DOI QR코드

DOI QR Code

황칠나무 추출물의 고초균 발효물로 제조된 가쓰오부시 단백가수분해물의 Lactobacillus plantarum 발효를 통한 고농도 GABA 생산

Production of highly enriched GABA through Lactobacillus plantarum fermentation of katsuobushi protein hydrolyzate made from Dendropanax morbiferus extract fermented by Bacillus subtilis

  • Yu-Jeong An (Department of Food Science and Technology, Keimyung University) ;
  • Nak-Ju Sung (Department of Food Science and Nutrition, Gyeongsang National University) ;
  • Sam-Pin Lee (Department of Food Science and Technology, Keimyung University)
  • 투고 : 2023.01.11
  • 심사 : 2023.02.06
  • 발행 : 2023.02.28

초록

참치액 소스를 가공할 때 가쓰오부시(가다랑어 훈연 숙성) 열수 추출물을 주원료로 사용하며, 이때 부산물로 생산되는 가쓰오부시 단백질을 이용하여 가수 분해산물인 peptide 강화소재를 제조하며 이를 젖산발효를 통한 기능성 물질 GABA가 증진된 기능성 발효소재를 개발하였다. 황칠나무 추출물 25%에 glucose 2%, skim milk 1%를 첨가한 제한배지에서 1차 고초균 발효 1일 동안 B. subtilis HA의 생균수는 8.83 log CFU/mL로 크게 증가하였고, pH 7.53과 산도 0.06%를 나타내었다. 고초균 발효물의 protease 활성은 발효 전 1.55 unit/mL에서 발효 후 102.22 unit/mL로 크게 증가하였고, 이에 가쓰오부시 부산물을 10% 첨가하여 60℃에서 3시간 동안 단백질 분해한 결과 tyrosine 함량이 분해 전 72.94 mg%에서 분해 3시간 후 156.85 mg%로 2배 이상 증가하였다. 고초균 발효물을 이용하여 가수분해시킨 가쓰오부시 단백질 분해물에 추가적으로 MSG 10%, glucose 3%, yeast extract 0.5%를 첨가하여 정치배양으로 젖산균 발효를 수행하였다. L. plantarum KS2020의 생균수는 0일 차 7.65 log CFU/mL에서 발효 7일 차 9.33 log CFU/mL까지 크게 증가하였고, B. subtilis HA의 생균수는 0일 차 5.98 log CFU/mL에서 1일 차부터 생균수가 검출되지 않았다. 젖산균 발효물의 pH 및 산도의 변화는 0일 차 pH 7.14, 산도 0.19%에서 1일 차에 pH는 5.77로 감소하고 산도는 1.92%로 증가한 후 발효 3일 차에 산도는 0% 및 pH 8.00 이상을 나타내었다. 젖산발효 1일 차부터 GABA의 전환이 보이면서 발효 3일 차에 고농도 GABA 생성을 나타내었다. GABA 및 glutamic acid의 함량을 HPLC로 분석한 결과, GABA의 함량은 3,139.58 mg/100 g으로 매우 높은 값을 나타냈으며, glutamic acid의 함량은 83.44 mg/100 g이었다. 결과적으로 황칠나무 추출물을 이용한 고초균 발효물을 효소원으로 이용하여 가쓰오부시 부산물의 단백질을 효과적으로 고온에서 단기간에 분해하여 peptide를 생성시켰으며, 연속적으로 젖산발효를 통해 약 3.1%의 GABA가 생성되었다. 최종 가쓰오부시 단백질의 가수분해물을 이용한 젖산균 발효물은 산도가 0%이며 풍미, probiotics, peptides, 고농도 GABA를 함유한 복합 기능성 발효소재로써 소스 등 다양한 식품의 건강소재로 활용이 기대된다.

To develop a multi-functional ingredient, the bioconversion of katsuobushi protein was optimized using Bacillus subtilis HA and Lactobacillus plantarum KS2020. The Dendropanax morbiferus extract (DME) culture with protease activity (102 unit/mL) was prepared by B. subtilis with 2% glucose and 1% skim milk through one day of alkaline fermentation. Katsuobushi protein was effectively hydrolyzed by the DME culture at 60℃ for 3 hours, resulting in a tyrosine content of 156.85 mg%. Subsequently, a second lactic acid fermentation was carried out with 10% monosodium glutamate (MSG) using L. plantarum KS2020 to produce higher levels of GABA. Following co-cultivation for three days, DME exhibited a pH of 8.3 (0% acidity). After seven days, the viable cell count of L. plantarum increased to 9.33 CFU/mL, but viable Bacillus cells were not detected. Taken together, a multi-functional ingredient with enriched GABA, peptides, probiotics, and umami flavor was developed through lactic acid fermentation using hydrolyzed katsuobushi protein. These results indicate that katsuobushi protein could be used as a byproduct to produce a palatable protein hydrolysate using alkaline-fermented DME culture as a proteolytic enzyme source.

키워드

과제정보

본 연구는 교육부와 한국연구재단의 재원으로 지원을 받아 수행된 3단계 산학연협력 선도대학 육성사업(LINC 3.0)의 연구결과입니다.

참고문헌

  1. An NY. Antidiabetic and antioxidative effects of Dendropanax morbifera in streptozotocin-induced diabetic rats. MS Thesis, Busan University, Busan, Korea, p 21-34 (2014)
  2. Chang JS, Lee BS, Kim YC. Changes in γ-amino butyric acid (GABA) and the main constituents by a treatment conditions and of anaerobically treated green tea leaves. Korean J Food Sci Technol, 24, 315-319 (1992)
  3. Choe JY, Lee SP. High production of GABA in Pyrus ussuriensis Maximowicz fruit extract by mixed fermentation of lactic acid bacteria. Korean J Food Preserv, 26, 642-649 (2019) https://doi.org/10.11002/kjfp.2019.26.6.642
  4. Choi JW. Optimized production of poly-γ-glutamic acid and γ-aminobutyric acid from Dendropanax morbifera extracts by Bacillus subtilis and Lactobacillus plantarum. MS Thesis, Keimyung University, Daegu, Korea, p 16-17 (2017)
  5. De Vries MC, Vaughan EE, Kleerebezem M, de Vos WM. Lactobacillus plantarum - Survival, functional and potential probiotic properties in the human intestinal tract. Int Dairy J, 16, 1018-1028 (2006) https://doi.org/10.1016/j.idairyj.2005.09.003
  6. Diana M, Quilez J, Rafecas M. Gamma-aminobutryic acid as a bioactive compound in foods: A review. J Funct Foods, 10, 407-420 (2014) https://doi.org/10.1016/j.jff.2014.07.004
  7. Fonda ML. L-Glutamate decarboxylase form bacteria. Methods Enzymol, 113, 11-16 (1985) https://doi.org/10.1016/S0076-6879(85)13005-3
  8. Hassessian H, Prat A, De Champlain J, Couture R. Regulation of cardiovascular sympathetic neurons by substance P and γ-aminobutyric acid in the rat spinal cord. Eur J Pharmacol, 202, 51-60 (1991) https://doi.org/10.1016/0014-2999(91)90252-L
  9. Heo S, Joo HK, Lee SK. Isolation and identification of fibrinolytic bacteria from Korean traditional Chungkookjang. J Appl Biol Chem, 41, 119-124 (1998)
  10. Kang HM, Lim JS, Lee SP. Higher production of γ-aminobutyric acid in Toxicodendron vernicifluum extract by co-fermentation using yeast and lactic acid bacteria. Korean J Food Preserv, 28, 129-140 (2021) https://doi.org/10.11002/kjfp.2021.28.1.129
  11. Kang TJ, Oh SH. GABA production and use. BRIC BioWave, 9, 1-18 (2007)
  12. Kawaguchi H. Analysis of dried bonito stock and application to development of food product. Jpn J Taste Smell Res, 12, 123-130 (2005)
  13. Kim GY, Lim JS, Lee SP. Fortification of γ-aminobutyric acid and bioactive compounds in whey by co-fermentation using Bacillus subtilis and Lactobacillus plantarum. Korean J Food Sci Technol, 50, 572-580 (2018)
  14. Kim HR. Chemical composition and antimicrobial activities in the leaves of Dendropanax morbifera Lev. MS Thesis, Chonnam National University, Chonnam, Korea, p 32 (1997)
  15. Kim JE, Lee SP. Production of bioactive components and anti-oxidative activity of soybean grit fermented with Bacillus subtilis HA according to fermentation time. Korean J Food Sci Technol, 41, 179-185 (2009)
  16. Kim W, Choi K, Kim Y, Park H, Choi J, Lee Y, Oh H, Kwon I, Lee S. Purification and characterization of a fibrinolytic enzyme produced from Bacillus sp. strain CK 11-4 screened from Chungkookjang. Appl Environ Microbiol, 62, 2482-2488 (1996)
  17. Kwon HY, Kim YS, Kwon GS, Kwon CS, Sohn HY. Isolation of immuno-stimulation strain Bacillus pumilus JB-1 from Chungkookjang and fermentational characteristics of JB-1. Kor J Microbial Biotechnol, 32, 291-296 (2004)
  18. Lee SY, Choi EJ, Bae DH, Lee DW, Kim S. Effects of 1-tetradecanol and β-sitosterol isolated from Dendropanax morbifera Lev. on skin whitening, moisturizing and preventing hair loss. J Soc Cosmet Sci Korea, 41, 73-83 (2015)
  19. Lim JS, Lee SP. Production of set-type yogurt fortified with peptides and γ-aminobutyric acid by mixed fermentation using Bacillus subtilis and Lactococcus lactis. Korean J Food Sci Technol, 46, 167-172 (2014)
  20. Lim JS, Park YH, Jo BS, Kim JE, Lee SP. Increased production of γ-aminobutyric acid using the extract of Phellinus linteus fruiting body by Lactobacillus plantarum KS2020. Korean J Food Preserv, 29, 472-481 (2022)
  21. Mungal MA, Surendra SK. An improved micromethod for tyrosine estimation. Z Naturforsch, 59, 897-900 (2004) https://doi.org/10.1515/znc-2004-11-1222
  22. Ningzi G, Long L. Microbial response to acid stress: Mechanisms and applications. Appl Microbiol Biotechnol, 104, 51-65 (2020)
  23. Rhee YK, Han MJ, Park SY, Kim DH. In vitro and in vivo antitumor activity of the fruit body of Phellinus linteus. Korean J Food Sci Technol, 32, 477-480 (2000)
  24. Sanders ME, In't Veld JH. Bringing a probiotic-containing functional food to the market: Microbiological, product, regulatory and labeling issues. Antonie van Leeuwenhoek, 76, 293-315 (1999) https://doi.org/10.1023/A:1002029204834
  25. Sepahy AA, Jabalameli L. Effect of culture conditions on the production of an extracellular protease by Bacillus sp. isolated from soil sample of Lavizan jungle park. Enzyme Res, 2011, 1-7 (2011) https://doi.org/10.4061/2011/219628
  26. Yoon KH, Lee MS, Park BW, Park YH, Kim H, Kim JH, Kim MS. Enzyme production of a protease-producing strain, Bacillus sp. SH-8 isolated from insect-eating plant. Korean J Microbiol Biotechnol, 34, 323-328 (2006)