• Microbiology and Biotechnology Letters
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• v.11 no.3
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• pp.175-180
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• 1983

Thialysine significantly inhibited the growth of wild type strain Gondida utilis NCYC-359. In the absence of thialysine, the culture reached stationary phase after 24hr, however, in the presence of 0.5% thialysine, the culture reached stationary phase after 40hr, respectively. Effect of amino acid or vitamin was investigated on recovery of the growth of wild type strain from thialysine inhibition. Glycine, methionine, arginine and tryptophan recovered growth inhibition by thialyslne to some extent. However, vitamins were inert. Especially, lysine at one eighth concentration of thialysine recovered almost fully the growth inhibition. Thialysine resistant mutants were induced from the parent strain of Condida utilis NCYC-359 by NTG treatment. Colonies of thialysine resistant mutants were obtained on agar minimal medium supplemented with 0.1-0.5% thialysine. The frequency of thialysine resistant mutants induced by the first mutation was the highest at 0.1% The wild strain produced no appreciable lysine extracellularly. However, almost thialysine resistant mutants excreted appreciably. Lysine excretion increased after repeated mutation. Finally, of the thialysine resistant mutants induced by NTG, Condida utilis TRN-4006 was obtained. This strain excreted lysine (400$\mu\textrm{g}$/$m\ell$) into the medium with a concomitant decrease of lysine in the intracellular pool.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.723-726
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• 2001

This study was performed to improve the yield of bacterial cellulose(BC) by UV and NTG mutagenesis of strain KJ-1 which produced largely BC. some mutants showed high BC productivity with twice elevation compared to that the wild strain KJ-1. A difference was found in production and bioconversion phase of synthesized organic acid, such as gluconic acid, 2-keto gluconic acid, and 5-keto gluconic acid between mutants and strain KJ-1 in the static culture. The organic acid produced in secondary metabolism phase, were more rapidly consumed in the culture with the mutants than that the parent strain after glucose in the broth was conversed to a limiting substrate. Therefore, we suggested the reason for increasing of BC production that the mutant strain consumed more efficiently synthesized acids as substrates than that of the parent strain.

• Archives of Pharmacal Research
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• v.23 no.4
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• pp.418-423
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• 2000

Oka strain VR-795 (Varicella Zoster Virus, VZV) of American Type Culture Collection (ATCC) has been used for chickenpox vaccine production. In order to use this strain for vaccine production, the strain must be identified and its stability must be confirmed. The identification of the Oka strain has been confirmed using Restriction Fragment Length Polymorphism (RFLP) and DNA sequence analysis of glycoprotein-II (gp-II). The amino acid sequences of Oka deduced from the DNA sequence of gp-II have changed at three amino acids against Ellen and at one amino acid against Webster. To prove the stability of the Oka strain during the passage, RFLP and DNA sequence analyses were also used with 11, 15 and 23 times of virus passage. We found that the Oka strain was stable at passages of up to 23 times, based on the RFLP and DNA sequence analyses. The confirmed Oka strain was renamed as BR-Oka for the purposes of chickenpox vaccine production.

• Journal of Aquaculture
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• v.10 no.4
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• pp.403-407
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• 1997

Intraspecific sequence variation was detected by polymerase chain reaction (PCR) and direct sequencing of a 350-nucleotide region of the mitochondrial 12S rRNA gene of two natural populations (Han River and Nakdong River) and one hatchery stock (Jinhae Inland Fisheries Institute) of local strain common carp, one Israeli strain of common carp stock from Pukyong National University (PKU), and one hybrid between Israeli strain of common carp female and local strain common carp male from PKU stock. There is little variation in 350 bases of the mitochondrial 12S rRNA gene sequences among 2 natural and 1 hatchery local strain common carp populatins, representing abut 7 to 20 nucleotide differences (less than 6%). The sequence of specimens from Han River was more similar to that from Nakdong River (identity=98.0%) than to that from Jinhae Inland Fisheries Institute (identity=96.3%). Sequence variation between Israeli strain and wild local strain common carp was higher than the variation within natural stocks. The level of variation was ranged from 15.7 to 17.7%. The hybrid showed very similar nucleotide4 sequence of 12S rRNA gene to the sequence of Israeli strain with the identity of 98.9%.

• ALGAE
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• v.28 no.1
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• pp.121-129
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• 2013

Intact phycobilisomes from a wild-type red Chondrus crispus and its vegetatively derived green mutant were isolated by centrifugation through a discontinuous sucrose density gradient. Pigment composition was subsequently characterized by spectrophotometry. Vegetative thalli of the two strains grown together for six months in the laboratory resulted in different pigment profiles. Two pigmented phycobilisome bands appeared in the sucrose gradient of the wild-type alga, a purple coloured one, and a pink one, whereas only a single blue band appeared in the gradient of the green mutant. Spectrophotometric and fluorescence analyses identified the phycobiliprotein composition of the purple band as the typical phycoerythrin-phycocyanin-allophycocyanin complement in the wild-type, but there was no detectable phycoerythrin present in the blue band of the green mutant. Sodium dodecyl sulphate, preparative polyacrylamide gel electrophoresis analysis confirmed the presence of allophycocyanin subunits in all extracts, but firm evidence of an R-phycoerythrin linker polypeptide in the blue band was missing. These results highlight the ability of C. crispus to adapt to a phycoerythrin deficiency by adjusting light harvesting pigment ratios.

• Journal of Microbiology and Biotechnology
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• v.9 no.1
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• pp.84-90
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• 1999

The growth of Bifidobacterium breve strain HP2 was completely inhibited by the addition of lactate higher than 4.0％ but not by the addition of acetate. Two kinds of lactate-tolerant mutants were isolated by the nitrosoguanidine treatment, enrichment on a liquid medium with 5％ lactate, and selection on agar plates with 5％ lactate. The mutants were not only able to grow in the presence of 5％ lactate but also improved in viable cell stability in the acidic pH range. In a pH-controlled fermentor, mutant N-1-5 grew at a rate slower than that of the wild type but its growth yield was higher. Notably, mutants were more halotolerant and more osmotolerant than the wild type and they were able to grow in the presence of 3％ NaCl or 25％ lactose at which the wild type entirely stopped the growth. The enzyme activities involved in the lactose metabolism in B. breve were measured to elucidate the biochemical basis for lactate tolerance. In the mutants, activities of several enzymes including phosphoglucomutase decreased compared to the wild-type, which may explain their lower growth rate. However, the activity of lactate dehydrogenase or its nature of inhibition by lactate was not altered.

• Microbiology and Biotechnology Letters
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• v.22 no.5
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• pp.538-543
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• 1994

Effect of partial oxygen pressure on the cell growth and the activities of oxidative defense enzymes were measured in the continuous culture of Streptomyces coelicolor. Both the wild type and the mutant strain resistant to hydrogen peroxide were cultured and the dry cell weight of the two cultures were measured at different oxygen tensions. Growth of the wild type was inhibited by oxygen at above 0.5 vvm. Growth of the hydrogen peroxide resistant mutant was stimulated by pure oxygen at 0.5 vvm but was inhibited by oxygen at 1.0 vvm. Therefore, growth of the hydrogen peroxide resistant mutant was less affected by the deleterious oxidative stress of oxygen. Activities of the several defense enzymes were also measured at different oxygen tensions. Activities of catalase and glucose-6-phosphate dehydrogenase increased significantly as oxygen pressure increased in the wild type culture. In the mutant, however, increase in those enzyme activities was not obvious whereas the uninduced levels of the above enzymes were higher than those of wild type. As judged by Western blotting, the amount of the major catalase increased as the oxygen pressure increased. This indicates that the induction of the catalase activity by oxygen pressure is mostly due to the increase in the expression level for the major catalase.

• Pediatric Infection and Vaccine
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• v.7 no.1
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• pp.83-93
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• 2000

Purpose : Urabe AM-9 strain was known to be associated with increased aseptic meningitis. The reason for high incidence of vaccine-associated meningitis was known that nucleotide(nt) substituted form G to A at position 1081 of the hemagglutinin-neuraminidase(HN) gene and therefore, glutamic acid changed to lysine at amino acid 335. We assessed by comparing nt sequence of the HN gene form Urabe AM-9 strain with wild strain and documented the correlation between nt substitution and vaccine-associated meningitis. Methods : Two lots of Urabe AM-9 vaccine distributed in Korea and mumps wild strains isolated from 1998 through 1999 were analysed. Analysis was made by nt sequencing following amplification of HN gene by RT-PCR. Results : Nucleotide substitution at position 343, 1476, 1570 was not found in both Urabe AM-9 vaccines and wild strains. But analysis of vaccine strains and wild strains isolated from patients revealed substitution from G to A at nt 1081 of the HN gene. Therefore, it encodes lysine instead of glutamic acid at amino acid 335. There was no mixture from of G and A at nt 1081. Nt at 1470 of one lot of Urabe AM-9 vaccines changed from C to A after Vero cell passage. Nt at 1727 of vaccines and wild strains was substituted A to G, so it encodes glycine instead of aspartic acid. Conclusion : Nucleotide analysis of HN gene revealed that nt 1081 of Urabe AM-9 vaccines and wild strains had wild type AAA($Lys^{335}$) instead of variant type GAA($Glu^{335}$). The results of this study suggest that there was a probability of vaccine-associated meningitis due to Urabe AM-9 in Korea before. But incidence of actual side effect was not evaluated because there was no reporting system in Korea.

• 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.

• Korean Journal of Food Science and Technology
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• v.36 no.4
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• pp.662-668
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• 2004

Ames test revealed most methanol extracts of 13 Korean wild mushroom species have strong antimutagenic effects against N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and benzo(a)pyrene[B(a)P]. Methanol extracts of Coriolus versicolor and Phaeolepiota aurea showed 74-94 and 83-88% antimutagenic effects against MNNG and B(a)P in Salmonella typhimurium TA100 strain, while 89 and 91% inhibitions were observed against B(a)P in TA98 strain, respectively. Most water extracts of wild mushrooms did not show antimutagenic activeiy on MNNG and B(a)P. Wild mushrooms extracts inhibited human colon carcinoma cells (HT29), human hepatoma cell (HepG2), and humann histiocytic lymphoma cell (U937) dose-dependently, with most methanol extracts exhibiting stronger effect than water extracts, Highest toxicity was observed against HT-29 cells in methanol extracts of Coriolus versicolor and Phaeolepiota aurea, showing 84% inhibition at 1 mg/mL, whereas C. versicolor water extract showed 53-65% inhibition against HepG2 and U937. These extracts did not show cytotoxic effects against human lymphocyte. Results revealed wild mushrooms have strong antimutagenic and in vitro cytotoxic effects.