Biological Activity of Viola mandshurica Fermented with Bacillus methylotrophicus CBMB205 and Leuconostoc pseudomesenteroides NRIC1777

미생물 분리주를 이용한 제비꽃 발효액의 생리활성 변화

Abstract

The aim of this study was to improve the efficacy and functionality of Viola mandshurica (VM). A water suspension of VM power was fermented for 72 hr with Bacillus methylotrophicus CBMB205 (BM) and Leuconostoc pseudomesenteroides NRIC1777 (LP) isolated from kimchi. The antioxidant activity and reducing power of fermented VM, its total phenolic and flavonoid compounds, as well its inhibitory activity on ${\alpha}$-amylase, ${\alpha}$-glucosidase, and pancreatic lipase were determined and compared to those of non-fermented VM (NVM), a negative control. The total phenolic and flavonoid compounds of VM fermented with BM and LP were higher than those of NVM by 1.4, 1.17, and about 3 times. There was no difference in 2, 2'-diphenyl-1-picryl hydrazyl (DPPH) radical scavenging activity between fermented VM and NVM. However, there was a 2.1- and 1.6-fold increase in 2, 2'-azino-bis-(3-ethylbenzothzoline-6-sulfonic acid) (ABTS) radical scavenging activity in VM fermented with BM and LP, respectively. The reducing power of BM was 1.6 times as high as NVM, but no significant difference was found between LP and NVM. Fermented VM's inhibitory activity on ${\alpha}$-amylase, ${\alpha}$-glucosidase, and pancreatic lipase was much higher than that of NVM. Fermenting VM with BM was superior to fermenting it with LP, except flavonoid content. Taken together, VM fermented with BM could be used as a functional food and as an additive to cosmetics.

미생물의 발효를 이용하여 제비꽃의 효능과 기능성을 증진시켜 응용을 높이기 위한 목적으로 발효식품에서 분리하고 동정한 B. methylotrophicus CBMB205와 L. pseudomesenteroides NRIC1777 두 미생물을 이용하여 72시간 제비꽃을 발효하면서 12시간 마다 발효액의 총 폴리페놀과 총 플라보노이드 함량, 항산화력, 환원력, 혈당조절과 관련이 있는 ${\alpha}$-amylase와 ${\alpha}$-glucosidase 저해활성, 비만관련 췌장 리파제 저해활성 등을 측정하고 대조구인 비발효 제비꽃과 비교, 분석하였다. 총 폴리페놀 함량의 경우 Bacillus와 유산균은 대조구에 비해 각각 1.4배 및 1.17배 증가되었고, 총 플라보노이드 함량은 각각 3.2배와 3.4배 증가되었다. 항산화력 비교에서 DPPH 라디칼 소거능은 모두 변화가 없었으나 ABTS 라디칼 소거능의 경우 2.1배 및 1.6배 증가된 것으로 나타났다. 환원력의 경우 Bacillus는 1.6배 높게 나타났으나 유산균은 대조구와 유의적인 차이점이 없었다. ${\alpha}$-Amylase의 경우 Bacillus와 유산균은 대조구보다 각각 6배와 3배의 저해율을 보였고 ${\alpha}$-glucosidase는 2.9배와 2.6배의 저해활성을 나타내었다. 췌장 리파제에 대해서 Bacillus와 유산균은 대조구에 비해 3.4배와 2.8배 높은 저해활성을 보였다. Bacillus와 유산균으로 발효한 제비꽃은 DPPH 라디칼 소거능을 제외하고 시험한 모든 항목에서 대조구보다 우수한 결과를 얻었다. 따라서 미생물 발효를 통해 항산화력과 관련이 있는 총 폴리페놀과 총 플라보노이드 함량이 증가되고 ABTS 라디칼 소거능, ${\alpha}$-amylase, ${\alpha}$-glucosidase 및 췌장 리파제에 대한 저해능 등이 대조구에 비해 높아진 결과를 토대로 건강기능식품이나 화장품 첨가제 등으로 이용될 수 있을 것으로 기대된다.

Fig. 1. Cell growth of B. methylotrophicus CBMB205 and L. pseudomesenteroides NRIC1777 during fermentation of V. mandshurica.

Fig. 2. Changes in total phenolic compounds of V. mandshurica fermented with B. methylotrophicus CBMB205 and L. pseudomesenteroides NRIC1777. Contents of total phenolic compounds of non-fermented and fermented V. mandshurica were determined using gallic acid as a standard.

Fig. 3. Changes in total flavonoid compounds of V. mandshurica fermented with B. methylotrophicus CBMB205 and L. pseudomesenteroides NRIC1777. Contents of total flavonoid compounds of non-fermented and fermented V. mandshurica were determined using catechin as a standard.

Fig. 4. DPPH radical scavenging activity of V. mandshurica fermented with B. methylotrophicus CBMB205 and L. pseudomesenteroides NRIC1777. DPPH radical scavenging activity was measured at 520 nm.

Fig. 5. ABTS radical scavenging activity of V. mandshurica fermented with B. methylotrophicus CBMB205 and L. pseudomesenteroides NRIC1777. ABTS radical scavenging activity was measured at 734 nm.

Fig. 6. Reducing power of V. mandshurica fermented with B. methylotrophicus CBMB205 and L. pseudomesenteroides NRIC1777. The reducing power activity was measured at 700 nm.

Fig. 7. Effect of fermented V. mandshurica on α-amylase activity. α-Amylase activity was measured at 540 nm.

Fig. 8. Effect of fermented V. mandshurica on α-glucosidase activity. α-Glucosidase activity was measured at 405 nm.

Fig. 9. Changes in lipase inhibition activity of V. mandshurica fermented with B. methylotrophicus CBMB205 and L. pseudomesenteroides NRIC1777. Inhibitory activity on pancreatic lipase was measured at 412 nm.