• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
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• pp.154-154
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• 2001

• 한국미생물·생명공학회지
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• 제29권3호
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• pp.155-161
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• 2001

Lactrococcus sp. HY449균줄르 M17-glucose broth에 배양하여 배양 상등액으로부터 propanol-actone 침전 ion-exchange chromatography gel-filtration chromatography 및 reverse-phase chroamtography 등을 통하여 비활성 25,600,000 BU/mg 인 순수한 bacteriocin 을 정제하였다. 정제 과정 주에서 ion-exchange chromatography 단계에 서는 35.3%의회수율이 7.3%로 감소하였다. Reverse-Phase chromatography에선 3.3%의 회수율을 보였고 활성도는 413.5배로 증가하였다. Tricine-SDS 전기영도 결과 bacteriocin 은 단일 밴드로 나타났으며, N-말단 아미노산 서열 분석을 수행한 결과 $NH_2$-IIe-Leu-Pro-GIn로 확인되었다. 아미노산조정 분석결과를 바탕으로 분자량을 예측한 결과 본 bacteriocin은 32개의 아미노산으로 이루어져 있으며 분자량은 3.6kDa인 것으로 추정되었다.

• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.1052-1056
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• 2004

A secretion signal sequence-selection vector (pGS40) was constructed based on an $\alpha$-amylase gene lacking a secretion signal and employed for selecting secretion signals from Lactococcus lactis ssp. cremoris LM0230 chromosomal DNA. Six fragments were identified based on their ability to restore $\alpha$-amylase secretion in E. coli, and among these, a fragment, S405, conferred the highest secretion activity (84%) in E. coli. Meanwhile, S407, which conferred poor secretion activity in E. coli, was quite active in L. lactis. The results suggested that the efficiency of a secretion signal depended on the host. All six fragments had an open reading frame (ORF) fused to the reporter gene, and the potential Shine-Dalgamo (SD) sequence and putative promoter sequences were located upstream of the ORF. Deduced amino acid sequences from the six fragments did not show any homology with known secretion signals. However, they contained three distinguished structural features and cleavage sites, commonly found among typical secretion signals. The characterized secretion signals could be useful for the construction of food-grade secretion vectors and gene expression in LAB.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.1152-1157
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• 2005

The effect of carbon sources on nisin Z biosynthesis in Lactococcus lactis subsp. lactis A164 was studied in batch culture using M17 broth containing different carbon sources. Among the eleven carbon sources tested, glucose, sucrose, and lactose were suitable carbon sources for cell growth of L. lactis A164. In particular, cells grown on lactose produced at least 3-fold greater amount of nisin Z than those on other carbon sources. Galactose resulted in less amount of cell mass than did sucrose or glucose, but gave a higher level of nisin Z activity. Northern blot analysis revealed. that lactose increased the transcription of the nisZ pre-peptide gene. Although galactose was less efficient than lactose, it increased the transcription of nisZ along with a higher level of nisin Z than did sucrose and glucose. These results suggest that the increased nisin Z production is correlated with the induction of nisZ by lactose and galactose. Among all the carbon sources tested, no remarkable differences were observed in nisRK and nisFEG transcripts, indicating that the lactose- or galactose-mediated induction is unique to the nisZ promoter.

• Journal of Microbiology and Biotechnology
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• 제16권11호
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• pp.1799-1808
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• 2006

A food-grade integration vector based on site-specific recombination was constructed. The 5.7-kb vector, pIMA20, contained an integrase gene and a phage attachment site originating from bacteriophage A2, with the ${\alpha}$-galactosidase gene from Lactobacillus plantarum KCTC 3104 as a selection marker. pIMA20 was also equipped with a controllable promoter of nisA ($P_{nisA}$) and a signal peptide-encoding sequence of usp45 ($SP_{usp45}$) for the production and secretion of foreign proteins. pIMA20 and its derivatives mediated site-specific integration into the attB-like site on the Lactococcus lactis NZ9800 chromosome. The vector-integrated recombinant lactococci were easily detected by the appearance of blue colonies on a medium containing $X-{\alpha}-gal$ and also by their ability to grow on a medium containing melibiose as the sole carbon source. Recombinant lactococci maintained these traits in the absence of selection pressure during 100 generations. The ${\alpha}-amylase$ gene from Bacillus licheniformis, lacking a signal peptide-encoding. sequence, was inserted downstream of $P_{nisA}\;and\;SP_{usp45}$ in pIMA20, and the plasmid was integrated into the L. lactis chromosome. ${\alpha}-Amylase$ was successfully produced and secreted by the recombinant L. lactis, controlled by the addition and concentration of nisin.

• Journal of Microbiology and Biotechnology
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• 제9권2호
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• pp.201-205
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• 1999

The effects of growth temperature and a carbon source on the expression of $\beta$-galactosidase gene of Lactococcus lactis ssp. lactis ATCC 7962 (L. lactis 7962) were investigated. At $25^{\circ}C$, L. lactis 7962 had a higher $\beta$-galactosidase activity than cells grown at $30^{\circ}C$ or $37^{\circ}C$, although cells grew most quickly at $37^{\circ}C$ The highest $\beta$-galactosidase activity was observed in cells grown in M17 with lactose (l %) followed by cells grown in a galactose (1 %) medium. L. lactis 7962 exhibited the minimum $\beta$-galactosidase activity in glucose media, indicating catabolite repression. When the cellular levels of $\beta$-galactosidase mRNA were examined using slot blot hybridization, no significant differences were observed between cells grown at $25^{\circ}C$ and cells at $30^{\circ}C$ or $37^{\circ}C$ in the same media. This suggests that the quantity of $\beta$-galactosidase mRNA may not be the reason for the higher $\beta$-galactosidase activities of L. lactis 7962 at $25^{\circ}C$ The level of ccpA (Catabolite Control Protein) transcript remained almost constant during the exponential growth phase irrespective of a carbon sourse.

• 한국축산식품학회지
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• 제37권3호
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• pp.402-409
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• 2017

A novel peptide having free radical scavenging activity was separated, using an on-line high-performance liquid chromatography (HPLC) - ABTS screening method, from bovine skim milk fermented by Lactococcus lactis SL6 (KCTC 11865BP). It was further purified using reverse phase-HPLC (RP-HPLC) and sequenced by RP-HPLC-tandem mass spectrometry. The amino acid sequence of the identified peptide was determined to be Phe-Ser-Asp-Ile-Pro-Asn-Pro-Ile-Gly-Ser-Glu-Asn-Ser-Glu-Lys-Thr-Thr-Met-Pro-Leu-Trp (2,362 Da), which is corresponding to the C-terminal fragment of bovine ${\alpha}_{s1}$-casein (f179-199). The hydroxyl radicals scavenging activity ($IC_{50}$ $28.25{\pm}0.96{\mu}M$) of the peptide chemically synthesized based on the MS/MS data showed a slightly lower than that of the natural antioxidant Trolox ($IC_{50}$ $15.37{\pm}0.52{\mu}M$). Furthermore, derivatives of the antioxidant peptide were synthesized. The antioxidative activity of the derivatives whose all three proline residues replaced by alanine significantly decreased, whereas replacement of two proline residues in N-terminal region did not affect its antioxidative activity, indicating that $3^{rd}$ proline in C-terminal region is critical for the antioxidative activity of the peptide identified in this study. In addition, N-terminal region of the antioxidant peptide did not show its activity, whereas C-terminal region maintained antioxidative activity, suggesting that C-terminal region of the peptide is important for antioxidative activity.

• 한국어병학회지
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• 제11권2호
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• pp.137-141
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• 1998

TSB 해수배지와 숙주로서 L. garvieae No. 44를 배양조건으로 사용한 경우, L. garvieae 111균주중 용원화되었다고 추정되어진 96균주의 cells에서 temperate Phage가 효과적으로 분리되었다. 하지만 동일한 배양조건에서 보통의 TSB 배지를 사용한 경우에서는 temperate phage는 전혀 나타나지 않았다. 이 temperate phages는 TSB 해수배지와 ultraviolet irradiation를 병용한 경우 역시 효과적으로 분리되었다. 분리된 모든 temperate Phages는 기존의 phage(PLgY, PLgW, PLgS) 의 lytic nature와는 달리 오직 L.garvieae No. 44만을 lysis하였다. 배양후 약 1시간후 phage가 나타났으며 12시간 후 virulent phage의 최고농도($10^{10}$ PFU/ml) 보다 훨씬 낮은 농도인 $10^6$ PFU/ml까지 증가하였다.

• 한국축산식품학회지
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• 제40권2호
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• pp.155-171
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• 2020

Fermented products, including sausages, provide several health benefits, particularly when probiotics are used in the fermentation process. This study aimed to examine the cytotoxicity (against Caco-2 and MCF-7 cell lines), antihypertensive activity via angiotensin-converting enzyme (ACE) inhibition, antioxidant capacity, antidiabetic activity via α-amylase and α-glucosidase inhibition, proteolysis rate, and oxidative degradation of fermented camel and beef sausages in vitro by the novel probiotic Lactococcus lactis KX881782 isolated from camel milk. Moreover, camel and beef sausages fermented with commercial starter culture alone were compared to those fermented with commercial starter culture combined with L. lactis. The degree of hydrolysis, antioxidant capacity, cytotoxicity against Caco-2 and MCF-7, α-amylase, α-glucosidase, and ACE inhibitory activities were higher (p<0.05) in fermented camel sausages than beef sausages. In contrast, the water and lipid peroxidation activity were lower (p<0.05) in camel sausages than beef sausages. L. lactis enhanced the health benefits of the fermented camel sausages. These results suggest that camel sausage fermented with the novel probiotic L. lactis KX881782 could be a promising functional food that relatively provides several health benefits to consumers compared with fermented beef sausage.

• 한국식품영양과학회지
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• 제41권8호
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• pp.1066-1071
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• 2012

본 연구에서는 3가지 서로 다른 미생물을 이용하여 콩을 발효시킨 후 제품의 알레르기성 변화를 살펴보았다. 증자 후 가열 살균된 콩은 유산균(Lactococcus actis subsp. lactis) 그리고/또는 황국균(Aspergillus oryzae) 그리고/또는 고초균(Bacillus subtilis)을 이용하여 발효되었다. 콩 단백질이 발효과정에서 분해됨을 확인하였다. 발효과정 후 콩 단백질의 알레르기성과 항원성의 변화는 콩 단백 특이 다클론 항체 또는 콩에 대해 알레르기 반응을 일으키는 환자들의 혈청을 이용하여 면역반응검사법으로 확인되었다. 콩 단백질은 니아신 생성균주 L. lactis subsp. lactis IFO12007와 황국균 그리고 고초균을 이용한 3단 발효에 의해 가장 강력한 분해가 이루어졌다. 3단 발효 후 콩의 알레르기성 및 항원성도 크게 감소되었다. 본 연구에서 확인된 3단 발효법은 콩의 알레르기성을 저하시키는 강력한 방법 중의 하나이다.