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Transcript of Microbiology in food processing presentation
Microbiology in Food Processing.
‘A study of detrimental and beneficial microbes in
Food Process Industries’
-Jaysing Parade
-Gargi Ramtirtha
www.alfalaval.com
Contents:
� Micro-organisms-Definition ,types.
� Growth of microbes :
• Growth phases of microbes.• Factors affecting growth of microbes.
� Micro-organisms in-
• Milk Powder technology.• Beverages and viscous foods.• Vegetable oil technology.
� Designing a microbiologically safe plant.
� Fermentation Technology.
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Microbiology-Introduction
Definition: Micro-organisms are microscopic organisms, invisible to naked eye that may be unicellular or
multicellular.
Why do we study Micro-0rganisms in Food Processing?
Microbial Terminology:
• Cell
• Colony-CFU/ml or CFU/gm.
• Media
• Incubation
• Spores
SPORE STRUCTURE
COLONIES IN A MEDIA
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Microbiology-Introduction
Classification of Microbes:
Micro-organism
Prokaryotes
Bacteria
Eukaryotes
Fungi Protozoa Algae
Non-living
Viruses
Yeasts Protozoa-Amoeba Molds
Virus
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Growth Cycle of Micro-organisms:
All microbes undergo similar growth patterns.
Each ‘Growth Curve’ has 4 Phases:
� Lag phase: Cells don’t multiply as its still dormant and acclimatizing itself to media.
� Log phase: Microbes grow at an Exponential rate utilizing all nutrients in the media & producing toxic metabolic waste products.
� Stationary phase: Here Growth Rate = Death Rate, due to depletion of nutrients, increase in waste products.
� Death Phase: Death rate exceeds growth rate due to very low conc. of nutrients & high conc. of metabolic wastes.
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Growth of Micro-organisms- Factors affecting growth of microbes in Food
i Temperature:
� Psychrophiles: -5 to -14 °C
� Psychotrophs: -15 to +20 °C
� Mesophiles: +20 to +44 °C
� Thermophiles: +45 to +60 °C
� Thermoduric : +70°C
ii pH:
� Acidophiles (pH < 5.4),preferred by yeasts and molds.
� Neutrophiles (pH 5.4 - 8.5), preferred by bacteria
� Alkaliphiles (pH 7.0 - 11.5).
iii Oxygen requirements: Anaerobes ,aerobes ,facultative aerobes.
iv Osmotic pressure: Higher -Cell lysis.
v Presence or absence of moisture: Water activity 0.7 (Aw)
Food Product Aw
Skim milk Powder 0.3
Whole milk Powder 0.2
Fruit/Veg. Juices 0.97
Edible Oil 0.3
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Micro-organisms in food processing :
Commonly conducted tests in Food Plants :
� Total Count –It helps to enumerate total no. of MO’s in a media. The number of bacteria present can be
indicative of the sanitary condition of material. Mo’s tested Bacillus Cereus, S. Aureus , Salmonella
� Coliforms-Their presence is sign of pollution problems & therefore is particularly significant from a public
health standpoint. MO tested E. Coli.
� Yeast & Mold-They are indicative of poor CIP and sanitation, atmosphere etc. MO tested –Saccharomyces
sp. , mycelium sp.
TPC:Yeast & Mold:
COLIFORMS
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Micro-organisms in food processing :
� Methods by which Microbes are tested:
� Pour Plate Technique
� Membrane Filtration
� Swab Method.
Membrane Filtration apparatus:
Pour Plate technique:
Swab Method
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Micro-organisms in food processing :
Clean in place (CIP) system:
A system used in the preparation, distribution, delivery, and subsequent removal of cleaning solutions to soiled equipment and piping systems without dismantling them.
� CIP cycle: The executed recipe of rinses, washes, and air blows used to clean soiled equipment.
� CIP circuit: The sum of paths within a process unit operation that are cleaned as part of a single CIP cycle.
� Variables to be considered while designing CIP System:
(1) Time of exposure (contact time) to wash and rinse solutions
(2) CIP flow rate (3 - 5 ft./Sec)
(3) Temperature of wash and rinse solutions
(4) Chemical concentration of wash solutions
� Efficiency of CIP Design:
� Spray ball test – Riboflavin test.
� Cleanability – Checked using the full CIP protocol at the facility where the vessel is installed including cleansers and temperatures
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Micro-organisms and their role in-
• Milk Powder technology.
• Beverages and viscous foods.
• Vegetable oil technology.
– Microbial Profile
– Microbial Criteria
– Recommended CIP Cycle.
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Microbial Profile during Milk Powder Production:
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Microbial aspect of Milk Powder Production:
Milk has a pH closer to alkaline range i.e. 6.4-6.9 (favorable for microbial growth) and it is a rich source of nutrients like carbohydrates , proteins , water an excellent media for microbial growth ,hence its CIP is very critical.
Typical CIP Program in a Milk Processing plant:
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Microbial aspect of Milk Powder Production:
Microbiological limit for Milk Powder (Reference: Microbial standards specified by Codex Commission (1983),ICMSF-International Commission for Microbial Specifications for food (1986).
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Microbiology in Vegetable oil processing:
Vegetable oil processing involves very low use of water and veg oils have a low Aw hence microbial contamination is a low concern in these plants.
Typical CIP in Vegetable oil Plant: Only Deodorizer and Alfa Laval VHE Economizer (High steam sparging heat exchanger) is CIP’ed with Water pre-rinse+caustic soln of 8-10%+Hot water final rinse at temp 70 to 80 degree C ,each cycle being of 5-10 minutes.
Frequency-Once in a year.
Microbiological limit for Fats & oil (Reference: Microbial standards specified by Codex (1983),ICMSF(1986):
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Microbiology in Beverages & Viscous foods:
Pasteurization: Partial sterilization of foods at a temperature that destroys harmful microorganisms without major
changes in the chemistry of the food.
Sterilization: Total destruction of all microorganisms (whether or not pathogenic) and their spores
Destruction of all micro-organisms and spores in viscous foods, involves sterilization, usually at temperatures between 110°C and 115°C for 60-90 seconds.
Alfa Laval supplies sterilizer (Steritherm) or pasteurizer units (SS PHE) for viscous fruits or vegetable pulps or
juices .It is mostly aseptic process (Asepto) with zero chance of contamination due to no contact with air or water.
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Microbiology in Beverages & Viscous foods:
� Fruit and vegetable juices or pulps are low acid foods, mainly harboring acidophiles (pH < 5.4)
such as Clostridium botulinum, which are completely destroyed in sterilization
� CIP in Beverage plant: CIP Cycle involves Hot water pre-rinse+caustic soln of 2-5 % + a final rinse at temp 70 to 80 degree C. Frequency-After each production batch.
� Microbial testing - Total plate count post sterilization and cooling: Standard- Not detectable or 5-10cfu/100gm.
� Fruit Juice Pasteurizer:
� Pasteurization of juice not only destroys spoilage organisms, but also inactivates the pectic enzymes, which cause chemical changes. Lactic acid and acetic acid bacteria do not grow in juice at temperatures above 54°C, and temperatures above 71°C are sufficient to destroy most other spoilage organisms including most fermentation organisms.
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Designing a Microbiologically safe Equipment & Plant :
A.) Drainablity:
� Equipment like tanks , vessels, hoppers, with discharge openings must be fully drainable .For good drainablity & cleanability sharp corners must be avoided. Horizontal surfaces must have a slope of > 3 °
� Designing properly sloping floors to drains (¼ inch per foot)
� Design floor drains to prevent the accumulation of water in or around the drains and making drains accessible for cleaning with drain covers to avoid pest entry.
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Designing a Microbiologically safe Equipment & Plant :
B.)Top Rims and equipment covers:
� Top rims of tanks, hoppers etc. must avoid ledges or crevices where product can lodge & are difficult to clean.
� Covers should be completely detachable for cleaning, if hinged covers are used, the hinge must be designed
such that hinge is easily cleanable & accumulation of dust/dirt is avoided.
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Designing a Microbiologically safe Equipment & Plant :
C.)Smooth food-contact surfaces:
� There should not be any unreachable crevices which can be formed by unsealed joints, tack welds &threaded fittings. Ideally product contact surfaces should have surface finish of 0.8 µm Ra or better.
� All food contact surfaces be smoothly bonded (e.g., free of pits, folds, cracks, crevices, open seams, exposed threads, and piano hinges) to avoid harboring pathogens.
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Designing a Microbiologically safe Equipment & Plant :
D.)Installation:
� Elevating food-contact surfaces sufficiently above the floor or away from wall, ideally 300 mm for cleaning & inspection.
� Un-clad framework should be constructed with hollow square or round section members.
� Supports for piping or equipment must be fabricated & installed such that no water or soils can remain on surface or within the supports.
� Insulation must be clad with Stainless steel which must be fully welded.
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Designing a Microbiologically safe Equipment & Plant :
F.)Piping:
• Dead leg : An area of entrapment in a vessel or piping that could lead to contamination.
Dead legs will be measured by the term L/D, where L is the leg extension from the I.D. wall perpendicular to the flow pattern or direction, and D is the I .D. of the extension or leg of a tubing fitting or the nominal dimension of a valve or instrument.
As a standard currently in Hygienic process industry L/D of 2:1 is preferred.
� Cross contamination of product streams shall be physically prevented by:(a) removable spool piece(b) U-bend transfer panel(c) double block-and-bleed valve system –Mix proof valve cluster
� Prevent use of by-pass as much as possible.
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Designing a Microbiologically safe Equipment & Plant :
� Orifice plates, shall be installed in a drainable position.
� Eccentric reducers shall be used in horizontal piping to eliminate pockets in the system.
� Ball valves are not recommended in fluid hygienic piping systems.
� Piping and tubing design should have routing and location priority over process and mechanical support
systems.
� Product hold-up volume should be minimized.
Dead leg in Ball ValvesDead leg at equipment attachment points
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Designing a Microbiologically safe Equipment & Plant :
E.) Layout:
• Designing the plant for one direction of personnel traffic, product, and air flow
• Locating catwalks with open grating so they do not pass over areas of food-contact
surfaces.
� References:
• EHEDG Standards for Hygienic design for food processing equipment.
• ASME-BPE for Bioprocessing Equipment.
• 3-A Sanitary Standards and 3-A Accepted Practices.
• Essentials of Food Science - 3rd Ed Springer 2007
• European Union (EU) Directives and Statutory Commission of England & Wales.
• Giese J. Ultra pasteurized liquid systems earn 1994 IFT Food Technology Industrial Achievement
Award. Food Technology 1994; 48(9): 94–96.
• Department of Food Science, North Carolina State University, Raleigh, NC.
www.alfalaval.com© Alfa Laval Slide 25
FERMENTATION PROCESS
What is Fermentation?
• Fermenation is the process in which complexorganic substance (molecules) is converted in tothe simpler organic substance (molecules) by theaction of enzymes secreated by the micro-organisms.
–Micro-organism+substrate= more microbial cells+ metabolic products
Two types:- -Aerobic fermentation
�Production of Acetic Acid, citric acid
- Anaerobic fermentation
�Alcohol,acetone,butanol,lactic acid
www.alfalaval.com© Alfa Laval Slide 26
a. Rapid growth in suitable organic substrates with easy cultivation in large quantities.
b. Ability to maintain physiological costancy under fermentation condition and to produce necessary enzymes readialy in order to bring about desired chemical changes.
c. Ability to carry out the transformations under simple and workable modifications of enviornmental conditions.
Characterstics of M.O
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Selection of Micor-organism
-Carry out desired reaction efficienty.
-Stock cultures
-Individual organism may be isolated from natural sources
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Industrially important micro –organisms may be classified under yeast,bacteria & molds
Micro-organism Uses
YEAST BACTERIA MOLDS
Ethyl Alcohol Lactic Acid -lactobacillus
Penicillin
Backers Yeast Acetone &Butanol-Clastidium
Streptomycin
Beer Vinegar&-Acetobactor
Whisky Dextran-Leconostoc
Wine (Saccharomycesellipsoidens)
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Industrially important yeasts
© Alfa Laval Slide 29
YEAST PRODUCT
Distiller’s Yeast:-Saccharomyces cerevisiae
Ethyl Alchol, potable liquors, backers yeast, pharmaceutical yeast, Invertase, Flavors enhancing agents
Schizosaccharomyces pombe Ethyl Alcohol (Alfa Laval Fermetation)
Saccharomyces cerevisiae & Saccharomyces carlsbergensis
Beer_Brewers Yeast
Saccharomyces cerevisiaevar.ellipsoideus
Wine
Candida species Food & fooder yeast
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YEAST
• They are generally defined as fungi, which in astage of their life cycle occurs as a single cells,reproducing commonly by budding or lessfrequently by fission
• They are >1500 strains of yeast, 500 species & 50genera.
• Asexual reproduction of yeast- doubling 1 to 2hours
• Occurrence:- nature soil, marine, many forms oforg-matter-particulearly with carbohydrates.
• Isolation:- Soil of wine yards, fruits etc© Alfa Laval Slide 30
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Yeast• Shapes :- Ovidal, elipsoid,
Spherical
• Size :Depends upon
– age/culture conditions etc.
– Width 1 to 9 mic,length 2-10 mic
– Much larger than bacteria.
– Budding:-
– Fission:-
© Alfa Laval Slide 31
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Yeast structure & function of cellular components-Cell wall:- Responsible for the shape
Bud scares are formedCan produce avg.24 buds
Schizosaccharomyces – reproduce by fission,exibit scare rings at the point where crsoo-wall divide after septum fomation.
Cytoplasmic membrance:- betn cell wall layers & cytoplasm
Nucleus :- Generic & Heriditarycontrol.
Vacuoles :- Contains amino acids
Endoplasmic Reticulum:-
Mitorchondria:- Power house of the cell.
Lipid globules:-
© Alfa Laval Slide 32
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Growth requirements of yeast
• Other organic carbon sources:-
– Nitrogen
– Aminoacids
– Other organic sources
– Vitamins
– Minerals
Carbohydrates (carbon &
Engergy sources)
Monosaccharides:- All yeast are
able to metabolise
D-glucose,D-fructose & D-mannose
Di,tri & polysaccharides are first hydrolysed to their respective hexose & then metaboliszed,Hydrolysis is carried out only if the species can utilise the hexose & then metabolised,
Glucose,fructose,Cellulose and melezitose
Glucose,fructose,mannose, sucrose,galactose, maltose,raffinose,mellibiose,lactose & trehalose are most commonly utilised sugars© Alfa Laval de 33
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Fundamentals of yeast growth(Facultative-Vegetative re-production, td=2hrs.)
Aerobic-Respiration
• EMP-TCA-Oxidative Phosphorylation(Mito.)
• O2 is final e acceptor
• Complete degradation
• 36ATP/glucose
• Cell membrane with UFA & sterols (Nicotinic acid)
Anaerobic- Fermentation
• Glycolytic(EMP) Pathway (cytoplasm)
• EtOh is final elec.acceptor
• Partial gedradation
• 2ATP/glucose
• Weak cell membrance
© Alfa Laval Slide 34
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Aerobic & Anaerobic growth
Aerobic(respiration) Anaerobic (fermentation
© Alfa Laval Slide 35
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Metabolism of yeast
Glycolytic Pathway(EMP)
© Alfa Laval Slide 36
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Metabolism of yeast
© Alfa Laval Slide 37
SUCROSE
INVERTASE
GLUCOSE+
FRUCTOSE
EMP
C2H5OH+CO2
• How Yeast is working
• - Sucrose (Invertase)
• - Glucose
• EMP pathway
• C2H5OH+Co2+Heat
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Basic Chemistry of Ethanol
C12H22O11+ H20 → C6H12O6+ C6H12O6 → 2(C2H5OH) + 2(CO2)
Sucrose Water Glucose Ethanol Carbon Dioxide
Fermentation(anaerobic, glycolysis)
S. cerevisciae
342 mw 18 mw 2 x 46 mw 2 x 44 mw
S. Pombe
Fructose
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The Wine making process
© Alfa Laval Slide 39
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The beer-brewing process
© Alfa Laval Slide 40
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DISTILLERY
© Alfa Laval Slide 41
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TYPES OF FERMENTATION IN DISTILLERY• MOLASSES BASED - BATCH TYPE
- CONTINUOUS FERMENTATION
» ALFA LAVAL CONTINUOUS FERMENTATION-BIOSTILL
» CASCADE TYPE
» SEMI CONTINUOUS
• GRAIN/STARCH BASE - BATCH TYPE
© Alfa Laval Slide 42
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Typical view of distillery plant
© Alfa Laval Slide 43
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Continuous fermentation plant
© Alfa Laval Slide 44
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GRAIN BASED FERMENTATION-BATCH TYPE
Ferm. #1 Ferm. #2 Ferm. #3 Ferm. #4
Mash from
Liquefaction
Yeast
Propagator
Mash
Coolers
To Beerwelland Distillation
Spirizyme Fuel/Plus
Tech.,Yeast
Water
CO2 to Scrubber
90 - 93oF (32 – 34 C)pH 3.8 - 5.0
88 - 90oF (31 - 32C)
pH 3.6 - 4.0
Batch
Fermentation
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Molasses based fermentation
© Alfa Laval Slide 46
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The Molasses Process
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Propagation System :
100/ 1000 ML
90 Liter2000 Litre
�Complete Sterile operation
�Propagation under controlled conditions: Air, Temp, Pressure
�Minimum cell count of 250 X 106 per ml in each stage
�Normally takes 10 to 12 hours in each stage
LAB Prop I Prop II
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100/ 1000 ML
Propagation System : Alfa Laval System
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Air
Cooling WaterOut
Mash Headerto Fermenters
Cooling Water In
Steam
Process WaterMash
Yeast Product Yeast Propogation ion ProcessYeast Propagation
yeast
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FERMENTATION PROCESS
© Alfa Laval Slide 51
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Parameters for Control :
Yeast Cell Count
Checked under microscope
Controlled by the air flow rate
to the fermenter
Residual Sugars
Checked by titration
Control of unfermented sugars
in molasses
Fermenter Dissolved Solids
Checked by Oven Maintained for check on infection
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Parameters for Control….
Yeast + Sludge concentration
Checked by spin test in lab centrifuge
To have better performance on Yeast Separation
Alcohol Concentration
Checked by Ebulliometer / Distillation
To have consistent feed to distillation plant
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Yeast Recycling1. Maintains cell concentration
(Efficient conversion)
1. Saves Sugars (More Ethanol Yield)
2. Quick conversion to ethanol (Less residence time)
Exhausted Wash Recycle
1. Maintains dissolved solids concentration
(Fights infection)
2. Decreases water requirement (Savings in Water)
3. Decreases spent wash quantity (Less effluent problem)
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Parameters Batch Fermentation
Cascade
4 Ferm. Sys
Alfa Laval cont.
2 Ferm System
Fermentation Efficiency
84% to 85% 86% to 87% 90% guaranteed
Yield at 46%FS @ 96%v/v
258 Liters/Mt 261 Litres/Mt 273 Litres/Mt
Sturdiness Prone to infection
Prone to Infection 100% infection free
Fermenter Alcohol %v/v
7.5 to 8.5% 7.5 to 8% 7.5 % - 8.5 %
Fermenter DS%w/w
9 to 10% 10 to 11% 14 to 16%
Yeast Count 100 X 106 100 X 106 500 X 106
Effluent Generation
15 Litre/Litre 10 to 12 Litre/Litre 5 Litre/Litre
Technology Comparison:
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Continuous Fermentation Plant
Flexibility: the GREATEST Advantage of Alfa laval Process�Raw Material : Molasses
�Process Water : NO treatment
�Yeast Strain : Schizosaccharomyces Pombe OR Saccharomyces Cerevisease.
�Fermenter Alcohol: 6.5 to 8%
�Dissolved Solids : 10% to 16%
�Spent Wash Generation: 11 – 5 litres
�Turn down ratio : below 40%
�Shutdowns & Start-up’s : Even for period of 15 days
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Stress factors affecting yeast & fermentation efficiency.-Sugar Concentration
-PH
-Temperature
-Alcohol Tolerance
-Neutrients
-Yeast concentration
© Alfa Laval Slide 57
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Bacterial Contamination
What Happens if the Fermenter is Infected?
Effects�Low yield�High impurities –fusel oil formation�Bad quality of product�Excessive foaming�Loss in production due to stoppages.�……
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Bacterial Contamination
• SOURCES
• Stock culture
• Raw Material
• Process water
• Environment
• Air
• Additives
• Hygienic conditions
• Defective designing
• Yeast & Bacteria
© Alfa Laval Slide 59
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Common contaminants in yeast fermentationsContaminants Effects
Acetobacter Prod.of Acetic Acid,Robiness
Lactobacillus Production of lactic acid,Mousy occur
Leconostoc mesenteroidesLeuconostoc dextranicm
Production of Dextran, a Polysaccharide from Sucrose
Saccharomyces pastorianus Turbidity in Beer
Saccharomyces turbidians Turbidity in Beer
Candida mycoderrna Formation of film on the surface, Breakdown of alcohol,organicacidsand other substances
Achromobacter anaerobicum Turbidity,Off flavour
Streptococci Sourness,turbidity,Ropiness
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Bacterial Growth Path
Bacteria Vs Yeast• Normally Bacterial
growth is faster than the Yeast.
• Temp factor
© Alfa Laval Slide 61
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Methods to control Contamination• Find out sources- Like Molasses,water,air supply
• Good yeast culture
• Propagation process errors/backers yeast.
• Unattended fermentation process.
• Unhygienic conditions: Immediately complete cleaning of
spillage.
• Sterilization of mash – practically not possible but during
propagation stages proper sterilization & care needs to be taken
• Good microbiological practices- Keen observation on the
process in microbiological point of view, like microscopic observation.
• CIP
• GOOD DESIGNS © Alfa Laval Slide 62
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Methods to control Contamination
© Alfa Laval Slide 63
“Cleaners bag of tricks” – Four T’s of cleaning
& Sanitizing.
-TIME
- TEMPERATURE
- TITRATIONS
-TURBULENCE
CIP- Cleaning in Place
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Methods to control Contamination
© Alfa Laval Slide 64
CIP- Cleaning in Place
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Designs Precautions
• Yeast propagators – good finishing
• Fermenters- finished surface
• Drainage layout
• Dead spots/crevices
• Piping/valves- no retention
• Sources include, but not limited to
– Tanks
– Transfer lines
– Heat Exchangers
© Alfa Laval Slide 65
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Turbulence
• Effective cleaning requires velocity min 1-1.5 to 3 m/s. forward flow & backward flow
• Flow 1.5 times the normal process flow.
• Dead ends
• Bypass valve© Alfa Laval Slide 66
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Cont….
• Elimination of dead end- branch lines from the
main line should have the valve installed not more than one dia form the wall of main pipe.(ensurence turbulancein branch)
© Alfa Laval Slide 67
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• Tees : Dead end should be minimum.
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Fermenter Design
• Mash Filling Pipe line loop
- min velocity should be 3m/s
© Alfa Laval Slide 69
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• Fermenter CIP Sprayhead(pressure requirement)
© Alfa Laval Slide 70
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• External Heat Exchanger loop
- 1.5 times normal process flow.
© Alfa Laval Slide 71
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• Fermented product emptying pipeline.
– Min 3m/s velocity.
© Alfa Laval Slide 72
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Thank you!
© Alfa Laval Slide 73