전통・발효식품 유래 기능성 유산균 분리 및 활용 연구
저자
발행사항
삼례: 우석대학교, 2012
학위논문사항
학위논문(석사)-- 우석대학교 대학원: 생명공학 2012.2
발행연도
2012
작성언어
한국어
주제어
KDC
574.4 판사항(4)
발행국(도시)
전북특별자치도
형태사항
x, 113p.: 삽도; 27cm.
일반주기명
참고문헌: p.98-102
소장기관
Isolation, characterization and utilization of Weissella koreensis OK1-6 strains with high ornithine producing capacity from kimchi.
Abstract
Two lactic acid bacteria (LAB) with high ornithine-producing capacity were isolated from kimchi. Examination of the biochemical features using an API kit indicated that the strains belonged to the members of Weissella genus. They were gram positive, short rod-type bacteria, and able to grow anaerobically with CO2 production. The isolates grew well on MRS broth at 25∼37℃ and pH of 6.0∼7.0. The optimum temperature and pH for growth are 30℃ and pH 6.5. The isolates fermented arabinose, ribose, xylose, glucose but not cellobiose, galactose, raffinose, or trehalsoe. The 16S rDNA sequences of isolates showed 99.6% and 99.7% homology with the Weissella koreensis S5623 16S rDNA (access no. AY035891). They were accordingly identified and named as Weissella koreensis OK1-4 and Weissella koreensis OK1-6. Ariginine deaminase (ADI) pathway by the strains could produce ornithine from MRS broth supplemented with 1% of arginine at a productivity of 27.01 and 31.41 mg/L/h, respectively.
In order to explore the commercial application of high ornithine-producing strain, elastase inhibition activity and antioxidant activity of W. koreensis OK1-6 strain were measured. The elastase inhibiting activities of cell extracts from the culture in MRS medium (CE) and cell-free medium from the culture in MRS medium (CFM) measured at 6.71~26.16% and 10.92~38.57%, respectively, while the inhibiting activities of cell extracts from the culture in MRS medium with 1% arginine (CEA) and cell-free medium from the culture in MRS medium with 1% arginine (CFMA) measured at 5.87~10.19% and 41.42~9.01%, respectively, and the ornithine-producing capacity made little difference to the elastase inhibiting activities. The antioxidant activities of the intact cell (IC) samples of W. koreensis OK1-6 and W. koreensis KCCM 41517 strains scored activity levels of 71.71~73.98% and 72.64~79.34%, respectively, regardless of the existence of arginine added.
To produce kimchi with higher ornithine contents by using these strains, four different kimchi groups were made: the kind of kimchi made with salted yellow corvine and no added starter (HN), kimchi added with salted yellow corvine and W. koreensis OK1-6 starter (HO), kimchi made with refined salt and no added starter (SN), kimchi added with refined salt and W. koreensis OK1-6 starter (SO). Changes in ornithine contents of kimchi were investigated during fermentation for a 35 day period at -1℃ (condition of kimchi refrigerator).
The kimchi made with added starter (HO, SO) had ornithine contents of 207.04 mg and 117.06 mg each per 100 g after the 7th day of fermentation, which increased to 295.20 mg and 272.30 mg by the 28th day and peaked at the 35th day of fermentation. The kimchi made with no added starter (HN, SN) showed a slow increase in ornithine content per 100 g of kimchi and showed total ornithine contents of 96.89 mg and 92.75 mg respectively at the 28th day of fermentation. There were no significant differences resulting from the addition of starter in the color, smell, taste, or texture of the kimchi based on the sensory evaluations.
These results are the first report on the ornithine production activities and functionality of W. koreensis OK1-6 strain, and such results are expected to be applied to kimchi manufacturing to further boost or ornithine production while also carrying implications of further applicability to a wider range of sector as a food and cosmetics ingredient.
Chapter Ⅱ
Isolation, characterization and activity screening of Weissella koreensis strains with ornithine producing capacity from natural sea salt
Abstract
Lactic acid bacteria (LAB) having high ornithine-producing capacities were isolated from natural sea salt. The 16S rDNA sequences of two isolates showed 99.5%와 99.6% homology with the Weissella koreensis S5623 16S rDNA (Access No. AY035891). They were gram positive, short rod-type bacteria, and able to grow anaerobically with CO2 production. The optimum temperature and pH for growth of the LAB are 37℃ and pH 7.0. High cell growth was maintained at a NaCl concentration of 0∼6%. The isolates fermented D-ribose, D-galactose, D-lactose, D-maltose, cellobiose, D-tagatose, D-trehalose, sucrose, D-melezitose, gentiobiose, D-glucose but not D-melibiose, inositol, D-sorbose. They were accordingly identified and named as Weissella koreensis MS1-3 and Weissella koreensis MS1-14, and could contain ornithine at levels of 72 mg/100 g cell F.W. and 105 mg/100 g cell F.W. and extracellular ornithine at levels of 4.5 mg/100 mL and 4.6 mg/100 mL medium, respectively, by culturing in MRS broth supplemented with 1% of arginine. High cell growth was maintained in MRS broth with a NaCl concentration of 0∼6%.
Although there was a time difference, through a 48h culture, W. koreensis strains metabolized almost all the 1% arginine added. W. koreensis strains isolated from natural sea salt produced citrulline and ornithine in the metabolism process with arginine which implied that the ornithine was being produced through ADI (arginine deaminase) metabolism.
Of the isolated strains, the W. koreensis MS1-14 was tested separately for its functionality due to its high potential for ornithine production, and was examined for its elastase inhibition activity and antioxidant activity. The elastase inhibition activity of W. koreensis MS1-14 strain for CE sample and CFM sample from the culture in MRS medium measured at 15.21~24.99% and 4.08~6.08% respectively. The elastase inhibition activity for CEA and CFMA from the culture in MRS medium with 1% arginine measured at 15.51~26.18% and 14.10~25.33%, respectively. The antioxidant effects of the intact cell (IC) samples from the culture in MRS medium and MRS medium with 1% arginine showed activity levels of 83.61%, and 77.09% respectively. The antioxidant effects of the cell wall (CW) samples from the culture in MRS medium and MRS medium with 1% arginine scored activity levels of 8.30~42.75% and 10.40~46.13% respectively, which means there was no significant correlation between ornithine production and antioxidant effectiveness of the cultures.
In conclusion, these results show that apart from the ornithine producing strains previously discovered in kimchi, the natural sea salt contains a separate W. koreensis strain that can independently produce ornithine while the biochemical similarities discovered in these cultures allow us to predict further discoveries of W. koreensis strains with diverse characteristics.
Furthermore, the discovery of such functional microorganism will further invigorate the jeotkal (salted-fermented sea foods) fermented food industry and natural sea salt industry which will require a correlating management of hygiene and safety, and the natural sea salt maybe able to develop as a global food ingredient.
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