Abstract
Purpose
The level of higher alcohols on top-fermentation determines the flavor profile and is one of the most important elements dictating the favorable top-fermented wheat beer (Ale beer) development. The optimization of the pitching rate has been shown to be crucial for industrial beer brewing. This study focused on understanding the effect of the variable inoculum size on the synthesis of higher alcohols.
Methods
We utilized sequencing to investigate the transcript changes under different inoculum sizes and link the results to fermentation performance.
Results
Variable cell inoculum density levels were linked with differences in higher alcohol production. Specifically, we observed significantly less higher alcohols produced at lower cell inoculum density during the stationary phase. Importantly, the accumulation of higher alcohols during the exponential growth phase was overall similar between different pitching rates. Moreover, free amino nitrogen (FAN) consumption and yeast cell viability were significantly decreased during stationary phase at the lowest inoculum density. Transcriptomic analysis revealed that amino acid metabolism genes ALD4, ALD6, ARO9, ARO10, and PUT1 were differentially expressed once the cells entered the declining growth phase at the lowest inoculum size.
Conclusion
The results suggest that the variable accumulation of higher alcohols in the top-fermenting yeast at different inoculum sizes is mostly accounted for in the stationary phase. We discovered that lower pitching rate was associated with a negative effect on amino acid metabolism and synthesis of higher alcohols during the stationary phase, leading to the decrease in higher alcohol concentration at low inoculum densities. Overall, our study provides valuable insights that could benefit wheat beer production.
Similar content being viewed by others
References
Avbelj M, Zupan J, Kranjc L, Raspor P (2015) Quorum-sensing kinetics in Saccharomyces cerevisiae: a symphonyof ARO genes and aromatic alcohols. J Agric Food Chem 63:8544–8550
Bevers J, Verachtert H (1976) Synthesis of higher alcohols in the genus zymomonas. J Inst Brew 82:35–40
Carrau F, Medina K, Fariña L, Boido E, Dellacassa E (2010) Effect of Saccharomyces cerevisiae inoculum size on wine fermentation aroma compounds and its relation with assimilable nitrogen content. Int J Food Microbiol 143:81–85
Chen EH (1978) Effects on flavour of innovations in brewery equipment and processing: a review. J Inst Brew 107:271–286
Chen H, Fink GR (2006) Feedback control of morphogenesisin fungi by aromatic alcohols. Genes Dev 20:1150–1161
Chen H, Fujita M, Feng Q, Clardy J, Fink GR (2004) Tyrosol is a quorum-sensing molecule in Candida albicans. Proc Natl Acad Sci USA 101:5048–5052
Edelen CL, Miller JL, Patino H (1996) Effects of yeast pitching rates on fermentation performance and beer quality. Tech Q Master Brew Assoc Am 33:30–32
Engan S (1981) In: Pollock JRA (ed) Brewing science, vol. Academic Press, London, p 2
Erten H, Tanguler H, Cakiroz H (2007) The effect of pitchinging rate on fermentation and flavour compounds in high gravity brewing. J Inst Brew 113:75–79
European Brewery Convention (1998) Analytica–EBC. Fachverlag Hans Carl, Nürnberg
Gabriel P, Dienstbier M, Matoulková D, Kosař K, Sigler K (2012) Optimised acidification power test of yeast vitality and its use in brewing practice. J Inst Brew 114:270–276
Hammond JRM (1993) In: Rose AH, Harrison JS (eds) The yeasts, vol 5: yeast technology. Academic Press, London
Hazelwood L, Daran J, Van Maris A et al (2008) The Ehrlich pathway for fusel alcohol production: a century of research on Saccharomyces cerevisiae metabolism. Appl Environ Microbiol 74:2259–2266
Horton CE, Huang KX, Bennett GN, Rudolph FB (2003) Heterologous expression of the Saccharomyces cerevisiae alcohol acetyltransferase genes in Clostridium acetobutylicum and Escherichia coli for the production of isoamyl acetate. J Ind Microbiol Biotechnol 30:427–432
Huang C, Li YY, Liu LP, Wu H et al (2014) Kinetics and mechanism analysis on microbial oil production by Trichosporon fermentans in rice straw hydrolysate. Ind Eng Chem Res 53:19034–19043
Ingraham JL, Guymon JF (1960) The formation of higher alcohols by mutant strains of Saccharomyces cerevisiae. Arch Biochem Biophys 88:157–166
Kara BV, Simpson WJ, Hammond JRM (1988) Prediction of the fermentation performance of brewing yeast with the acidification power test. J Inst Brew 94:153–158
Kim D, Langmead B, Salzberg SL (2015) HISAT: a fast spliced aligner with low memory requirements. Nat Methods 12:357–360
Kobayashi M, Nagahisa K, Shimizu H, Shioya S (2014) Simultaneous control of apparent extract and volatile compounds concentrations in low-malt beer fermentation. Appl Microbiol Biotechnol 73:549–558
Kumar S, Dheeran P, Singh SP, Mishra IM, Adhikari DK (2014) Kinetic studies of ethanol fermentation using Kluyveromyces sp. IIPE453. J Chem Technol Biotechnol 88:1874–1884
Landaud S, Latrille E, Corrieu G (2001) Top pressure and temperature control the fusel alcohol/Ester ratio through yeast growth in beer fermentation. J Inst Brew 107:107–117
Li J, Feng R, Wen Z, Zhang A (2017) Overexpression of ARO10 in pdc5ΔMutant resulted in higher isobutanol titers in Saccharomyces cerevisiae. Biotechnol Bioprocess Eng 22:382–389
Luedeking R, Piret EL (1959) A kinetic study of the lactic acid fermentation. Batch process at controlled pH. J Biochem Microbiol Technol Eng 4:393–412
Ma LJ, Huang SY, Du LP, Tang P, Xiao DG (2017) Reduced production of higher alcohols by Saccharomyces cerevisiae in red wine fermentation by simultaneously overexpressing BAT1 and deleting BAT2. J Agric Food Chem 65:6936–6942
Meilgaard MC (1975) Flavor chemistry of beer: part II: flavor threshold of 239 aroma volatiles. Tech Q Master Brew Assoc Am 12:151–168
Meilgaard MC (2001) Effects on flavour of innovations in brewery equipment and processing: a review. J Inst Brew 107:271–286
Okabe M, Katoh M, Furugoori F, Yoshida M, Mitsui S (1992) Growth and fermentation characteristics of bottom brewer’s yeast under mechanical stirring. J Ferment Bioeng 73:148–152
Opekarová M, Sigler K (1982) Acidification power: Indicator of metabolic activity and autolytic changes in Saccharomyces cerevisiae. Folia Microbiol 27:395–403
Pank SH, Kim S, Hahn JS (2014) Metabolic engineering of Saccharomyces cerevisiae for the production of isobutanol and 3-methyl-1-butanol. Appl Microbiol Biotechnol 98:9139–9147
Pires EJ, Teixeira JA, Branyik 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
Powell CD, Quain DE, Smart KA (2003) Quain DE and Smart KA, The impact of brewing yeast cell age on fermentation performance, attenuation and flocculation. FEMS Yeast Res 3:149–157
Procopio S, Qian F, Becker T (2011) Function and regulation of yeast genes involved in higher alcohol and ester metabolism during beverage fermentation. Eur Food Res Technol 233:721–729
Rotar R, Stoicescu AG (2006) Modified acidification power test applied to evaluate temperature and ethanol stress impact on yeast fermentation performance. J Agroaliment Process Technol 2:481–488
Saerens SM, Verbelen PJ, Vanbeneden N, Thevelein JM, Delvaux FR (2008) Monitoring the influence of high-gravity brewing and fermentation temperature on flavour formation by analysis of gene expression levels in brewing yeast. Appl Microbiol Biotechnol 80:1039–1051
Saintprix F, Bönquist L, Dequin S (2004) Functional analysis of the ALD gene family of Saccharomyces cerevisiae during anaerobic growth on glucose: the NADP+-dependent Ald6p and Ald5p isoforms play a major role in acetate formation. Microbiology 150:2209–2220
Sen R, Swaminathan T (2004) Response surface modeling and optimization to elucidate and analyze the effects of inoculum age and size on surfactin production. Biochem Eng J 21:141–148
Urrestarazu A, Vissers S, Iraqui I, Grenson M (1998) Phenylalanine and tyrosine-auxotrophic mutants of Saccharomyces cerevisiae impaired in transamination. Mol Gen Genet 257:230–237
Vanderhaegen B, Neven H, Coghe S, Verstrepen KJ, Verachtert H, Derdelinckx G (2003) Evolution of chemical and sensory properties during aging of top-fermented beer. J Agric Food Chem 51:6782–6790
Verbelen PJ, Mulders SV, Saison D, Laere SV, Delvaux et al (2008) Characteristics of high cell density fermentations with different lager yeast strains. J Inst Brew 114:127–133
Verbelen PJ, Dekoninck TM, Saerens SM, Van Mulders SE, Thevelein JM et al (2009) Impact of pitchinging rate on yeast fermentation performance and beer flavour. Appl Microbiol Biotechnol 82:155–167
Wang D, Xu Y, Hu J, Zhao G (2015) Fermentation kinetics of different sugars by apple wine yeast Saccharomyces cerevisiae. J Inst Brew 110:340–346
Wuster A, Babu MM (2009) Transcriptional control of the quorum sensing response in yeast. Mol BioSyst 6:134–141
Zhang CY, Liu YL, Qi YN, Zhang JW, Dai LH (2013) Increased esters and decreased higher alcohols production by engineered brewer’s yeast strains. Eur Food Res Technol 236:1009–1014
Funding
This work was supported by the National Natural Science Foundation of China (No. 31771969), the National Key Research and Development Program of China (No. 2016YFD0400505), the China Postdoctoral Science Foundation (2017M611169), the Hebei Province Postdoctoral Research Projects (No. B2018003031) and the Public Service Platform Project for Selection and Fermentation Technology of Industrial Microorganisms (No. 17PTGCCX00190).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Research involving human participants and/or animals
This article 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.
Rights and permissions
About this article
Cite this article
Wang, M., Sun, Z., Wang, Y. et al. The effect of pitching rate on the production of higher alcohols by top-fermenting yeast in wheat beer fermentation. Ann Microbiol 69, 713–726 (2019). https://doi.org/10.1007/s13213-019-01463-w
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13213-019-01463-w