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There are three samples used to do this experiment. One is a fermented Kimchi (weight except water content), the next one is a room temperature-dried material of the fermented Kimchi which is prepared by a heat drying at 45’C and a cold keeping at 8’C. And the third one is a freeze-dried material of the fermented Kimchi having been kept at 8’C. and lastly the triplex coated Kimchi lactic acid bacteria (weight except coating materials) obtained after freeze drying and having been kept at 8’C. One gram of each sample is suspended in a sterilized buffer solution at pH 1.2 and put into a capsule dissolver equipped with a up and down reciprocating device in a water bath set at 35’C, and shaken for 2 hours, and then spread on the MRS plate culture medium finally to compare and analyze the appearing colonies.

The ways of analysis are divided into three, and the number of Kimchi lactic acid bacteria both right after and one year after the manufacture is counted as the following Table 1. 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

As shown in Table 1, with the criteria of 9 x 1010 of fermented Kimchi, the RT-dried scored only 3 x 104 compared with the freeze-dried, and the freeze-dried recorded 1 x 106, which mean that lots of Kimchi lactic acid bacteria were killed by stomach acid. And after one year of storage the number was reduced by 108 and 107, respectively. However, the triplex coated Kimchi lactic acid bacteria survived by as many numbers as 6 x 1010. Therefore, the Kimchi lactic acid bacteria can be told to have been remarkably killed by exposing to room temperature, water and air, but in the case of the triplex coated Kimchi lactic acid bacteria, though some of bacteria were killed during the freeze drying, most of the bacteria have been protected from stomach acid by the first coating of Guar gum and Gellan gum, the second one of casein and the last third of sea tangle stuff. After 1 year of storage, the survived bacteria of the triplex coat were 3 x 1010, the number of which is far better than that of freeze-dried lactic acid bacteria products generally selling on the market. Comparing that the survival rate of the lactic acid bacteria in the previously launched products on the market usually stays under 5%, the result is very much valuable and inspiring. This is supporting the result that triplex coated Kimchi lactic acid bacteria might be possible to preserve for a long time as shown in ‘food industry and nutrition 6<1>, P 71-77 in 2001’. 

 

 

•  Experimental results on the bile salt tolerance of triplex coated Kimchi lactic acid bacteria 

 

Considering the condition in the stomach and duodenum in our body, as done in Table 1, one gram of three samples each is suspended in a sterilized buffer solution at pH1.2 and put into a capsule dissolver equipped with a up and down reciprocating device in a water bath set at 35’C, and shaken for 2 hours, and then a hygienically-controlled bile salt is added at the amount of 500 ug/g (W/W) and after the pH is calibrated to 7.5 the mixture is cultured for 2 hours in a shaking water bath set at 35’C. And the mixtures are spread on the MRS plate culture medium finally to compare and analyze the appearing colonies as shown in Table 2.

 

 

 

 

 

 

 

 

 

 

 

 

 

As shown in Table 2, comparing the death rates by stomach acid and bile salt, that of bile salt is remarkably lower. This proves that most of bacteria are killed by stomach acid than by bile salt.

In the case of triplex coated Kimchi lactic acid bacteria, right after the triplex coating the number of survived bacteria was 6 x 1010 in a stomach acid solution and 3 x 1010 in a bile salt.

This means that the death rate by bile salt is lower than by stomach acid. Likewise, the similar result was also obtained after a year of storage, where the number of stomach acid tolerable bacteria was 3 x 1010 and that of bile salt tolerable ones 2 x 1010. This result supports that the coating materials also protect the bacteria from killing by bile salt, because uncoated bacteria were killed in a large scale.

 

Only when the triplex coated Kimchi lactic acid bacteria are released after the coats are dissolved in the guts, normally settle and proliferate on the small and large intestines, the usefulness of the triplex coats could be proven. For this experiment, the triplex coated Kimchi lactic acid bacteria are suspended in a solution 1 (2 grams of sodium chloride and diluted hydrogen chloride are mixed and made into 1 L with the addition of water, and then the pH becomes about 1.2) and processed with a up and down reciprocating motion for 2 hours, and then it is expected that the opening, exfoliation or damage of the gut-soluble triplex coat might be derived and the inside contents might be exude. The triplex coated Kimchi lactic acid bacteria are also suspended in a solution 2 (a 250 ml of 0.2 N potassium two-hydrogen phosphate and a 118 ml of 0.2 N sodium hydroxide are mixed and made into 1L with the addition of water, and then the pH becomes about 6.8) and processed with a up and down reciprocating motion for 1 hour, and then, if any remnants are observed, further processed until the remnants are completely removed. And the mixtures are spread on the MRS plate culture medium finally to compare and analyze the appearing colonies as shown in Table 3.

 

In the case of triplex coated Kimchi lactic acid bacteria, the right-after-the manufacture group showed 2 x 1010 of bacteria similar to bile salt tolerance, and even after 1 year of storage the number reached 1 x 1010. Among the Kimchi lactic acid bacteria, L. plantarum and L. brevis were 3 x 109 and 2 x 109, respectively, and Leuc. Mesenteriodes was detected 7 x 103 in a relatively smaller numbers. Likewise, the reduction of the bacteria was low even after 1 year of storage. Therefore, it is expected that besides the intake of a lot of useful materials in the Kimchi, the triplex coated Kimchi lactic acid bacteria can settle and proliferate on the guts and exert several positive functions such as the normalization of guts and the other beneficial effects by a variety of Kimch lactic acid bacteria. And it is also possible to derive various types of functional health foods such as tablets, capsules, pills, granules, and syrups.