Abstract
Oleaginous yeast Trichosporon cutaneum CGMCC 2.1374 was found to utilize inulin directly for microbial lipid fermentation without a hydrolysis step. The potential inulinase-like enzyme(s) in T. cutaneum CGMCC 2.1374 were characterized and compared with other inulinase enzymes produced by varied yeast strains. The consolidated bioprocessing (CBP) for lipid accumulated using inulin was optimized with 4.79 g/L of lipid produced from 50 g/L inulin with the lipid content of 33.6 % in dry cells. The molecular weight of the enzyme was measured which was close to invertase in Saccharomyces cerevisiae. The study provided information for inulin hydrolyzing enzyme(s) in oleaginous yeasts, as well as a preliminary CBP process for lipid production from inulin feedstock.
Similar content being viewed by others
References
Zhao, C. H., Cui, W., Liu, X. L., Chi, Z. M., & Madzak, C. (2010). Expression of inulinase gene in the oleaginous yeast Yarrowia lipolytica and single cell oil production from inulin-containing materials. Metabolic Engineering., 6(12), 510–517.
Meng, X., Yang, J. M., Xu, X., Zhang, L., Nie, Q. J., Xian, M. (2009). Biodiesel production from oleaginous microorganisms. Renewable Energy 1(34): 1–5.
Zhao, C. H., Chi, Z., Zhang, F., Guo, F. J., Li, M., Song, W. B., & Chi, Z. M. (2011). Direct conversion of inulin and extract of tubers of Jerusalem artichoke into single cell oil by co-cultures of Rhodotorula mucilaginosa TJY15a and immobilized inulinase-producing yeast cells. Bioresource Technology., 10(102), 6128–6133.
Donot, F., Fontana, A., Baccou, J. C., Strub, C., & Schorr-Galindo, S. (2014). Single cell oils (SCOs) from oleaginous yeasts and moulds: production and genetics. Biomass and Bioenergy, 68, 135–150.
Lynd L. R, Laser M. S, Brandsby D., Dale B. E., Davison B. (2008). How biotech can transform biofuels. Nature Biotechnology. 2(26): 169–172.
Chi, Z. M., Chi, Z., Zhang, T., Liu, G. L., & Yue, L. X. (2009). Inulinase-expressing microorganisms and applications of inulinases. Applied Microbiology Biotechnology., 2(82), 211–220.
Marzena, J. K., Karolina, L. T., & Stanislaw, B. (2003). Identification of the gene for β-fructofuranosidase of Bifidobacterium lactis DSM10140T and characterization of the enzyme expressed in Escherichia coli. Current Microbiology., 6(46), 391–397.
Yuna, B., Wang, S. A., & Li, F. L. (2013). Improved ethanol fermentation by heterologous endoinulinase and inherent invertase from inulin by Saccharomyces cerevisiae. Bioresource Technology., 139, 402–405.
An, K. H., Hu, F. X., & Bao, J. (2013). Simultaneous saccharification of inulin and starch using commercial glucoamylase and the subsequent bioconversion to high titer sorbitol and gluconic acid. Applied Microbiology Biotechnology., 8(171), 2093–2104.
Schorr-Galindo, S., Ghommidh, C., & Guiraud, J. P. (2000). Influence of yeast flocculation on the rate of Jerusalem artichoke extract fermentation. Current Microbiology, 2(41), 89–95.
Dao, T. H., Zhang, J., & Bao, J. (2013). Characterization of inulin hydrolyzing enzyme(s) in commercial glucoamylases and its application in lactic acid production from Jerusalem artichoke tubers (Jat). Bioresource Technology., 148, 157–162.
Zhao, C. H., Zhang, T., Li, M., & Chi, Z. M. (2010). Single cell oil production from hydrolysates of inulin and extract of tubers of Jerusalem artichoke by Rhodotorula mucilaginosa TJY15a. Process Biochemistry., 7(45), 1121–1126.
Lane, M. M., & Morrissey, J. P. (2010). Kluyveromyces marxianus: a yeast emerging from its sister’s shadow. Fungal Biology reviews, 1-2(24), 17–26.
Wang, Y. M., Liu, W., & Bao, J. (2012). Repeated batch fermentation with water recycling and cell separation for microbial lipid production. Frontiers of Chemical Science and Engineering., 4(6), 453–460.
Wang, S. A., & Li, F. L. (2013). Invertase SUC2 is the key hydrolase for inulin degradation in Saccharomyces cerevisiae. Applied Environment Microbiology., 1(79), 403–406.
Wang, Z. P., Fu, W. J., Xu, H. M., & Chi, Z. M. (2014). Direct conversion of inulin into cell lipid by an inulinase-producing yeast Rhodosporidium toruloides 2F5. Bioresource Technology., 161, 131–136.
Chu, D. Q., Zhang, J., & Bao, J. (2012). Simultaneous saccharification and ethanol fermentation of corn Stover at high temperature and high solids loading by a thermotolerant strain Saccharomyces cerevisiae DQ1. Bioenerg. Res., 4(5), 1020–1026.
Folch, J., Lees, M., & Sloane-Stanley, G. H. (1957). A simple method for the isolation and purification of total lipids from animal tissues. Journal of biological Chemistry., 226, 497–509.
Gao, Q. Q., Cui, Z. Y., Zhang, J., & Bao, J. (2014). Lipid fermentation of corncob residues hydrolysate by oleaginous yeast Trichosporon cutaneum. Bioresource Technology., 152, 552–556.
Wang, J., Jin, Z. Y., Bo, J., & Adamu, A. (2003). Production and separation of exo- and endoinulinase from Aspergillus ficuum. Process Biochemistry, 1(39), 5–11.
Westphal, V., Marcusson, E. G., Winther, J. R., Emr, S. D., & van den Hazel, H. B. (1996). Multiple pathways for vacuolar sorting of yeast proteinase A. Journal of Biological Chemistry., 271, 11865–11870.
Guo, L. H., Zhang, J., Hu, F. X., & Bao, J. (2013). Consolidated bioprocessing of highly concentrated Jerusalem artichoke tubers for simultaneous saccharification and ethanol fermentation. Biotechnology Bioengineering., 10(110), 2606–2615.
Kim, S., Park, J. M., & Kim, C. H. (2013). Ethanol production using whole plant biomass of Jerusalem artichoke by Kluyveromyces marxianus CBS1555. Applied Biochemistry and Biotechnology., 5(169), 1531–1545.
Olmea, O., Chinea, G., Beldarrain, A., Marquez, G., Acosta, N., Rodriguez, L., & Valencia, A. (1998). Structural model for family 32 of glycosyl-hydrolase enzymes. Proteins Structure Function & Bioinformatics., 3(33), 383–395.
Zhang, L. L., Tang, M. J., Liu, G. L., Wang, G. Y., & Chi, Z. M. (2015). Cloning and characterization of an inulinase gene from the marine yeast Candida membranifaciens subsp. flavinogenie W14-3 and its expression in Saccharomyces sp. W0 for ethanol production. Molecular Biotechnology, 4(57), 337–347.
Goldman, D., Lavid, N., Schwartz, A., Shoham, G., Danino, D., & Shoham, Y. (2008). Two active forms of Zymomonas mobilis levansucrase: an ordered microfibril structure of the enzyme promotes levan polymerization. Journal of Biological Chemistry., 47(283), 32209–32217.
Chaudhary, A., Gupta, L. K., Gupata, J. K., & Banerjee, U. C. (1996). Purification and properties of levanase from Rhodotorula sp. Journal of Biotechnology., 1(46), 55–61.
Ali, S., & Haq, I. (2007). Kinetics of improved extracellular beta-D-fructofuranosidase fructohydrolase production by a derepressed Saccharomyces cerevisiae. Letters in Applied Microbiology., 2(45), 160–167.
Kushi, R. T., Monti, R., & Cotiero, J. (2000). Production, purification and characterization of an extracellular inulinase from Kluyveromyces marxianus var. bulgaricus. Journal of Industrial Microbiology & Biotechnology, 2(25), 63–69.
Laloux, O., Cassart, J. P., Delcour, J., Van, B. J., & Vandenhaute, J. (1991). Cloning and sequencing of the inulinase gene of Kluyveromyces marxianus var. marxianus ATCC 12424. FEBS Letters, 1(289), 64–68.
Yu, X. J., Guo, N., Chi, Z. M., Gong, F., Sheng, J., & Chi, Z. (2009). Inulinase overproduction by a mutant of the marine yeast Pichia guilliermondii using surface response methodology and inulin hydrolysis. Biochemical Engineering Journal., 3(43), 266–271.
Williams, R. S., Trumbly, R. J., MacColl, R., Trimble, R. B., & Maley, F. (1985). Comparative properties of amplified external and internal invertase from the yeast SUC2 gene. Journal of Biological Chemistry., 24(260), 13334–13341.
Dujon, B., Sherman, D., Fischer, G., et al. (2004). Genome evolution in yeasts. Nature, 6995(430), 35–44.
Agaphonov, M. O., Packeiser, A. N., Chechenova, M. B., Choi, E. S., & Ter-Avanesyan, M. D. (2001). Mutation of the homologue of GDP-mannose pyrophosphorylase alters cell wall structure, protein glycosylation and secretion in Hansenula polymorpha. Yeast, 5(18), 391–402.
Bommareddy, R. R., Sabra, W., Maheshwari, G., & Zeng, A. P. (2015). Metabolic network analysis and experimental study of lipid production in Rhodosporidium toruloides grown on single and mixed substrates. Microbial Cell Factories, 14, 36.
Vajpeyi, S., & Chandran, K. (2015). Microbial conversion of synthetic and food waste-derived volatile fatty acids to lipids. Bioresource Technology., 188, 49–55.
Sestric, R., Munch, G., Cicek, N., Sparling, R., & Levin, D. B. (2014). Growth and neutral lipid synthesis by Yarrowia lipolytica on various carbon substrates under nutrient-sufficient and nutrient-limited conditions. Bioresource Technology., 164, 41–46.
Ratledge C., Wynn J. P. (2002). The biochemistry and molecular biology of lipid accumulation in oleaginous microorganisms. Advances in Applied Microbiology. 51: 1–51.
Acknowledgments
This research was supported by the National Basic Research Program of China (2011CB707406) and the National High-Tech Program of China (2012AA022301, 2014AA021901).
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Wang, J., Zhang, H. & Bao, J. Characterization of Inulin Hydrolyzing Enzyme(s) in Oleaginous Yeast Trichosporon cutaneum in Consolidated Bioprocessing of Microbial Lipid Fermentation. Appl Biochem Biotechnol 177, 1083–1098 (2015). https://doi.org/10.1007/s12010-015-1798-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-015-1798-5