• Title/Summary/Keyword: mutant strain

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Physiological Response of a White Mutant of Ganoderma lucidurn Induced by Light and Temperature (영지버섯 백색변이주의 광 및 온도에 의한 생리적 반응)

  • Cho, Soo-Muk;Seo, Geon-Sik;Yoo, Ick-Dong;Shin, Gwan-Chull
    • Microbiology and Biotechnology Letters
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    • v.22 no.2
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    • pp.115-119
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    • 1994
  • White mutant of Ganoderma lucidum(G4142) induced the non-basediocarpous basidiospores(NBB) from the aerial mycelia on agar media by the light illumination. Light was found to be necessary for NBB formation, but it also inhibited the growth of mycelium. The best sporulation was obtained at the periodic exposure of 16 hour light and 8 hour dark. Blue and yellow light were the most effective on sporulation, however, near UV and red light did not induce any spores. Effective light intensity for NBB bearing was about 1,000 lux as white light. Even after 16 days of culture, this strain did not form the pinhead nor chlamydospore. Optimum temperature for the mycelial growth and NBB formation were 30$\circ $C. Ganoderma lucidum G4142 exhibited the formation of stroma after five days of incubation at 30$\circ $C.

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Production of L-Ornithine by Citrulline Auxotrophic Mutants of Glutamate-Producing Bacteria

  • CHOI, DAE KEON;WUK SANG RYU;BONG HYUN CHUNG;SOO WAN NAM;YOUNG HOON PARK
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.102-107
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    • 1992
  • For the purpose of producing L-ornithine by microbial fermentation, mutant strains were developed from glutamate-producing bacteria by mutagenesis using N-methyl-N'-nitro-N-nitrosoguanidine (NTG) and UV irradiation. Brevibacterium ketoglutamicum BK1046, a L-citrulline auxotroph which is also resistant to arginine hydroxamate (Arghx), was isolated and selected as the best producer of L-ornithine. This strain was capable of producing L-ornithine at a concentration of 24 g/l after 69 hours of cultivation in the 21 jar fermentor. The optimum supplementary level of L-arginine, a substitute for L-citrulline, was found to be about 0.2 g/l in the shake-flask fermentation.

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Purification of Cyclodextrin Glucanotransferase by Affinity Chromatography (Affinity Chromatography를 이용한 Cyclodextrin Glucanotransferase의 정제)

  • 안중훈;황진봉;김승호;김경은
    • Microbiology and Biotechnology Letters
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    • v.19 no.3
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    • pp.313-314
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    • 1991
  • - The cyclodextrin glucanotransferase (CGTase) of a mutant of Bacillus stearothermophilus was purified in one step by affinity chromatography. The recovery was 95%. The specific activity of the CGTase increased from 26.2 U/mg protein to 485.5 U/mg protein. The purified CGTase was almost homogeneous by SDS-polyacrylamide gel electrophoresis. The one-step purification proved to be feasible with the mutant in contrast to the parent strain, which required pre-purification step of ammonium sulfate precipitation.

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The Relationship between Virginiae Butanolide C(VB-C) and Receptor in Virginiamycin Production (Virginiamycin 생산유도에 관여하는 Virginiae Butanolide C(VB-C) 및 Receptor의 상관관계)

  • Kim, Hyun-Soo;Hyun, Ji-Sook;Yu, Tae-Shick
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.59-66
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    • 1996
  • Virginiae butanolide C(VB-C) is one of the butyrolactone autoregulators, which triggers the productin of virginiamycin in Streptomyces virginiae. To further understand the mechanism of virginiamycin induction, we isolated three mutants from S. virginiae by N-methyl-N'-nitrosoguanidine (NTG) treatment. The characteristics of the three mutants were confirmed as follows: the mutant No. 1 delayed the production of the VB-C, receptor and antibiotics; the mutant No.3 hyperproduced receptor; the mutant No.4 failed to produce the VB-C. The addition of synthetic VB-C couldn't induce the production of antibiotics in the mutant No.1 due to delayed production of receptor, could provoke the production of larger amount of antibiotics than parental wild type strain in the mutant No.3 due to the presence of large amount of receptor, and could induce production of very small amount of antibiotics in the mutant No.4 due to the absence of VB-C. Antimicrobial spectrum and HPLC analysis of the mutant No.1 and No.3 suggested that the VB-C might have a specific ability to induce the production of virginiamycin M and S. These results imply that the VB-C has an ability to trigger the production of virginiamycin under receptor existence in S. virginiae.

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Production of Purplish-red Pigment in Mixed Culture of Streptomyces propurpuratus ATCC 21630 and Bacillus sp. R-89 (Streptomyces propurpuratus ATCC 21630 Bacillus sp. R-89의 혼합배양에 의한 적자색 색소의 생산)

  • Ho, Ryu-Beung;Park, Bub-Gyu;Chi, Young-Eh;Lee, Ju-Hwa
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.327-333
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    • 1989
  • The purplish-red pigment was formed on agar plate by superimposed streaking of Streptomyces propurpuratus ATCC 21630 and strain R-89, The strain No.89 was ascribable to the genus Bacillus and designated as Bacillus sp. R-89. Both strain did not produced such pigment when cultivated independently. For hyperpigment production, we selected the mutant S.P-6 from Streptomyces propurpuratus ATCC 21630 by MNNG (N-methyl-N'-nitro-N-nitrosoguanidine) treatment. Maximum purplish-red pigment 1420 mg/$m\ell$ were produced, when the mutant of R-16 and Bacillus sp. R-89 were mixed cultured at 3$0^{\circ}C$ for 72 hr.

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Streptomyces griseus HH1, An A-factor Deficient Mutant Produces Diminished Level of Trypsin and Increased Level of Metalloproteases

  • Kim, Jung-Mee;Hong, Soon-Kwang
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.160-168
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    • 2000
  • A-factor I a microbial hormone that can positively control cell differentiation leading to spore formation and secondary metabolite formation in Streptomyces griseus. to identify a protease that is deeply involved in the morphological and physiological differentiation of Streptomyces, the proteases produced by Streptomyces griseus IFO 13350 and its A-factor deficient mutant strain, Streptomyces griseus HH1, as well as Streptomyces griseus HH1 transformed with the afsA gene were sturdied. In general Streptomyces griseus showed a higher degree of cell growth and protease activity in proportion to its ability to produce a higher amount of A-factor. In particular, the specific activity of the trypsin of Streptomyces griseus IFO 13350 was greatly enhanced more than twice compared with that of Streptomyces griseus HH1 in the later stage of growth. The specific activity of the metalloprotease of Streptomyces griseus HH1 was greatly enhanced more than twice compared with that of Streptomyces griseus IFO 13350, and this observation was reversed in the presence of thiostreptione, However, Streptomyces griseus HH1 transformed with the afsA gene showed a significantly decreased level of trypsin and metalloprotease activity compared with that of the HH1 strain. There was no significant difference between Streptomyces griseus IFO 13350 and HH1 strain in their chymotrypsin and thiol protease activity, yet the level of leu-amionpeptidase activity was 2 times higher in Streptomyces griseus HH1 than in strain IFO 13350 . Streptomyces griseus HH1 harboring afsA showed a similar level of enzyme activity , however, all the three protease activities sharply increased and the thiol protease activity was critically increased at the end of the fermentation. When a serine protease inhibitor, pefabloc SC, and metalloprotease inhibitor, EDTA, were applied to strain IFO 13350 to examine the in vivo effects of the protease inhibitors on the morpholofical differentiation, the formation of aerial meycelium and spores was delayed by two or three days.

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Isolation, Production, and Characterization of Protease from Bacillus subtilis IB No. 11

  • Lee, Min-Hyang;Lee, Kang-Moon;Choi, Yong-Jin;Baek, Yeon-Soo
    • Journal of Animal Science and Technology
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    • v.51 no.6
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    • pp.527-536
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    • 2009
  • A potent protein degrading bacterium was isolated from soil samples of different environments. Polyphasic taxonomic studies and phylogenetic 16S rRNA sequence analyses led to identify the isolate IB No. 11 as a strain of Bacillus subtilis. The isolated strain was recognized to produce protease constitutively, and the maximum production (1.64 units/ml) was attained in a shake flask culture when the isolate was grown at $40^{\circ}C$, for 32 h in basal medium supplemented with starch (0.25%) and gelatin (1.25%) as sole carbon and nitrogen source, respectively. The optimum pH and temperature for the protease activity were determined to be pH 7.0 and $50^{\circ}C$, respectively. $Ca^{2+}$ and $Mn^{2+}$ enhanced remarkably the protease activity but neither showed positive effect on the protease's thermal stability. In addition, it was observed that the protease was fairly stable in the pH range of 6.5-8.0 and at temperatures below $50^{\circ}C$, and it could be a good candidate for an animal feed additive. The inhibition profile of the protease by various inhibitors indicated that the enzyme is a member of serine-proteases. A combination of UV irradiation and NTG mutagenesis allowed to develop a protease hyper-producing mutant strain coded as IB No. 11-4. This mutant strain produced approximately 3.23-fold higher protease activity (6.74 units/mg) than the parent strain IB No. 11 when grown at $40^{\circ}C$ for 32h in the production medium. The protease production profile of the selected mutants was also confirmed by the zymography analysis.

Scratching Stimuli of Mycelia Influence Fruiting Body Production and ROS-Scavenging Gene Expression of Cordyceps militaris

  • Liu, Gui-Qing;Qiu, Xue-Hong;Cao, Li;Han, Ri-Chou
    • Mycobiology
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    • v.46 no.4
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    • pp.382-387
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    • 2018
  • The entomopathogenic fungus Cordyceps militaris is a valuable medicinal ascomycete, which degenerates frequently during subsequent culture. To avoid economic losses during industrialized production, scratching stimuli of mycelia was introduced to improve the fruiting body production. The present results indicated that higher yields and biological efficiency were obtained from two degenerate strains (YN1-14 and YN2-7) but not from g38 (an insertional mutant in Rhf1 gene with higher yields and shorter growth periods). Furthermore, the growth periods of the fruiting bodies were at least 5 days earlier when the mycelia were scratched before stromata differentiation. Three ROS-scavenging genes including Cu/Zn superoxide dismutase (CmSod1), Glutathione peroxidase (CmGpx), and Catalase A (CmCat A) were isolated and their expression profiles against scratching were determined in degenerate strain YN1-14 and mutant strain g38. At day 5 after scratching, the expression level of CmGpx significantly decreased for strain g38, but that of CmSod1 significantly increased for YN1-14. These results indicated that scratching is an effective way to promote fruiting body production of degenerate strain, which may be related at least with Rhf1 and active oxygen scavenging genes.

Inhibition of Verticillium Wilt in Cotton through the Application of Pseudomonas aeruginosa ZL6 Derived from Fermentation Residue of Kitchen Waste

  • Qiuhong Niu;Shengwei Lei;Guo Zhang;Guohan Wu;Zhuo Tian;Keyan Chen;Lin Zhang
    • Journal of Microbiology and Biotechnology
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    • v.34 no.5
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    • pp.1040-1050
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    • 2024
  • To isolate and analyze bacteria with Verticillium wilt-resistant properties from the fermentation residue of kitchen wastes, as well as explore their potential for new applications of the residue. A total of six bacterial strains exhibiting Verticillium wilt-resistant capabilities were isolated from the biogas residue of kitchen waste fermentation. Using a polyphasic approach, strain ZL6, which displayed the highest antagonistic activity against cotton Verticillium wilt, was identified as belonging to the Pseudomonas aeruginosa. Bioassay results demonstrated that this strain possessed robust antagonistic abilities, effectively inhibiting V. dahliae spore germination and mycelial growth. Furthermore, P. aeruginosa ZL6 exhibited high temperature resistance (42℃), nitrogen fixation, and phosphorus removal activities. Pot experiments revealed that P. aeruginosa ZL6 fermentation broth treatment achieved a 47.72% biological control effect compared to the control group. Through activity tracking and protein mass spectrometry identification, a neutral metalloproteinase (Nml) was hypothesized as the main virulence factor. The mutant strain ZL6ߡNml exhibited a significant reduction in its ability to inhibit cotton Verticillium wilt compared to the strain P. aeruginosa ZL6. While the inhibitory activities could be partially restored by a complementation of nml gene in the mutant strain ZL6CMߡNml. This research provides a theoretical foundation for the future development and application of biogas residue as biocontrol agents against Verticillium wilt and as biological preservatives for agricultural products. Additionally, this study presents a novel approach for mitigating the substantial amount of biogas residue generated from kitchen waste fermentation.

Characterization of Nonhypersensitive Mutant and Nonpathogenic Mutant of Xanthomonas campestris pv. campestris (Xanthomonas campestris pv. campestris의 비과민성 돌연변이주와 비병원성 돌연변이주의 특성)

  • Kim, Mee-Hyang;Bae, Dong-Won;Lee, Jun-Teak;Yun, Han-Dae;Kim, Hee-Kyu
    • Korean Journal Plant Pathology
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    • v.13 no.3
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    • pp.179-183
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    • 1997
  • We have screened hypersensitive responses of 18 cultivars of Nicotiana tabacum to Xanthomonas campestris pv. campestris. TC500 cultivar produced the most strong hypersensitive response (HR) to Xanthomonas campestris pv. campestris. By NTG mutagenesis, nonhypersensitive mutants (XHN 514-774, XHN 620-831) were generated, which does not induce hypersensitive response on tobacco leaves (Nicotiana tabacum cv. TC500). Also nonpathogenic mutant (XPN 1001), which does not incite any of the black rot symptoms on leaves was generated. We observed that HR mutants were still pathogenic on cabbage leaves producing black rot symptoms and nonpathogenic mutant induced HR in tobacco leaves. The inplanta growth of wild type and HR mutants were examined for up to 120 hrs after inoculation : population of wild type strain increased to $10^{8}$ in 24hrs, but rapidly declined thereafter; HR-mutants increased to more than $10^{6}$ in 48 hrs after inoculation but subsequently stabilized and slowly decreased. We observed that wild type and these mutans produced similar amounts of degradative enzymes such as protease, pectate lyase, cellulase and amylase.

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