Fermented fish products-fish sauce Fish sauce is very popular as a condiment around Southeast Asia regieon also known as ngapi (Burma), nouc-man (cambodia and vietnam), nam-pla (Laos and Thailand), ketjapikan (indonesia) 3/14/2017 82
Mixing salt with fish (anchovies) at 1:3 ratio This creates environment that is suitable for starter culture (from salt and environment) 3/14/2017 83
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Alcoholic beverage Wine Beer Sake Sato Converting sugar to alcohol However, if the starting raw material is starch, steps for converting starch to sugar are required. 3/14/2017 87
ท มา: senior project ของนางสาวจ ราพร ผาส ข และ นางสาวพรพ มล หม นอาจย ม FT#3 3/14/2017 88
Sato (Thai rice wine) rice wine = 2 processes (koji preparation and alcoholic fermentation) These two processes can occur simultaneously. koji production: enzyme production using Aspergillus sp. / Rhizopus / Mucor (amylase, protease, lipase) 5-7 days at 30 ๐ C Continue mixing the bed (aeration and heat reduction) 3/14/2017 89
สาโท Then mix koji with cooked rice and water (to extract enzymes) and yeast This is called alcoholic fermentation Amylases in koji convert starch to fermentable sugar and fermentable sugar is then converted to alcohol take 7-21 days depending on temperature 3/14/2017 90
By Sitthipong Chotpattarasumon, Sutchai Ruenchit, Pitiporn Ritthiruangdej and Thanut Amatayakul* at The 3rd International Conference on Fermentation Technology for Value Added Agricultural Products Kosa Hotel, Khon Kaen, Thailand 3/14/2017 91
Experimental plan Size reduction using a food processor Waxy rice Size classification using sieves No. 7, 8, 10, 12 Full kernel (control) 2.0-2.3 mm. 1.7-2.0 mm. 1.4-1.7 mm. Sato production Koji preparation Aspergillus niger TISTR 3257 for 7 days at 30 o C Alcoholic fermentation Saccharomyces cerevisiae SC90 for 7 days at 30 o C Measurement - Crude amylase activity - TSS - percentage of starch digestion Measurement - Yeast growth - Alcohol content - 8 volatile compounds - percentage of starch digestion 3/14/2017 92
Crude amylase activity and total soluble solid of koji Figure 1. Crude amylase activity and total soluble solid of koji made from waxy rice reduced to size 1.4-1.7, 1.7-2.0, 2.0-2.3 mm and full kernel. Bars show the same letter (a, b, c or A, B) did not show significantly different (p < 0.05) 3/14/2017 93 One unit of crude amylase activity was defined as the amount of reducing sugar produced per reaction time
Solid State Fermentation Limitation Oxygen transfer Heat transfer Moisture content Limiting Enzyme production -amylase and protease Udo Ho lker and Ju rgen Lenz (2005) Solid-state fermentation are there any biotechnological advantages?. Current Opinion in Microbiology, 8: 301-306 3/14/2017 94
Table 3 Volatile compounds in Sato made using waxy rice reduced to size 1.4-1.7 mm, 1.7-2.0 mm, 2.0-2.3 mm and full kernel Volatile compounds Ethyl butyrate Ethyl decanoate Diethyl succinate Phenethyl acetate 3-methyl-1- butanol Full kernel Concentration (mg/l) 2.0-2.3 mm 1.7-2.0 mm 1.4-1.7 mm Odor threshol d (mg/l) Odor description 18.00 b 54.75 a 12.62 b 14.54 b 0.4 Strawbery, apple, banana 7.10 b 60.19 a 9.41 b 6.83 b 0.5 Fruity, wine 1.25 b 6.54 a 1.56 b 1.62 b 1.8 Fruity 1.05 b 2.98 a 2.78 a 1.82 a,b 0.25 Fruity, green apple, banana 146.25 b 235.01 a 180.95 b 156.20 b 60 Solvent Pentanol 5.30 a 4.04 b 0.70 c 0.95 c 0.1 Phenolic, medicinal 2-methyl-1- propanol Phenethyl alcohol 49.35 b 86.90 a 55.03 b 61.05 b 75 Alcohol, nail polish 39.05 c 61.46 a 67.09 a 53.40 b 200 Rose, honey 3/14/2017 95
Fatty acid ethyl and acetate Fatty acid ethyl esters: Condensation of acyl-coa and higher alcohols esters formation Acetate esters: Condensation of Acetyl-CoA and higher alcohols Verstrepen et al. (2003) Journal of bioscience and bioengineering, 96(2): 110-118 3/14/2017 96
Ehrlich pathway formation of fusel alcohols by Sacchariomyces cerevisiae 3/14/2017 97
Sake production 3/14/2017 98
Sake video 3/14/2017 99
Use of mould originate from China famous place: Kyoto, Kobe Steaming sterile rice and make starch and protein more easier for enzyme from koji to digest Koji (Aspergillus oryzae) = Malt Liquefaction and saccharification Sake mashing : Moromi (parallel fermentation) slowly add rice to moromi helps yeast survival and increase ethanol to 20% 3/14/2017 100
Several types of Sake 3/14/2017 101
Wines vin ENOLOGY Enology = Winemaking British version of the word is Oenology 3/14/2017 106
What is wine? Water ~ 85% Alcohol ~ 12% Everything else ~ 3% 3/14/2017 107
Wineries 3/14/2017 108
Classification Table wine (Natural wine) Sparkling wine (contain CO 2 ) champagne Fortified wine (added alc.): 14-24% alc. Red or white 9-14% alc. Sweet wine: reducing sugar 10-100 g/l Dry wine: reducing sugar < 10 g/l Called by variety of grapes Vinification 3/14/2017 109
Example grape varieties Red Carbernet Sauvignon Shiraz Durif Merlot Pinot Noir Grenache White Chardonnay Chennin Blanc Riesling Sauvignon Blanc Semillion 3/14/2017 110
Wine vinification Grapes Crushed and stemmed Fining and filtration SO 2 Press Bottling Saccharomyces cerevisiae Juice Fermentation Maturation 3/14/2017 111
Total Soluble Solids Use as an indicator for maturation of the grapes As an approximate basis for calculating the alcohol yield ~ 90% of soluble solids are fermentable sugar 3/14/2017 112
How can total soluble solids be measured? 2 ways Hydrometers Refractometry Brix, Baume % Brix represent weight of sugar in solution (g of sucrose/ 100 g of liquid) 1.8 Brix = 1.0 Baume Baume represent final expected alcohol content of wine 3/14/2017 113
Harvesting Hand or machine picking Damaging skin can result in enzymati 3/14/2017 115
Simple anatomy of the grape 3/14/2017 117
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Pressing Directly for white wine Red wine after maceration 3/14/2017 121
Preparing juice for fermentation Polyphenol oxidase browning in white wine Sulphur dioxide (SO 2 ) commonly add to white wine Prevent browning by inactivate enzyme Inhibit activity of undesirable microorganisms 30 to 50 ppm (mg/l) or 70 to 100 ppm (highly contaminate) 3/14/2017 122
SO 2 addition Often used sodium or potassium metabisulphite (containing 58% SO 2 ) Want grape juice 500 ml to have 30 mg/l of SO 2 from potassium metabisulphite (KMBS): 58 g SO 2 come from 100 g KMBS Need 0.03 g use (100*0.03)/58 = 0.0517 g/500 ml 3/14/2017 123
Yeast Fermentation Pure culture of Saccharomyces cerevisiae; bought in dried powder Activation with grape juice several hours before fermentation (10 8 cell / ml) Yeast use O 2 first few hours, follow by produce CO 2 and alcohol Adding nutrients: Diammonium phosphate (DAP) 3/14/2017 124
C 6 H 12 O 6 2C 2 H 5 OH + 2CO 2 (1 Glucose 2 ethanol + 2 CO 2 ) ~ 50% conversion In reality < 50% by weight 3/14/2017 127
Temperature of fermentation Opt. temp. for yeast fermentation is between 10-15 o C (white wine) Red wine are fermented at higher temperature ~ 25 o C (pigments extraction) 3/14/2017 128
Stuck fermentation premature arrest of fermentation high level of sugar Cause fermentation at too high temperature nutritional deficiency osmoregulation ethanol toxicity low temperature fermentation too high SO 2 presence of inhibitory substance 3/14/2017 129
Maturation (continued) Flavor: vanillin, Oak: fill with water purged with SO 2 for disinfection Wine racking (standing) : to separate wine from lees (Lees - The solids left at the bottom of a fermentation) 3/14/2017 132
Fining of Wines (clearing of wines) Fining is the process of removing suspended particles from wines and will sometimes be used for softening of wines of phenolic compounds Common types of fining agents are Bentonite, and egg whites 3/14/2017 133
Barrels (cont.) French barrels average $700 each American barrels average $300 each Oak species (commonly used) Allier Limousin American Nevers 3/14/2017 134
Oak barrel Barrel (cask) 3/14/2017 137
Glass Air Lock - twin -S - Wine Thie Bubble 3/14/2017 138
Precision Wine Hydrometers Ebulliometer 3/14/2017 139
Ebulliometer http://www.monashscientific.com.au/ebulli 3/14/2017 140
PRINCIPLE The boiling temperature of wines is based on their alcoholic content Note the boiling temperature of the wine to be tested Consult a calculation disk that gives directly the alcoholic strength by volume. 3/14/2017 calculation 141 disk
Distillation temperature and concentration relationship Temperature (C) 100 98 96 94 92 90 88 86 84 82 80 78 Vapor Liquid Alcohol ( % by volume) 3/14/2017 142
Color The color of red wine is due to red and purple anthocyanins (first come out from fermentation)--> dark red over 24 months of aging Old wine change from red to red/brown brick White wine pale (light) green/yellow, over 24 months become yellow and over 10 years become brown 3/14/2017 143
Stabilization of wine Wine stabilization involves one or more of the following: Removal of tartarate Complexing agents: Carboxylmethyl cellulose (CMC) Removal of metal ions and some tannins Ion exchange (cation exchange column) Membrane filtration 3/14/2017 144
Removal of particulate material Aged wine : separation of clear liquid and lees still contain polysaccharides, large molecular weight tannin, protein-complexed with fine tartrate crystal filter (1-5 micron), filter aids (diatomaceous earth), polyvinyl polypyrolidone (PVPP, insoluble) 3/14/2017 145
Filtration Filtration is carried out just before bottling Centrifugation follow by sterile membrane filtration Sterile filtration with membrane 0.45 1.2 micron 3/14/2017 146
Bottling Purge with inert gas or nitrogen to prevent oxidation 3/14/2017 147
Closures Corks; both synthetic and natural Natural cork has risk of contamination with mold Screw caps Proven to be a better seal for wine bottles than corks. 3/14/2017 148
Capsules Lead-based capsules very popular until it is banned to use in the late 1980 s. Now wineries use tinaluminum or aluminum polylaminate. It is better to have no capsule rather than a bad one. Very important part of packaging, primarily for consumer appeal. 3/14/2017 149
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Main ingredients of beer Barley Yeast Hops Water 3/14/2017 155
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Barley germination beta-amylase Malt 3/14/2017 157
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Color of malt Light Amber Dark Milling to Grits This makes the malt easier to wet at the mashing stage and aids faster extraction of the soluble components from the malt during the enzymic conversion. 3/14/2017 160
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Adjunct cereal To produce a more stable beer, as they contain less protein To produce a different flavor maize, for example, is said to give a fuller flavor To produce a better beer foam due to lower fat (lipid) levels and different proteins To improve the ease of processing in the brewhouse To produce beer at lower cost 3/14/2017 162
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Enzyme The alpha enzymes break up the long chains of starches by splitting them in half. The beta enzymes break down the starches by chopping them off a couple at a time from the ends of the chain. The alpha enzymes are most active at 65 to 67 oc, and the beta enzymes are most active at 126 to 144 F 52 to 62 oc. So the temperature and duration of the mash must be carefully controlled to get a good conversion. 3/14/2017 164
Wort separation The principal objectives are to produce bright wort and to collect the maximum amount of sugars (extract) from the residual solid materials. These solids are sold as animal feed, draff, or spent grains for milk and beef cattle herds. 3/14/2017 165
Wort boiling Sterilization of the wort to eliminate all bacteria, yeasts, and molds that could compete with the brewing yeast and possibly cause off-flavors Extraction of the bittering compounds from hops added early to the boil and oils and aroma compounds from late additions Coagulation of excess proteins and tannins to form solid particles (trub) that can be removed later. This is important for beer stability and foam Color and flavor formation Removal of undesirable volatiles, such as dimethyl sulfide, by evaporation Concentration of the sugars by evaporation of water 3/14/2017 166
Wort cooling, aeration and yeast pitching The wort is cooled from almost boiling point to fermentation temperature through a heat exchanger using water as the main cooling medium. The temperature for fermentation is typically 8 to 13oC for lager and 14 to 17oC for ale. Typically, 5 to 20 million yeast cells are pitched per milliliter of wort. Air or oxygen is also added to the wort as an essential factor for the production of yeast membranes and, hence, new cells. 3/14/2017 168
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Ageing Chill haze formation Clarification Carbonation (to a limited extent) Flavor maturation (again to a limited extent) Stored capacity for demand smoothing 3/14/2017 171
Filtration Clarification filter press with filter aids (bentonite) then filling pasteurization 3/14/2017 172