Abstract
Kluyveromyces marxianus yeasts represent a valuable industry alternative due to their biotechnological potential to produce aromatic compounds. 2-phenylethanol and 2-phenylethylacetate are significant aromatic compounds widely used in food and cosmetics due to their pleasant odor. Natural obtention of these compounds increases their value, and because of this, bioprocesses such as de novo synthesis has become of great significance. However, the relationship between aromatic compound production and yeast’s genetic diversity has yet to be studied. In the present study, the analysis of the genetic diversity in K. marxianus isolated from the natural fermentation of Agave duranguensis for Mezcal elaboration is presented. The results of strains in a haploid and diploid state added to the direct relationship between the mating type locus MAT with metabolic characteristics are studied. Growth rate, assimilate carbohydrates (glucose, lactose, and chicory inulin), and the production of aromatic compounds such as ethyl acetate, isoamyl acetate, isoamyl alcohol, 2-phenylethyl butyrate and phenylethyl propionate and the diversity in terms of the output of 2-phenylethanol and 2-phenylethylacetate by de novo synthesis were determinate, obtaining maximum concentrations of 51.30 and 60.39 mg/L by ITD0049 and ITD 0136 yeasts respectively.
Graphical abstract
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
Adame-Soto PJ, Aréchiga-Carvajal ET, López MG, González-Herrera SM, Moreno-Jiménez MR, Urtiz-Estrada N, Rutiaga-Quiñones OM (2019) Potential production of 2-phenylethanol and 2-phenylethylacetate by non-saccharomyces yeasts from Agave duranguensis. Annal Microbiol 69(9):989–1000. https://doi.org/10.1007/s13213-019-01489-0
Alonso-Vargas M, Téllez-Jurado A, Gómez-Aldapa CA, Ramírez-Vargas MDR, Conde-Báez L, Castro-Rosas J, Cadena-Ramírez A (2022) Optimization of 2-phenylethanol production from sweet whey fermentation using kluyveromyces marxianus. Fermentation 8(2):39. https://doi.org/10.3390/fermentacion8020039
Arrizon J, Morel S, Gschaedler A, Monsan P (2012) Fructanase and fructosyltransferase activity of non-saccharomyces yeasts isolated from fermenting musts of mezcal. Bioresour Technol 110:560–565. https://doi.org/10.1016/j.biortech.2012.01.112
Bi E, Park HO (2012) Cell polarization and cytokinesis in budding yeast. Genetics 191(2):347–387. https://doi.org/10.1534/genetics.111.132886
Beckner Whitener ME, Carlin S, Jacobson D et al (2015) Early fermentation volatile metabolite profile of non-saccharomyces yeasts in red and white grape must: a targeted approach. LWT - Food Sci Technol 64:412–422. https://doi.org/10.1016/j.lwt.2015.05.018
Calvo-Gómez O, Morales-López J, López MG (2004) Solid-phase microextraction-gas chromatographic-mass spectrometric analysis of garlic oil obtained by hydrodistillation. J Chromatogr A 1036:91–93. https://doi.org/10.1016/j.chroma.2004.02.0724
Cayré M, Vignolo G, Garro A (2007) Selección de un modelo primario para describir la curva de crecimiento de bacterias lácticas y Brochothrix thermosphacta sobre emulsiones cárnicas cocidas. Información Tecnológica 18(3):23–29. https://doi.org/10.4067/S0718-07642007000300004
Celińska E, Kubiak P, Białas W et al (2013) Yarrowia lipolytica: the novel and promising 2-phenylethanol producer. J Ind Microbiol Biotechnol 40:389–392. https://doi.org/10.1007/s10295-013-1240-3
Cernak P, Estrela R, Poddar S, Skerker JM, Cheng YF, Carlson AK, Chen B, Cate JH (2018) Engineering Kluyveromyces marxianus as a robust synthetic biology platform host. MBio 9(5):e01410–e01418. https://doi.org/10.1128/mBio.01410-18
Chambi-Rodriguez AD, Torres-Jimenez AM (2021) Kinetic models applied to the growth of saccharomyces boulardii. Rev investig Altoandin 21:127–141. https://doi.org/10.18271/ria.2021.213
Chi ZM, Zhang T, Cao TS, Liu XY, Cui W, Zhao CH (2011) Biotechnological potential of inulin for bioprocesses. Bioresour Technol 102(6):4295–4303. https://doi.org/10.1016/j.biortech.2010.12.086
Conde-Báez L, Castro-Rosas J, Villagómez-Ibarra JR, Páez-Lerma JB, Gómez-Aldapa C (2017) Evaluation of waste of the cheese industry for the production of aroma of roses (phenylethyl alcohol). Waste Biomass Valoriz 8:1343–1350. https://doi.org/10.1007/s12649-016-9654-6
Cordente AG, Curtin CD, Varela C, Pretorius IS (2012) Flavour-active wine yeasts. Appl Microbiol Biotechnol 96:601–618. https://doi.org/10.1007/s00253-012-4370-z
Cubillos FA, Billi E, Zorgo E, Parts L, Fargier P, Omholt S, Blomberg A, Warringer J, Louis EJ, Liti G (2011) Assessing the complex architecture of polygenic traits in diverged yeast populations. Mol Ecol 20(7):1401–1413. https://doi.org/10.1111/j.1365-294X.2011.05005.x
de Lima LA, Ventorim RZ, Bianchini IA, de Queiroz MV, Fietto LG, da Silveira WB (2021) Obtainment, selection and characterization of a mutant strain of Kluyveromyces marxianus that displays improved production of 2-phenylethanol and enhanced DAHP synthase activity. J Appl Microbiol 130(3):878–890. https://doi.org/10.1111/jam.14793
Eshkol N, Sendovski M, Bahalul M et al (2009) Production of 2-phenylethanol from L-phenylalanine by a stress tolerant Saccharomyces cerevisiae strain. J Appl Microbiol 106:534–542. https://doi.org/10.1111/j.1365-2672.2008.04023.x
Etschmann MMW, Bluemke W, Sell D, Schrader J (2002) Biotechnological production of 2-phenylethanol. Appl Microbiol Biotechnol 59:1–8. https://doi.org/10.1007/s00253-002-0992-x
Etschmann MMW, Sell D, Schrader J (2003) Screening of yeasts for the production of the aroma compound 2-phenylethanol in a molasses-based medium. Biotechnol Lett 25:531–536. https://doi.org/10.1023/A:1022890119847
Etschmann MMW, Sell D, Schrader J (2004) Medium optimization for the production of the aroma compound 2-phenylethanol using a genetic algorithm. J Mol Cat B: Enzymatic 29:187–193. https://doi.org/10.1016/j.molcatb.2003.10.014
Fasoli G, Tofalo R, Lanciotti R, Schirone M, Patrignani F, Perpetuini G, Grazia L, Corsetti A, Suzzi G (2015) Chromosome arrangement, differentiation of growth kinetics and volatile molecule profiles in Kluyveromyces marxianus strains from italian cheeses. Int J Food Microbiol 214:151–158. https://doi.org/10.1016/j.ijfoodmicro.2015.08.001
Fasoli G, Barrio E, Tofalo R, Suzzi G, Belloch C (2016) Multilocus analysis reveals large genetic diversity in Kluyveromyces marxianus strains isolated from Parmigiano Reggiano and Pecorino di Farindola cheeses. J Food Microbiol 233:1–10. https://doi.org/10.1016/j.ijfoodmicro.2016.05.028. Int
Fonseca GG, Heinzle E, Wittmann C, Gombert AK (2008) The yeast Kluyveromyces marxianus and its biotechnological potential. Appl Microbiol Biotechnol APPL- 79(3):339–354. https://doi.org/10.1007/s00253-008-1458-6
Fu X, Li P, Zhang L, Li S (2019) Understanding the stress responses of Kluyveromyces marxianus after an arrest during high-temperature ethanol fermentation based on integration of RNA-Seq and metabolite data. Appl Microbiol Biotechnol 103:2715–2729. https://doi.org/10.1007/s00253-019-09637-x
Goddard MR, Greig D (2015) Saccharomyces cerevisiae: a nomadic yeast with no niche? FEMS Yeast Res https://doi.org/10.1093/femsyr/fov009
Gao J, Yuan W, Li Y, Xiang R, Hou S, Zhong S, Bai F (2015) Transcriptional analysis of Kluyveromyces marxianus for ethanol production from inulin using consolidated bioprocessing technology. Biotechnol Biofuels. https://doi.org/10.1186/s13068-015-0295-y
Garcia BE, Rodriguez E, Salazar Y, Valle PA, Flores-Gallegos AC, Rutiaga-Quiñones OM, Rodriguez-Herrera R (2021) Primary model for biomass growth prediction in batch fermentation. Symmetry 13(08):1468. https://doi.org/10.3390/sym13081468
Gethins L, Guneser O, Demirkol A, Rea MC, Stanton C, Ross RP, Yuceer Y, Morrissey JP (2015) Influence of carbon and nitrogen source on production of volatile fragrance and flavour metabolites by the yeast Kluyveromyces marxianus. Yeast 32:67–76. https://doi.org/10.1002/yea.3047
Gonzalez-Hernandez JC, Esparza MJH, Madrigal-Perez LA, Valencia RT, Ramírez JIA (2018) Comparative analysis of the kinetic growth parameters and ethanol production in non-saccharomyces yeasts at the bioreactor level using Agave cupreata juice. J Biochem Technol 7(3):1132–1138
Groeneveld P, Stouthamer AH, Westerhoff HV (2009) Super life–how and why ‘cell selection’ leads to the fastest-growing eukaryote. FEBS J 276:254–270. https://doi.org/10.1111/j.1742-4658.2008.06778.x
Hall TA (1999) BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41:95–98
Hanson SJ, Wolfe KH (2017) An evolutionary perspective on yeast mating-type switching. Genetics 206(1):9–32. https://doi.org/10.1534/genetics.117.202036
Ha-Tran DM, Nguyen TTM, Huang CC (2020) Kluyveromyces marxianus: current state of omics studies, strain improvement strategy and potential industrial implementation. Fermentation 6(4):124. https://doi.org/10.3390/fermentation6040124
Hazelwood LH, Daran J-MG, van Maris AJA et al (2008) The Ehrlich pathway for fusel alcohol production: a century of research on Saccharomyces cerevisiae metabolism. Appl Environ Microbiol 74:2259–2266. https://doi.org/10.1128/AEM.02625-07
Holyavka M, Artyukhov V, Kovaleva T (2016) Structural and functional properties of inulinases: a review. Biocatal Biotransform 34:1–17. https://doi.org/10.1080/10242422.2016.1196486
Inokuma K, Ishii J, Hara KY, Mochizuki M, Hasunuma T, Kondo A (2015) Complete genome sequence of Kluyveromyces marxianus NBRC1777, a nonconventional thermotolerant yeast. Genome Announc 3:e00389–e00315. https://doi.org/10.1128/genomeA.00389-15
Karim A, Gerliani N, Aïder M (2020) Kluyveromyces marxianus: an emerging yeast cell factory for applications in food and biotechnology. Int J Food Microbiol y 333:108818. https://doi.org/10.1016/j.ijfoodmicro.2020.108818
Lane MM, Morrissey JP (2010) Kluyveromyces marxianus: a yeast emerging from its sister’s shadow. Fungal Biol Rev 24:17–26. https://doi.org/10.1016/j.fbr.2010.01.001
Lane MM, Burke N, Karreman R, Wolfe KH, O’Byrne CP, Morrissey JP (2011) Physiological and metabolic diversity in the yeast Kluyveromyces marxianus. Antonie Van Leeuwenhoek 100:507–519. https://doi.org/10.1007/s10482-011-9606-x
Lertwattanasakul N, Kosaka T, Hosoyama A, Suzuki Y, Rodrussamee N, Matsutani M, Murata M, Fujimoto N, Suprayogi, Tsuchikane K, Limtong S, Fujita N, Yamada M (2015) Genetic basis of the highly efficient yeast Kluyveromyces marxianus: complete genome sequence and transcriptome analyses. Biotechnol Biofuels 8:47. https://doi.org/10.1186/s13068-015-0227-x
Liu GL, Chi Z, Chi ZM (2013) Molecular characterization and expression of microbial inulinase genes. Crit Rev Microbiol 39:152–165. https://doi.org/10.3109/1040841X.2012.694411
Löbs AK, Schwartz C, Wheeldon I (2017) Genome and metabolic engineering in non-conventional yeasts: current advances and applications. Synth Syst Biotechnol 2:198–207. https://doi.org/10.1016/j.synbio.2017.08.002
Loser C, Urit T, Bley T (2014) Perspectives for the biotechnological production of ethyl acetate by yeasts. Appl Microbiol Biotechnol 98:5397–5415. https://doi.org/10.1007/s00253-014-5765-9
Lu X, Wang Y, Zong H, Ji H, Zhuge B, Dong Z (2016) Bioconversion of L-phenylalanine to 2-phenylethanol by the novel stress-tolerant yeast Candida glycerinogenes WL2002–5. Bioeng 5979:1–6. https://doi.org/10.1080/21655979.2016.1171437
Miller G (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analitycal Chem 31(3):426–428. https://doi.org/10.1021/ac60147a030
Martell Nevárez MA, Córdova Gurrola EE, López Miranda J et al (2011) Effect of fermentation temperature on chemical composition of mescals made from Agave duranguensis juice with different native yeast genera. Afr J Microbiol Res 5:3669–3676. https://doi.org/10.5897/AJMR11.467
Martínez O, Sánchez A, Font X, Barrena R (2018) Bioproduction of 2-phenylethanol and 2-phenethyl acetate by Kluyveromyces marxianus through the solid-state fermentation of sugarcane bagasse. Microbiol Biotechnol Appl. https://doi.org/10.1007/s00253-018-8964-y
Martins DB, De Souza CG Jr, Simoes DA, De Morais MA Jr (2002) The beta-galactosidase activity in Kluyveromyces marxianus CBS 6556 decreases by high concentrations of galactose. Curr Microbiol 44:379–382. https://doi.org/10.1007/s00284-001-0052-2
Montgomery DC, Runger GC (2013) Applied Statistics and Probability for Engineers, 6th edn. Wiley, Hoboken, p 920
Morrissey JP, Etschmann MMW, Schrader J, De Billerbeck GM (2015) Cell factory applications of the yeast Kluyveromyces marxianus for the biotechnological production of natural flavour and fragrance molecules. Yeast 32:3–16. https://doi.org/10.1002/yea.3054
Nurcholis M, Nitiyon S, Rodrussamee N, Lertwattanasakul N, Limtong S, Kosaka T, Yamada M (2019) Functional analysis of Mig1 and Rag5 as expressional regulators in thermotolerant yeast Kluyveromyces marxianus. Appl Microbiol Biotechnol 103:395–410. https://doi.org/10.1007/s00253-018-9462-y
Ortiz-Merino RA, Varela JA, Coughlan AY, Hoshida H, da Silveira WB, Wilde C, Kuijpers NGA, Geertman JM, Wolfe KH, Morrissey JP (2018) Ploidy variation in Kluyveromyces marxianus separates dairy and non-dairy isolates. Front Genet 9:0–16. https://doi.org/10.3389/fgene.2018.00094
Pfeiffer T, Morley A (2014) An evolutionary perspective on the Crabtree effect. Front Mol Biosci 1:17. https://doi.org/10.3389/fmolb.2014.00017
Padilla B, García-Fernández D, González B et al (2016) Yeast biodiversity from DOQ priorat uninoculated fermentations. Front Microbiol 7:930. https://doi.org/10.3389/fmicb.2016.00930
Pires EJ, Teixeira JA, Brányik T, Vicente AA (2014) Yeast: the soul of beer’s aroma—a review of flavour-active esters and higher alcohols produced by the brewing yeast. Appl Microbiol Biotechnol 98:1937–1949. https://doi.org/10.1007/s00253-013-5470-0
Rubio-Texeira M (2006) Endless versatility in the biotechnological applications of Kluyveromyces LAC genes. Bio technol Adv 24(2):212–222. https://doi.org/10.1016/j.biotechadv.2005.10.001
Rojas V, Gil JV, Piaga F, Manzanares P (2001) Studies on acetate ester production by non-saccharomyces wine yeasts. Int J Food Microbiol 70:283–289. https://doi.org/10.1016/S0168-1605(01)00552-9
Rotureau B, Subota I, Buisson J, Bastin P (2012) A new asymmetric division contributes to the continuous production of infective trypanosomes in the tsetse fly. Development 139(10):1842–1850. https://doi.org/10.1242/dev.072611
Rouwenhorst RJ, Visser LE, van der Baan AA, Scheffers WA, van Dijken JP (1988) Production, distribution, and kinetic properties of inulinase in continuous culture of Kluyveromyces marxianus CBS 6556. Appl Environm Microbiol 54:1131–1137. https://doi.org/10.1128/aem.54.5.1131-1137.1988
Sambrook J, Russell RW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold spring harbor, New York
Sunchu B, Cabernard C (2020) Principles and mechanisms of asymmetric cell division. Development 147(13):dev167650. https://doi.org/10.1242/dev.167650
Salvado Z, Arroyo-Lopez FN, Guillamon JM, Salazar G, Querol A, Barrio E (2011) Temperature adaptation markedly determines evolution within the Saccharomyces genus. Appl Environ Microbiol 77(7):2292–2302. https://doi.org/10.1128/AEM.01861-10
Sandoval-Nuñez D, Romero-Gutiérrez T, Gómez-Márquez C, Gshaedler A, Arellano-Plaza M, Amaya-Delgado L (2023) Physiological and transcriptome analyses of Kluyveromyces marxianus reveal adaptive traits in stress response. Appl Microbiol Biotechnol. https://doi.org/10.1007/s00253-022-12354-7
Schabort DTWP, Letebele PK, Steyn L, Kilian SG, du Preez JC (2016) Differential RNA-seq, multi-network analysis and metabolic regulation analysis of Kluyveromyces marxianus reveals a compartmentalized response to xylose. PLoS ONE 11:e0156242. https://doi.org/10.1371/journal.pone.0156242
Styger G, Prior B, Bauer FF (2011) Wine flavor and aroma. Microbiol Biotechnol 38:1145–1159. https://doi.org/10.1007/s10295-011-1018-4
Tong Z (2012) Spot assay for yeast. Bio-Protocol 2:e16. https://doi.org/10.21769/BioProtoc.16
Tjørve KM, Tjørve E (2017) The use of Gompertz models in growth analyses, and new Gompertz-model approach: an addition to the Unified-Richards family. PLoS ONE 12(6):e0178691. https://doi.org/10.1371/journal.pone.0178691
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30(12):2725–2729
Tittarelli F, Varela JA, Gethins L, Stanton C, Ross RP, Suzzi G, Grazia L, Tofalo R, Morrissey JP (2018) Development and implementation of multilocus sequence typing to study the diversity of the yeast Kluyveromyces marxianus in italian cheeses. Microb genomics. https://doi.org/10.1099/mgen.0.000153
Tofalo R, Fasoli G, Schirone M, Perpetuini G, Pepe A, Corsetti A, Suzzi G (2014) The predominance, biodiversity and biotechnological properties of Kluyveromyces marxianus in the production of Pecorino di farindola cheese. Int J Food Microbiol 187:41–49. https://doi.org/10.1016/j.ijfoodmicro.2014.06.029
Varela JA, Montini N, Scully D, Van der Ploeg R, Oreb M, Boles E, Hirota J, Akada R, Hoshida H, Morrisey J (2017) Polymorphisms in the LAC12 gene explain lactose utilisation variability in Kluyveromyces marxianus strains. FEMS Yeast Res 17:fox021. https://doi.org/10.1093/femsyr/fox021
Vigneault F, Lachance D, Cloutier M, Pelletier G, Levasseur C, Séguin A (2007) Members of the plant NIMA-related kinases are involved in organ development and vascularization in poplar, Arabidopsis and rice. Plant J 51:575–588. https://doi.org/10.1111/j.1365-313X.2007.03161.x
Wittmann C, Hans M, Bluemke W (2002) Metabolic physiology of aroma-producing Kluyveromyces marxianus. Yeast 19:1351–1363. https://doi.org/10.1002/yea.920
Yarimizu T, Nonklang S, Nakamura J, Tokuda S, Nakagawa T, Lorreungsil S, Sutthikhumpha S, Pukahuta C, Kitagawa T, Nakamura M, Cha-Aim K, Limtong S, Hoshida H, Akada R (2013) Identification of auxotrophic mutants of the yeast Kluyveromyces marxianus by non-homologous end joining-mediated integrative transformation with genes from Saccharomyces cerevisiae. Yeast 30:485–500. https://doi.org/10.1002/yea.2985
Zhang H, Cao M, Jiang X, Zou H, Wang C, Xu X, Xian M (2014) De novo synthesis of 2-phenylethanol by Enterobacter sp. CGMCC 5087 BMC Biotechnol 14:30. https://doi.org/10.1186/1472-6750-14-30
Zhang G, Lu M, Wang J, Wang D, Gao X, Hong J (2017a) Identification of hexose kinase genes in Kluyveromyces marxianus and thermo-tolerant one step producing glucose-free fructose strain construction. Sci Rep 7:45104. http://doi.org/10.1038/srep45104
Zhang L, Liu Q, Pan H et al (2017b) Metabolic engineering of Escherichia coli to high efficient synthesis phenylacetic acid from phenylalanine. AMB Expr 7:105. https://doi.org/10.1186/s13568-017-0407-0
Zhang B, Ren L, Zeng S, Zhang S, Xu D, Zeng X, Li F (2020) Functional analysis of PGI1 and ZWF1 in thermotolerant yeast Kluyveromyces marxianus. Appl Microbiol Biotechnol 104:7991–8006. https://doi.org/10.1007/s00253-020-10808-4
Zhou J, Zhu P, Hu X, Lu H, Yu Y (2018) Improved secretory expression of lignocellulolytic enzymes in Kluyveromyces marxianus by promoter and signal sequence engineering. Biotechnol Biofuels 11:235. https://doi.org/10.1186/s13068-018-1232-7
Funding
This work was supported Proyect: 317235 the Consejo Nacional de Ciencia y Tecnología (CONACyT), Fortalecimiento de infraestructura-CONACyT 2021, grant awarded to Pablo Jaciel Adame-Soto 435680.
Author information
Authors and Affiliations
Contributions
All authors contributed to the study conception and design. PA: investigation, methodology, validation, writing—original draft, writing—review and editing. EA: conceptualization, resources, supervision-formal analysis,-validation, writing—review and editing. SG: supervision, formal analysis, writing—review and editing. MM: methodology, validation, writing—review and editing. OR: conceptualization, formal analysis, resources-supervision, validation, writing—original-draft, writing—review and editing.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Ethical approval
This study does not contain any studies with human participants or animals performed by any of the authors.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
11274_2023_3659_MOESM1_ESM.xlsx
Supplementary material 1: Characterization of mating type on aroma production and metabolic properties wild Kluyveromyces marxianus yeasts (XLSX 15.0 kb)
11274_2023_3659_MOESM2_ESM.jpg
Supplementary material 2: Characterization of mating type on aroma production and metabolic properties wild Kluyveromyces marxianus yeasts (JPG 2259.0 kb)
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Adame-Soto, P.J., Aréchiga-Carvajal, E.T., González-Herrera, S.M. et al. Characterization of mating type on aroma production and metabolic properties wild Kluyveromyces marxianus yeasts. World J Microbiol Biotechnol 39, 216 (2023). https://doi.org/10.1007/s11274-023-03659-4
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11274-023-03659-4