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Fermentation Process

Fermentation Process

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18 2.203
© Copy Right: Rai University
BIOPROCESS ENGINEERING
Objectives
After this session you will be able
To prepare, initiate, observe, and understand fermentation,out of all the Natural Sciences, modern biology is the mostdiversified exhibiting a varied array of subdisciplines. Thisdiversity can be attributed to the introduction of otherscientific disciplines such as physics, chemistry, andmathematics. This has resulted in a more profounddescription of life at the cellular and nuclear level. Thenewly acquired knowledge in the field of biology hasresulted in great contributions to the health and welfare of man.
Know many areas of application of biotechnology. Thetable below lists the main ones from which all others stem:
1)Fermentation Technology
This is, historically, the most important area inbiotechnology. There has been extensive development inprogress with new products such as medically importantdrugs, solvents, protein enhanced foods, etc. This alsoincludes research on different types of fermentationdesigns to optimize the process.
2)Enzyme Engineering
This area is used for the catalysis of extremely specificchemical reactions, for the immobilization of enzymes, andto create specific molecular converters (bioreactors).Products formed include L-amino acids, high fructosesyrup, semi-synthetic penicillins, starch and cellulosehydrolysis, etc.
3)Waste Technology
This has a long array of historical importance, but nowemphasis is on the coupling of this field with theconservation and recycling of resources. Examples wouldinclude foods, fertilizers, biological fuels.
4)Environmental Technology
Problems like pollution control, removing toxic wastes,recovery of metals from mining wastes and low grade ores,are just some of the categories that fall under this field.
5)Renewable Resources Technology
The use of renewable energy sources, in particular lignocelluloseto generate new sources of chemical raw material and energy -ethanol, methane, and hydrogen.Each of these fields utilizes knowledge from Biochemistry,Genetics, Chemistry, Applied Microbiology, Chemical andProcess Engineering, and Mathematics and Computer Technol-ogy. Also, these areas of biotechnology attempt to use the bestpossible catalysts in optimum environment to carry out variouschemical reactions. In the pages to come, some important areasof biotechnology will be considered in order to achieve a broadoverall understanding of basic principles.
Define Fermentation ?
Introduction
: Fermentation is a common process in bothnatural and man-made environments. In fermentation, yeastcells metabolize sugars, producing carbon dioxide and alcohol.Some products which result from fermentation include, amongmany others: alcoholic beverages, bread, soy sauce, sauerkraut,pickles, and kimchee.Observing fermentation provides an interesting platform fromwhich to study and understand chemical and biologicalprocesses.
Fermentation
is defined as an energy-yielding metabolicpathway that involves no net change in oxidation state.Anaerobic glycolysis is a type of fermentation. The lactic acidfermentation (conversion of glucose to lactate) is important inthe manufacture of cheese. Another important fermentationinvolves cleavage of pyruvate to acetaldehyde and CO2, thatfollows: with the acetaldehyde then reduced to ethanol byalcohol dehydrogenase in the reactionAcetaldehyde + NADH + H
+
<-> Ethanol + NAD
+
As carried out by yeasts, this fermentation generates the alcoholin alcoholic beverages. alcoholic fermentation
What is necessary to begin fermentation ?
All that is necessary to begin fermentation is to mix theactivated yeast and the cooled, pH-adjusted mash in thefermentation tank. Aside from the considerations of pH asdiscussed earlier, the most important thing during the fermen-tation is temperature control. When the fermentation begins,carbon dioxide gas will be given off. At the height of fermenta-tion, the mash will literally “boil” from the carbon dioxideproduced. The reaction also produces some heat. The optimumtemperature for the fermentation process is between 70-85 degF., and it is desirable not to let the temperature go much above90-95 deg F. Cooling is readily done with the use of ice bags, asdiscussed earlier, or by the use of a cooling coil. A less desirablemethod of controlling temperature is to dilute the mash.The actual time required to ferment a mash varies with thematerial being fermented, the pH, temperature, and severalother factors. It can take from one to four days. You will knowthat the fermentation is complete when the mash ceasesbubbling and the yeast cake, which forms on top, sinks to thebottom. At this point, the fermented liquor is known as “beer”and it is ready to be distilled.It is advantageous to distill the beer as soon as possible.Occasionally, if it is allowed to sit, it will turn to vinegar.Vinegar is alcohol that has been oxidized to acetic acid. Certainenzymes present after fermentation act as catalysts and allow anyair present in the mash solution to react with the alcohol toform acetic acid. In fact, if you want to produce vinegar, all you
UNIT 2FERMENTATION PROCESS:GENERAL REQUIREMENTS OFFERMENTATION PROCESS
LESSON 8FERMENTATION PROCESS
 
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BIOPROCESS ENGINEERING
have to do to start the reaction is to bubble air through thefermented mash. Once the vinegar reaction has set in, the mashis lost. There is no cure. The only prevention is to separate thebeer from the mash sediment and distill it as soon afterfermentation is complete as possible.It is also advantageous to use a fermentation lock to preventalcohol vapors from escaping the fermenter. Otherwise, theCO2 gas can carry with it a considerable amount of alcohol.Note that the small, glass fermentation locks available fromwine-making supply houses are suitable, at most) for a 5-galloncontainer. Larger containers must have proportionately largerfermentation locks or a dangerous amount of pressure willbuild and the vessel could explode.It is permissible to open the fermenter to check progress andtake samples for pH analysis, etc. as long as care is taken not tointroduce bacteria that could contaminate the mash.
Fermentation By-products
The principle products of fermentation are alcohol, carbondioxide, and fermentation residue. The alcohol is distilled fromthe beer and used as fuel . The carbon dioxide gas in largedistilleries is usually compressed or made into dry ice. Anotheruse for the gas would be to pipe it into a greenhouse. Theplants will then use it in the photosynthesis cycle, removing thecarbon and giving off oxygen. Lacking a use for the carbondioxide, it can be simply vented into the air as it is totally non-polluting and non-toxic.What will be left is a lot of water and solids. A portion of thewater can be used for backslopping. The remaining solidscontain proteins, vitamins, minerals, fats, and yeast cells. All of the nutrition value of the original feedstock, except the starch orsugar that has been turned into alcohol, survives intact. It maybe fed to cattle, or if suitably processed it can be used forhuman consumption. However, in the wet state, it will keep fora maximum of 3-5 days depending on conditions. After this itwill begin to rot. Therefore, for long term storage these residues(stillage) must be dried. This can be done by straining out thesolids and spreading them in a thin layer to dry in the sun, byuse of rotary grain dryers, or similar equipment.
Note of Caution
Alcohol produced for human consumption is made underspecial conditions and purified to a high degree. Ethanol that isproduced according to the procedures in this book will containfusel oils (high boiling alcohols), aldehydes, and ketones. Noneof these chemicals affect fuel performance but, if ingested,could cause fatal poisoning at worst or a horrible hangover atbest. In addition, if the distillation equipment used later on isnot tinned copper or stainless steel, many toxic metal oxides canbe introduced to the alcohol. Solder, for example, contains a lotof lead and can react to form poisonous lead oxides. So besidesbeing illegal, drinking your fuel could be hazardous to yourhealth!What is lactic acid fermentation
?
Lactate Fermentation
Lactate fermentation occurs in anaerobic organisms or in aerobiccells that are undergoing very high rates of glycolysis. In thesecells, NADH generated in glycolysis cannot be reoxidized toNAD
+
. When this situation happens, NADH is oxidized toNAD
+
by reducing pyruvate to lactate. The enzyme catalyzingthis reaction is lactate dehydrogenase. The equilibrium for thisreaction lies far in favor of formation of lactate.Until recently it was thought that lactate accumulation in skeletalmuscle was largely a consequence of anaerobic metabolism,which occurs when the need for tissues to generate energyexceeds their capacity to oxidize the pyruvate produced inglycolysis. Recent metabolic studies, including
31
P NMR analysesof the levels of phosphorylated intermediates in living musclecells during exercise, suggest that lactate is actually an intermedi-ate and not a metabolic “dead end,” whose only fate isreconversion to pyruvate. These studies show that even in fullyoxygenated muscle tissue, as much as 50% of the glucosemetabolized is converted to lactate. This may represent a meansfor coordinating energy-storing and energy-generating pathwaysin different tissues, but the mechanisms involved are not yetclear.
Lactate dehydrogenase
was the first enzyme that establishedthe structural basis for the existence of isoenzymes (differentforms of an enzyme resulting from variations in amino acidsequence). Most tissues contain five isoenzymes of lactatedehydrogenase that can be resolved electrophoreticallyYEASTYeast is an organism belonging to the vegetable family. Theyeast itself does not take a direct part in the fermentationprocess, but it secretes a complex of enzymes that act upon thesugar and convert it to alcohol and carbon dioxide gas.The yeast used in alcoholic fermentation is a special strain bredto be tolerant to variations in pH and resistant to alcohol. Inthe past, distilleries bred and propagated their own yeast strains.The yeast was kept alive in cultures and grown in batches of ever-increasing size to be used in the fermenters. Keeping yeastalive and growing cultures is a tricky business that requiresprecise control of temperature, nutrients, and the like. However,a simplified method is described later. Fortunately, special activedry yeast is available. To use it, you merely add warm water toreactivate it and then add it to the mash in the fermenter. Twopounds is sufficient for 1000 gallons of mash. It is availablefrom Universal Foods Corporation as listed in the appendix.This yeast should be rehydrated for 15 minutes prior to use at atemperature of 100-105 deg Fahrenheit, or it can be added dryto the fermentation tank prior to filling.In a pinch, it is possible to use ordinary baker’s yeast from yourgrocer’s shelf. However, this yeast is not bred for alcoholtolerance, and you will probably not get the yields associatedwith the distiller’s yeast.
Yeast Propagation
It is possible to grow and propagate your own yeast cultures if you observe certain precautions. Above all, the conditions mustbe absolutely sterile. Ordinary boiling water does not kill all of the bacteria present. It is necessary to use a pressure cooker.Make a solution of (proportionately) one cup sugar, one cupflour and two quarts water. Place the solution in a pressurecooker and boil at elevated pressure for at least 45 minutes.Without opening the pressure cooker, cool the solution to
 
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BIOPROCESS ENGINEERING
about room temperature. Then open the container and add acake of baker’s or distiller’s yeast. Close the container and keep itin the refrigerator. The yeast will slowly grow. Some carbondioxide will be given off, so be sure to leave the vent open. If desired, the yeast slurry can be transferred to jars. Just be surethey are sterile and remember to poke a small hole in the lid tolet the carbon dioxide escape.To use the yeast culture, merely remove a teaspoon or so, place itin another (sterile) container, feed it some sugar and warm it toroom temperature. When it becomes active, it is ready for thefermenter. If at any time your refrigerated culture goes bad (dueto bacterial contamination) it must be thrown out and theprocedure started again. Also, yeast cultures should not befrozen.Yeast convert pyruvate to acetaldehyde in a reaction catalyzed bythe enzyme pyruvate decarboxylase. This is followed byreduction of acetaldehyde to ethanol catalyzed by alcoholdehydrogenase. The reaction uses NADH and releases NAD
+
,which is subsequently used in glycolysis.Yeasts used in baking also carry out the alcoholic fermentation;the CO2 produced by pyruvate decarboxylation causes bread torise, and the ethanol produced evaporates during baking.Among the dozens of other useful fermentations are thoseleading to acetic acid (manufacture of vinegar) and propionicacid (manufacture of Swiss cheese).
Quiz Questions
1.Alcoholic fermentation is caused by the interaction of 
a)sugar and carbon dioxideb)sugar and acidc)sugar and yeastsd)yeast and pectin
2.The products of the alcoholic fermentation are
a)alcohol, carbon dioxide, and heatb)carbon monoxide and sugarc)tannin and sugard)sugar and higher pH
3.As the sugar content of the grapes increases during thegrowing season, the acids
a)stay the sameb)fallc)rised)change to sugar
4.Grapes grown in cooler climates generally producewine with
a)high alcohol and low acidsb)high alcohol and high acidsc)low to moderate alcohol and low acidsd)low to moderate alcohol and high acids
5.The scale used to measure sugar in grape juice in theU.S. is the
a)Richter scaleb)Metric scalec)Fish scaled)Brix scale
6.The color of wines made from red grapes can be
a)red, white, and roséb)red onlyc)red or rosé onlyd)white only
7.The process of adding sugar to grape juice before orduring fermentation to achieve higher alcohol levels iscalled
a)Consternationb)Pasteurizationc)Finingd)Chaptalization
8.A full-bodied white wine with deep yellow color andcomplex aromas and flavors would most likely havebeen produced by
a)fermentation and aging in small wooden barrelsb)fermentation and aging in large, cool stainless steeltanksc)carbonic macerationd)adding red grapes to the process
9.The color of red wines comes from
a)wooden barrelsb)adding caramelc)the pigments in red grape skinsd)using beet sugar
10. Tannins are extracted from
a)grape skinsb)wooden barrelsc)grape stemsd)all of the above
11.A wine labelled as Zinfandel will be
a)whiteb)redc)roséd)clear
12.The term “dry” indicates that a wine is
a)not sweetb)low in alcoholc)astringentd)in powder form
13. The bubbles in sparkling wines are
a)oxygen that permeates the cork and settles in the wineb) carbon monoxide produced by a second fermentationin the bottle

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