• Korean Journal of Microbiology
• /
• v.44 no.3
• /
• pp.264-269
• /
• 2008

To develop an amylolytic industrial yeast strain producing $\beta$-amylase, the BAMY gene encoding Achlya bisexualis $\beta$-amylase was constitutively expressed under the control of the alcohol dehydrogenase gene promoter (ADC1p) in an industrial strain of Saccharomyces cerevisiae. Yeast transformation was carried out by an integration system containing $\delta$-sequences as the recombination site. The integrative cassette devoid of bacterial DNA sequences was constructed that contains the BAMY gene and $\delta$-sequences. Industrial S. cerevisiae transformed with this integrative cassette secreted 45 kDa $\beta$-amylase into the culture medium. The $\beta$-amylase activity of the transformant was approximately 18.5-times higher than that of A. bisexualis. The multi-integrated BAMY genes in the transform ant were stable after 100 generations of growth in nonselective medium. Hydrolysis of soluble starch and various starches with the enzyme released maltose but not glucose or oligosaccharides.

• Korean Journal of Microbiology
• /
• v.44 no.3
• /
• pp.169-177
• /
• 2008

Global gene expression of Deinococcus radiodurans, a highly radiation resistant bacterium, in response to excess copper was analyzed by using oligonucleotide microarray chip. Among 3,187 open reading frames of D. radiodurans, seventy genes showed a statistically significant expression ratio of at least 2-fold changes under growth conditions of excess copper; 64 genes were induced and 6 genes were reduced. Especially, two operons ($DRB0014{\sim}DRB0017$ and $DRB0125{\sim}DRB0121$) presumably involved in the iron transport and utilization were the most highly induced genes by excess copper. A quantitative real-time PCR assay revealed that DRB00l4 and DRB0125 are highly transcribed responding to excess copper and 2,2'-dipyridyl, an iron chelator. In addition, the transcription of both genes was not changed by excess iron and bathocuproine disulphonate, a copper chelator. These results suggested that the copper metabolism may be closely connected with the iron transport and utilization in D. radiodurans. However, the disruption of each gene, DRB00l4 and DRB0125, did not affect the copper and radiation resistance, the most well-known character of this organism.

• Korean Journal of Microbiology
• /
• v.44 no.3
• /
• pp.193-198
• /
• 2008

ErmSF is one of the proteins which are produced by Streptomyces fradiae to avoid suicide by its autogenous macrolide antibiotic, tylosin and one of ERM proteins which are responsible for transferring the methyl group to $A_{2058}$ (Escherichia coli coordinate) in 23S rRNA, which reduces the affinity of MLS (macrolide-lincosamide-streptogramin B) antibiotics to 23S rRNA, thereby confers the antibiotic resistance on microorganisms ranging from antibiotic producers to pathogens. ErmSF contains an extra N-terminal end region (NTER), which is unique to ErmSF and 25% of amino acids of which is arginine known well to interact with RNA. Noticeably, arginine is concentrated in $^{58}RARR^{61}$ and functional role of each arginine in this motif was investigated through deletion and site-directed mutagenesis and the activity of mutant proteins in cell R60 and R61 was found to play an important role in enzyme activity through the study with deletion mutant up to R60 and R61. With the site-directed mutagenesis using deletion mutant of 1 to 59 (R60A, R61A, and RR60, 61AA), R60 was found more important than R61 but R61 was necessary for the proper activity of R60 and vice versa. And these amino acids were presumed to assume a secondary structure of $\alpha$-helix.

• Korean Journal of Materials Research
• /
• v.7 no.1
• /
• pp.69-75
• /
• 1997

Thin films of $Si0_2(1000{\AA})/CoNiCr(400{\AA})/Cr$ were fabricated as a function of Cr thickness by KF magnetron sputtering. The saturation magnetization, coercive force and squareness with annealing temperature for these films were investigated. The values of saturation magnetization of $SiO_2/CoNiCr/Cr$ thin films decreased as the thickness of Cr underlayer increased, whereas coercive force increased as the thickness of Cr underlayer increased. The value of Ms was 600 emu/cc and the maximum value of Hc was 550 Oe. Especially, the value of saturation magnetization was rapidly decreased $SiO_2/CoNiCr/Cr(1700{\AA})$ thin films as the annealing temperature increased And the coercive force increased as the annealing temperature increased When annealing temperature was $650^{\circ}C$, the Ms was reduced to 90 % of the as-deposited film. And the Hc was showed maximum 1600 Oe. It was thought that Cr diffusion into CoNiCr layer reduced the magnetic moment of CoKiCr layer. In addition. Hc might he increased due to grain growth perpendicular to the film plane.

• Korean Journal of Microbiology
• /
• v.52 no.3
• /
• pp.254-259
• /
• 2016

Acinetobacter sp. B-W producing siderophore, 2, 3-dihydroxybenzoic acid (DHB) was analyzed for plasmid content. Strain B-W harbored plasmid of 20 kb in size. Growth at $43^{\circ}C$ was effective in producing mutant cured of plasmid of strain B-W. This mutant lost the ability to produce 2, 3-DHB. Formation of siderophore halos on the chrome azurol S (CAS) agar medium was not detected by cured strain B-W. pHs of supernatants of wild type strain B-W and cured mutant grown in glucose and $MnSO_4$ containing medium at $28^{\circ}C$ for 3 days were 4.5 and 8.5, respectively. Antibiotic resistance against ampicillin, actinomycin D, bacitracin, lincomycin, and vancomycin was lost in cured mutant. Plasmid curing of strain B-W resulted in drastic reduction of minimal inhibitory concentration (MIC) of several antibiotics. E. coli $DH5{\alpha}$ was transformed with plasmid isolated from strain B-W. The transformant E. coli $DH5{\alpha}$ harbored a plasmid of the same molecular size as that of the donor plasmid. Transformant E. coli $DH5{\alpha}$ produced 2, 3-DHB and contained antibiotic resistant ability. Thus a single plasmid of 20 kb seemed to be involved in 2, 3-DHB production. Genes encoding resistance to antibiotics were also supposed to be located on this plasmid.

• Korean Journal of Microbiology
• /
• v.52 no.3
• /
• pp.352-364
• /
• 2016

In this study, we examined in vitro the potential probiotic properties of lactic acid bacteria (LAB) obtained from the fish intestine and their ability to degrade histamine through the production of diamine oxidase (DAO) enzymes and bacteriocin. Among 97 LAB strains isolated from the intestine of croaker, flatfish, pollack, and rockfish, CIL08, CIL16, FIL20, FIL31, PIL45, PIL49, PIL52, and RIL60 isolates exhibited excellent survival rates under simulated gastrointestinal tract conditions, high adhesion ability to HT-29 epithelial cells, and resistance to the antibiotics such as amoxicillin, ampicillin, erythromycin, penicillin G, streptomycin, tetracycline, or vancomycin. In addition, these strains did not produce histamine in decarboxylating broth containing histidine. In particular, 4 strains (CIL08, FIL20, PIL52, and RIL60) that may produce DAO were significantly able to degrade histamine. The bacteriocins produced by FIL20, FIL31, and PIL52 LAB inhibited the growth and histamine production of Enterococcus aerogenes CIH05, Serratia marcescens CIH09, Enterococcus faecalis FIH11, Pediococcus halophilus FIH15, Lactobacillus sakei PIH16, Enterococcus faecium PIH19, Leuconostoc mesenteroides RIH25, or Aeromonas hydrophilia RIH28. Histamine-producing strains isolated from fish intestine were found to reduce histamine accumulation during co-culture with CIL08, FIL20, PIL52, and RIL60 LAB showing histamine degradation or bacteriocin production ability. The probiotic strains preventing histamine formation were identified as Pediococcus pentosaceus CIL08, Lactobacillus plantarum FIL20, Lactobacillus paracasei FIL31, Lactobacillus sakei PIL52, and Leuconostoc mesenteroides RIL60 with high similarity based on 16S rRNA gene sequencing.

• Korean Journal of Microbiology
• /
• v.52 no.3
• /
• pp.243-248
• /
• 2016

RNase E (Rne) is an essential enzyme involved in the processing and degradation of a large portion of RNAs in Escherichia coli. The enzymatic activity of RNase E is controlled by regulators of ribonuclease activity, namely, RraA and RraB. Gram-positive bacterium Streptomyces coelicolor also contains homologs of Rne and RraA, designated as RNase ES (Rns), RraAS1, and RraAS2. In the present study, we investigated the effect of S. coelicolor RraAS1 on the ribonucleolytic activity of RNase E in E. coli. Coexpression of RraAS1 with Rne resulted in the decreased levels of rpsO, ftsZ, and rnhB mRNAs, which are RNase E substrates, and augmented the toxic effect of Rne overexpression on cell growth. These in vivo effects appeared to be induced by the binding of RraAS1 to Rne, as indicated by the results of co-immunoprecipitation analysis. These results suggested that RraAS1 induces ribonucleolytic activity of RNase E in E. coli.

• The Korean Journal of Mycology
• /
• v.24 no.1 s.76
• /
• pp.67-80
• /
• 1996

Eleven fruit bodies of Ganoderma sp. were collected from eight locations throughout the forest of Kangwon province and Kyunggi province in Korea. The hosts in forest were cut trunks of Quercus dentata, Q. variabilis, Prunus peria and Alnus japonica that was newly surveyed but 5 isolates were collected at the farms of Ganoderma mushroom. Most fruit bodies were formed solitarily on the cut trunks but GS-106 isolate grown in crowds on cut trunk of Alnus japonica. Optimal temperature ranges for isolates of species studied were: G. applanatum $28^{\circ}C{\sim}30^{\circ}C$, G. lucidum $28{\sim}30^{\circ}C$, G. neo-japonicum $28^{\circ}C$, and G. tsuage $26^{\circ}C$ and all the species grew slowly at the $32^{\circ}C$. Hamada medium adjusted with pH 5.4 and 6.2 is better than other media for mycelial growth. Mycelial morphological characteristics of six species were studied: G. applanatum, G. lucidum and G. neo-japonicum produced typical type of staghoru hyphae but G. oregonens and G. valeosiacum produced staghoru hyphae with a branch of grape form. Clamp connection was observed on hypha of G. applanatum, G. lucidum, G. oregonense and G. valeosiacum except G. neo-japonicum with node type. Chlamydospore was produced by G. applanatum, G. neo-japonicum. and cuticular cells were present on hyphae of G. lucidum, G. neo-japonicum, G.oregonense and G. tsugae.

• Korean Journal of Materials Research
• /
• v.14 no.12
• /
• pp.893-899
• /
• 2004

Even though Ti-6Al-4V has gained popularity as an implant material, the possible dissolution of Al and V ions in body fluids remains a matter of concern. Though commercially pure Ti (Cp-Ti) overcomes this problem, the mechanical strength of pure titanium remains very low. Thus, in this experiment Cp-Ti was processed by Equal channel angular processing (ECAP), in order to increase the mechanical strength. The biocompatibility of ECAP-Ti, Cp-Ti and Ti-6Al-4V was examined by the apatite formation on each sample surface, after treating the surface with 5M NaOH and soaking in Simulated body fluids (SBF). Initially, the samples were mechanically polished on silicone carbide paper (#2000). The polished samples were treated with 5M NaOH solution at $60^{\circ}C$ for 24 hours. The NaOH treated samples were washed gently with distill water and dried at $40^{\circ}C$ for 1 day. The dried samples were heat treated in air at $600^{\circ}C$ for 1 hour. The surface morphology of these samples were studied using SEM and XRD. The SEM studies showed network of pores in all samples. The XRD showed oxide layer formation on Cp-Ti and Ti-6Al-4V. samples. However the oxide layer in ECAP-Ti was not substantial. These samples were immersed in SBF, kept at $36.5^{\circ}C$ for seven days period. At the end of 7 days, the apatite formation was confirmed only on Cp-Ti and was not observed in Ti-6Al-4V and ECAP-Ti. These observations of apatite formation relate to the fact that Cp-Ti showed greater oxide layer than other samples. The apatite examined was confirmed as tricalcium phosphate (TCP) using EDS and XRD.

• The Korean Journal of Mycology
• /
• v.19 no.2
• /
• pp.136-142
• /
• 1991

Colonial morphology of the various yeasts grown on the yeast morphology agar me­dium containing orange-yellow pigments extracted from the fruits of Gardenia jasminoides (GJPM) was investigated in hopes of the differential identification of yeasts on primary cultures. Colonies of Candida lusitaniae and Ca. guilliermondii on GJPM turned to prussian blue within three days of incubation and Ca. tropicalis and Ca. viswanathii turned to bluish gray but the latter species turned to deep blue after 7 days. Ca. krusei, Saccharomyces cerevisiae, and Torulopsis glabrata showed neutral gray, grayish green, and baby blue respectively after one or two weeks. However, the colonies of Ca. albicans and parapsilosis remained unchanged even after 20 days. Colonial color of Cryptococ­cus neoformans showing brown to purple brown was distinguishable not only from buff color of Cr. laurentii after one or two weeks incubation but also from those of Candida spp. Growth of certain species was promoted on GJPM. The findings clearly showed that Ga. jasminoides pigments medium was useful to the morphological differentiation of medically important yeasts which were often encountered in sputum or other clinical specimens.