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| − | '''''------------- Work in progress ------------'''''
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| − | In mashing the milled grain (grist) is mixed with water to create the mash. It is an essential process in the production of beer and a continuation of the malting process on the way to sweet wort. During the mash soluble malt compounds like enzymes, proteins and sugars are dissolved by the mash water and insoluble malt compounds like starch and some long chained proteins are converted into soluble compounds and dissolved into the water. The latter happens through a combination of physical and biochemical process which can be controlled by the brewer to achieve a sweet wort of desired quality. The biochemical processes are catalyzed by various malt enzymes. Their function and behavior is dependent on the conditions in the mash (e.g. temperature, pH, concentration, etc.) and the brewer should be familiar with that behavior in order to control the quality of the sweet wort that is run off from the mash in the lautering process.
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| − | =Enzymes=
| + | In mashing the milled grain (grist) is mixed with water to create the mash. It is an essential process in the production of beer and a continuation of the malting process on the way to sweet wort. During the mash already soluble malt compounds like enzymes, proteins and sugars are dissolved by the mash water and insoluble malt compounds like starch and some long chained proteins are converted into soluble compounds and also dissolved into the water. The latter happens through a combination of physical and biochemical process which can be controlled by the brewer to achieve a sweet wort of desired quality. The biochemical processes are catalyzed by various malt enzymes. Their function and behavior is dependent on the conditions in the mash (e.g. temperature, pH, concentration, etc.) and the brewer should be familiar with that behavior in order to control the quality of the sweet wort that is run off from the mash in the lautering process. |
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| − | [[Image:Enzymatic_reaction_diagram.gif|frame|right|Figure 1 - A simplified illustration of beta amylase splitting maltotetraose into two maltose molecules. A - a beta amylase molecule bonds to the non reducing end of the glucose chain. Because of its shape it fits only there. B - The intermediary bond reacts with a water molecule. C - The reaction releases 2 maltose molecules and frees the beta amylase molecule]] | + | * [[Enzymes]] |
| | + | * [[Carbohydrates]] |
| | + | * [[Starch Conversion]] |
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| − | Enzymes are very important to mashing. They catalyze conversion reactions which break down malt compounds (the largest one being starch). In the case of starch this conversion is necessary to form water soluble dextrines and sugars. The latter of which can be metabolized by the yeast. Enzymes are proteins (chains of amino acids) which have the ability to lower the energy needed for a chemical chemical reaction which allows that reaction to occur faster and at lower temperatures. Most enzymes are very specific to the reaction they catalyze and work only with a specific substrate and produce only a specific product. In case of the beta amylase enzyme, for example, the substrate is a glucose chain and the product is maltose. The reaction that is catalyzed the the split of a glucose chain link while a molecule of water is consumed. The highly specific nature to the reaction that is catalyzes stems form the shape of the enzyme which is just right for reacting with the substrate and releasing the product and residual product shortly after. The enzyme itself is not consumed in the reaction. I.e. once free again it can catalyze another reaction. | + | The article [[Enzymes]] explains the dynamics of enzymatic reactions. In particular how parameters like substrate concentration, enzyme concentration, pH, temperature and time affect the speed of the enzymatic reaction and the amount of product that is produced. |
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| − | Without an enzyme a chemical reaction may look like this [rsc.org]:
| + | The article [[Carbohydrates]] sheds some light on the largest group of substrate and products that is involved in mashing. It details the structure of simple and complex carbohydrates all the way to the structure of the starch granule. |
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| − | :'''substrate1 + substrate2 -> product'''
| + | The article [[Starch Conversion]] explains in detail how in mashing the starch found in malt or mash tun adjuncts is converted into fermentable and unfermentable sugars. It covers the process of gelatinization and the enzymatic hydrolyzation of starch. |
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| − | But the initial energy (i.e. temperature) needed for this reaction may be so high that it rarely ever happens. With the presence of an enzyme this reaction changes to
| + | [[Image:Work_in_progress.jpg]] |
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| − | :'''substrate1 + enzyme -> enzyme-substrate-intermediate'''
| + | * Proteins |
| − | :'''enzyme-substrate-intermediate + substrate2 -> product + enzyme'''
| + | * Protein degradation |
| | + | * other mash enzymes |
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| − | The energy needed for these enzymatic reactions are now low enough for the reaction to happen at practical temperatures (e.g. mash temperatures).
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| − | Here is an eample of an actual enzymatic reaction in the mash. The enzyme is maltase which can split a maltose molecule into two glucose molecules. The reaction consumes one water molecule:
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| − | : '''C<sub>12</sub>H<sub>22</sub>O<sub>11</sub> + H<sub>2</sub>O -> 2 C<sub>6</sub>H<sub>12</sub>O<sub>6</sub>'''
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| − | With the enzyme maltase the reaction is actually 2 reactions:
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| − | : '''C<sub>12</sub>H<sub>22</sub>O<sub>11</sub> + <Maltase> -> C<sub>12</sub>H<sub>22</sub>O<sub>11</sub>-<Maltase>'''
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| − | : '''C<sub>12</sub>H<sub>22</sub>O<sub>11</sub>-<Maltase> + H<sub>2</sub>O -> C<sub>6</sub>H<sub>12</sub>O<sub>6</sub> + <Maltase>'''
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| − | The same is true for other enzymatic reactions in the mash. But for simplicity sake the shorter form of:
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| − | :'''substrate1 + substrate2 -> product'''
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| − | Is also used to show the reaction that is happening. The involvement of the enzyme in this reaction is implied
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| − | As mentioned earlier. Enzymes are generally very specific in the type of substrate they work on and which reaction they catalyze. A simple model for that behavior is the lock-and-key hypothesis [rsc.org]. In that theory, the enzyme has a shape that fits only a particular substrate. Like the beta amylase which fits only on the non reducing end of a starch chain (Figure 1). If the shape of the enzyme changes significantly due to temperature or pH or inhibitors block the receptor site of the enzyme it cannot react with the substrate and the enzyme becomes inactive.
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| − | ==How temperature effects enzymatic reactions==
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| − | [[Image:Enzyme_reaction_rate.gif|frame|right|Figure 2 - The rate of enzymatic reaction increases exponentially as the temperature is increased]]
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| − | The rate of most chemical reactions, including enzymatic reactions, follows the Arrhenius equation [wikipedia.org]:
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| − | :'''reaction rate = A * e<sup>(-ΔG*/RT)</sup>'''
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| − | Where A is the Arrhenius constant, a simple non exponential factor for the reaction rate. ΔG* is the activation energy of the reaction and determines at what temperature the reaction will become "significant". R is the universal gas constant and T is the absolute temperature in Kelvin. To convert a temperature in Celsius into Kelvin 273.15 needs to be added to the Celsius value (I.e. the absolute temperature (0 Kelvin) is -273.15 Celsius). But all that doesn't really matter for understanding this subject. What matters is that the reaction rate of an enzymatic reaction follows an exponential curve (Figure 2). For every 10 C (18F) increase the reaction rate increases 1.2 - 2.5 fold [lsbu.ac.uk].
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| − | ''But if the enzymatic reaction rate follows an exponential curve, why are there different temperature optima for different enzymes?''
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| − | To answer this question we need to look at another effect that temperature has on enzymes. As mentioned earlier, enzymes are proteins (long chains of amino acids) that are shaped in a particular way which allows them to hold onto their substrate during the reaction. This shape is held in place by weak bonds between different amino acids of the chain. But these bonds are easily broken if the temperature gets too high and the enzyme molecule starts to vibrate too much. This doesn't cause the chain of amino acids to get split, these bonds are much stronger and require much higher temperatures, but the enzyme looses its shape and becomes unable to catalyse the reaction it was made for. It is said to have denatured. Above a critical temperature the rate of denature increases 6 - 36 fold for every temperature increase of 10 C [lsbu.ac.uk]. This is much higher than the increase in the reaction rate and as a result, the productivity of the enzyme will decrease is the temperature is raised past the temperature optimum. Figure 3 shows how the enzyme concentration of a hypothetical enzyme drops for 5 different temperatures.
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| − | [[Image:Enzyme_activity_time.gif|frame|Figure 3 - The decrease of active enzyme concentration over time for 5 different temperatures for a hypothetical enzyme. The higher the temperature the steeper the decline in the number of enzymes that are able to catalyze reactions]]
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| − | [[Image:Enzyme_product_time.gif|frame|Figure 4 - The increase of the product concentration for 5 different temperatures. The temperature that yields the most product depends on the duration of the reaction]]
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| − | * (X) what is an enzyme?
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| − | * (X) structure of an enzyme
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| − | * (X) how does it work
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| − | * temperature affects on enzymes
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| − | * idealized model for enzymatic activity
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| − | * optimum temperature vs. time
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| − | * pH
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| − | * [http://www.rsc.org/education/teachers/learnnet/cfb/enzymes.htm]
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| − | * [http://www.worthington-biochem.com/introbiochem/default.html]
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| − | * [http://www.lsbu.ac.uk/biology/enztech/ph.html pH effects are reversible if pH shift is low]
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| − | * [http://www.lsbu.ac.uk/biology/enztech/temperature.html the effect of temperature on enzymes]
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| − | : [rsc.org] [http://www.rsc.org/education/teachers/learnnet/cfb/enzymes.htm Chemistry for Biologists - How enzymes work]
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| − | : [lsbu.ac.uk] [http://www.lsbu.ac.uk/biology/enztech/temperature.html The effect of temperature on enzymes], London Southbank University
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| − | : [wikipedia.org] [http://en.wikipedia.org/wiki/Arrhenius_equation Arrhenius Equation]
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| − | =Starch=
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| − | * where it is found
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| − | * glucose and 1-4/1-6 branches
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| − | * amylose
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| − | * amylopectin
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| − | * starch granule structure
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| − | =Starch conversion=
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| − | * gelatinization
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| − | * active enzymes
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| − | * emzyme temperature and pH optima
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| − | * mash parameters affecting conversion
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| − | =Proteins=
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| − | * [http://www.rsc.org/education/teachers/learnnet/cfb/proteins.htm]
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| − | =Other enzymes in mashing=
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| − | =Sources=
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