THE EFFECT OF DIFFERENT MEDIUM COMPOSITIONS ON PRODUCTION OF BACTERIOCIN OF LACTIC ACID BACTERIA ISOLATED FROM YELLOW BETUNG BAMBOO SHOOT (Dendrocalamus asper) PICKLE UNDER DIFFERENT FERMENTATION CONDITIONS - Unika Repository

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7. APPENDICES Appendix 1. Species Identification of LAB Isolates using API 20 strep

  a b b

  Key : a = before incubation, b = after 24 hours incubation

  Figure 12. Identification of Isolate A43 using API 20 strep

  Appendix 2. Species Identification of LAB Isolates using API 50 CH

Key : a = before incubation, b = after 24 hours incubation, c = after 48 hours incubation

  Figure 13. Identification of Isolate D44 using API 50 CH

  a b c b c

  

Appendix 3. Media Used for Growth and Bacteriocin Inhibitory Activity Test of

LAB Isolated from Yellow Betung Bamboo Shoot Pickle

  3.1. MRS Broth Medium (Merck Brand)

  MRS Broth medium was prepared by dissolving 52.2 grams of powdered MRS Broth into 1 liter of distilled water. Stir until the powder dissolve evenly. MRS Broth medium then

• 1

  sterilized using an autoclave at 121°C for 15 minutes. Composition MRS Broth: 20 gL

• 1 -1 -1

  D(+)-glucose, 10 gL of casein/meat peptone, 8 gL of meat extract, 5 gL of sodium

• 1 -1 -1

  acetate, 4 gL of extract of yeast, 2 gL of dipotassium hydrogen phosphate, 2 gL of di-

• -1 -1

  ammonium hydrogen citrate, 1 gL of tween 80, 0.2 gL of magnesium sulfate, and 0.04

• 1 gL of manganese sulfate.

  3.2. Nutrient Broth Medium (Merck Brand)

  Nutrient Broth (NB) medium was prepared by suspending 8 grams of powdered NB into 1 litre of distilled water. Stir until the powder dissolve evenly. NB medium then sterilized

• 1

  using an autoclave at 121°C for 15 minutes. Composition NB: peptone from meat 5 gL ,

• 1 meat extract 3 gL .

  3.3. Nutrient Agar Medium (Merck Brand)

  Nutrient Agar (NA) medium was prepared by dissolving 20 grams of powdered NA into 1 liter of distilled water. Stirred using a stirrer while heated until dissolved powder evenly and boil first. Then sterilized using an autoclave at 121°C for 15 minutes. NA

• -1 -1 -1

  Composition: 12 gL of gelatin, 5 gL of meat peptone, and 3 gL of meat extract.

  Appendix 4. Composition of Standard Solution McFarland No. 2 and 3

  McFarland No. 2 standard solution was prepared by mixing 0.2 mL of BaCl

  2 1% with 9.9

  2

  4

  mL of 1% H SO . McFarland No. 3 standard solution was prepared by mixing 0.3 mL of

  2

  2

4 BaCl 1% with 9.7 mL of 1% H SO . McFarland No. 5 standard solution was prepared

  by mixing 0.5 mL of BaCl

  2 1% with 9.5 mL of 1% H

  2 SO 4 . McFarland No. 2 standard

  6

  solution equivalent to the number of bacteria 600 CFU (x10 /mL), McFarland No. 3

  6

  standard solution equivalent to the number of bacteria 900 CFU (x10 /mL) and McFarland No. 5 standard solution equivalent to the number of bacteria 1500 CFU

  6

  (x10 /mL). In this test, a solution of McFarland No.2 standard was used in the identification of species, while McFarland No.3 and Mc Farland No. 5 were used in the test of bacteriocin inhibitory activity.