• 한국미생물·생명공학회지
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• 제30권3호
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• pp.258-263
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• 2002

E. faecalis RKYl을 이용한 젖산발효에서 세포성장과 젖산 생산에 대한 배양온도 및 pH의 영향을 조사한 결과온도가 낮을수록 세포성장은 향상되었으나 발효시간이 지연되어 젖산생산이 저하되었으며$ 50^{\circ}C$에서는 세포성장이 거의 없었다. pH 7.0-9.0에서 거의 비슷한 세포성장 및 젖산생산을 보였으나 pH 6.0에서는 세포성장이 저하되었고 발효시간이 지연되었으며, pH 10.0에서는 세포성장이 거의 없었다. E. faecalis RKY에 의한 젖산발효는 비교적 넓은 범위의 배양온도($34- 46^{\circ}C$)및 pH(7.0-9.0)에서 안정하였으며 최적 배양온도 및 pH 는 각각 $42^{\circ}C$, 7.0이었다. 또한 일반적으로 일인산염에 비해 이인산염을 첨가해 주었을 때 젖산생산성이 향상되었으며, 효모추출물 10g·$L^{-1}$을 질소원으로 사용하여 배양하였을때 $Mg^{2+}$ 과 $Mn^{2+}$의 첨가효과는 없는 것으로 나타났다.

• 한국지반공학회논문집
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• 제20권8호
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• pp.135-145
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• 2004

본 연구에서는 화강풍화토 지반상 unpropped diaphragm wall의 거동을 연구하기 위하여 과재하중의 이격거리를 변화시키면서 원심모형실험을 수행하였다. 원심모형실험시 지반굴착은 흙과 동일한 밀도로 혼합된 zinc chloride 용액이 배수되도록 밸브를 조작하여 실시하였으며, 굴착에 따라 발생하는 지반의 변형과 벽체의 변위 및 휨모멘트를 측정하였다. 수치해석은 대부분의 지반공학 문제에 적용할 수 있는 FLAC 프로그램을 이용하였다. 수치해석에서 모형지반은 Mohr-Coulomb 모델, diaphragm wall은 탄성모델을 사용하여 2차원 평면변형률 조건으로 해석을 수행하였다. 모형실험 결과 파괴면의 직선적인 형태로 파괴면내의 배면측 지반은 벽체를 향하여 하향의 변위를 일으키면서 벽체의 회전에 의해 파괴되었으며, 파괴면의 각도는 67∼74$^{\circ}$정도로 이론적인 파괴면의 각도보다 크게 평가되었다. 실험 및 해석 결과 지반의 최대침하량이 발생하는 위치는 잘 일치하였으며, 깊이에 따른 벽체변위는 선형적인 관계를 나타내었다.

• 한국미생물·생명공학회지
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• 제16권6호
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• pp.489-493
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• 1988

Starch로부터 ethanol을 직접적으로 발효 생산할 수 있는 새로운 효모 균주를 개발하고자 glucoamylase 생성균으로 알려진 Saceharomyces diastaticus의 glucoamylase gene을 cloning vector를 사용하지 않고 S, cerevisiae에 transformation시켰다. Li$_2$SO$_4$, 처리로써 competent화 한 S, cerevisiae의 Intact cells을 recipient로 하여 BamHI으로 partial digestion한 S, diastaticus의 chromosomal DNA를 transformation시키고 starch를 유일한 탄소원으로 함유한 최소 배지상에서 starch 자화능을 marker로 하여 transformant를 선별한 결과, 8.5$\times$$10^{-7}$ 빈도로 transformant를 얻었다. Transformant의 특성을 recipient 및 donor와 비교하기 위해 copper resistance와 당 발효능을 조사한 결과, donor인 S, diastaticus와 동일한 성질로서 표현된 maltose와 starch 발효능을 제외하고는 800ppm 농도까지 생육 가능한 copper resistance와 galactose 발효능 등에 있어서는 recipent와 동일하게 나타났다. 또한 transformant가 생성하는 glucoamylase의 그 작용에 있어서의 최적온도와 최적pH를 조사하여 본 바 각각 pH5.0, 50C로서 donor의 glucoamylase와 동일함을 알 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제19권1호
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• pp.108-112
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• 2009

A simplified method for the purification of cholera toxin was developed. The 569B strain of Vibrio cholerae, a recognized hyper-producer of cholera toxin, was propagated in a bioreactor under conditions that promote the production of the toxin. The toxin was separated from the bacterial cells using 0.2-${\mu}m$ crossflow microfiltration, the clarified toxin was passed through the membrane into the permeate, and the bacterial cells were retained in the retentate. The 0.2-${\mu}m$ permeate was then concentrated 3-fold and diafiltered against 10 mM phosphate buffer, pH 7.6, using 30-kDa crossflow ultrafiltration. The concentrated toxin was loaded onto a cation exchange column, the toxin was bound to the column, and most of the impurities were passed unimpeded through the column. The toxin was eluted with a salt gradient of phosphate buffer, pH 7.0, containing 1.0 M NaCl. The peak containing the toxin was assayed for cholera toxin and protein and the purity was determined to be 92%. The toxin peak had a low endotoxin level of $3.1\;EU/{\mu}g$ of toxin. The purified toxin was used to prepare antiserum against whole toxin, which was used in a $G_{M1}$ ganglioside-binding ELISA to determine residual levels of toxin in an oral inactivated whole-cell cholera vaccine. The $G_{M1}$ ganglioside-binding ELISA was shown to be very sensitive and capable of detecting as little as 1 ng/ml of cholera toxin.

• 한국식품과학회지
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• 제40권6호
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• pp.649-655
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• 2008

한방발효음료를 제조하기 위하여 한방감주의 젖산발효를 유도하였다. 이를 위해 각종 시료로부터 다양한 젖산균을 순수 분리하여 발효적성을 검토한 결과 최적균주로서 LAB19 균주를 선별하였다. LAB19 균주는 곶감으로부터 분리하였으며 생리 화학적 특성 및 16S rRNA 염기서열 분석을 통하여 Leuconostoc mesenteroides로 동정되었다. 한방감주에 종배양한 LAB19 균주를 2.5%(v/v) 접종하고 $25^{\circ}C$에서 60시간 발효시켰을 때, 한방감주는 141.3 g/L의 환원당과 5.33 g/L의 유기산, 그리고 1.19 g/L의 가용성 페놀을 함유하고 있었다. 당과 유기산 구성을 살펴보면 당의 약 90%는 맥아당이었으며, 유기산의 58%는 젖산이었다. 수용성 phenolics 성분 등에 기인하는 라디칼 소거 활성은 L-ascorbic acid의 92.4% 보다 낮은 76.6-75.7% 범위를 유지하고 있었다.

• KSBB Journal
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• 제15권6호
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• pp.600-604
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• 2000

다양한 식품의 미생물에의한 변패를 억제하고 저장성을 높 일수 있는 포장필름을 개발하기위하여, 미생물이 생산하는 천연 항균성 물질을 홉착시킨 합성 세라믹올 첨가하여 세라 믹 LDPE 필륨을 제조하였다. 천연 항균제로는 methanol자화 방선균 MO-16 및 MO-17균이 생산한 항세균제와 새로이 분 리한 방선균 No. 31이 생산한 항진균제를 사용하였다. 방선 균 No. 31이 생산한 항진균성 불질은 배양 4일째 항균효과가 우수하였으며, $121^{\circ}C$에서 15분간 열처리시에도 항균력이 유 지되는 내열성이 확인되었다. 제조한 세라믹 LDPE 필름의 미생불 생육 억제효과를 검토한 결과 조분쇄한 돈육의 경우 포장하여 실온 및 $4^{\circ}C$에서 보존시 시판필름에 비해 미생물 생육 억제효과가 우수하였다. 고제배지에 제조필름을 첨가하 여 E. coli에 대한 항균효과를 검토한 결과 첨가량에 따라 항 균효과가 우수한 것이 입증되었다 제조펼름으로 포장한 6종 류의 식품에 대한 저장성을 검토한 결과 시판필름에 비해 저 장성이 우수한 것이 확인되었으며 특히 양송이에 대한 저장 효과가 우수하였다.

• Journal of Microbiology and Biotechnology
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• 제17권2호
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• pp.287-296
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• 2007

A Lactobacillus paraplantarum strain producing a bacteriocin was isolated from kimchi using the spot-on-the lawn method and named L. paraplantarum C7 [15]. The bacteriocin, paraplantaricin C7, was found to inhibit certain Lactobacillus strains, including L. plantarum, L. pentosus, and L. delbrueckii subsp. lactis. It also inhibited Enterococcus faecalis, yet did not inhibit most of the other LAB (lactic acid bacteria) tested. The maximum level of paraplantaricin C7 activity was observed under the culture conditions of $25^{\circ}C$ and a constant pH of 4.5. Paraplantaricin C7 retained 90% of its activity after 10 min of treatment at $100^{\circ}C$ and remained stable within a pH range of 2-8. Based on a culture supernatant, paraplantaricin C7 was purified by DEAE-Sephacel column chromatography and $C_{18}$ reverse-phase HPLC. SDS-PAGE and activity staining were then conducted using the purified paraplantaricin C7, and its molecular mass determined to be about 3,800 Da. The 28 N-terminal amino acids from the purified paraplantaricin C7 were determined, and the structural gene encoding paraplantaricin C7, ppnC7, was cloned by PCR using degenerate primers based on the N-terminal amino acid sequence. The nucleotide sequences for ppnC7 and other neighboring orfs exhibited a limited homology to the previously reported plantaricin operon genes. Paraplantaricin C7 is a novel type II bacteriocin containing a double glycine leader sequence.

• Journal of Microbiology and Biotechnology
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• 제22권10호
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• pp.1388-1394
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• 2012

A xylanase-producing thermophilic strain, Geobacillus sp. TC-W7, was isolated from a hot spring in Yongtai (Fuzhou, China). Subsequently, the xylanase gene that encoded 407 amino acids was cloned and expressed. The recombinant xylanase was purified by GST affinity chromatography and exhibited maximum activity at $75^{\circ}C$ and a pH of 8.2. The enzyme was active up to $95^{\circ}C$ and showed activity over a wide pH range of 5.2 to 10.2. Additionally, the recombinant xylanase showed high thermostability and pH stability. More than 85% of the enzyme's activity was retained after incubation at $70^{\circ}C$ for 90 min at a pH of 8.2. The activity of the recombinant xylanase was enhanced by treatment with 10 mM enzyme inhibitors (DDT, Tween-20, 2-Me, or TritonX-100) and was inhibited by EDTA or PMSF. Its functionality was stable in the presence of $Li^+$, $Na^+$, and $K^+$, but inhibited by $Hg^{2+}$, $Ni^{2+}$, $Co^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Pb^{2+}$, $Fe^{3+}$, and $Al^{3+}$. The functionality of the crude xylanase had similar properties to the recombinant xylanase except for when it was treated with $Al^{2+}$ or $Fe^{2+}$. The enzyme might be a promising candidate for various industrial applications such as the biofuel, food, and paper and pulp industries.

• 한국축산식품학회지
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• 제33권1호
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• pp.75-82
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• 2013

This study aimed to isolate lactic acid bacteria (LAB) from Kimchi and to identify suitable probiotic strain for application in fermented dairy product as a commercial starter culture. A total of 106 (LAB) strains were isolated from Kimchi collected from different regions in Korea and their phenotypic characteristics were assayed. Four isolates from MRS agar plates were selected and designated as DKL109, DKL119, DKL121 and DKL128. They were identified first by API 50 CHL kit and then 16S rRNA gene sequencing. DKL121 and DKL128 were identified as Lactobacillus paracasei and Lactobacillus casei, respectively. Other two isolates (DKL109 and DKL119) were identified as Lactobacillus plantarum. To estimate their applicability in dairy products, the characteristics including acid and bile tolerance, cold shock induced cryotolerance and enzymatic activities were determined. There was wide variation in ability of strains to acid tolerance, but no significant differences in bile tolerance, cold shock induced cryotolerance within selected strains. DKL119 and DKL121 showed the highest resistance to acid and bile and the highest ${\beta}$-galactosidase activity, respectively. When these two strains were used for yogurt preparation as a single starter culture, their viable cell counts reached to $1.0{\times}10^9CFU/mL$. Lactobacillus plantarum DKL119 showed faster acid development than commercial starter culture. Also storage trials at $10^{\circ}C$ showed that the viability of these strains was retained over 15 d. With these results, it was indicated that probiotics isolated from Kimchi can be used in yogurt manufacturing as a starter culture.

• 한국미생물·생명공학회지
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• 제48권4호
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• pp.491-505
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• 2020

A newly isolated strain, Proteus vulgaris OR34, from olive mill waste was found to secrete an alkaline extracellular lipase at 11 U·ml-1 when cultivated on an optimized liquid medium. This lipase was purified 94.64-fold with a total yield of 9.11% and its maximal specific activity was shown to be 3232.58 and 1777.92 U·mg-1 when evaluated using the pH-stat technique at 55℃ and pH 9 and Tributyrin TC4 or olive oil as the substrate. The molecular mass of the pure OR34 lipase was estimated to be around 31 kDa, as revealed by SDS-PAGE and its substrate specificity was investigated using a variety of triglycerides. This assay revealed that OR34 lipase preferred short and medium chain fatty acids. In addition, this lipase was stable in the presence of high concentrations of bile salt (NaDC) and calcium ions appear not to be necessary for its activity. This lipase was inhibited by THL (Orlistat) which confirmed its identity as a serine enzyme. In addition, the immobilization of OR34 lipase by adsorption onto calcium carbonate increased its stability at higher temperatures and within a larger pH range. The immobilized lipase exhibited a high tolerance to organic solvents and retained 60% of its activity after 10 months of storage at 4℃. Finally, the OR34 lipase was applied in biodiesel synthesis via oleic acid mediated esterification of methanol when using hexane as solvent. The best conversion yield (67%) was obtained at 12 h and 40℃ using the immobilized enzyme and this enzyme could be reused for six cycles with the same efficiency.