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The science of malting, brewing, and fermenting beer

The science of malting, brewing, and fermenting beer. Oct 6, 2010. Protein : a chain made up of 20 different amino acids from a few to as many as 34,350 residues. α -amylase. DNA. transcription. translation. RNA. Temperature effects on biology and chemistry.

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The science of malting, brewing, and fermenting beer

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  1. The science of malting, brewing, and fermenting beer Oct 6, 2010

  2. Protein: a chain made up of 20 different amino acids from a few to as many as 34,350 residues α-amylase DNA transcription translation RNA

  3. Temperature effects on biology and chemistry

  4. Water is the universal solvent of life The Structure and Properties of Water D. Eisenberg and W. Kauzmann 308 pages Oxford Press 1969 : : + : : : :

  5. Overview of metabolism • Most efficient way of getting energy is by combining reduced carbon with oxygen. The more reduced the carbon, the more energy it has, the more oxygen added, the more energy released. • In the absence of oxygen, fermentation is a suitable alternative. glucose and other sugars fatty acid (-2880 kJ/mol) lactic acid (-198 kJ/mol) ethanol + CO2 (-235 kJ/mol) pyruvate

  6. Overview of metabolism = = = = = β α

  7. Overview of metabolism R = some chemical group

  8. Overview of metabolism

  9. Barley: A member of the grass family. It is a self-pollinating, diploid species with 14 chromosomes. The wild ancestor of domesticated barley, Hordeum vulgare subsp. spontaneum, is abundant in grasslands and woodlands throughout the Fertile Crescent and has been cultivated for millennia. Malting barley is usually lower protein which leads to more uniform germination, with shorter steeping. The lower protein content also reduces the haze that results from precipitated protein. Two-row barley generally has a lower protein content compared to six-row.

  10. -A single mutation is responsible for the difference between two-row and six-row barley. -Two mutations of wild barley prevent the spike from shattering. Traditionally barley was classified by morphological differences and were considered to be different species. -Two-rowed barley with shattering spikes (wild barley) is classified as HordeumspontaneumK.Koch. -Two-rowed barley with non-shattering spikes is classified as H. distichum L. -Six-rowed barley with non-shattering spikes as H. vulgare L. (or H. hexastichum L.). -Six-rowed with shattering spikes as H. agriocrithonÅberg. Recent cytological and molecular evidence has led most recent classifications to consider all forms as a single species, H. vulgare L. Maris Otter is a 2-row, "winter" variety bred by researchers at Cambridge and introduced in 1966 possessing low nitrogen (protein) and superior malting characteristics. It is a cross of Proctor and Pioneer.

  11. Germinating/maltingbreaking down chains of stuff http://plantphys.info/plant_physiology/gibberellin.shtml

  12. Drying and Kilning During drying and or kilning, some enzymes become denatured. Generally darker grains are roasted longer and at higher temperatures and thus have less active enzymes then pale malt. Crystal malt is kilned without drying which denatures all enzymes. Importantly, during the drying phase, most lipase and lipoxygenase enzymes are destroyed. These enzymes are implicated in the formation of off flavors in beer as it ages. Kilning also roasts the grain. During the roasting process a glorious reaction called the Maillard reaction occurs. Modification: the degree of breakdown of the starch-protein matrix during the malting, drying, and kilning process

  13. The Maillard reaction A general reaction between an amino acid and a reducing sugar and contributes to the color and flavor of browned bread, chocolate, seared meat, caramel and deep fried death. sugar amino acid Strecker degradation melanoidins (dark color and toasty aroma) pyrazines thiophenes pyrroles furans isobutyraldehyde (wet cereal/straw)

  14. The modern history of enzymes began in 1833 when French chemists described the isolation of an amylase complex from germinating barley and named it diastase.  Sugar enzymology I http://blog.targethealth.com/?p=9586

  15. Mashing and resting breaking down more chains of stuff Generally only needed when using >25% unmalted wheat or barley, corn, or rye β-1,4 glycosidic linkage cellulose

  16. ~122 °F Protein rest: lower temperatures (122ºF (50ºC) ) yield shorter peptides and single amino acids (free amino nitrogen aka FAN) which aren’t as good for head retention. Temperatures closer to 133ºF (55ºC) leave longer peptide chains and more available amino acids for yeast. . ~133 °F Generally only needed when using minimally modified malt or a lot (>25%) of adjuncts Some recent research suggests that most proteases are destroyed during kilning and that there is no significant reduction in the molecular weight spectrum of the mash.

  17. Sugar enzymology II amylopectin O O O amylose O O O O O O O O O O O O O β O O O O O O O O limit dextrinase (debranching enzyme) O . O O O O O… β O O α O O O O O . . . . O O α

  18. Sugar enzymology II α β

  19. General enzymology Enzyme activity is affected mainly by temperature, but also pH, presence of metals or cofactors, substrate concentration, viscosity, etc The thicker the mash, the more active the enzymes. Maximum rate possible reaction rate ([producet]/second) [Substrate] KM

  20. Humulus lupulus ΔT humulone isohumulone lupulones Lupulin16% Soft Resins 13% Alpha Acids 8%Beta Acids 4%Other Soft Resins 1% Hard Resins 2%Essential Oils 1% Hydrocarbons 0.75%Oxidation Products 0.2%Sulphur containing compounds 0.05% Vegative Matter 84% Linalool (spicy)Geraniol MyrceneCaryophyleneFareseneSelinene

  21. trans-isohumulone cis-isohumulone

  22. Further adventures in stereochemistry carvone thalidomide D-form amino acids tend to taste sweet, L-form amino acids are generally tasteless. Proteins use L-amino acids Most sugars we digest and incorporate are D (R) (S) caraway spearmint

  23. Antibiotic properties of hops Hop compounds act as ionophores that exchange protons for cellular divalent cations. This decreases the intracellular pH and dissipates the transmembrane proton gradient (ΔpH) and the proton motive force (pmf). Bacteria have evolved a number of ways to resist killing by hops. HorA (a) and probably also by a pmf-dependent transporter (b) overexpressed H+-ATPase increases the pumping of protons released from the hop compounds (c) Galactosylated glycerol teichoic acid in the cell wall and a changed lipid composition of the cytoplasmic membrane of beer spoilage lactic acid bacteria may increase the barrier to hop compounds. - + H+ antibacterial form trans-isohumulone

  24. Saccharomyces cerevisiaea -Single-celled fungus from the phylum Ascomycota -One of the most well characterized organisms -Genome sequenced in 1996 -Capable of sexual and asexual reproduction -Found in wild on fruit surfaces Lager yeast is more complex. First called S. carlsbergensis or S. pastorianus, then considered to be S. cerevisiae, are now recognized as a hybrid of S. cerevisiae and S. bayanus

  25. S. cerevisiae life cycle Gene expression in lag phase and early log phase Time Brejning et al. J Appl Microbiol. 2005.

  26. If fermentation is anaerobic why is so much oxygen needed when pitching?

  27. S. cerevisiae life cycle Fermentation profiles with various sugar supplements Piddocke et al. Applied Microbiology and Biotechnology 2009

  28. S. cerevisiaelife cycle Stationary phase is more complex than it seems Extending healthy life span--from yeast to humans. Fontana L, Partridge L, Longo VD. Science. 2010 Apr 16;328(5976):321-6. Review. Insulin/IGF-I and related signaling pathways regulate aging in nondividing cells: from yeast to the mammalian brain. Parrella E, Longo VD. ScientificWorldJournal. 2010 Jan 21;10:161-77. Review. Genetic links between diet and lifespan: shared mechanisms from yeast to humans. Bishop NA, Guarente L. Nat Rev Genet. 2007 Nov;8(11):835-44. Review. Gray et al. Microbiology and Molecular Biology Reviews. 2004.

  29. Yeast metabolism

  30. Why is S. cerevisiaeso good at making beer? “Make-accumulate-consume” Yeast can suppress respiration in the presence of glucose and oxygen

  31. Yeast settling to the bottom is not a passive process

  32. What goes wrong when beer goes bad? diacetyl rest: yeast convert acetolactic acid into valine instead of diacetyl (butanedione) and converts any butanedione into butanediol which is neutral as far as beer flavoring Lagering: beer stored at 34-40 F for a few weeks. levels of diacetyl, acetaldehyde and sulfur compounds decrease. ethanol valine butanediol acetaldehyde pyruvate acetoin diacetyl acetolactate The dynamics of the Saccharomyces carlsbergensis brewing yeast transcriptome during a production-scale lager beer fermentation. Olesen K, Felding T, Gjermansen C, Hansen J. FEMS Yeast Res. 2002 Dec;2(4):563-73. Two-dimensional gel analysis of the proteome of lager brewing yeasts. Joubert R, Brignon P, Lehmann C, Monribot C, Gendre F, Boucherie H. Yeast. 2000 Apr;16(6):511-22.

  33. Kobayashi et al. J. Biosci. and Bioengr. 2008.

  34. Kobayashi et al. J. Biosci. and Bioengr. 2008.

  35. Kobayashi et al. J. Biosci. and Bioengr. 2008.

  36. nd Not determined, SMM S-methyl methionine, and 3-MTP 3-methylthiopropionaldehyde aUnless stated otherwise, odor threshold values were determined in beer. bIn alcohol-free beer Landaud et al. App. Microbiol. Biotechnol. 2008

  37. Landaud et al. App. Microbiol. Biotechnol. 2008

  38. How can we make even better beer? Breeding an Amylolytic Yeast Strain for Alcoholic Beverage Production. Cheng MC, Chang RC, Dent DF, Hsieh PC. Appl Biochem Biotechnol. 2010 Sep 5. [Epub ahead of print] The potential of genetic engineering for improving brewing, wine-making and baking yeasts. Dequin S. Appl Microbiol Biotechnol. 2001 Sep;56(5-6):577-88. Review. Improvement of Saccharomyces yeast strains used in brewing, wine making and baking. Donalies UE, Nguyen HT, Stahl U, Nevoigt E. Adv Biochem Eng Biotechnol. 2008;111:67-98. Review. Genetic improvement of brewer's yeast: current state, perspectives and limits. Saerens SM, Duong CT, Nevoigt E. Appl Microbiol Biotechnol. 2010 May;86(5):1195-212. Epub 2010 Mar 2. Review. Multiobjective optimization and multivariable control of the beer fermentation process with the use of evolutionary algorithms. Andrés-Toro B, Girón-Sierra JM, Fernández-Blanco P, López-Orozco JA, Besada-Portas E. J Zhejiang Univ Sci. 2004 Apr;5(4):378-89. Use of a modified alcohol dehydrogenase, ADH1, promoter in construction of diacetyl non-producing brewer's yeast. Onnela ML, Suihko ML, Penttilä M, Keränen S. J Biotechnol. 1996 Aug 20;49(1-3):101-9.

  39. Saerens et al. Appl Microbiol Biotechnol. 2010.

  40. Yeast and human health Tumor cell energy metabolism and its common features with yeast metabolism. Diaz-Ruiz R, Uribe-Carvajal S, Devin A, Rigoulet M. Biochim Biophys Acta. 2009 Dec;1796(2):252-65. Epub 2009 Aug 12. Review. Yeast cell wall polysaccharides as antioxidants and antimutagens: can they fight cancer? Kogan G, Pajtinka M, Babincova M, Miadokova E, Rauko P, Slamenova D, Korolenko TA. Neoplasma. 2008;55(5):387-93. Review. Brewer's/baker's yeast (Saccharomyces cerevisiae) and preventive medicine: Part II. Moyad MA. Urol Nurs. 2008 Feb;28(1):73-5. Review. Combined yeast-derived beta-glucan with anti-tumor monoclonal antibody for cancer immunotherapy. Liu J, Gunn L, Hansen R, Yan J. Exp Mol Pathol. 2009 Jun;86(3):208-14. Epub 2009 Jan 21. Review. Protein folding diseases and neurodegeneration: lessons learned from yeast. Winderickx J, Delay C, De Vos A, Klinger H, Pellens K, Vanhelmont T, Van Leuven F, Zabrocki P. Biochim Biophys Acta. 2008 Jul;1783(7):1381-95. Epub 2008 Feb 11. Review. Saccharomyces cerevisiae: a useful model host to study fundamental biology of viral replication. Alves-Rodrigues I, Galão RP, Meyerhans A, Díez J. Virus Res. 2006 Sep;120(1-2):49-56. Epub 2006 May 15. Review.

  41. References and further reading National Institutes of Health digital archive of biomedical and life sciences journal literature http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed http://www.wikipedia.org http://homebrewandchemistry.blogspot.com/ Site of Brew Your Own magazine http://www.byo.com/ http://forum.northernbrewer.com/ http://www.brewingtechniques.com/ Site of UC-Davis Anheuser-Busch Endowed Professor of Brewing Science Charles Bamforth http://foodscience.ucdavis.edu/bamforth/ Rensselaer Polytechnic Institute brewing class http://www.rpi.edu/dept/chem-eng/Biotech-Environ/beer/index1.htm

  42. Tools of the trade High performance liquid chromatograph (HPLC) Gas chromatograph (GC) Fermenter Electrospray ionization m mass spectrometer (ESI-MS)

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