethod for the determination of acyl phosphates. J Biol Chem 159:21–28
Liu XY, Chi Z, Liu GL, Wang F, Madzak C, Chi ZM (2010) Inulin hydrolysis and citric acid production from inulin using the surface-engineered Yarrowia lipolytica displaying inulinase. Metab Eng 12:469–476
Liu GL, Wang DS, Wang LF, Zhao SF, Chi ZM (2011) Mig1 is involved in mycelial formation and expression of the genes encoding extracellular enzymes in Saccharomycopsis fibuligera A11. Fung Genet Biol 48:904–913
Lotfy WA, Ghanem KM, El-Helow ER (2007) Citric acid production by a novel Aspergillus niger isolate: I. Mutagenesis and cost reduction studies. Biores Technol 98:3464–3469
Papagianni M (2007) Advances in citric acid fermentation by Aspergillus niger: biochemical aspects, membrane transport and modeling. Biotechnol Adv 25:244–263
Papanikolaou S, Fakas S, Fick M, Chevalot I, Galiotou-Panayotou M, Komaitis M, Marc I, Aggelis G (2008a) Biotechnological valorisation of raw glycerol discharged after bio-diesel (fatty acid methyl esters) manufacturing process: production of 1,3-propanediol, citric acid and single cell oil. Biom Bioenerg 32:60–71
Papanikolaou S, Galiotou-Panayotou M, Fakas S, Komaitis M, Aggelis G (2008b) Citric acid production by Yarrowia lipolytica cultivated on olive-mill wastewater-based media. Biores Technol 99:2419–2428
Rywinska A, Juszczyk P, Wojtatowicz M, Rymowicz W (2011) Chemostat study of citric acid production from glycerol by Yarrowia lipolytica. J Biotechnol 152:54–57
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor Laboratory Press, Beijing, pp. 367–370 (Chinese translating ed.)
Slightom JL, Metzger BT, Luu HT, Elhammer AP (2009) Cloning and molecular characterization of the gene encoding the Aureobasidin A biosynthesis complex in Aureobasidium pullulans BP-1938. Gene 431:67–79
Spiro RG (1966) Analysis of sugars found in glycoproteins. Meth Enzymol 8:3–26
Venter T, Kock JIF, Botes PJ, Smit MS, Hugo A, Joseph M (2004) Acetate enhances citric acid production by Yarrowia lipolytica when grown on sunflower oil. Syste Appl Microbiol 27:135–138
Wang F, Yue LX, Wang L, Madzak C, Li J, Wang XH, Chi ZM (2009) Genetic modification of the marine-derived yeast Yarrowia lipolytica with high-protein content using a GPI-anchor-fusion expression system. Biotechnol Prog 25:1297–1303
Watanabe T, Suzuki A, Nakagawa H, Kirimura K, Usami S (1998) Citric acid production from cellulose hydrolysate by a 2-deoxy-D-glucose-resistant mutant strain of Aspergillus niger. Biores Technol 66:271–274
Xuan JM, Fournier P, Gaillardin C (1988) Cloning of the LYS5 gene encoding saccharopine dehydrogenase. Curr Genet 14:15–21
Yue LX, Chi ZM, Wang L, Liu J, Madzak C, Li J (2008) Construction of a new plasmid for surface display on cells of Yarrowia lipolytica. J Microbiol Meth 72:116–123
Zhang BZ (1988) Determination of isocitrate lyase activity in seeds. Plant Physiol Comm 3:68–70
Zhao CH, Cui W, Liu XY, Chi ZM, Madzak C (2010) Expression of inulinase gene in the oleaginous yeast Yarrowia lipolytica and single cell oil production from inulin-containing materials. Metab Eng 12:510–551
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This research was supported by Grant 31070029 from the National Natural Science Foundation of China and Hi-Tech Research and Development Program of China (863) (grant no. 2012AA021205).
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Liu, XY., Chi, Z., Liu, GL. et al. Both Decrease in ACL1 Gene Expression and Increase in ICL1 Gene Expression in Marine-Derived Yeast Yarrowia lipolytica Expressing INU1 Gene Enhance Citric Acid Production from Inulin. Mar Biotechnol 15, 26–36 (2013). https://doi.org/10.1007/s10126-012-9452-5
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DOI: https://doi.org/10.1007/s10126-012-9452-5