• Mycobiology
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• v.42 no.4
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• pp.368-375
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• 2014

Red ginseng (Panax ginseng), a Korean traditional medicinal plant, contains a variety of ginsenosides as major functional components. It is necessary to remove sugar moieties from the major ginsenosides, which have a lower absorption rate into the intestine, to obtain the aglycone form. To screen for microorganisms showing bioconversion activity for ginsenosides from red ginseng, 50 yeast strains were isolated from Korean traditional meju (a starter culture made with soybean and wheat flour for the fermentation of soybean paste). Twenty strains in which a black zone formed around the colony on esculin-yeast malt agar plates were screened first, and among them 5 strains having high ${\beta}$-glucosidase activity on p-nitrophenyl-${\beta}$-D-glucopyranoside as a substrate were then selected. Strain JNO301 was finally chosen as a bioconverting strain in this study on the basis of its high bioconversion activity for red ginseng extract as determined by thin-layer chromatography (TLC) analysis. The selected bioconversion strain was identified as Candida allociferrii JNO301 based on the nucleotide sequence analysis of the 18S rRNA gene. The optimum temperature and pH for the cell growth were $20{\sim}30^{\circ}C$ and pH 5~8, respectively. TLC analysis confirmed that C. allociferrii JNO301 converted ginsenoside Rb1 into Rd and then into F2, Rb2 into compound O, Rc into compound Mc1, and Rf into Rh1. Quantitative analysis using high-performance liquid chromatography showed that bioconversion of red ginseng extract resulted in an increase of 2.73, 3.32, 33.87, 16, and 5.48 fold in the concentration of Rd, F2, compound O, compound Mc1, and Rh1, respectively.

• Journal of Yeungnam Medical Science
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• v.2 no.1
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• pp.183-190
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• 1985

The glucose nonfermenting gram-negative bacilli encountered about 10% of all gram-negative bacilli isolated from clinical material. Therefore, a rapid and correct identification of glucose nonfermenting gram-negative bacilli is impostent for a better management of infectious disease. There are many conventional systems for the Identification of glucose nonfermenting gram-negative bacilli but most of them have problems and difficulties. Commercial Kit Systems exist and they are too expensive for dally use 10 Korea because of high cost. Based on 12 selected tests we propose a new code system, MCRCODE-N for rapid and 10-expensive identification of glucose nonfermenting gram-negative bacilli. The selective 12 tests are oxidase, glucose oxidation motility, urease, DNase arginine dehydrolase, nitrate reduction, gelatin Liquefaction, esculin hydrolysis, mannitol oxidation, maltose oxidation, Lactose oxidation. The 12 tests are divided 4 group and then each group has 3 tests. The result of each group is expressed by the number as below. The positive test is given by specific number (1st test = 1, 2nd test = 2, 3rd test = 4), while any negative result is 0. Each 3 numbers of one group are added and make number of 1 digit. Four digit number is refered to the code book of MCRCODE-N system or MCRCODE system using computer (Apple-II model) created by authors. This MCRCODE-N system is suitable ones for our use 10 Korea. We propose the MCRCODEN-N system for clinical use.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.901-907
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• 2008

Thermotoga neapolitana $\beta$-glucosidase (BglA) was subjected to site-directed mutagenesis in an effort to increase its ability to synthesize arbutin derivatives by transglycosylation. The transglycosylation reaction of the wild-type enzyme displays major ${\beta}(1,6)$ and minor ${\beta}(1,3)$ or ${\beta}(1,4)$ regioselectivity. The three mutants, N291T, F412S, and N291T/F412S, increased the ratio of transglycosylation/hydrolysis compared with the wild-type enzyme when pNPG and arbutin were used as a substrate and an acceptor, respectively. N291T and N219T/F412S had transglycosylation/hydrolysis ratios about 3- and 8-fold higher, respectively, than that of the wild-type enzyme. This is due to the decreased hydrolytic activity of the mutant rather than increased transglycosylation activity. Interestingly, N291T showed altered regioselectivity, as well as increased transglycosylation products. TLC analysis of the transglycosylation products indicated that N291T retained its ${\beta}(1,3)$ regioselectivity, but lost its ${\beta}(1,4)$ and ${\beta}(1,6)$ regioselectivity. The altered regioselectivity of N291T using two other acceptors, esculin and salicin, was also confirmed by TLC. The major transglycosylation products of the wild type and N291T mutant were clearly different. This result suggests that Asn-291 is highly involved in the catalytic mechanism by controlling the transglycosylation reaction.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.99-104
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• 2009

The $\beta$-glucosidase gene from Streptomyces coelicolor A3(2) was cloned and expressed in Escherichia coli. The ORF consisted of 1377 nucleotides encoding 51 kDa in a predicted molecular weight. Effects of pH indicated that the $\beta$-glucosidase showed similar activity using $\alpha$-pNPG($\rho$-nitrophenyl-$\alpha$-D-glucopyranoside), $\beta$-pNPG($\rho$-nitrophenyl-$\beta$-D-glucopyranoside), and $\beta$-pNPF($\rho$-nitrophenyl-$\beta$-D-fucopyranoside) at range of pH 3 to 10, and high activity using $\beta$-pNPGA ($\rho$-nitrophenyl-$\beta$-D-galactopyranoside) from pH 5 to 10, especially, 3.3 times higher activity at pH 9. Effects of temperature indicated that the $\beta$-glucosidase showed low activity using $\alpha$-pNPG, $\beta$-pNPG, and $\beta$-pNPF from $20^{\circ}C$ to $70^{\circ}C$, and increased activity using $\beta$-pNPGA from $30^{\circ}C$ to $50^{\circ}C$, 1.8 times higher activity at $50^{\circ}C$ than at $30^{\circ}C$. According to activity determination of other substrates, the enzyme was active on daidzin, genistin, and glycitin, inactive on esculin and apigenin-7-glucose. The EDTA and DTT as reducing agents inhibited $\beta$-glucosidase activity, but SDS and mercaptoethanol did not inhibit. Monovalent or divalent metal ions such as $MnSO_4$, $CaCl_2$, KCl, and $MgSO_4$ did not inhibited $\beta$-glucosidase activity. $CuSO_4$ and NaCl showed low inhibition, and $ZnSO_4$ inhibited 3.3 times higher than control.

• Journal of the korean academy of Pediatric Dentistry
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• v.27 no.2
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• pp.274-282
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• 2000

Stainless steel crowns are widely used for restoration for primary molars. The material used for the crowns is an alloy of $70\sim80%$ nickel and $5\sim15%$ chromium. Nickel has been known to cause allergic reaction, cancer and cell toxicity. Little is known about nickel with respect to the relationship between Ni-contained SS crown and graining of Ni-resistance in oral microorganisms. The purpose of this study is to examine whether use of Ni-contained SS crown leads to occurrence of Ni-resistant microorganism, especially enterococci. The gingival crevicular fluid of two different groups was taken. Experimental group included patients wearing SS crown, and control group comprised individuals without SS crown. The samples were plated in BHI agar, BHI agar supplemented with nickel chloride at the concentration of 3mM and bile esculin azide (BEA) agar. The cultured enterococci on BEA agar medium were tested their Ni-resistance in nickel-containing media increasing concentrations from 3mM to 50mM. The results were as follows: 1. In experimental group, a total of 507,350 strains were isolated on BHI agar, of which 53,864(10.62%) strains were found to be resistant to 3mM nickel. In control group, of 414,590 isolates on BHI agar, 37,523 isolates were resistant to 3mM nickel. 2. A total of 95 enterococci were isolated on BEA agar in experimental group, while 20 were isolated in control group. of the enterococci, 68 and 12 isolates were found to be nickel-resistant in experimental and control group, respectively. 3. Of 68 nickel-resistant isolates in experimental group, one survived 50mM nickel. In contrast, none of the isolates in control group was observed to grow at the concentrations over 30mM nickel.

• Korean Journal of Microbiology
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• v.52 no.3
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• pp.319-326
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• 2016

An antimicrobial bacterium to pathogenic microorganisms, strain $W5-1^T$ was isolated from Korean fermented-food Kimchi. The isolate was Gram-staining-variable, strictly aerobic, rod-shaped, endospore-forming, and motile with peritrichous flagella. It grew at $15-40^{\circ}C$, at pH 6.0-10.0, and in the presence of 0-4% NaCl. Strain $W5-1^T$ could hydrolyze esculin and xylan, and assimilate $\small{D}$-mannose, but not $\small{D}$-mannitol. Strain $W5-1^T$ showed antimicrobial activity against Listeria monocytogens, Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella typhi. The G+C content of the DNA of strains $W5-1^T$ was 52.6 mol%. The predominant respiratory quinone was menaquinone-7 (MK-7) and the major cellular fatty acids were $C_{16:0}$, antieiso-$C_{15:0}$, $C_{18:0}$, and $C_{12:0}$. The strain contained meso-diaminopimelic acid in cell-wall peptidoglycan. On the basis of 16S rRNA gene sequence and phylogenetic analysis, the strain W5-1 was shown to belong to the family Paenibacillaceae and was most closely related to Paenibacillus pinihumi $S23^T$ (98.4% similarity) and Paenibacillus tarimensis $SA-7-6^T$ (96.4%). The DNA-DNA relatedness between the isolate and Paenibacillus pinihumi $S23^T$ was 8.5%, indicating that strain $W5-1^T$ represented a species in the genus Paenibacillus. On the basis of the evidence from this polyphasic study, it is proposed that strain $W5-1^T$ is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus kimchicus sp. nov. is proposed. The type strain is $W5-1^T$ (=KACC $15046^T$ = $LMG 25970^T$).

• Food Science of Animal Resources
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• v.37 no.5
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• pp.735-742
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• 2017

This study was conducted to isolate and characterize Paenibacillus sp. MBT213 possessing ${\beta}$-glucosidase activity from raw milk, and examine the enzymatic capacity on the hydrolysis of a major ginsenoside ($Rb_1$). Strain MBT213 was found to have a high hydrolytic ability on ginsenoside $Rb_1$ by Esculin Iron Agar test. 16S rDNA analysis revealed that MBT213 was Paenibacillu sp. Crude enzyme of MBT213 strain exhibited high conversion capacity on ginsenoside $Rb_1$ into ginsenoside Rd proven by TLC and HPLC analyses. The API ZYM kit confirmed that Paenibacillu sp. MBT213 exerted higher ${\beta}$-glucosidase and ${\beta}$-galactosidase activity than other strains. Optimum pH and temperature for crude enzyme were found at 7.0 and $35^{\circ}C$ in hydrolysis of ginsenoside $Rb_1$. After 10 d of optimal reaction conditions for the crude enzyme, ginsenoside $Rb_1$ fully converted to ginsenoside Rd. Ginseng roots (20%) were fermented for 14 d, and analyzed by HPLC showed that amount of ginsenoside $Rb_1$ significantly decreased, while that of ginsenoside Rd was significantly increased. The study confirmed that the ${\beta}$-glucosidase produced by Paenibacillus sp. MBT213 can hydrolyze the major ginsenoside $Rb_1$ and convert to Rd during fermentation of the ginseng. The ${\beta}$-glucosidase activity of this novel Paenibacillus sp. MBT213 strain may be utilized in development of variety of health foods, dairy foods and pharmaceutical products.

• Microbiology and Biotechnology Letters
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• v.37 no.1
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• pp.55-61
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• 2009

Ginseng saponins (a secondary metabolite, termed ginsenosides) are the principal bioactive ingredients of ginseng, and modification of the sugar chains may markedly change the its biological activity. One of soil bacteria having $\beta$-glucosidase (to transform ginsenoside $Rb_1$) activity was isolated from soil of a ginseng field in Daejeon. 16S rRNA gene sequence analysis revealed that the isolate belonged to the genus Cellulosimicrobium, with highest sequence similarity (99.7%) to Cellulosimicrobium funkei ATCC BAA-$886^T$. The strain, Gsoil 235, could transform ginsenoside $Rb_1$ into Rd, $Rg_3$ and 3 of un-known ginsenosides by the analyses of TLC, HPLC. By investigating its deglycosylation progress, the optimal broth for, $\beta$-glucosidase was nutrient broth (In 48 hours, almost ginsenoside $Rb_1$ could be transformed into minor ginsenosides). On the contrary, the optimal broth for growth was determined as trypic soy broth (TSB).

• Korean journal of applied entomology
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• v.14 no.3 s.24
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• pp.111-131
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• 1975

The study has been carried out to investigate the occurrence, damage, characteristics of the pathogen, environmental conditions affecting the disease outbreak, varietal resistance, forecasting, and chemical control of bacterial leaf blight of rice in Korea since 1964. Bacterial leaf blight of rice became a major disease in Korea since 1960. A correlation was found between the annual increase of epidemics and increase of cultivation area of susceptible varieties, Jinheung, Keumnampung etc. Areal damage within the country showed that the more was at southern province, Jeonnam, Gyeongnam and western coast, and at flooded rice paddy. Yield reduction directly related with the amount of infection on upper leaves at heading stage. Fifty per cent of reduction resulted when the lesion area was more than 60 per cent. Less than 20 per cent of lesion area, however, was not affected so much on yield loss One hundred and six isolates collected from all over the country were classified as 8 strains by using 4 different bacteriophages in 1973. It was, however, only two in 1965. There were some specificities on varietal distributions among the strains such as that the Jinheung attacked mainly by strain A, B, C and I, those attack Kimmaze were A, B, H and I. Most strains were found from Tongil except D and E, whereas Akibare was only variety that attacked by strain E. Low temperature, high humidity, heavy rainfall and insutficient daylight favored the disease epidemics. Especially, typhoon and flooding at heading stage were critical factors. The earlier transplanting the more disease was resulted, and more nitrogen fertilizer application accerelated the diseased development in general. The resistance to the disease varied by growing stage of the sane plants. All of recommended varieties in Korea were susceptible to the disease except Norm No. 6 and Sirogane which moderately resistant. The pathogen, Xanthomonas oryzae, was detectable from extract of healthy seedlings that were grown in the field with an heavy infection previous year. The more bacteriophage in irigation water resulted the more disease outbreak, and the existence of more than 50 bacteriophages in 1ml. of irrigation water were necessary to initiate the disease out break. The curves representing occurrence of bacteriophages and disease outbreak were similar with 15 days interval. The survey of bacteriophage occurrence can be utilized in forecasting of the disease two weeks ahead of disease outbreak. Three applications of chemicals, Phenazin and Sangkel, in weekly intervals at the early satage of out-break depressed the symptom development, and increased yield by 20per cent. Proper period for the chemical application was just before the number of bacteriophage reaches 50 in 1ml. of irrigation water.