• 한국생물공학회:학술대회논문집
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• 2005.04a
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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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• v.43 no.8
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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.

• Food Science of Animal Resources
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• v.27 no.4
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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.

• Korean Journal of Environmental Biology
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• v.25 no.3
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• pp.260-266
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• 2007

A thermophilic bacterium capable of poly (L-lactide)(PLLA) degradation was isolated from cultivating soil in Korea. The isolate was Gram positive rod-shaped bacterium, and was identified as Geobacillus caldoxylosilyticus based on the 16S rDNA sequence analysis. The strain proved to be a new PLLA degrading bacterium which has not been reported in the open literatures yet. The degradation activity of the strain was assessed in a sterilized compost inoculated with the strain under controlled compost condition at $58^{\circ}C$ for 40 days. The strain mineralized 66%, 57%, 41% and 40% of PLLA5000, PLLA11000, PLLA34000 and PLLA256000 whose weight average molecular weights were 5000, 11000, 34000 and 256000, respectively. Incorporation of 0.1% each of gelatin, yeast extract and ammonium sulfate in the compost containing PLLA256000 as a nutritional supplement raised the biodegradation activity by 27%, 13% and 10%, respectively. Increase of the inoculum size from $10^9cfu\;g^{-1}\;to\;10^{10}cfu\;g^{-1}\;and\;10^{11}cfu\;g^{-1}$ also enhanced the biodegradation activity by 14% and 20%, respectively.

• Korean journal of applied entomology
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• v.41 no.1
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• pp.49-53
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• 2002

Paenibacillus larvae is a gram-positive, spore-forming bacterium that is etiological agent for american foulbrood disease (AFB), which is the most severe disease in honey bee. To detect P. larvae from infected honeybee-comb or larvae, polyclonal antibody against whole bacterium was produced from guineapig and its specificity was evaluated. After optimization of ELISA-based detection system using these antibodies, a number of different P. larvae strains were analysed. Polyclonal antibody against P. larvae ATCC 25747 showed high affinity to most strains of P. larvae including P. larvae. strain ATCC 9545 (type strain), ATCC 25747 and other korean strain, SJl5 but exhibited no cross-reaction with other bacterial species. Additionally, this type of ELISA system was used for the detection of AFB in field-application The results have shown that this antibody could be useful for the rapid identification and monitoring of P. larvae in honeybee-comb.

• Applied Biological Chemistry
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• v.43 no.3
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• pp.169-173
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• 2000

For the development of enzyme detergent capable of effectively washing at low temperature, a bacterium producing alkaline protease was isolated from soil samples, and properties of the enzyme were investigated. The selected strain was Gram negative, rod shape$(0.6{\sim}0.7{\times}1.3{\sim}2.6\;{\mu}m\;in\;size)$ and motile. It had the degradation activity of aesculin, gelatin and casein, and was catalase-positive. The cell wall components was meso-DAP, and G+C mole contents was 43.3%. From these results, the strain was identified as Acinetobacter sp. KN-27. The activity of alkaline protease by this strain peaked with 3,300 D.U/mL after 36 hours in the liquid culture at $40^{\circ}C$. The optimal pH and temperature of the enzyme were pH 9 and $60^{\circ}C$, respectively. Alkaline protease produced by Acinetobacter sp. KN-27 has shown two active bands on the electrophoresis of native gel.

• Restorative Dentistry and Endodontics
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• v.23 no.1
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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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• v.17 no.6
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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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• v.25 no.7
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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.

• Microbiology and Biotechnology Letters
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• v.17 no.6
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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.