본 연구에서는 다양한 식품유해미생물에 대한 길항작용 및 항산화활성이 우수한 B. subtilis SRCM102046 균주를 식품보존 소재로서 이용하기 위해 반응표면분석법을 이용하여 균체 증량을 통한 항균활성 및 항산화활성 증대를 도모하고자 하였다. SRCM102046의 산업적 활용을 위한 성장조건을 최적화하기 위해 배양시간에 따른 균체 성장을 조사하였으며, 통계학적 기법인 반응표면분석법을 사용하였다. SRCM102046의 최적 성장을 위한 배지 성분을 선별하기 위해 Plackett-Burman design을 이용하였으며, PBD 결과 선별된 배지 성분으로 molasses, sucrose, peptone으로 예측되었다. 각 배지 성분의 최적농도를 결정하기 위한 방법으로 central composite design을 이용하였으며, 최종적으로 예측된 각 배지 성분의 농도는 molasses 7 g/L, sucrose 7 g/L, peptone 2 g/L로 예측되었다. 이때의 균체량은 22.03±1.30 g/L로 예측되었으며, 통계분석을 통해 실험모델의 적합성을 확인하였다. 또한, 실험 모델을 수행하여 건조균체량을 측정한 결과 22.02±0.35 g/L로 측정되어 실험모델에 의해 예측된 값이 오차범위 내에 존재하여 모델의 신뢰성이 매우 높음을 확인하였으며, 이는 실험모델에 의해 예측된 최적배지 사용시 최적화 이전의 LB 배지에서의 균체량(2.47±0.03 g/L)대비 약 9배의 균체량이 증가함을 확인하였다. 또한 최적배지에서 B. subtilis SRCM102046 배양 시 항균활성은 대조구로 사용된 LB 배지에서의 항균활성 대비 최대 140% 향상되었으며, 항산화활성은 약 100.41% 증가함을 확인하였다. 본 연구를 통하여 식품보존제로서B. subtilis SRCM102046의 산업화를 위한 배지최적화를 수행하였으며, 추후 박테리오신의 정제 및 특성 등에 대한 세부적인 연구가 필요하지만 본 연구를 통해 확립된 배양조건을 기반으로 식품보존 소재의 측면에서 매우 유용한 자료로 활용될 수 있을 것으로 기대된다.

When used as a starter in the manufacture of Meju, it is expected that the quality of the soup products can be improved. In this study, we isolated Lactobacillus strain having possible safety and food-industrial benefits as a starter. Four hundred and seven isolates were screened from Meju, and chemically characterized for their antibacterial activity against Bacillus cereus, non-productivity of biogenic amine, and hemolysis. Eight of the isolates were selected upon chemical characterization, and their antioxidant and β-glucosidase activity was measured. Finally, we selected, and measured its enzyme activity and antibiotic resistance. Next, we investigated its cell growth, showed maximum biomass of 3.5 g/L after 28 h of culture. The ingredients of the medium to improve biomass were selected using the Plackett-Burman design (PBD) and central composite design (CCD). The results obtained using PBD revealed molasses, yeast extract, and maltose to be significant factors determining the biomass of the L. brevis SCML 432 strain. The CCD was then applied with three variables found from PBD and the optimum values were predicted to be 5.5% molasses, 1.5% yeast extract, and 2.0% maltose, and the maximum biomass was predicted to be 11.2 g/L. Through model verification, we confirmed that the predicted and actual results were similar, with about 3.2-fold improvement in the biomass from 3.5 g/L to 11.3 g/L when compared to that obtained in basal medium. These results suggest that SCML 432 has high potential in the food industry as a starter.

In this study, we tried to screen the Bacillus strain having safety probability by isolation of strains from traditional fermented food, measurement of probiotic properties, and the fermentative characteristics of Cheonggukjang. We isolated 400 Bacillus-like isolates from traditional fermented foods. Selected strains examined on the prevalent characteristic such as extracellular enzyme and antibacterial activities, and their safety probability was confirmed by biogenic amine productivity, hemolytic, and harmful substances and enzyme productivity. We selected the 5 strains by analysis of biogenic amine, antibacterial and B. cereus toxic associated gene. Five selected strains were examined on cell surface hydrophobicity, and bile and acid tolerance, and we selected the SRCM100730 as the final strain. SRCM100730 was confirmed B. amyloliquefaciens by 16S rRNA sequencing, and named the B. amyloloquefaciens SRCM100730 (KCCM11966P). Finally, we manufactured Cheonggukjang using SRCM100730 for confirmation of fermentation properties. Manufactured Cheonggukjang did not contain B. cereus, and showed that γ-PGA and extracellular enzyme activities were superior to commercial Chunggukjang. Amino nitrogen content was 544.02 mg% and 26 free amino acid were detected, and the bitterness-related amino acid content was lower than commercial Cheonggukjang. Especially, the amount of GABA was 3 fold higher than commercial Cheonggukjang. These results suggest that SRCM100730 have high availability in commercial probiotics market and fermented food industry.