• Title/Summary/Keyword: wild strain

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Identification of Wild Yeast Strains and Analysis of Their ${\beta}$-Glucan and Glutathione Levels for Use in Makgeolli Brewing

  • Kang, Sun Hee;Kim, Hye Ryun;Kim, Jae Ho;Ahn, Byung Hak;Kim, Tae Wan;Lee, Jang-Eun
    • Mycobiology
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    • v.42 no.4
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    • pp.361-367
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    • 2014
  • Makgeolli, also known as Takju, is a non-filtered traditional Korean alcoholic beverage that contains various floating matter, including yeast cells, which contributes to its high physiological functionality. In the present study, we assessed the levels of ${\beta}$-glucan and glutathione in various yeast strains isolated from traditional Korean Nuruk and selected a ${\beta}$-glucan- and glutathione-rich yeast strain to add value to Makgeolli by enhancing its physiological functionality through increased levels of these compounds. Yeast ${\beta}$-glucan levels ranged from 6.26% to 32.69% (dry basis) and were strongly species-dependent. Dried Saccharomyces cerevisiae isolated from Nuruk contained $25.53{\mu}g/mg$ glutathione, $0.70{\mu}g/mg$ oxidized glutathione, and $11.69{\mu}g/g$ and $47.85{\mu}g/g$ spermidine and L-ornithine monohydrochloride, respectively. To produce functional Makgeolli, a ${\beta}$-glucan- and glutathione-rich yeast strain was selected in a screening analysis. Makgeolli fermented with the selected yeast strain contained higher ${\beta}$-glucan and glutathione levels than commercial Makgeolli. Using the selected yeast strain to produce Makgeolli with high ${\beta}$-glucan and glutathione content may enable the production of functional Makgeolli.

Changes in Lipids- and Fatty Acids Compositions in Response to Growth Temperature of Streptomyces viridochromogenes (배양온도 변화에 따른 Streptomyces viridochromogenes의 지질과 지방산 조성변화)

  • 김재헌;김우상
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.155-160
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    • 2003
  • The wild type and two morphological variants of Streptomyces viridochromogenes were studied for their lipidand fatty acid compositions at different incubation temperatures. It showed that a decrease in triacylglycerol content was closely linked to the aerial mycelium formation. Phospholipids showed no characteristic changes, except that the contents of phosphatidylethanolamine were clearly high for aerial mycelium deficient strain BR2 grown at $20^{\circ}C$. The strain BR2 also presented unidentified aminolipids with various $R_{f}$ values. Among the aminolipids, ornithinolipid increased gradually during the cultivation for all strains. The changes in fatty acid compositions showed a temperature dependency that the proportion of unsaturated acids decreased as the growth temperature increased. The proportion of straight chain saturated fatty acids decreased as the aerial mycelium developed, and it was most evident for the mutant strain M13 with more extensive aerial mycelium. The mutant strain BR2 presented significantly higher level of iso branched odd numbered saturated fatty acids.

Mutagenesis of Slow Growing Rhizobium japonicum by Transposon Tn5 (Transposon Tn5를 이용한 Slow growing Rhizobium japonicum의 돌연변이 유도)

  • Kim, Sung-Hoon;Rhee, Yoon;Sun, Dae-Kyu;Yoo, Ick-Dong
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.305-311
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    • 1988
  • The spectinomycin resistant strain of slow growing R. japonicum R-168 was selected to be participated in conjugation with E. coli WA803/pGS9. Tn5 was introduced from suicide vector pGS9 into R. japonicum R-168 $spr^{r}$ chromosome at the frequency of $1.0\times 10^{-5}-5.0\times 10^{-7}$ and the transconjugante were selected on the yeast extract-mannitol plate containing kanamycin ($50{\mu}$g/ml) and spectinomycin ($100{\mu}$g/ml) after 8-9 days incubation. All transconjugants we tested were found to contain Tn 5 DNA on their genome, which was confirmed by Southern hybridization experiments. R. japonicum RNa75, which had been selected through plant test, was found to be defective in symbiotic nitrogen fixing ability and the production of leghemoglobin in soybean nodules formed by the inoculation of this mutant. In addition, this mutant strain hardly developed nitrogenase activity asymbiotically in contrast with the wild type strain, indicating that some nitrogen fixing gene might be blocked in this strain and the production of leghemoglobin could be decreased by the interference in nitrogen fixing genes.

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Increased mRNA Stability and Expression Level of Croceibacter atlanticus Lipase Gene Developed through Molecular Evolution Process

  • Jeong, Han Byeol;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.6
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    • pp.882-889
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    • 2021
  • In order to use an enzyme industrially, it is necessary to increase the activity of the enzyme and optimize the reaction characteristics through molecular evolution techniques. We used the error-prone PCR method to improve the reaction characteristics of LipCA lipase discovered in Antarctic Croceibacter atlanticus. Recombinant Escherichia coli colonies showing large halo zones were selected in tributyrin-containing medium. The lipase activity of one mutant strain (M3-1) was significantly increased, compared to the wild-type (WT) strain. M3-1 strain produced about three times more lipase enzyme than did WT strain. After confirming the nucleotide sequence of the M3-1 gene to be different from that of the WT gene by four bases (73, 381, 756, and 822), the secondary structures of WT and M3-1 mRNA were predicted and compared by RNAfold web program. Compared to the mean free energy (MFE) of WT mRNA, that of M3-1 mRNA was lowered by 4.4 kcal/mol, and the MFE value was significantly lowered by mutations of bases 73 and 756. Site-directed mutagenesis was performed to find out which of the four base mutations actually affected the enzyme expression level. Among them, one mutant enzyme production decreased as WT enzyme production when the base 73 was changed (T→ C). These results show that one base change at position 73 can significantly affect protein expression level, and demonstrate that changing the mRNA sequence can increase the stability of mRNA, and can increase the production of foreign protein in E. coli.

Studies on Mild Mutants of Tabacco Mosaic Virus I. Induction of Mild Mutants and Their Characteristics (약독 담배 모자이크 바이러스 I. 약독바이러스의 유기 및 특성)

  • Choi Jang Kyung;Son Kyung Ok
    • Korean Journal Plant Pathology
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    • v.1 no.1
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    • pp.3-11
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    • 1985
  • Three mild mutant strains of tobacco mosaic virus (TMV) were isolated from Nicotiana tabacum var. Samsun incubated at $38^{\circ}C$ for 10 days after inoculation with a wild type of TMV-OM strain. They were designated into Tg 5272, Tw 227 and Tw 333. All mild strains could be distinguished from TMV-OM by their reactions on different indicator plants. The mild strains induced the mild mottling without distinct symptoms, whereas the wild strain produced severe mosaic, rugose and stunting on tobacco and red pepper plants. Tw 227 and Tw 333 produced smaller necrotic spots than those of Tg 5272 and TMV-OM on N. glutinosa and Datura stramonium. The former two strains also produced ring spots and mosaic on Gomphrena globosa compared with necrotic spots by the latter strains. Three mild strains were serologically identical to TMV-OM. Their physical properties were thermal inactivation point $80-85^{\circ}C$, dilution end point between $10^{-4}\;and\;10^{-6}$, and longevity in vitro 7days or longer. Ultraviolet absorption spectra of purified preparations of the mild strains and TMV-OM were identical, with a minimum at 247nm, a maximum at 260nm, and a slight shoulder at 290nm. Electrophoresis of the strains in polyacrylamide-agarose gel showed that all the strains formed one major band and two minor bands, except for one minor band of Tw 333. However, when sodium dodecyl sulfate was added to the purified viruses before electrophoresis, each strain formed only one major band.

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Expression Profiles of Streptomyces Doxorubicin Biosynthetic Gene Cluster Using DNA Microarray System (DNA Microarray 시스템을 이용한 방선균 독소루비신 생합성 유전자군의 발현패턴 분석)

  • Kang Seung-Hoon;Kim Myung-Gun;Park Hyun-Joo;Kim Eung-Soo
    • KSBB Journal
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    • v.20 no.3
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    • pp.220-227
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    • 2005
  • Doxorubicin is an anthracycline-family polyketide compound with a very potent anti-cancer activity, typically produced by Streptomyces peucetius. To understand the potential target biosynthetic genes critical for the doxorubicin everproduction, a doxorubicin-specific DNA microarray chip was fabricated and applied to reveal the growth-phase-dependent expression profiles of biosynthetic genes from two doxorubicin-overproducing strains along with the wild-type strain. Two doxorubicin-overproducing 5. peucetius strains were generated via over-expression of a dnrl (a doxorubicin-specific positive regulatory gene) and a doxA (a gene involved in the conversion from daunorubicin to doxorubicin) using a streptomycetes high expression vector containing a strong ermE promoter. Each doxorubicin-overproducing strain was quantitatively compared with the wild-type doxorubicin producer based on the growth-phase-dependent doxorubicin productivity as well as doxorubicin biosynthetic gene expression profiles. The doxorubicin-specific DNA microarray chip data revealed the early-and-steady expressions of the doxorubicin-specific regulatory gene (dnrl), the doxorubicin resistance genes (drrA, drrB, drrC), and the doxorubicin deoxysugar biosynthetic gene (dnmL) are critical for the doxorubicin overproduction in S. peucetius. These results provide that the relationship between the growth-phase-dependent doxorubicin productivity and the doxorubicin biosynthetic gene expression profiles should lead us a rational design of molecular genetic strain improvement strategy.

Detection of single-nucleotide polymorphism in RPB2 of Wolfiporia hoelen strains and assessment of its applicability for strain breeding (복령 균주의 RPB2 유전자 내 단일염기다형성 및 육종 활용성 분석)

  • Su Yeon, Kim;Mi-Jeong, Park;Seong Hwan, Kim;Kang-Hyeon, Ka
    • Journal of Mushroom
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    • v.20 no.4
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    • pp.199-207
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    • 2022
  • The demand for novel strains has been rising in the domestic market to increase the production of sclerotia from Wolfiporia hoelen. To improve strain breeding efficiency, we investigated whether single-nucleotide polymorphisms (SNPs) in the RNA polymerase II subunit (RPB2) gene, which may be linked to the mating type locus, are useful for distinguishing monokaryons from dikaryons in Korean W. hoelen strains. We designed a specific primer set to efficiently amplify a region of RPB2 using PCR with the genomic DNA of 12 cultivated strains and 31 wild strains of W. hoelen collected from Korea. Nucleotide sequences of the PCR-amplified RPB2 genes were determined and analyzed for the presence of SNPs among the 43 W. hoelen strains. Previously reported SNP loci were detected in the RPB2 gene of all W. hoelen strains tested. However, these previously reported SNP loci could not be applied to differentiate monokaryons from dikaryons in approximately one-third of Korean wild strains with homozygous genotypes. Three additional SNPs in the RPB2 gene, which may improve the ability to distinguish monokaryons from dikaryons, were identified by searching through the multiple sequence alignments of the 43 W. hoelen strains. The applicability of these three novel SNPs, together with the previously known SNPs, in the RPB2 gene to W. hoelen strain breeding was verified by examining the hybrid strains and their parental strains.

A High-Throughput Method Based on Microculture Technology for Screening of High-Yield Strains of Tylosin-Producing Streptomyces fradiae

  • Zhiming Yao;Jingyan Fan;Jun Dai;Chen Yu;Han Zeng;Qingzhi Li;Wei Hu;Chaoyue Yan;Meilin Hao;Haotian Li;Shuo Li;Jie Liu;Qi Huang;Lu Li;Rui Zhou
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.831-839
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    • 2023
  • Tylosin is a potent veterinary macrolide antibiotic produced by the fermentation of Streptomyces fradiae; however, it is necessary to modify S. fradiae strains to improve tylosin production. In this study, we established a high-throughput, 24-well plate screening method for identifying S. fradiae strains that produce increased yields of tylosin. Additionally, we constructed mutant libraries of S. fradiae via ultraviolet (UV) irradiation and/or sodium nitrite mutagenesis. A primary screening of the libraries in 24-well plates and UV spectrophotometry identified S. fradiae mutants producing increased yields of tylosin. Mutants with tylosin yield 10% higher than the wild-type strain were inoculated into shake flasks, and the tylosin concentrations produced were determined by high-performance liquid chromatography (HPLC). Joint (UV irradiation and sodium nitrite) mutagenesis resulted in higher yields of mutants with enhanced tylosin production. Finally, 10 mutants showing higher tylosin yield were re-screened in shake flasks. The yield of tylosin A by strains UN-C183 (6767.64 ± 82.43 ㎍/ml) and UN-C137 (6889.72 ± 70.25 ㎍/ml) was significantly higher than that of the wild-type strain (6617.99 ± 22.67 ㎍/ml). These mutant strains will form the basis for further strain breeding in tylosin production.

RNase P-dependent Cleavage of Polycistronic mRNAs within Their Downstream Coding Regions in Escherichia coli

  • Lee, Jung-Min;Kim, Yool;Hong, Soon-Kang;Lee, Young-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.29 no.6
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    • pp.1137-1140
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    • 2008
  • M1 RNA, the catalytic subunit of Escherichia coli RNase P, is an essential ribozyme that processes the 5' leader sequence of tRNA precursors (ptRNAs). Using KS2003, an E. coli strain generating only low levels of M1 RNA, which showed growth defects, we examined whether M1 RNA is involved in polycistronic mRNA processing or degradation. Microarray analysis of total RNA from KS2003 revealed six polycistronic operon mRNAs (acpP-fabF, cysDNC, flgAMN, lepAB, phoPQ, and puuCBE) showing large differences in expression between the adjacent genes in the same mRNA transcript compared with the KS2001 wild type strain. Model substrates spanning an adjacent pair of genes for each polycistronic mRNA were tested for RNase P cleavage in vitro. Five model RNAs (cysNC, flgMN, lepAB, phoPQ, and puuBE) were cleaved by RNase P holoenzyme but not by M1 RNA alone. However, the cleavages occurred at non-ptRNA-like cleavage sites, with much less efficiency than the cleavage of ptRNA. Since cleavage products generated by RNase P from a polycistronic mRNA can have different in vivo stabilities, our results suggest that RNase P cleavage may lead to differential expression of each cistron.

Characteristics of Korean native strains in the domesticated silkworm, Bombyx mori (현재하는 한국 재래성의 형질특성)

  • 노시갑;이재만
    • Journal of Sericultural and Entomological Science
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    • v.42 no.1
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    • pp.10-13
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    • 2000
  • The objective of this research was to characterize of the silkworm that might be recognized to the Korean native strains. The eleven strains of Korean race used in this study, which is cultured in Korea and Japan seri-cultural research organs. Most of Korean varieties were three molting and univoltine, comparatively longer larval duration than the trimolter general. The egg characters of varieties showed short-eliptic shape and dark brown egg color except for a few varieties which shows greenish dark brown. Also, most of varieties were showed the plain(p) and moricaud( $p_{M}$) in larval markings. Moric marking of the varieties also consist of innumerable dark grayish brown lines and dots, though somewhat darker and lighter than that of the wild silkworm, Bombyx mandarina. Cocoon characters variations of varieties were seen in the size, color and shape. 8 varieties were colored cocoon, i.e., yellow, greenish yellow and light green etc., the others were white cocon. The shape of cocons were consisted of constricted shallowly in the middle and attenuated at one or both ends of cocoon, i.e., spindle. The results of this study is sufficient evidence that Korean strains were shown as the one of regional origin strain of domesticated silkworm such as the Chinese and Japanese etc.c.

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