EXPERIMENT: FERMENTATION OF PLANT OBJECTIVES To study the affect of salt during sauerkraut processing To determine the pH value of sauerkraut

To determine microbiology growth in sauerkraut within 14 days INTRODUCTION Preservation of vegetables has been in our world for centuries and it have been passed down to many generation like a social customs and traditions like the kimchi from Korea. Even though some individuals regard the foods as spoiled most people who already grown custom regard it as fermented food. Fermentation can be defined as anaerobic breakdown of an organic substance by an enzyme system, where the organic compound is the final hydrogen acceptor. There are three things to understand in fermented foods. Firsts, they are less vulnerable to spoilage. Second, there less likely to vector foodborne illness. Lastly, only at certain places it is accepted it is where the origins of the fermented places. There are many types of food fermentation products such as sauerkraut, pickles, mine and milk. Sauerkraut is an acidic cabbage which results from natural fermentation by bacteria indigenous to cabbage in the presence of 2% to 3% salt. The addition of salt restricts the activities of Gram negative bacteria, while the growth of lactic acid bacteria involved in production are Leuconostoc mesenteroides, leuconostoc fallax and Lactobacillus plantarum. Sauerkraut is also known as sour cabbage in German. The lactic acid fermentation of vegetables originated from the preserving effect of the product. The growth of the lactic acid bacteria during the fermentation of sauerkraut will result in the restriction of the growth of the undesirable organisms and slows down or limit the normal spoilage. It also result in the production of the various flavours caused by the accumulation of organic acids or by-products that give the characteristic and distinct finished product. The fermentation yields lactic acid as the major product. The brine forms as the fermentation reaction begins. The salt is mixed with the shredded cabbage to restrict the activities of gram-negative bacteria while the lactic acid and cocci are favoured. The cabbage is protected from air (oxygen) in a manner to permit gases produced during the fermentation to escape. The presence of oxygen would permit the growth of some spoilage microorganisms particularly mold and yeast. Fermentation of sauerkraut is also known as wild fermentation because there is no culture added as mentioned earlier. The normal flora of the cabbage leaves is relief upon to

include the organisms responsible for a desirable fermentation that will enhance the preservation and also the organoleptic acceptability. The potential and application of fermentation of plants is to extend it shelf life and to prevents hazard microbial growth for the consumer. APPARATUS AND MATERIAL 1. Bunsen burner. 2. Universal bottle. 3. Petri dish. 4. Saline solution 5. Micropipette. 6. Distilled water. 7. Nutrient agar. 8. Chopping board 9. Cabbage. 10. Electronic weight scale. 11. Phenolphthalein. 12. Salt. 13. 0.1M NaOH. 14. Burette. 15. Sterilized knives. 16. Hotplate. 17. Beakers. 18. Sterilized conical flask 19. Sterilized pipette. 20. Micropipette. 21. Sterilized spoon. 22. Sterilized glass bottles 23. Retort stand. METHOD 1. The cabbage is cut into portions and its central core is discarded. The outer leaves and all bruised or soiled tissues are removed. The leaves are shred and washed with water. Then some of the larger leaves are cut into smaller pieces. 2. The shredded leaves are weighed and 1.5% of salt is added. Then the salt is mixed well and put into the container provided. The mixtures are compressed moderately while avoiding crushing or bruising the cabbage tissue. The container is closed tightly. Two separate containers of sauerkraut are prepared. One is for analysis purpose and the other is kept unopened throughout the fermentation. 3. Next, there is examination or analysis on the products. Examination/analysis on the product. 1. Examined colour, aroma, texture and taste of the fermented cabbage. Microbiology analysis 1. The suspension is being taken 1.0 ml and dilution is made according to the table. 2. Based on experience make dilution from higher or lower dilution.

3. By using pour plate technique the number viable organisms on certain period was determined. Determination of juice acidity 1. 10ml of sauerkraut juice was pipetted from the container and transferred to the conical flask. 2. After that, it was mixed with 10ml of distilled water and boiled for 1 minute to drive off the dissolved carbon dioxide. 3. Then it was cooled down and added 5ml of phenolphthalein 1%. 4. It was then titrated with 0.1 NaOH until a light pink colour persists. 5. The percentage of lactic acid was calculated. RESULT Microbiology analysis Days 0 Group (1.5%) 5.7 3.89 3.65 3.13 3.30 0.018 0.279 0.324 0.657 0.504 Table 1: percentage of lactic acid and pH of sauerkraut by different percentage of salt. 2 pH 5 7 14 0 2 Acidity (%) 5 7 14

Days 0 Salt % 1.5% UNC 9.6 x105 35 x106 FAIL FAIL 10

-2

Dilution factor 2 10
-4

5 10
-4

7
-5

14 10
-6

10

-3

10

-3

10

-4

10

10

-6

10

-5

10-7

FAIL FAIL

TNTC TNTC

TNTC TNTC

17 X 108

FAIL

FAIL

FAIL

Table 2: Colony Forming Unit (CFU) of microorganisms Sensory analysis Day 0 2 Colour Yellowish green Smell/Aroma smell Brown on top, pale yellow Wilted cabbage Salty crunchy Taste Texture crunchy Strong cabbage Salty

at the bottom 5 7 the bottom The sauerkraut at become brown while the bottom part remain in pale green-yellow colour 14

smell cabbage smell top Sour smell little bit Sour and salty Crunchy bite when

Brown on top, yellow at A little bit of Salty and sour a crunchy

Brown at top, while at the Sour smell bottom part, milky colour

Sour and salty

Crunchy

Table 3: Sensory evaluation DISCUSSION In the present investigation, the pH of sauerkraut brine ranged between 3.13 and 5.70. There were change in pH during the first two days of the sauerkraut fermentation from 5.70 to 3.89 and continued declining until the 14th day were slight increase from 3.13 to 3.30. The total acidity expressed as percent lactic acid of sauerkraut ranged between 0.015 and 0.504. the Jay et el who suggested that the final acidity of sauerkraut approximately lies between 1.6 and 1.8% and pH in the range of 3.1 and 3.7 (Jay, Loessner, & Golden, 2005). The floral succession is governed mainly by the pH of the growth medium. To stabilized the shelf life of pickle there are two measure factors one is low moisture and the others is pH reduced. In the present study, the naturally occurring microbial load was found to vary between 9.6 x 105 and 17 x 108 CFU/ml. The bacterial counts increased from time to time. According to Doyle et el increase of bacterial counts are selectively favoured by sauerkraut fermentation by the complete lack of oxygen, lowered pH and elevated salt conten (Doyle, Beuchat, & Montville, 2001) t. The initial population, growth rate of microorganisms, salt and acid tolerance are the important factors that influence the sequential development of various lactic acid bacteria in most vegetables fermentation. The presence of the lactic acid bacteria count in the olive and sauerkraut brine is important not only to assure acidification of the medium, inhibiting the gram-negative bacteria and some heterofermentive lactic acid bacteria, but also to inhibits fermentative metabolism of yeast that produce bloaters in olives and sauerkraut reported by Leroi and Pidoix (Leroi & Pidoux, 1993). The control of pH and salt concentration in brine, mostly practiced in the olive industry, does not reduce the incidence of this spoilage in fruits and it gas an influence on the fermentation (Balatsouras, 1985). The present study of high

acidification to pH of 3.13 and the addition of 1.5% salt would have inhibitory effect on the lactic acid bacterial flora involving in this fermentation process. Jay et el report when cabbage packed tightly in the beginning of fermentation, the aerobic bacteria decreases instantly because of the lack oxygen and overgrown by the facultatively anaerobic lactic acid bacteria (Jay, Loessner, & Golden, 2005). Sensory analysis of the sauerkraut of the present study the cabbage texture throughout the experiment from 0 to 14 days the texture still crunchy, while the taste the first 2 days it was salty then come the 5th day where the taste have both salty and sour until 14th day. From 0 to 5th day the small of cabbage decreasing from strong smell to wilted smell and the a little bit cabbage smell. Until then from 7th to 14th day the smell becoming sour smell. By the colour of the cabbage throughout the study we can see the discolouration from yellowish to brown. In study from Pundir and Jain sauerkraut is especially subjected to spoilage at its surface, where it is exposed to air. The surface film yeasts and molds destroy the acidity permitting other microorganisms to grow and causing softening, darkening and bad flavours (Pundir & Jain, 2010). The troubled of overseeing the microbial growth at certain days examples day 2, 7 and 14 may caused by the handling of samples where when the pour plate procedure the broth is too hot causing no microbial growth on the agar. For precautions and safety issues the handling of the sample must be precise and the handling of the hot broth must be cooled down a little bit before pour it into the plate. CONCLUSION In conclusion salt was the most effective for sauerkraut process. The 3.13 pH value of the sauerkraut is in reliable range of sauerkraut production. The microbiology growth was not reliable as some misfortune occurs, overall the fermentation process was conducted successfully. REFERENCES Balatsouras, G. (1985). Taxonomical and physiological characteristics of the facultative rod

type lactic acid bacteria isolated from fermenting green and black olives. 239-249:
Grasasy Aceites, 36. Doyle, M. P., Beuchat, L. R., & Montville, T. J. (2001). Food Microbiology: Fundamentals and

Frontiers 2nd ed. Washington D.C.: ASM Press.

Jay, J. M., Loessner, M. J., & Golden, D. A. (2005). Modern Food Microbiology 7th ed. New York: Springer.

Leroi, F., & Pidoux, M. (1993). Characterization of interactions between Lactobacillus

hilgardii and Saccharomyces florentinus isolated from sugary kefir grains. J. Appl. Bacteriol., 74 , 54-60.
Pundir, R. K., & Jain, P. (2010). Change in Microflora of Sauerkraut During Fermentation and Storage. World Journal of Dairy and Food Sciences 5 (2) , 221-225.