EXPERIMENTAL%20DESIGN%20FOR%20IMPROVEMENT%20OF%20FERMENTATIONS-%20PLACKETT-BURMAN%20DESIGNS

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EXPERIMENTAL DESIGN FOR IMPROVEMENT OF
FERMENTATIONS-PLACKETT-BURMAN DESIGNS

• JASMN LABORATORY,JERF
• PERUNGUDI, CHENNAI- 97

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• MICROBIOLOGIST
• MICROBE
• MEDIUM
• SET OF CONDITIONS
• TEMP
• pH
• METABOLITE/ACTIVITY OF INTEREST
• ANTIBIOTIC
• ANTICANCER AGENT
• ENZYMES

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INTRODUCTION

• WE UNDERSTAND MANY THINGS IN MICROBIAL PHYSIOLOGY
  AND MOLECULAR BIOLOGY
• BUT IMPROVEMENT OF FERMENTATIONS REMAINS LARGELY
  ON EMPERICAL PROCESS
• THE RELEVANT LITERATURE AND PRIOR TECHNIQUES
  SERVE ONLY AS A STARTING POINT

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• WHEN A NEW STRAIN IS ISOLATED AND THE DETAILS OF
  IT IS NOT WELL KNOWN, EMPERICAL APPROCHES TO
  FERMENTATION IMPROVEMENT ARE THE ONLY WAY
• THESE ARE VERY COMMON IN INDUSTRIAL SETTINGS
• EXPERIMENTAL DESIGNS AND STATISTICAL ANALYSIS
  WILL HELP IMPROVE FERMENTATIONS

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• MICROBIOLOGIST STARTS WITH SOME MEDIUM AND SET OF
  CONDITIONS FOR THE EXPRESSION OF METABOLITE OR
  ACTIVITY OF INTEREST.

• MICROBIOLOGIST
• MICROBE
• MEDIUM
• SET OF CONDITIONS
• TEMP
• pH
• METABOLITE/ACTIVITY OF INTEREST
• ANTIBIOTIC
• ANTICANCER AGENT
• ENZYMES

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• NEXT TASK IS TO IMPROVE THE EXPRESSION TO A LEVEL
  SUFFICIENT FOR THE ISOLATION AND CHARACTERIZATION
  OF THE DESIRED PRODUCT

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Fermentation improvement programme- begins with

• Measuring the product yield
• In response to factors
• Medium strength
• Increase conc. by 25
• Incubation temperature
• Control of pH
• Replace original carbon and nitrogen source
• Alternative source of P and S may be considered

• Classical method
• One factor changed at a time when others held
  constant

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Example single factorial design

• Microbe ? Antibiotic
• Grown on 4glucose 2 Soybean flour?150 µg/ml
• First test for alternative carbon source
• Glucose is replaced by glycerol
• 2 glycerol2 Soybean flour? 180 µg/ml

• Next Soy bean is replaced by cotton seed
• 4glycerol 2cotton seed ? 200 µg/ml
• Based on this further study will be carried out

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NATURE OF THE STUDY

• THIS STUDY IS CALLED NONNUMERIC OR CATEGORICAL
  COTTON SEED/ SOYABEAN
• ANOTHER STUDY IS KNOWN AS NUMERICAL OR
  CONTINUOUS
• 2CS, 4CS,8CS ETC
• 2SB,4SB,8SB,ETC

• A FACTOR IS SAID TO BE CATEGORICAL IF ITS FACTOR
  LEVELS OR SETTINGS , REPRESENT DISJOINT ENTITIES
  WHICH FOLLOW NO NUMERICAL HIERARCHY
• EX COTTON SEED, SOY BEAN ETC

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FULL FACTORIAL DESIGN

• WHEN CATEGORICAL FACTORS NEED TO BE TESTED, A
  FULL FACTORIAL IS THE EXPERIMENTAL DESIGN OF
  CHOICE
• (EX EFFECT OF Glucose, lactose, NO3, NH3 ETC)
• AS THE NUMBER OF FACTORS INCREASES, FULL
  FACTORIAL DESIGN RESULTS IN EXESSIVELY LARGE
  NUMBER OF EXPERIMENTAL TRIALS

• MOST EXPERIMENTAL SITUATIONS DEAL WITH CONTINOUS
  FACTORS WHICH ARE AMEABLE TO SCREENING DESIGNS
  REQUIRING FEWER TRIALS.
• EXPERIMENTS CONTAINING BOTH CATEGORICAL AND
  CONTINUOUS FACTORS CAN BE DESIGNED

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NUTRITIONAL FACTORS IMPORTANT IN COMMERCIAL
FERMENTATION
FACTORS RANGE OF FACTOR SETTING CON. (WT/VOL) EXAMPLES
CARBON SOURCE(S) 0.5-20 GLUCOSE,SUCROSE, STARCH, WHEY CORN MEAL, GLYCEROL, ALCOHOL, LIPIDS
NITROGEN SOURCE(S) 0.1-10 AMMONIA GAS, AMM.SALTS, CASEIN HYDROLYSATES, CORN GLUTEN MEAL, CORN STEEP LIQUOR, COTTON SEED FLOUR, FISH MEAL, GLUTAMIC ACID, NITRATES, PEPTONES, SOYBEAN FLOUR, UREA, WHOLE YEAST, YEAST EXTRACT
PHOSPHORUS 0.1-2 CORN STEEP LIQUOR, PHOSPHATES
SULFUR 0.1-1 METHIONINE, PROTEINS, SULFATES
OTHER NUTRIENTS lt1 Iron salts, Mg salts, oxygen,K, trace elements, vitamines
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For further details

• Contact
• JASMN laboratory
• 9444117141