• Title/Summary/Keyword: mutant strain

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Development of Strain Fermenting the Glucose/Cellbiose Mixed Sugar for Simultaneous Saccharification of Fermentation of Cellulosic Materials (섬유소 물질의 동시당화발효에 적합한 Glucose/Cellbiose 혼합당 발효균주의 개발)

  • 박승원;홍영기;김승욱;홍석인
    • Microbiology and Biotechnology Letters
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    • v.27 no.2
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    • pp.145-152
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    • 1999
  • Brettanomyces custersii CBS 5512 which has reported as a thermotolerant glucose-cellobiose co-fermentable yeast strain was mutated with UV and NTG to improve ethanol yield at higher than 4$0^{\circ}C$ B. custersii H1-23, H1-39, H1-55 and H1062 were finally selected for hyper-fermentable strains at higher than 4$0^{\circ}C$ from thermotolerant 7510 colonies through 5th selection. Among the selected strains, H1-39 mutant had better fermentability at 4$0^{\circ}C$ and 43$^{\circ}C$ from different concentrations of glucose. H1-39 and H1-23 mutants yielded more than 70% of the theoretical ethanol yield in 4 and 8% mixed sugars at above 4$0^{\circ}C$, which was 5-11% higher than those by original strain. Especially, H1-39 mutant had better fermentability in 4% mixed sugar. It showed 78.5% of the theoretical yield at 4$0^{\circ}C$ and 72.2% of the theoretical yield at 43$^{\circ}C$. On the other hand, theoretical yield of ethanol by H1-39 mutant in 8% mixed sugar at 4$0^{\circ}C$ and 43$^{\circ}C$ were 75.2% and 70.2%, respectively. Theses values increased up to 7-11% as compared to those by orginal strain. By the simultaneous saccharification and fermentation, ethanol production by H1-39 mutant increased up to more than 23% as compared to that by original strain.

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$^{31}p$ Nuclear Magnetic Resonance Studies of Acetic Acid Inhibition of Ethanol Production by Strains of Zymomonas mobilis

  • Kim, In-Seop;Barrow, Kevin D.;Rogers, Peter L.
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.90-98
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    • 2003
  • In vivo $^31p$ Nuclear Magnetic Resonance ($^31p$NMR) and metabolic studies were carried out on an acetic acid tolerant mutant, Zymomonas mobilis $ZM4/Ac^R$, and compared to those of the parent strain, Z. mobilis ZM4, to evaluate possible mechanisms of acetic acid resistance. This investigation was initiated to determine whether or not the mutant strain might be used as a suitable recombinant host far ethanol production from lignocellulose hydrolysates containing various inhibitory compounds. $ZM4/Ac^R$ showed multiple resistance to other lignocellulosic toxic compounds such as syringaldehyde, furfural, hydroxymethyl furfural, vanillin, and vanillic acid. The mutant strain was resistant to higher concentrations of ethanol or lower pH in the presence of sodium acetate, compared to ZM4 which showed more additive inhibition. in vivo $^31p$ NMR studies revealed that intracellular acidification and de-energization were two mechanisms by which acetic acid exerted its inhibitory effect. For $ZM4/Ac^R$, the internal pH and the energy status were less affected by sodium acetate compared to the parent strain. This resistance to pH change and de-energization caused by acetic acid is a possible explanation for the development of resistance by this strain.

Improved Production, and Purification of Aclacinomycin A from Streptomyces lavendofoliae DKRS

  • Kim, Wan-Seop;Youn, Deok-Joong;Cho, Won-Tae;Kim, Myung-Kuk;Kim, Hak-Ryul;Rhee, Sang-Ki;Choi, Eui-Sung
    • Journal of Microbiology and Biotechnology
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    • v.5 no.5
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    • pp.297-301
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    • 1995
  • An anthracycline antibiotic, aclacinomycin A (aclarubicin), was produced from a mutant strain of Streptomyces lavendofoliae. The mutant strain which showed a 4-fold higher productivity of aclacinomycin A compared with the parent strain was also found to produce a significantly higher amount of aclacinomycin A than the reported production strain, Streptomyces galilaeus. The aclacinomycin A was produced up to 125 mg/l using potato starch and soybean meal as carbon and nitrogen sources, respectively, on a 3 liter scale fermentation in a 5 liter jar fermentor. The mutant strain also produced significant amount of aclacinomycins Band Y. Aclacinomycin A was isolated from the culture broth by solvent extractions and further purified by silica gel column chromatography. The yield of aclacinomycin A with over 99$%$ purity was found to be over 60$%$ starting from 3 liters of culture broth.

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Production of Xylitol by Catabolite Derepressed Mutant of Candida sp. (Candida sp.의 Catabolite Derepressed Mutant에 의한 Xylitol 생산)

  • 한완옥;서진호;유연우
    • KSBB Journal
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    • v.13 no.1
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    • pp.6-12
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    • 1998
  • In order to produce xylitol from hemicellulose hydrolysate which is widely used as a substrate, the development of strain such as catabolite derepressed mutant is required. After treatment of Candida sp. with EMS, GM-17 and PM-34 as catabolite derepressed mutant were isolated from Candida guilliermondii and Candida parapsilosis, respectively. Mutant GM-17 and PM-34 simultaneously assimilated xylose and glucose during the fermentation. The specific xylose reductase and xylitol dehydrogenase activities of mutant strains were also higher than those of wild strains in glucose medium and mixed medium of glucose and xylose. The xylitol productivity and yield of mutant GM-17 and PM-34 were improved as compared to the wild types in the mixed medium. The xylitol productivity and yield of mutant GM-17 were 0.09 g/L·hr and 0.56 g-xylitol/g-xylose, and those of mutant PM-34 were 0.21 g/L·hr and 0.52 g-xylitol/g-xylose in the mixed medium, respectively.

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Contribution of the murI Gene Encoding Glutamate Racemase in the Motility and Virulence of Ralstonia solanacearum

  • Choi, Kihyuck;Son, Geun Ju;Ahmad, Shabir;Lee, Seung Yeup;Lee, Hyoung Ju;Lee, Seon-Woo
    • The Plant Pathology Journal
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    • v.36 no.4
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    • pp.355-363
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    • 2020
  • Bacterial traits for virulence of Ralstonia solanacearum causing lethal wilt in plants were extensively studied but are not yet fully understood. Other than the known virulence factors of Ralstonia solanacearum, this study aimed to identify the novel gene(s) contributing to bacterial virulence of R. solanacearum. Among the transposon-inserted mutants that were previously generated, we selected mutant SL341F12 strain produced exopolysaccharide equivalent to wild type strain but showed reduced virulence compared to wild type. In this mutant, a transposon was found to disrupt the murI gene encoding glutamate racemase which converts L-glutamate to D-glutamate. SL341F12 lost its motility, and its virulence in the tomato plant was markedly diminished compared to that of the wild type. The altered phenotypes of SL341F12 were restored by introducing a full-length murI gene. The expression of genes required for flagella assembly was significantly reduced in SL341F12 compared to that of the wild type or complemented strain, indicating that the loss of bacterial motility in the mutant was due to reduced flagella assembly. A dramatic reduction of the mutant population compared to its wild type was apparent in planta (i.e., root) than its wild type but not in soil and rhizosphere. This may contribute to the impaired virulence in the mutant strain. Accordingly, we concluded that murI in R. solanacearum may be involved in controlling flagella assembly and consequently, the mutation affects bacterial motility and virulence.

Xanthomonas oryzae pv. oryzae RpfE Regulates Virulence and Carbon Source Utilization without Change of the DSF Production

  • Cho, Jung-Hee;Yoon, Joo-Mi;Lee, Sang-Won;Noh, Young-Hee;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.29 no.4
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    • pp.364-373
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    • 2013
  • It has been known that most regulation of pathogenicity factor (rpf ) genes in xanthomonads regulates virulence in response to the diffusible signal factor, DSF. Although many rpf genes have been functionally characterized, the function of rpfE is still unknown. We cloned the rpfE gene from a Xanthomonas oryzae pv. oryzae (Xoo) Korean race KACC10859 and generated mutant strains to elucidate the role of RpfE with respect to the rpf system. Through experiments using the rpfE-deficient mutant strain, we found that mutation in rpfE gene in Xoo reduced virulence, swarm motility, and production of virulence factors such as cellulase and extracellular polysaccharide. Disease progress by the rpfE-deficient mutant strain was significantly slowed compared to disease progress by the wild type and the number of the rpfE-deficient mutant strain was lower than that of the wild type in the early phase of infection in the inoculated rice leaf. The rpfE mutant strain was unable to utilize sucrose or xylose as carbon sources efficiently in culture. The mutation in rpfE, however, did not affect DSF synthesis. Our results suggest that the rpfE gene regulates the virulence of Xoo under different nutrient conditions without change of DSF production.

Selection and Characterization of Catabolite Repression Resistant Mutant of Bacillus firmus var. alkalophilus Producing Cyclodextrin Glucanotransferase

  • Do, Eun-Ju;Shin, Hyun-Dong;Kim, Chan
    • Journal of Microbiology and Biotechnology
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    • v.3 no.2
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    • pp.78-85
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    • 1993
  • In order to elucidate the mechanism which regulates the production of cyclodextrin glucanotransferase (CGTase) and to achieve overproduction of CGTase by releasing catabolite (glucose) repression, several catabolite repression resistant mutants were selected from newly screened Bacillus firmus var. alkalophilus H609, after NTG (N-methyl-N -nitro-N-nitrosoguanidine) treatment, using 2-deoxyglucose as a nonmetabolizable analog of catabolite glucose and as a selection marker. Five catabolite repression resistant mutants were selected from about 30, 000 2-deoxyglucose resistant colonies. Relative catabolite repression indices of the selected mutants were in the range of 8~80% assuming 100% for parent strain. The amount of CGTase produced by the mutant strain CR41, which was 250 units/ml, was three times larger than that produced by its parent strain. The mutation seems to have occurred in the regulatory region of CGTase gene and not in the structural region or the glucose transporting system in cell membrane. The enzymatic properties of CGTase excreted from parent and mutant strains were also compared.

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Improvement of a Fungal Strain by Repeated and Sequential Mutagenesis and Optimization of Solid-State Fermentation for the Hyper-Production of Raw-Starch-Digesting Enzyme

  • Vu, Van Hanh;Pham, Tuan Anh;Kim, Keun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.718-726
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    • 2010
  • A selected fungal strain, for production of the raw-starchdigesting enzyme by solid-state fermentation, was improved by two repeated sequential exposures to ${\gamma}$-irradiation of $Co^{60}$, ultraviolet, and four repeated treatments with Nmethyl-N'-nitrosoguanidine. The mutant strain Aspergillus sp. XN15 was chosen after a rigorous screening process, with its production of the raw-starch-digesting enzyme being twice that of usual wild varieties cultured under preoptimized conditions and in an unsupplemented medium. After 17 successive subculturings, the enzyme production of the mutant was stable. Optimal conditions for the production of the enzyme by solid-state fermentation, using wheat bran as the substrate, were accomplished for the mutant Aspergillus sp. XN15. With the optimal fermentation conditions, and a solid medium supplemented with nitrogen sources of 1% urea and 1% $NH_4NO_3$, 2.5 mM $CoSO_4$, 0.05% (v/w) Tween 80, and 1% glucose, the mutant Aspergillus sp. XN15 produced the raw-starch-digesting enzyme in quantities 19.4 times greater than a typical wild variety. Finally, XN15, through simultaneous saccharification and fermentation of a raw rice corn starch slurry, produced a high level of ethanol with $Y_{p/s}$ of 0.47 g/g.

Role of Dual Flagella in the Pathogenesis of Vibrio parahaemolyticus

  • Lee, Hwa-Gyu;Jeong, Byung-Gon;Park, Kwon-Sam
    • Fisheries and Aquatic Sciences
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    • v.14 no.2
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    • pp.73-78
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    • 2011
  • Vibrio parahaemolyticus possesses two flagella systems: polar and lateral flagella for swimming in liquid and swarming on solid surfaces or in viscous environments. To elucidate the pathogenic role of these dual flagella systems, we constructed single- and double-deletion mutants of the lafA and flhAB flagellum genes and investigated their biofilm formation, cell adhesion, and colonization of the small intestine of suckling mice. The double-mutant strain was more impaired in biofilm formation than either of the single-mutant strains. In addition, the lafA, flhAB, and double-mutant strains showed 40%, 45%, and 60%, respectively, lower adherence to HeLa cells than the wild-type strain. Moreover, the lafA, flhAB, and double-mutant strains exhibited 49%, 5.6 and 6.7 times, respectively, lower colonization in a competition assay than the wild-type strain. These findings indicated that polar flagella were more important than lateral flagella for the pathogenesis of V. parahaemolyticus.

Biosyntheses of Nucleic Acids and Proteins of Bacillus sphaericus ts-Dl290 Lethal Mutant (Bacillus sphaericus ts-D1290 치사돌연변이체의 핵산과 단백질합성)

  • 서정희;이형환;이희무
    • Microbiology and Biotechnology Letters
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    • v.18 no.5
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    • pp.535-540
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    • 1990
  • Bacitlus sphaericus ts-Dl290 was characterized comparatively with the wild type strain 1593 by themeasurements of the biosynthesis of total DNA, RNA and protein on the temperature-shift culturesat permissive temperature of $30^{\circ}C$ and at nonpermissive temperature of $42^{\circ}C$. The growth patterns of the wild type strain and ts-Dl290 were similar at $30^{\circ}C$, but at 4Z C the mutant almost did not grow (temperature-sensitivity). When the growth temperatures of both stains were shifted-up from $30^{\circ}C$ to $42^{\circ}C$ after a 4 hour culture, their growths were normal, but when shifted-down from $42^{\circ}C$ to $30^{\circ}C$ after a 4 h culture, the mutant did not grow. When shifted up from $30^{\circ}C$ to $42^{\circ}C$ after a 4 hculture, the DNA syntheses of the two strains were at a normal rate for 1 h, but after 1 h the biosynthesesdecreased. The rate of DNA synthesis of the wild type strain at the nonpermissive temperature was about 93%, and that of the mutant was about 50% of the ratio of the wild type strain, and the RNA synthesis of the wild type strain was maintained for 3 h, and that of the mutant for 2 h. Thereafter the RNA synthesis decreased, and the synthesis of proteins in the both strains were similarlykept high for 8 h. The reversibility of the DNA synthesis of the mutant at $42^{\circ}C$ was lessened whenthe culture times were increased.re times were increased.

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