Journal of Microbiology

Volume 35(1); March 1997

Distribution of airborne microorganisms in yellow sands of Korea: Choi, Dae Sung , Park, Yong Keun , Oh, Sang Kon , Yoon, Hee Ju , Kim, Jee Cheon , Seo, Won Jun , Cha, Seung Hee; J. Microbiol. 1997;35(1):1-9.

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Abstract: Distribution of airborne microorganisms was determined with two different types of air samplers, the Anderson cascade sampler and the Aerobioscope sampler, in the vicinity of Taejon. The size distribution of particles carrying bacteria and fungi was concurrently measured. The concentration of detected viable airborne particles was greatly varied. It was observed that the number of microbial particles increased in April and October. The most size of particles carrying bacteria was larger than 4.7 um in mean aerodiameter, which made up 69.8% of the total particle fraction. About 63.2% of fungi-carrying particles were smaller than 4.7 um in aerodiameter. The distribution of particles on Yellow Sand Phenomena days was also analyzed. The number of fine particles having mass median aero-diameter from 1.0 to 10.mu.m increased on Yellow Sand Phenomena days to about 6 times that on normal days and the n umber of colony forming unit (CFU/㎥) of airborne bacteria also increased by 4.3 times in April. The results from the Anderson sampler showed that the concentration of bacteria increased greatly on the fraction of fine particles ranging from 0.6 um to 4.7 um in diameter. Unlike the increase in bacterial floraon Yellow Sand Phenomena days, the fungal concentration slightly decreased and showed a normal size distribution pattern. This study suggests that a long-range transmission of bacteria results form bacteria adsorbing onto the fine particles during the Yellow Sand Phenomena.

Identification of Leuconostoc strains isolated from kimchi using carbon -source utilization patterns: Lee, Jung Sook , Chun, Chang Ouk , Hector, Martin , Kim, Sam Bong , Kim, hong Joong , Park, Bong Keun , Joo, Yun Jung , Lee, Hun Joo , Park, Chan Sun , Ahn, Jong Seog , Park, Yong Ha , Mheen, Tae Ick; J. Microbiol. 1997;35(1):10-14.

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Abstract: The database of metabolic fingerprints generated using the GIolog system of lactic acid bacteria isolated from kimchi, described by Lee et al. (8), was used for the identification of 75 Leuconostoc isolates. The test strains were isolated using a selective isolation medium specific for the genus Leuconostoc and examined for their ability to oxidize carbon sources using the Biology system. The results show that the 75 test strains were identified to the known Leuconostoce clusters. It is suggested that the Biolog system can be applied for rapid identification of lactic acid bacteria isolated from kimchi.

Catalytic mechanism and inhibition studies of purine nucleoside phosphorylase (PNP) in micrococcus luteus: Choi , Hye Seon; J. Microbiol. 1997;35(1):15-20.

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Abstract: Kinetic studies were done to elucidate the reaction mechanism of purine nucleoside phosphorylase (PNP) in Micrococcus Luteus. PNP catalyzes the reversible phosphorolysis of ribonucleosides to their respective base. The effect of alternative competing substrates suggested that a single enzyme was involved in binding to the active site for all purine nucleosides, inosine, deoxyiosine, guanosine, deoxyguanosine, adenosine and deoxyadenosine. Affinity studies showed that pentose moiety reduced the binding capacity and methylation of ring N-1 of inosine and guanosine had little effect on binding to bacterial enzyme, whereas these compounds did not bind to the mammalian enzymes. The initial velocity and product inhibition studies demonstrated that the predominant mechanism of reaction was an ordered bi, bi reaction. The nucleoside bound to the enzyme first, followed by phosphate. Ribose 1-phosphate was the first product to leave, followed by base.

Structure-antifungal activity relationships of cecropin a hybrid peptides against trichoderma sp.: Shin, Song Yub , Lee, Dong Gun , Lee, Sung gu , Kim, Kil Lyong , Lee, Myung Kyu , Hahm, Kyung Soo; J. Microbiol. 1997;35(1):21-24.

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Abstract: The hybrid peptides, CA-ME, CA-MA and CA-BO, with the N-terminal sequence 1-8 of cecropin A and the N-terminal sequences 1-12 of melittin, magainin 2 and bombinin, respectively, have more improved antibacterial activities. CA-MA was found to have stronger antifungal activity against Trichoderma sp than other hybrid peptides and their parental peptides. In order to elucidate the relationships between the peptide structure and antifungal activity, several analogues of CA-MA or CA-BO were also designed and synthesized by the solid phase method. Antifungal activity was measured against T. reesei and T. viride, and hemolytic activity was measured by a solution method against human red blood cells. The residue 16 of CA-MA, Ser, was found to be important for antifungal activity. When the residue was substituted with Leu, showed powerful antifungal activity was dramatically decreased. CA-MA, P1, P4 and P5 designed in this study showed powerful antifungal activity against T. reesei and T. viride with low hemolytic activity against human red blood cells. These hybrid peptides will be potentially useful model to further design peptides with powerful antifungal activity for the effective therepy of fungal infection and understand the mechanisms of antifungal actions of hybrid peptides.

Reduction of hexavalent chromium by pseudomonas aeruginosa HP014: Oh, Young Sook , choi, Sung Chan; J. Microbiol. 1997;35(1):25-29.

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Abstract: Microbial reduction of hexavalent (VI) to trivalent (III) chromium decreases its toxicity by two orders of magnitude. In order to investigate the nature of Cr-reduction, Cr-resistant Pseudomonas aeruginosa HP014 was isolated and tested for its reduction capability. At the concentration of 0.5 mM Cr(VI), cell growth was not inhibited by the presence of Cr(VI) in a liquid medium, and Cr(VI) reduction was accompanied by ell growth. When cell-free extract was tested, the reduction of Cr(VI) showed a saturation kinetics with the maximum specific activity of 0.33 umol min^-1 mg^-1 cell protein, and an apparent K. of 1.73 mM Cr(VI). The activity required either NADH or NADPH as an electron donor. However, NADPH gave 50% as much activity as NADH. To locate the reductase activity, cell free extract was centrifuged at 150,000×g, and subsequently the supernatant and pelleted membrane fractions were tested for Cr(VI) reduction activity. The supernatant of the centrifugation showed almost the same Cr(VI) reduction activity as compared with that of the cell-free extract, indicating that the Cr(VI)-reducing activity of P. aeruginosa HP-14 is due to soluble enzyme. Moreover, the activity appeared to be the highest among the known activities, suggesting that the strain might be useful for remediation of Cr(VI)-contaminated sites.

Growth on methanol of a carboxydobacterium, acinetobacter sp. strain JC1 DSM 3803: Ro, Young Tae , Seo, Jae Goo , Lee, Joo Hun , Kim, Dae Myung , Chung, In Kwon , Kim, Tae Ue , Kim, Young Min; J. Microbiol. 1997;35(1):30-39.

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Abstract: Acinetobacter sp. strain JC1 DSM 3803, a carboxydobacterium, was found to grow methylotrophically at the expense of methanol and methlamine, but not of methane, formaldehyde, formate, dimethylamine, or trimethylamine, as the sole source of carbon and energy. The doubling times of the bacterium growing on methanol (0.5% v/v) and methylamine (0.5% w/v) at 30℃ and pH 6.8 were 4.8 h and 5.7 h respectively. Cells grown on methanol, however, failed to show typical methanol dehydrogenase and oxidase activities. The cell was found to contain no c-type cytochromes. Cells grown on methanol exhibited higher catalase activity than those grown on pyruvate or glucose. The catalase present in the cells also exhibited peroxidase activity. The catalase activity, growth on methanol of the cell, and oxygen consumption by methanol-grown maldehyde dehydrogenase, formaldehyde reductase, glucose-6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase activities were detected from cells grown on methanol.

Nucleotide sequence analysis of a second set of the polyketide synthase β-ketoacyl synthase and chain length factor genes from the salinomycin-producing streptomyces albus: Hyun, Chang Gu , Park, Kwan Hyung , Hutchinson, C. Richard , Suh, Joo Won; J. Microbiol. 1997;35(1):40-46.

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Abstract: The pWHM220 cosmid with a 24-kb insert cloned from Streptomyces albus ATCC 21838 induces the biosynthesis of a polyether antibiotic similar to salinomycin in Streptomyces invidans. We have analyzed this region by DNA sequencing as well as Southern blot hybridization with type I and type II polyketide synthase (PKS) probes. Surprisingly, we found another set of type II SKS genes only 10-kb from the original PKS genes, salABCDE. The DNA sequence revealed two complete open reading frames (ORFs) named salB2 and salC2, and one partial ORF that does not resemble any known DNA or deduced protein sequence. The salC2 should code for chain length determining factor while the deduced amino acid sequence encoded by salB2 exhibits high similarity to β-ketoacyl synthase from different PKS gene clusters. The highest identity was found for β-keetoacyl synthesis from S. argillaceus (MtmP. 59.1% identity), the mithramycin producer and from S. venezuelae ISP5230 (JadA, 52.3% identity), the jadomycin producer. The SalC2 protein clearly resembles its counterparts in order aromatic PKS gene clusters that are believed to influence the length of the polyketide chain. The highest identities observed were to that of S. argillaceus (MtmK, 62.3%) and S. venezuelae ISP 5230 (JadB, 55.1%) proteins, Moreover, the deduced amino acid sequences of the salB2 and salC2 products were 29.0% identical.

Formation of humus-bound residues in the course of BTX biodegradation in soil: Song , Hong Gyu; J. Microbiol. 1997;35(1):47-52.

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Abstract: To examine whether the xylene component of BTX (benzene, toluene, xylene) mixture is cometabolized and residues are produced in soil, ¹⁴C-labeled-0-xylene was added to sandy loam in combination with unlabeled benzene and toluene. After 4 weeks of incubation in a sealed system connected to an oxygen reservoir, 55.1% of the radiocarbon was converted to ¹⁴CO₂, 3.0% was to 95.8% radiocarbon recovery. Biomass incorporation of o-xylene radiocarbon which was detected by fumigation/extraction was usually low (5.6%), but 32.1% radiocarbon became associated with soil humus. Most of the numus-bound radiocarbon was found in humin fraction. In addition to o-xylene, p-xylene and toluene also showed similar results. The evidence shows that some of their reactive methylcatechol biodegradation intermediates attach to the humic metrix in soil in preference to mineralization and biomass incorporation.

Improvement of 4-chlorobiphenyl degradation bya recombinant strain, pseudomonas sp. DJ12-C: Kim, Ji Young , Kim, Young Chang , Lim, Jai Yun , Lee, Ki Sung , Ka, Jong Ok , Min, Kyung Hee , Kim, Chi Kyung; J. Microbiol. 1997;35(1):53-60.

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Abstract: Pseudomonas sp. P20 and Pseudomonas sp. DJ-12 isolated from the polluted environment are capable of degrading biphenyl and 4-chlorobiphenyl (4CB) to produce benzoic acid and 4-chlorobenzoic acid (4CBA) respectively, by pcbABCD-encoded enzymes. 4CBA can be further degraded by Pseudomonas sp. DJ-12, but not by Pseudomonas sp P20. However, the meta-cleavage activities of 2,3-dihydroxybiphenyl (2, 3-DHBP) and 4-chloro-2,3-DHBP dioxygenases (2, 3-DHBD) encoded by pcbC in Pseudomonas sp. P20 were stronger than Pseudomonas sp. DJ-12. In this study, the pcbC gene encoding 2, 3-DHBD was cloned from the genomic DNA of Pseudomonas sp. P20 by using pKT230. A hybrid plasmid pKK1 was constructed and E. coli KK1 transformant was selected by transforming the pKK1 hybrid plasmid carrying pcbC into E. coli XL1-Blue. By transferring the pKK1 plasmide of E. coli KK1 into Pseudomonas sp. DJ-12 by conjugation, a recombinant strain Pseudomonas sp. P20, Pseudomonas sp. DJ-12, and the recombinant cell assay methods. Pseudomonas sp. DJ12-C readily degraded 4CB and 2, 3-DHBP to produce 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoic acid (HOPDA), and the resulting 4CBA and benzoic acid were continuously catabolized. Pseudomonas sp. DJ12-C degraded 1 mM 4CB completely after incubation for 20 h, but Pseudomonas sp. P20 and Pseudomonas sp. DJ-12 showed only 90% and Pseudomonas sp. DJ-12 had, but its degradation activity to 2, 3-DHBP, 3-methylcatechol, and catechol was improved.

Novel strategy for isolating suppressors of meiosis-deficient mutants and its application for isolating the bcy1 suppressor: Shin, Deug Yong , Yun, Jean Ho , Yoo, Hyang Sook; J. Microbiol. 1997;35(1):61-65.

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Abstract: A novel strategy was developed for isolating suppressors from sporulation-deficient mutants. The mutation in the BCY1 gene, which codes for the regulatory subunit of cAMP-dependent protein kinase, when homozygous, results in diploids being meiosis and sporulation deficient. Two plasmids, YCp-MATα and YEp-SPOT7-lacZ, were introduced into MATα BCY1^+ or MATα bcy1 haploid cells. The transformant of the BCY1^+ haploid cell produced β-galactosidase under nutrient starvation, but the bcy1 transformant did not. Using this system, the mutagenesis experiment performed on the bcy1 transformant strain resulted in a number of sporulation mutants that produced β-galactosidase under nutrient starvation. One complementation group, sob1, was identified from the isolated suppressor mutants and characterized as a single recessive mutation by tetrad analysis. Genetic analysis revealed that the sob1 mutation suppressed the sporulation deficiency, the failure to arrest at the G1 phase of the cell cycle, and the sensitivity to heat or nitrogen starvation caused by the bcy1 mutation. However, the sob1 mutation did not suppress the sporulation deficiency of ime1 and of ime2 diploids. These results suggest that the sob1 mutation affects a gene which functions as a downstream regulator in both meiosis and cell cycle regulation.

Biodegradation of aromatic hydrocarbons by several white-rot fungi: Song , Hong Gyu; J. Microbiol. 1997;35(1):66-71.

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Abstract: To investigate the biodegrading capability of several white-rot fungi isolated in Korea, biodegradation of BTX (benzene, toluene, xylene), phenanthrene and pyrene were tested in fungal cultures. Phanerochaete chrysosporium removed 20-30% of BTX mixture during 21 days of incubation in serum bottle. Coriolus versicolor KR-11W and Irpex lacteus mineralized 10.02 and 8.26% of totla phenanthrene, respectively, which were higher than in other studies with P. chrysosporium. These two strains also showed high mineralization rates (9.2-10.1%) for 4-ring pyrene. I. lacteus metabolized most of the added pyrene and 23.29% was incorporate dinto fungal biomass. Almost 50% of the pyrene was converted to polar metabolites and recovered from aqueous phase of culture. These results indicated that some white- rot fungi have higher biodegradability than P. chrysosporium and could be used in bioremediation of aromatic hydrocarbon contaminants in soil.

Oligomerization of the substitution mutants of autographa californica nuclear polyhedrosis Virus (AcNPV) gp64 glycoprotein: Kim, Ki Nam , Poo, Ha Ryoung , Yang, Jai Myung; J. Microbiol. 1997;35(1):72-77.

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Abstract: The baculovirus gp64 glycoprotein is a major component of the envelope protein of budded virus (BV). It has been shown that the gp64 glycoprotein plays an essential role in the infection process, especialy fusion between virus envelope and cellular endosomic membrane. Recently we reported optimal conditions required for gp64-mediated membrane fusion in pGP64 DNA transfected Spodoptera frugiperda (Sf9) cells (H. J. Kim and J. M. Yang, Jour, Microbiology, 34, 7-14). In order to investigate the role of hydrophobicity within the fusion domain of the gp64 glycoprotein for membrane fusion, 13 mutants which have substitution mutation within hydrophobic region I were constructed by PCR-derived site-derected mutagenesis. Each mutated gp64 glycoproteins was transiently expressed by transfecting plasmid DNA into Spodoptera frugiperda (Sf9) cells. Oligomerization of the transisently expressed gp64 glycoproteins was a analysed by running them on SDS-polyacrylamide gel electrophoresis under non-reducing condition followed by immunoblotting. All of the mutant gp64 glycoproteins expect cysteine-228 were able to form trimers. These results suggest that hydrophobic region I of the gp64 may not be responsible for the oligomerization of the gp64 glycoprotein.

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