• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.82-85
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• 2005

Doxorubicin is a highly-valuable anthracycline-family polyketide drug with a very potent anticancer activity, typically produced by a Gram-positive soil bacterium called Streptomyces peucetius. Thanks to the recent development of Streptomyces genomics-based technologies, the random mutagenesis approach for Streptomyces strain improvement has been switched toward the genomics-based technologies including the application of DNA microarray systems. In order to identify and characterize the genomics-driven potential target genes critical for doxorubincin overproduction, three different types of doxorubicin overproducing strains, a dnrI(doxorubicin-specific positive regulatory gene)-overexpressor, a doxA (gene involved in the conversion from daunorubicin to doxorubicin)-overexpressor, and a recursively-mutated industrial strain, were generated and examined their genomic transcription profiles using Streptomyces DNA microarray systems. The DNA microarray results revealed several potential target genes in S. peucetius genome, whose expressions were significantly either up- or down-regulated comparing with the wild-type strain. A systematic understanding of doxorubicin overproduction at the genomic level presented in this research should lead us a rational design of molecular genetic strain improvement strategy.

• Molecules and Cells
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• 제43권8호
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• pp.694-704
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• 2020

HslUV is a bacterial heat shock protein complex consisting of the AAA+ ATPase component HslU and the protease component HslV. HslV is a threonine (Thr) protease employing the N-terminal Thr residue in the mature protein as the catalytic residue. To date, HslUV from Gram-negative bacteria has been extensively studied. However, the mechanisms of action and activation of HslUV from Gram-positive bacteria, which have an additional N-terminal sequence before the catalytic Thr residue, remain to be revealed. In this study, we determined the crystal structures of HslV from the Gram-positive bacterium Staphylococcus aureus with and without HslU in the crystallization conditions. The structural comparison suggested that a structural transition to the symmetric form of HslV was triggered by ATP-bound HslU. More importantly, the additional N-terminal sequence was cleaved in the presence of HslU and ATP, exposing the Thr9 residue at the N-terminus and activating the ATP-dependent protease activity. Further biochemical studies demonstrated that the exposed N-terminal Thr residue is critical for catalysis with binding to the symmetric HslU hexamer. Since eukaryotic proteasomes have a similar additional N-terminal sequence, our results will improve our understanding of the common molecular mechanisms for the activation of proteasomes.

• 한국축산식품학회지
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• 제27권4호
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• pp.509-516
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• 2007

Starters of lactic acid bacteria(LAB) were isolated from the commercial yoghurt products and the four isolates have been studied on their identification and some physiological characteristics. For the purpose of identification, microscopic examination, API test, and 16s rRNA gene sequencing were conducted. Isolate A from a yogurt product of local dairy company A was shown to be Gram-positive rod-shaped bacterium. All strains isolated were turned out to be as Lactobacillus paracasei by using a API 50 CHL kit. In contrast, isolate A was identified as a strain of Lactobacillus helveticus based on the 16S rRNA sequencing data, and L. casei ssp. casei for both B and D and L. paracasei for C. All the isolates survived the simulated gastric juice, pH 2.0 within 3 hours and sharply decreased in viability so that no viable cell was observed after 4.5 hours incubation. In addition, the four isolated strains were almost identical in antibiotic susceptibility to six different kinds of antibiotics including erythromycin ($15\;{\mu}g$), ampicillin ($10\;{\mu}g$), gentamycin ($10\;{\mu}g$), neomycin ($30\;{\mu}g$), but rather resistant to colistin ($10\;{\mu}g$) and streptomycin ($10\;{\mu}g$). It was noteworthy that four isolates were confirmed to produce antibacterial substance against foodborne pathogens of Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli 0157:H7 as test organisms based on the inhibitory zones on an MRS soft agar medium. At presence, the inhibitory factor is unknown so that further studies are required to ascertain the active factor responsible for the inhibitory activities.

• 환경생물
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• 제25권3호
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• pp.260-266
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• 2007

PLLA분해활성을 가진 고온성세균을 우리나라 경작토로부터 분리하였다. 분리된 균주는 그람양성 간균으로 16S rDNA 염기서열분석 결과 Geobacillus caldoxylosilyticus로 동정되었으며, 현재까지 보고되지 않은 새로운 PLLA분해세균으로 밝혀졌다. 조절화된 퇴비화조건에서 멸균퇴비에 분리균주를 접종하고 $58^{\circ}C$에서 40일간 분해활성을 측정하였다. 이 균주는 중량평균 분자량이 5000, 11000, 34000 및 256000인 PLLA5000, PLLA11000, PLLA34000 및 PLLA256000를 각각 66%, 57%, 41%및 40%생분해하였다. PLLA256000을 기질로 하여 gelatin, yeast extract 및 ammonium sulfate를 0.1%씩 첨가하여 분해활성 변화를 조사한 결과 대조군에 비하여 분해활성이 각각 27%, 13% 및 10%증진되었으며, 접종량을 $10^9cfu\;g^{-1}$로부터 $10^{10}cfu\;g^{-1}$ 및 $10^{11}cfu\;g^{-1}$로 증가시킴에 따라 PLLA256000의 생분해도는 각각 14% 및 20% 증가하였다.

• 한국응용곤충학회지
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• 제41권1호
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• pp.49-53
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• 2002

Paenibacillus larvae는 Gram양성의 내생포자 형성 세균으로, 꿀벌의 질병 중 가장 심각한 피해를 입히는 미국부저병(AFB)의 원인균이다. 이 P. larvae를 발병전인 봉군 또는 애벌레에서 보다 빠르게 검출하기 위하여 기니아피그를 이용하여 이 병원균에 대한 다클론 항체를 제작하였고 또한 그 항체의 성능을 평가하였다 제작된 항체를 이용한 ELISA검색법을 개발한 후, 다수의 P. larvae균주를 검사하였다. 그 결과 P. larvae ATCC 25748의 균체를 사용하여 제작된 항체는 ATCC 9545 (대표균주), ATCC 25747, 국내 분리 균주인 SJl5등과 높은 항체 친화성을 나타내었으며. 다른 세균 종들과는 반응하지 아니하였다. 또한 이 ELISA검색법은 미국 부저병의 현장검사에 적용하기 위하여 사용되었으며, 그 결과로 본 연구에서 제시하는 항체는 벌집 내 또는 애벌레 내에 존재하는 P. larvae의 빠른 동정과 모니터 링에 매우 유용함을 보여주었다.

• Applied Biological Chemistry
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• 제43권3호
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• pp.169-173
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• 2000

저온에서 높은 세척력을 갖는 호소세제의 개발을 위하여 토양으로부터 alkaline protease의 활성이 높은 균주를 분리, 동정하였으며, 호소의 성질을 조사하였다. 분리균주의 형태적 특징은 Gram 음성균 이고, 간균$(0.6{\sim}0.7{\times}1.3{\sim}2.6\;{\mu}m\;in\;size)$ 형태를 하고 있으며, 운동성을 보였다. 또한 catalase 양성, aesculin, gelatin 및 casein 분해능이 있었다. 분리균주의 세포벽 구성 성분은 meso-DAP를 함유하였으며, G+C mol 함량은 43.3%를 나타내었다. 이러한 형태적, 생리 생화학적 특성의 결과로부터 분리 균주는 Acinetobacter 속으로 동정되었다. 분리 균주를 이용한 alkaline protease의 생산은 초기 pH 10과 $40^{\circ}C$에서 36시간 배양하였을 때 3,300 D.U/mL로 최대 효소 활성을 보였으며, 최적 pH와 온도는 9와 $60^{\circ}C$이었다. 또한 본 균주에 의해 생산된 alkaline protease는 두개의 활성 band를 나타내었다.

• Restorative Dentistry and Endodontics
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• 제23권1호
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• pp.175-182
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• 1998

Polyphosphate has been used to prevent decomposition of foods and has been shown to have inhibitory effect on the growth of gram positive bacteria. The purpose of this study was to evaluate the effect of polyphosphate on the growth of Porphyromonas endodontalis, a gram negative endodontopathic bacterium. Porphyromonas endodontalis ATCC 35406 was grown in the presence of polyphosphates with different chain lengths. Inhibitory effect of each polyphosphate which was added at the beginning or during the culture, was determined by measuring the optical density of the bacterial cell at 540nm and by viable cell count. The results from this study were as follows : 1. Polyphosphates were shown the growth inhibition of the Porphyromonas endodontalis. 2. The minimal inhibitory concentration(MIC) of polyphosphate was observed to be 0.04%. 3. Polyphosphates with chain lengths of 25 and 75 demonstrated the greatest inhibitory effect on the growth of Porphyromonas endodontalis. 4. Polyphosphates are bactericidal to Porphyromonas endodontalis, demonstrating the growth inhibition of the bacterium. The overall results suggest that use of polyphosphate may affect the growth of Porphyromonas endodontalis. Further studies will be needed to confirm the effect of, polyphosphate.

• Journal of Microbiology and Biotechnology
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• 제17권6호
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• pp.913-918
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• 2007

A Gram-positive, aerobic, rod-shaped, nonmotile, endospore-forming bacterium, designated Gsoil $349^T$, was isolated from soil of a ginseng field and characterized using a polyphasic approach. Comparative analysis of 16S rRNA gene sequences revealed that the strain Gsoil $349^T$ belongs to the family Paenibacillaceae, and the sequence showed closest similarity with Cohnella thermotolerans DSM $17683^T$ (94.1%) and Cohnella hongkongensis DSM $17642^T$ (93.6%). The strain showed less than 91.3% 16S rRNA gene sequence similarity with Paenibacillus species. In addition, the presence of MK-7 as the major menaquinone and $anteiso-C_{15:0},\;iso-C_{16:0},\;and\;C_{16:0}$ as major fatty acids suggested its affiliation to the genus Cohnella. The G+C content of the genomic DNA was 53.4 mol%. On the basis of its phenotypic characteristics and phylogenetic distinctiveness, strain Gsoil $349^T$ should be treated as a novel species within the genus Cohnella for which the name Cohnella panacarvi sp. nov. is proposed. The type strain is Gsoil $349^T\;(=KCTC\;13060^T=\;DSM\;18696^T)$.

• Journal of Microbiology and Biotechnology
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• 제25권7호
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• pp.963-977
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• 2015

The well-characterized gram-positive bacterium Bacillus subtilis is an outstanding industrial candidate for protein expression owing to its single membrane and high capacity of secretion, simplifying the downstream processing of secretory proteins. During the last few years, there has been continuous progress in the illustration of secretion mechanisms and application of this robust host in various fields of life science, such as enzyme production, feed additives, and food and pharmaceutical industries. Here, we review the developments of Bacillus subtilis as a highly promising expression system illuminating strong chemical- and temperatureinducible and other types of promoters, strategies for ribosome-binding-site utilization, and the novel approach of signal peptide selection. Furthermore, we outline the main steps of the Sec pathway and the relevant elements as well as their interactions. In addition, we introduce the latest discoveries of Tat-related complex structures and functions and the countless applications of this full-folded protein secretion pathway. This review also lists some of the current understandings of ATP-binding cassette transporters. According to the extensive knowledge on the genetic modification strategies and molecular biology of Bacillus subtilis, we propose some suggestions and strategies for improving the yield of intended productions. We expect this to promote striking future developments in the optimization and application of this bacterium.

• 한국미생물·생명공학회지
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• 제17권6호
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• pp.634-644
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• 1989

Listeria monocytogenes, one of five species in the genus Listeria and the only one currently believed to be pathogenic for humans, is a small gram-positive, nonsporeforming, aerobic, motile and hemolytic rod-shaped bacterium. The bacterium is widespread in the environment, having been isolated from soil, dust, animal feed, water, sewage, almost every type of animal that has been cultured, and asymptomatic humans. L. monocytogenes causes listeriosis, a disease which most often affects humans with a compromised immune system. Included are pregnant woman, infants and adults suffering from such diseases as cancer, cirrhosis of liver or AIDS or are being treated with drugs such as corticosteroids. Listeriosis is manifested by such syndromes as pregnancy infections, granulomatosis infantiseptica, sepsis, meningoencephalitis, and focal infections. Infections, can be treated successfully with penicillin, ampicillin, or erythromycin. However, a mortality rate of about 30％ has occurred in outbreaks of listeriosis. Food-associated outbreaks of listeriosis have been attributed to coleslaw (Canada, 1981), pasteurized milk (U.S., 1983), and soft cheese (U.S., 1985). Presence of L. monocytogenes in various dairy foods has prompted recall of such products from the U.S. market-place. L. monocytogenes also has been found in raw meats and seafood.