[논문]Isolation of the Cholesterol-Assimilating Strain Pediococcus acidilactici LRCC5307 and Production of Low-Cholesterol Butter

Kim, Yunsik; Yoon, Seokmin; Shin, Hyejung; Jo, Miyoun; Lee, Sunmin; Kim, Sae-hun

doi:10.5851/kosfa.2020.e101

Isolation of the Cholesterol-Assimilating Strain Pediococcus acidilactici LRCC5307 and Production of Low-Cholesterol Butter 원문보기

Food science of animal resources, v.41 no.2, 2021년, pp.300 - 311

Kim, Yunsik (Department of Biosystems and Biotechnology, College of Life Science and Biotechnology, Korea University) , Yoon, Seokmin (Food-Biotech Team, Division of Basic Research, Lotte R&D Center) , Shin, Hyejung (Department of Food Bioscience and Technology, College of Life Science and Biotechnology, Korea University) , Jo, Miyoun (Food-Biotech Team, Division of Basic Research, Lotte R&D Center) , Lee, Sunmin (Food-Biotech Team, Division of Basic Research, Lotte R&D Center) , Kim, Sae-hun (Department of Food Bioscience and Technology, College of Life Science and Biotechnology, Korea University)

Abstract ▼ AI-Helper

The objective of the present study was to evaluate the cholesterol-assimilation ability of lactic acid bacteria (LAB), which were isolated from kimchi, a Korean traditional fermented cabbage. The isolated strain, using modified MRS medium, showed 30.5% cholesterol assimilation activity and was named Pediococcus acidilactici LRCC5307. Types and concentrations of bile were investigated for their effects on increasing the cholesterol assimilation ability of the LRCC5307 strain, a 74.5% decrease in cholesterol was observed when 0.2% bile salts were added. In addition, the manufacture of low-cholesterol butter using LRCC5307 was examined. After fermentation, LRCC5307 with butter showed 8.74 Log CFU/g viable cells, pH 5.43, and a 11% decrease in cholesterol. These results suggest that LRCC5307 could help in the production of healthier butter by decreasing cholesterol and including living LAB.

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표/그림 (7)

그림 Fig. 1. Cholesterol-MRS agar culture of bacterial strains isolated from traditional kimchi. P-CNT, positive control: peroxidase 1,000 U 10 μL; N-CNT, negative control: cholesterol-MRS broth 10 μL; Strains A, L. acidophilus ATCC43121; Strain B, E. faecalis EF5326 (isolated strain from kimchi); Strain C, Pediococcus acidilactici PA5296; Strain D, P. acidilactici LRCC5307; Strain E, L. plantarum LP5272; Strain F, Lactobacillus brevis LB5238 (isolated strain from kimchi) strains isolated from kimchi that formed yellow colonies on MRS agar.
표 Table 1. The decrease in cholesterol with the isolated bacterial strains
표 Table 2. Carbohydrate fermentation patterns analysis of PA5307
그림 Fig. 2. Phylogenetic tree of Pediococcus acidilactici LRCC5307. The analyzed sequences were compared against the GenBank database on the NCBI website using BLAST analysis (https://blast.ncbi.nlm.nih.gov/Blast.cgi).
그림 Fig. 3. The cholesterol clearing ability of LRCC5307 with the addition of different types of bile. Blank, media with cholesterol, not fermented; Non-bile, LRCC5307 fermented without bile; BE, LRCC5307 fermented with bile extract; OX, LRCC5307 fermented with oxgall; BA, LRCC5307 fermented with bile acids; BS, LRCC5307 fermented with bile salts. Results are expressed as mean±SE (n=3). ^a-d Means in different marks superscript letters are significantly different at p<0.05.
그림 Fig. 4. The cholesterol clearing ability of LRCC5307 at different concentrations of bile salts. Blank, media with cholesterol, not fermented; 0-0.5, cholesterol concentration in media, fermented with LRCC5307. Results are expressed as mean±SE (n=3). ^a-c Means in different marks superscript letters are significantly different at p<0.05.
표 Table 3. Butter before and after LRCC5307 fermentation

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제안 방법

Further, to investigate the effects of bile on cholesterol assimilation, various types and concentrations of bile were tested. The ultimate goal of this study was to produce butter with a reduced amount of cholesterol using bile and isolated LAB to provide butter, which otherwise is high in cholesterol despite its excellent flavor and nutritional value, as a healthy food.

데이터처리

Multiple comparisons between different groups were assessed using Duncan’s test.
0 (IBM, Armonk, NY, USA). The statistical differences between the mean values of test groups were analyzed by one-way analysis of variance and a paired sample t-test. Statistical significance was defined as p<0.

성능/효과

As shown in Fig. 1, when peroxidase was applied as a positive control, there was a clearly visible brown halo around the paper disc; however, when only the cholesterol-MRS broth was applied as a negative control, there was no significant change on the paper disc. When the isolated strains were added to the medium, although there were differences in size, the results could be divided into two major patterns: those that formed a clear halo (strains A and D) and those that did not (strains B, C, E, and F).
As excessive amounts of cholesterol can lead to hypercholesterolemia, the WHO and The American Heart Association recommend restricted intake of food with high levels of saturated fatty acids and cholesterol. Although butter has high calorie and fat contents for its high nutritional value and excellent flavor, it contains more than 2,000 mg/kg of cholesterol.
As excessive amounts of cholesterol can lead to hypercholesterolemia, the WHO and The American Heart Association recommend restricted intake of food with high levels of saturated fatty acids and cholesterol. Although butter has high calorie and fat contents for its high nutritional value and excellent flavor, it contains more than 2,000 mg/kg of cholesterol.
20 mg/kg after fermentation. Compared to that with cultivation in MRS medium, the pH was approximately 1.0 units higher and the viable cells were approximately 1 Log CFU/g lower; this was thought to be due to the lack of nutrients and a suitable environment for growth in butter (excessive fat content, etc.) compared to those in a medium that is optimized for LAB growth. Moreover, cell viability, pH, and cholesterol showed statistically significant differences compared to those with cultivation in butter without fermentation.
However, cholesterol was 2017.37±14.73 mg/kg, which was reduced by approximately 4.2% compared to that in butter before fermentation
Therefore, if the cholesterol assimilation rate could be further improved by optimizing the LRCC5307 fermentation conditions for butter, we anticipate that it would enable the production of healthier butter. However, due to properties of the LAB, as fermentation conditions improve, the unique sour taste imparted by LAB would also increase, thus making it important to ensure that this does not drastically alter the particular flavor of butter.
In butter fermented without bile salts, the number of viable cells and pH were similar to those of butter fermented with bile salt, and there was no statistically significant difference. However, cholesterol was 2017.
Although it varies depending on the region and diet, daily butter intake ranges 1–10 g/d, and as a result, cholesterol intake ranges from 2–20 mg/d. In this study, cholesterol in butter was reduced by 10% with LRCC5307. If this effect can be enhanced to produce butter without a risk of cholesterol, it would broaden options for consumers and greatly contribute to the growth of the butter industry.
In this study, methods using fermentation and LAB were studied to reduce cholesterol amounts in butter. The cholesterol- assimilating ability of P.
) compared to those in a medium that is optimized for LAB growth. Moreover, cell viability, pH, and cholesterol showed statistically significant differences compared to those with cultivation in butter without fermentation.
The cholesterol concentration decreased by approximately 230 mg/L, or 11% of the initial concentration, from 2, 105.21±27.99 mg/kg before fermentation to 1, 873.16±15.20 mg/kg after fermentation
The cholesterol concentration decreased to 65.64±1.76 mg/L when 0.1% bile salts were added and 51.54±1.65 mg/L when 0.2% bile salts were added
The cholesterol concentration in the medium in the absence of LRCC5307 was 201.73±1.44 mg/L, which decreased to 146.06±2.01 mg/L in the medium inoculated with LRCC5307 in the absence of bile
In this study, methods using fermentation and LAB were studied to reduce cholesterol amounts in butter. The cholesterol- assimilating ability of P. acidilactici LRCC5307 isolated from kimchi was evaluated, and the optimal conditions for cholesterol assimilation by this strain in the presence of different types and concentrations of bile in the cultures were determined.
When actual butter was fermented, the cholesterol concentration decreased by approximately 11%, showing potential for this strain to produce lower- cholesterol butter. Therefore, if the cholesterol assimilation rate could be further improved by optimizing the LRCC5307 fermentation conditions for butter, we anticipate that it would enable the production of healthier butter. However, due to properties of the LAB, as fermentation conditions improve, the unique sour taste imparted by LAB would also increase, thus making it important to ensure that this does not drastically alter the particular flavor of butter.
2% compared to that in butter before fermentation. Therefore, similar results to the increased cholesterol assimilation after the addition of bile salts to MRS medium containing cholesterol were also observed in butter.
Thus, of the 300 isolated bacterial strains, 54 formed red halos and halo sizes were measured to determine their cholesterol-assimilation ability. There were 33, 12, and nine strains with a halo diameter of 10–12 mm, 12–14 mm, and 15 mm and above, respectively.
Other strains showed a cholesterol concentration of ≥170 mg/L after cultivation, representing a decrease of <20%. Thus, the LRCC5307 strain was selected as the strain with the highest cholesterol-assimilation ability.
5%. When actual butter was fermented, the cholesterol concentration decreased by approximately 11%, showing potential for this strain to produce lower- cholesterol butter. Therefore, if the cholesterol assimilation rate could be further improved by optimizing the LRCC5307 fermentation conditions for butter, we anticipate that it would enable the production of healthier butter.
acidilactici LRCC5307 isolated from kimchi was evaluated, and the optimal conditions for cholesterol assimilation by this strain in the presence of different types and concentrations of bile in the cultures were determined. When cholesterol was added to general MRS broth as the only carbon source, there was a 30% reduction in cholesterol, but when 0.2% bile salt was added, the cholesterol concentration decreased by 74.5%. When actual butter was fermented, the cholesterol concentration decreased by approximately 11%, showing potential for this strain to produce lower- cholesterol butter.
When the culture was incubated under the same conditions but with the addition of different types of bile, the cholesterol concentration significantly decreased compared to that after cultivation without bile and produced a 68% decrease in cholesterol to 64.27±2.84 mg/L.
The cholesterol- assimilating ability of P. acidilactici LRCC5307 isolated from kimchi was evaluated, and the optimal conditions for cholesterol assimilation by this strain in the presence of different types and concentrations of bile in the cultures were determined. When cholesterol was added to general MRS broth as the only carbon source, there was a 30% reduction in cholesterol, but when 0.
com), they were similar to those of P. acidilactici standard strain (99.9% ID, T index 0.91), and the utilization of rhamnose and salicin were each 75% different. The next closest identified species was Lactococcus lactis ssp.

후속연구

In this study, cholesterol in butter was reduced by 10% with LRCC5307. If this effect can be enhanced to produce butter without a risk of cholesterol, it would broaden options for consumers and greatly contribute to the growth of the butter industry.
Therefore, in future studies, finding the optimal fermentation condition that does not affect the inherent flavor of butter while reducing cholesterol would be crucial, and this would allow for the manufacturing of healthy butter with consumer acceptance.

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