LABORATORY MANUAL BTY325 BIOPROCESS ENGINEERING LABORATORY

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TABLE OF CONTENTS

S. No. 1

Title of the Experiment To isolate lactic acid bacteria from curd

Page No.

2 3 4 5 6 7

Quantification of Cellulase Enzyme. Determination of Thermal Death Point Determination of Thermal Death Time To study the working and construction of fermenter Study of rheology of viscous polymers and microbial cultures Determination of mixing time in a fermenter

36 7 10 11 14 15 18 19 21 22 24 25 27 28 30

Measuring DO concentration in a fermenter 8 Determination of volumetric mass-transfer coefficient (KLa ) in a fermenter Production of alcohol using S. cerevisiae

9 10

31 35 36 39

Text Books:1. Aneja K. R., Experiments in Microbiology, Plant Pathology and Biotechnology, 4th edition, New Age international Ltd. References:2. Bioprocess Engineering Principles by Pauline M. Doran, Elsevier, 1st Edition (1995)

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Experiment No.1 1. Experiment: To isolate lactic acid bacteria from curd Equipment Requirement: Incubator, laminar air flow, autoclave. Material Requirement: inoculating loop, petri plate, MRS media, curd sample. 2. Learning Objective: a. To isolate lactic acid producing microorganisms from curd sample. 3. Outline of the Procedure: a. b. c. d. e. f. g. h. i. The given curd sample was diluted upto 10-6 dilution. Prepare 100 ml of MRS agar media. Autoclave it and pour into sterilized petri plates, allow it to solidify. Inoculate 1ml of the diluted curd sample by spreading aseptically. Incubate for 24 hours at 37 C. Observe for colony formation. Prepare 100ml MRS broth and set pH 6.5 and autoclave it, allow it to cool down. Inoculate the broth with colony appeared on MRS agar plate. Incubate for 24 hours at 37 C.

Determination of lactic acid from standard curve. a. For a standard curve we added different concentration of lactic acid to test tubes that is 20, 40, 60 and 80 l respectively and 1 test tube for control in which lactic acid is absent. b. Make up the final volume to 1000l by help of distilled water. c. Add conc.H2SO4(3ml) in each test tube and mixed on a vortex mixer. d. Add 0.5 ml of CuSO4 reagent(4% w/v ) and add 0.5 ml of p-phenylphenol(1.5% w/v in 99% ethanol) and mix well on vortex mixer. e. Leave the tubes for 15 min at room temperature. f. Mix well and read the absorbance at 570nm. 4. Required Results: Parameters: Compare the mixing efficiency at different rotation speed by using plot mixing time versus rotation speed. Plots: Draw a plot of mixing time versus rotation speed. Relationships: N.A. Error Analysis: N.A.

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5. Cautions: a. Prepare 4% w/v CuSO4 reagent 1 day prior to requirement. Requirement for Lab Technician: 1. Copper sulphate. 2. Sulphuric Acid (99%) 3. MRS Broth 4. MRS agar. 5. Curd sample. 6. p-phenyl phenol 7. Ethanol (99%)

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Worksheet of the student Date of Performance Registration Number:

Aim: To isolate lactic acid producing microorganisms from curd sample. Observation Table1: Latic acid concentration S. No. Water Lactic acid l l H2SO4 H2SO4 (98%), CuSO4 w/v), 0.5 ml (4% p-phenyl OD phenol (1.5% w/v in 99% 570nm ethanol) 0.5 ml

1 l=2.54 g 3ml

1. 2. 3. 4. 5. 6. 7.

1000 980 960 940 920 -

0 20 40 60 80 Sample1 1(((10) sample2

Graphs/ Plot : plot a graph between O.D. and Lactic acid conc. Take O.D. on Y-axis and Lactic acid conc on X-axis.

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Calculations: Conc. Of lactic acid in sample = O.D./ slope. Conversion formula : 1 l=2.54 g Result and Discussion

Error Analysis

Learning Outcomes (what I have learnt)

To be filled in by Faculty S.No. Parameter (Scale from 1-10, 1 for very poor and 10
excellent)

Marks obtained

Max. Marks 20 20 10

1 2 3

Understanding of the student about the procedure/apparatus. Observations and analysis including learning Outcomes Completion* of experiment, Discipline and Cleanliness Signature of Faculty

Total marks obtained

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Experiment No.: 2 1. Experiment: Production of Enzymes [Cellulase] using Microorganisms Quantification of Enzyme. Equipment Required Sterile Petri plates (2) Inoculating loop/ needle Glass rod Bunsen burner Wax marking pencil Materials Required Nutrient-agar plate cultures of B. cereus Potato-dextrose agar plate culture of T. viridae Modified Czapek-mineral salt medium(40ml) Carboxymethyl cellulose(CMC) Hexadecyltrimethyl ammonium bromide (1% solution)

2. Learning Objectives To induce and detect cellulose production by cellulolytic microorganisms 3. Outline of Procedure a. Preparation of the medium prepare 1 liter of the mineral Czapek-mineral salt agar medium whose constitutes are as followsSodium nitrate Potassium phosphate Magnesium sulphate Potassium chloride Carboxymethyl cellulose Peptone Agar Distilled water 2.0 g 1.0 g 0.5 g 0.5 g 5.0 g 2.0 g 20.0 g 1000 ml

b. Dissolve the agar in 400 ml of hot distilled water by adding in small amounts and stirring with a glass rod c. Dissolve the magnesium sulphate, Potassium chloride, peptone, sodium nitrate in 200 ml of water d. Dissolve potassium sulphate in 100 ml of water. e. Dissolve CMC in 200 ml of water with heat and mix in a warring blender f. Mix all the solutions and make up to 1000 ml volume g. Adjust the pH of the medium to 6.6 with the addition of acid or alkali. h. Dispense the medium in 250 ml c o n i c a l flasks plug them and autoclave a t 15 lb/in (121C) for 15 minutes i. Pour the autoclaved medium cooled to 45-50C into sterile Petri plates j. Allow the medium to solidify. k. Label the plates each with the organism to be inoculated. l. Inoculate the appropriately labeled plates with the respective organism.
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m. Incubate inoculated plates at 35C in an inverted position for 3-5 days. n. Flood the plates with 1% aqueous solution of hexadecyltrimethyl ammonium bromide. 4. Required Results: Parameters: Observe the plates for the formation of a zone around the growth. A clear zone will be observed around the colonies of B. cereus and T. viridae indicating degradation of CMC by the production of extracellular enzyme, i.e., cellulase (or CMCase). Relationships to be determined: N.A. Graphs/Plots: N.A. Error Analysis: N.A. 5. Cautions While preparing Czapek-mineral salt medium, phosphate should always be dissolved separately and be added to the mixture at the last. Requirement for Lab Technician: 1.Nutrient-agar plate cultures of B. cereus 2.Potato-dextrose agar plate culture of T. viridae 3. Modified Czapek-mineral salt medium(40ml) 4. Carboxymethyl cellulose(CMC) 5. Hexadecyltrimethyl ammonium bromide (1% solution)

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Worksheet of the student Date of Performance Registration Number:

Aim: To induce and detect cellulase production by cellulolytic microorganisms Write the principle of experiment in your own words.

Calculations: Result and Discussion

Error Analysis

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Learning Outcomes (what I have learnt)

To be filled in by Faculty S.No. Parameter (Scale from 1-10, 1 for very poor and 10
excellent)

Marks obtained

Max. Marks 20 20 10

1 2 3

Understanding of the student about the procedure/apparatus. Observations and analysis including learning Outcomes Completion* of experiment, Discipline and Cleanliness Signature of Faculty

Total marks obtained

10

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Experiment 3. 1. Experiment: Determination of Thermal Death Point (TDP). Equipments Required: Water bath, Laminar hood, Autoclave, Sterile empty tube with cup, Sterile cotton plugged 10 ml pipettes, Wax glass marking pencil Culture tube stand, Culture tube with 10 ml water, Centigrade thermometer Materials Required: Nutrient broth cultures of Bacillus subtilis (30-48 hours culture taken in flasks), Czapek-Dox spore suspension of Aspergillus niger (3-4 days culture taken in flasks), Nutrient agar plates, Czapek-Dox agar plates. 2. Learning Objectives: 1. To study the lethal effects of temperature on microorganisms and understand its relationship with time and temperature. 2. To determine the TDP of B. Subtilis and A.Niger required for an exposure time of 10 minutes.

3. Outline of Procedure: 1. Divide the bottoms of the 2 sterile nutrient agar plates into six sectors with the glass marking pencil. 2. Repeat procedure 1 for the Czapek Dox agar plate. 3. Label the 6 sectors of each plate as C, 40C, 50C, 60C, 70C, 80C. 4. Label bottom of nutrient agar plate with B.subtilis and Czapek-Dox plate with A.niger. 5. Inoculate each experimental organism in the sector marked C on the plates into its appropriately labelled plate by means of a streak inoculation. 6. Line up 5 sterile test tubes for each organism in a culture tube stand and label 15 tubes with E. Coli and temperatures 40,50,60,70,80C. 7. Repeat step 6 for B.subtilis and A.niger. 8. Aseptically transfer 2 ml cell/ spore suspension of each test organism into its appropriately labeled tubes using sterile pipettes 9. Suspend one each tube of E. coli, B. subtilis, and A. niger in the 40C warer bath ( be sure that the water bath is above the broth level) and keep for 10 minutes at a constant 40C. 10. Inoculate 40C sectors in each plate with a straight streak immediately after removing the tubes from the water bath and discard 0C C broth tube.
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11. Repeat the steps 9 and 10 for the remaining temperatures (50, 60, 70 and 80C) and make inoculations into the respective sectors. 12. Incubate the bacterial inoculated plates at 30 degree C for 24-48 hours and fungal inoculated plates at 25C for 3-4 days or until growth occurs in the control sectors in an inverted position. 4. Required Results: Examine the plates for the presence (+) or absence (-) of growth in all the sectors and compare with control. Parameter-NA Relationship to be determined-NA Graph/plots-NA Error analysis-NA 5. Cautions: a. While making streak inoculation of an organism, the inoculum should be at least 1-2 cm away from the plate center where the sectors converge. b. Inoculation of an organism may be made on the plate before cooling the broth. c. Constant attention should be given to the temperature of water bath so that adjustment can be made before a serious fluctuation in temperature occurs.

Requirement for Lab Technician: 1. Nutrient broth 2. nutrient agar 3. Czapek Dox Broth 4. Czapek Dox Agar

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Worksheet of the student Date of Performance Registration Number:

Aim: To determine the TDT of B. subtilis and A. niger required for an exposure time of 10 minutes Observation Table1: Estimation of TDP 40C B.subtilis A.niger 50C 60C 70C 80C

Calculations: NA Result and Discussion

Error Analysis

Learning Outcomes (what I have learnt)

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To be filled in by Faculty S.No. Parameter (Scale from 1-10, 1 for very poor and 10 1 2 3 4 5 6 7 8 9 10 excellent) Assistance required by the student while performing 1 experiment(more assistance less marks) Understanding of the student about the 2 procedure/apparatus. Observations and analysis including learning outcomes 3 Completion of experiment 4 Discipline and Cleanliness 5 Signature of Faculty Total marks obtained

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Experiment 4. 1. Experiment: Determination of Thermal Death Time (TDT). Equipment Required: Water bath, Laminar hood, Autoclave Materials Required: Nutrient broth cultures of Bacillus subtilis (30-48 hours culture taken in flasks), Czapek-Dox spore suspension of Aspergillus niger (3-4 days culture taken in flasks), Nutrient agar plates, Czapek-Dox agar plates, Hot water bath, Sterile empty tube with cup, Sterile cotton plugged 10 ml pipettes, Culture tube with 10 ml water, Centigrade thermometer, Wax glass marking pencil, Culture tube stand, Inoculating loop/ needle 2. Learning Objectives: 1. To study the lethal effects of temperature on microorganisms and understand its relationship with time and temperature. 2. To determine the TDT of B.subtilis and A.niger required for a particular temperature. 3. Outline of the Procedure: a. Divide the bottoms of each Petri plate into six sectors with the glass marking pencil. b. Label four sets of plates with the time intervals as 0, 5, 10, 15, 20 and 25 c. Pour melted and cooled nutrient agar medium into two sets and czapek-dox agar into the other two sets and allow the media to solidify. d. Label the bottom of nutrient agar plates with B. subtilis and Czapek-dox agar plate with A. niger. e. Inoculate 0 sector of NA plate with B. subtilis and of CDA plate with A. niger as streak inoculations. f. Suspend one each tube of B. subtilis and A. niger into the water bath set at a particular temperature( as determined in the TDP experiment ) say 60C water bath for 6 minutes (one minute for the temperature of the broth and the water equalize and 3 minutes time of exposure). g. Remove both of these tubes and cool these quickly under tap water. h. Make streak inoculations on sector 3 on NA plate and CDA plate with the B. subtilis and A. niger respectively. i. Replace the tubes in the water bath and allow 1 minute for the broth and water to become equalized before starting the next time interval, i.e. 6 minutes and repeat steps 7 and 8 quickly. j. Incubate the B. subtilis inoculated plates at 30C (24-48 hours ) and A. niger inoculated plates at 25C (3-5 days ) in an inverted position for the period-mention or until growth occurs in the 0 time segments.
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4. Required Results: Parameters: Examine the plates for the presence (+) or absence (-) of growth in all the sectors and compare with control. Relationships: NA Graphs/Plots: NA Error Analysis: Check whether you transferred the correct organism into labeled conical flask by double-checking the name of the organism on the stock culture.

5. Cautions: a. While making streak inoculation of an organism, the inoculum should be at least 1-2 cm away from the plate center where the sectors converge. b. Inoculation of an organism may be made on the plate before cooling the broth. c. Constant attention should be given to the temperature of water bath so that adjustment can be made before a serious fluctuation in temperature occurs.

Requirement for Lab Technician: 1. Nutrient broth 2. nutrient agar 3. Czapek Dox Broth 4. Czapek Dox Agar

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Worksheet of the student Date of Performance Registration Number:

Aim: To determine the TDT of B. subtilis and A. niger required for an exposure time of 10 minutes. Observation Table1: Estimation of TDT TDP B.subtilis A.Niger 0 mins 5 mins 10 mins 15 mins 20 mins 25 mins

Calculations: Result and Discussion

Error Analysis

Learning Outcomes (what I have learnt)

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To be filled in by Faculty S.No. Parameter (Scale from 1-10, 1 for very poor and 10
excellent)

Marks obtained

Max. Marks 20 20 10

1 2 3

Understanding of the student about the procedure/apparatus. Observations and analysis including learning Outcomes Completion* of experiment, Discipline and Cleanliness Signature of Faculty

Total marks obtained

18

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Experiment 5 1. Experiment: To study the working and construction of fermenter. Equipment Required: Fermenter Materials Required: NA 2. Learning Objective: I) To familiarize with the parts and working of a fermenter. 3. Outline of the Procedure: a. The teacher will explain the construction and working of fermenter. 4. Required Results: Hand on training on fermenter and other parameters. Parameters: N.A Relationships: N.A Plots: N.A 5. Cautions: handle the fermenter carefully. 6. Requirement for Lab Technician: NA

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Worksheet of the student Date of Performance Registration Number:

Aim: To familiarize with the parts and working of a fermenter. Draw a well labeled diagram of a fementer

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Calculations: NA Result and Discussion

Error Analysis

Learning Outcomes (what I have learnt)

To be filled in by Faculty S.No. Parameter (Scale from 1-10, 1 for very poor and 10
excellent)

Marks obtained

Max. Marks 20 20 10

1 2 3

Understanding of the student about the procedure/apparatus. Observations and analysis including learning Outcomes Completion* of experiment, Discipline and Cleanliness Signature of Faculty

Total marks obtained

21

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Experiment 6 1. Experiment: Study of Rheology of viscous polymers and microbial cultures Equipment Required: Ostwald Viscometer Materials Required: Thermostat at 25 deg C, microbial culture (fermented broth), chromic acid, pipettes, timer. 2.Learning Objective: I) To determine the viscosity of a microbial culture (fermented broth) in liquid media. 3. Outline of the Procedure: a. The viscosity of the fermented broth is to be measured by Ostwald Viscometer. b. Clean the viscometer with chromic acid and fill with a liquid of known viscosity (water = liquid Place the viscometer in a thermostatically controlled water bath at 30C for 3 minutes. Suck the liquid through the capillary until the surface of the liquid is above the upper gradation line. c. Allow the liquid to flow down and calculate the time required for the meniscus to drop from the upper to the lower gradation line. d. Thoroughly clean the apparatus with chromic acid and wash with distilled water. Repeat the steps 3 4 with the fermented broth (liquid B) and measure the time taken for the drop in meniscus. 4. Required Results: Parameters: B , A = Viscosity of the liquids B and A respectively, where found and A is known. Relationships: B / A = (tB x dB ) / ( tA x dA ) Poise Where tB , tA = The time taken for flow of liquid B and A respectively. dB , dA = The density of liquid B and A respectively. Plots: N.A Error Analysis: Use of Ostwald Viscometer not treated with chromic acid 5. Cautions: a. Attention should be given to the time of flow of fermentation broth. Requirement for Lab Technician: 1. Ethanol 2. Glycerol 3. Nutrient Broth 4. Chromic acid
B

is to be

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Worksheet of the student Date of Performance Registration Number:

Aim: To determine the viscosity of a microbial culture (fermented broth) in liquid media Observation Table1: Determination of viscosity. S.No. 1. 2. 3. Average = Calculations: Average = ta (time in sec) tb (time in sec)

Result and Discussion

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Error Analysis

Learning Outcomes (what I have learnt)

To be filled in by Faculty S.No. Parameter (Scale from 1-10, 1 for very poor and 10
excellent)

Marks obtained

Max. Marks 20 20 10

1 2 3

Understanding of the student about the procedure/apparatus. Observations and analysis including learning Outcomes Completion* of experiment, Discipline and Cleanliness Signature of Faculty

Total marks obtained

24

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Experiment 7. 1. Experiment: Determination of mixing time of a fermenter Equipment Requirement: Fermenter Material Requirement: LB Broth, Ink 2. Learning Objective: To determine the mixing time of any soluble substance in fermenter at different RPM. 3. Outline of the Procedure: a. Put ink into the LB Broth. b. Take reading of time when ink homogeneously distributed into the fermentation media. c. Repeat these steps at different RPM i.e. 100,125,150,200. 4. Required Results: Parameters: Compare the mixing efficiency at different rotation speed by using plot mixing time versus rotation speed. Plots: Draw a plot of mixing time versus rotation speed. Relationships: N.A. Error Analysis: N.A.

5. Cautions: a. Adjust the rotation speed precisely.

Requirement for Lab Technician: 1. LB broth 2. Ink

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Worksheet of the student Date of Performance Registration Number:

Aim: To determine the mixing time of any soluble substance in fermenter at different RPM. Observation Table1: Determination of mixing time S.No 1 2 3 4 RPM 100 125 150 200 Mixing time (in secs)

Calculations: Result and Discussion

Error Analysis

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Learning Outcomes (what I have learnt)

To be filled in by Faculty S.No. Parameter (Scale from 1-10, 1 for very poor and 10
excellent)

Marks obtained

Max. Marks 20 20 10

1 2 3

Understanding of the student about the procedure/apparatus. Observations and analysis including learning Outcomes Completion* of experiment, Discipline and Cleanliness Signature of Faculty

Total marks obtained

27

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Experiment 8. 1. Experiment: Measuring DO concentration in a fermenter Equipment Required: Fermenter, D.O probe Material Required: N.A. 2. Learning Objective: To determine the DO concentration in a fermenter.. 3. Outline of the Procedure: a. Fermenter is filled with nutrient broth (2.5 litre). b. Agitation is given at 400 rpm. c. Calibrate the D.O. probe. d. Start taking readings after every 2 mins interval. 4. Required Result: Parameters: Estimate the value of D O. Plots: N.A. Relationships: N.A. Error Analysis: N.A. 5. Cautions a. Attention should be given to the readings of dissolved oxygen concentration.

Requirement for Lab Technician: 1. Nutrient broth.

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Worksheet of the student Date of Performance Registration Number:

Aim: To determine the hardness of the given hard water sample by EDTA method. Provided Standard Hard Water (1 ml of SHW =1 mg of CaCo3). Observation Table1: Standardization of EDTA S.No Time (in mins) D.O conc

Calculations: Result and Discussion

Error Analysis

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Learning Outcomes (what I have learnt)

To be filled in by Faculty S.No. Parameter (Scale from 1-10, 1 for very poor and 10
excellent)

Marks obtained

Max. Marks 20 20 10

1 2 3

Understanding of the student about the procedure/apparatus. Observations and analysis including learning Outcomes Completion* of experiment, Discipline and Cleanliness Signature of Faculty

Total marks obtained

30

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Experiment 9. 1. Experiment: Determination of volumetric mass-transfer coefficient (KLa) in a fermenter. Equipment Required: Fermenter Material Required: 0.5M Na2SO3, 1% starch solution, 0.1M Na2S2O7, 0.003M CuSO4, 5H2O, Standard Na2SO3 (0.1M to 0.5M), 0.1M Iodine solution (20 g/l KI + 12.7 g/l I2) 2. Learning Objective: To determine the KLa in a fermenter.. 3. Outline of the Procedure: a. Fermentor is filled with 0.5M Na2SO3 and 0.003 M CuSO4. 5H2O (2.5 litre). b. Agitation is given at 400 rpm. c. After the aeration begins collect samples every 10 mins. d. Estimate the concentration of Na2SO3 by titration. Burette solution: 0.1 M Na2S2O7. Flask solution: 500l of Na2SO3 solution (sample or standard) + 15 ml iodine solution + 50l starch. e. During titration colour changes to straw yellow. Add the starch solution at this point only and not before. f. Estimate the concentration of Na2SO3 in sample from standard curve. 4. Required Result: Parameters: Estimate the value of kLa. Plots: The volumes of thiosulphate are plotted against sample time. OTR is obtained from the slope of the plot OTR= KLa (C* -CL) where C* is the saturated [D.O.] As O2 enters solution, it is immediately consumed so that OTR = KLa. C* i.e. CL = 0 Relationships: N.A. Error Analysis: N.A. 5. Cautions a. Attention should be given to the readings of dissolved oxygen concentration.

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Requirement for Lab Technician: 1. Sodium sulphite 2. Copper sulphate pentahydrate 3. NaSO 4. Starch 5. Potassium iodide 6. Iodine

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Worksheet of the student Date of Performance Aim: To determine the KLa in a fermenter. Observation Table1: Estimation of Kla S.No Time (in mins) Vol of Na2S2O7 consumed (in ml) Registration Number:

Graphs/ Plots : The volumes of thiosulphate are plotted against sample time

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Calculations:

Result and Discussion

Error Analysis

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Learning Outcomes (what I have learnt)

To be filled in by Faculty S.No. Parameter (Scale from 1-10, 1 for very poor and 10
excellent)

Marks obtained

Max. Marks 20 20 10

1 2 3

Understanding of the student about the procedure/apparatus. Observations and analysis including learning Outcomes Completion* of experiment, Discipline and Cleanliness Signature of Faculty

Total marks obtained

35

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Experiment No.: 10

1. Experiment: Production of alcohol using S. cerevisiae Fermentation Equipment required:Autoclave Laminar air hood Mortar and Pestle Weighing balance Funnel Distillation unit. Filter Paper Bottles Corks Hot Plate Measuring Cylinder

Material required:Inoculum:25g fresh yeast or 15 g of dried yeast Medium 1 kg sugar 40ml (8 tsp) ground ginger Juice of 2 lemons Distilled Water 2. Learning Objectives:To demonstrate fermentation by producing ginger beer from sugar and ground ginger. To estimate the amount of alcohol produced. 3. Outline of the Procedure:Starter Plant a. b. c. d. Put the yeast into a large clean jar. Pour in 275 ml warm water. Stir in 10 ml (2 tsp) sugar and 10ml (2 tsp) ground ginger. Cover and leave in a warm place for 24 hours. This is the starter plant.

Feeding the Plant a. On each of the following 6 days feed the plant with 5 ml (1 tsp) sugar and 5 ml (1 tsp) ground ginger. Stir and cover the jar each time. b. After the last addition leave the solution to stand covered for another 24 hours. c. Line a sieve with muslin. Strain the solution reserving both the liquid and the sediment. d. Over a low heat dissolve 900 g (2 lb sugar) in 575 ml water. Stir well. e. When the sugar has dissolved bring to the boil and boil for 3 minutes.
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f. Pour the syrup into a large bowl and stir in the lemon juice and liquid from the plant. g. Dilute with 3.5 L water, stir well and pour into clean, rinsed out bottles. Secure the bottles with corks (do not use screw tops as yeast produces carbon dioxide as it ferments liquid s which may cause the bottles to explode). h. Store for at least 1 week before using. % V/V Calculation a. Take out 5 ml of the solution from the previously prepared alcohol solution and measure its density. Say it dM. b. Say dW is the density of the media [prepared separately, but no fermentation is carried out]; dA is the density of pure alcohol. Then, %v/v can be calculated as % v/v = [(dM dW) / (dA dW)] * 100

c. Alternatively, alcoholmeter can be used directly as well. d. Distil about 50 ml of the liquid. Measure the % v/v strength of the alcohol again as elucidated in step b. 4. Required Results: The alcohol strength in crude and distilled sample. Parameters: % V/V alcohol in crude and distilled sample. Relationship: NA Graphs: NA Error Analysis: NA

5. Cautions:a. No step should be missed out. b. Care should be taken while measuring densities. c. Temperature of the environment should be kept constant during the whole experiment.

Requirement for Lab Technician: 1. 25g fresh yeast or 15 g of dried yeast Medium 2. 1 kg sugar 3. 40ml (8 tsp) ground ginger 4. Juice of 2 lemons

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Worksheet of the student Date of Performance Aim: To estimate the amount of alcohol produced. Registration Number:

Calculations:

Result and Discussion

Error Analysis

Learning Outcomes (what I have learnt)

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To be filled in by Faculty S.No. Parameter (Scale from 1-10, 1 for very poor and 10
excellent)

Marks obtained

Max. Marks 20 20 10

1 2 3

Understanding of the student about the procedure/apparatus. Observations and analysis including learning Outcomes Completion* of experiment, Discipline and Cleanliness Signature of Faculty

Total marks obtained

39

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