• Title/Summary/Keyword: wild strain

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Enhanced production of cellulase by a mutant strain of aspergillus phoenicis (Aspergillus phoenicis의 한 돌연변이주에 의한 cellulase의 생성 및 그 특성)

  • 이영록;고상균
    • Korean Journal of Microbiology
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    • v.20 no.3
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    • pp.125-133
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    • 1982
  • Mutational experiments were performed to imporve the cellulase productivity of Aspergillus phoenicis KU175, isolated from the southern part of Korea, as a high cellulase producer. By treatment ultra-violet light nad 4-NQO(4-Nitroquinoline-N-Oxide), mutation waas induced, and treatment ultra-violet light and 4-NQO (4-Nitroquinoline-N-Oxide), mutation was induced, and A.phoenicis KU175-115 was finally selected for its highest avicelase production. Avicelase production of the mutant was increased about 2 times compared with those of the wild strain. However, activities of other hydrolytic enzymes, such as amylase, protease and nuclease, of the mutant strain didn't show a marked difference compared with those of the nuclease, of the mutant strain didn't show a marked difference compared with the wild strain, except slight increase in ribonuclease activity and slight decrease in glucoamylase activity. Avicelases from the mutant strain selected were purified from wheat bran culture by successive salting out, followed by dialysis and column chromatography, and their charcteristics were compared with thosw of the wild strain. Avicelase was separated into three peaks in the mutant strain as well as in the case of wild strain. Avicelase II activity of the mutant strain was prominently higher than that of the wild strain, while avicelase I and III activities of those were equivalent. The optimal pH ranges and stability of avicelase II from the mutant strain were pH4-5 and pH3.5-6.0, respectively, as well as in the case of the wild strain. The optimal temperature and thermal stability of avicelase II from the mutant strain were $40{\sim}50^{\circ}C\;and\;20{\sim}55^{\circ}C$, respectively. These results were same as those of the wild strain. By the using of Eadie-Hofastee plot, $K_m\;and\;V_{max}$ of avicelase II from the mutant and the wild strain were calculated to be 2.29mg/ml and $4.84{\mu}g$ reducing sugar as glucose per min equally, from the line fitted to the data by the least square method. Activity of avicelase II from the mutant strain was slightly activated by $Mg^{++}\;but\;inhibited\;by\;Cu^{++}, \;Mn^{++}\;and\;Zn^{++}$, as well as in the case of the wild strain. Therefore, it was concluded that the mutant didn't induce the formation of another avicelase isozyme, or the changes in the properties of avicelase, but induce the changes in the productively of the same avicelase II by the action of regulatory gane.

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Strain Improvement of Candida tropicalis for the Production of Xylitol: Biochemical and Physiological Characterization of Wild-type and Mutant Strain CT-OMV5

  • Rao Ravella Sreenivas;Jyothi Cherukuri Pavanna;Prakasham Reddy Shetty;Rao Chaganti Subba;Sarma Ponnupalli Nageshwara;Rao Linga Venkateswar
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.113-120
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    • 2006
  • Candida tropicalis was treated with ultraviolet (UV) rays, and the mutants obtained were screened for xylitol production. One of the mutants, the UV1 produced 0.81g of xylitol per gram of xylose. This was further mutated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), and the mutants obtained were screened for xylitol production. One of the mutants (CT-OMV5) produced 0.85g/g of xylitol from xylose. Xylitol production improved to 0.87 g/g of xylose with this strain when the production medium was supplemented with urea. The CT-OMV5 mutant strain differs by 12 tests when compared to the wild-type Candida tropicalis strain. The XR activity was higher in mutant CT-OMV5. The distinct difference between the mutant and wild-type strain is the presence of numerous chlamvdospores in the mutant. In this investigation, we have demonstrated that mutagenesis was successful in generating a superior xylitol-producing strain, CT-OMV5, and uncovered distinctive biochemical and physiological characteristics of the wild-type and mutant strain, CT-OMV5.

Role of Surface Protective Antigen A in the Pathogenesis of Erysipelothrix rhusiopathiae Strain C43065

  • Borrathybay, Entomack;Gong, Feng-juan;Zhang, Lei;Nazierbieke, Wulumuhan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.206-216
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    • 2015
  • To clarify the role of surface protective antigen A (SpaA) in the pathogenesis of Erysipelothrix rhusiopathiae C43065 (serotype 2), the spaA deletion mutant of E. rhusiopathiae ${\Delta}spaA$ was constructed by homologous recombination. The virulence of the ${\Delta}spaA$ mutant decreased more than 76-fold compared with that of the wild-type strain C43065 in mice. The mutant strain was sensitive to the bactericidal action of swine serum, whereas the wild-type strain was resistant. The adhesion of wild-type strain to MEF cells was inhibited significantly by treatment with rabbit antiserum against recombinant SpaA (rSpaA) as compared with the treatment with normal rabbit serum, but the mutant strain was not affected. The mutant strain was readily taken up by mouse peritoneal macrophages in the normal rabbit serum, whereas the wild-type strain was resistant. Whereas the rabbit antiserum against rSpaA promoted the phagocytosis of wild-type strain by macrophages, the mutant strain was not affected. In addition, mice vaccinated with the formalin-killed mutant strain were provided 40% protection against challenge by the homologous virulent strain as compared with those with wild-type strain, NaOH-extracted antigen, or rSpaA, which provided more than 80% protection against the same infection. These suggested that SpaA has an important role in the pathogenesis of E. rhusiopathiae infection and could be a target for vaccination against swine erysipelas.

Further induction of amylase producing mutants from a highly proteolytic mutant strain of asppergillus flavus (돌연변이에 의한 Aspergillus flavus의 아밀라아제 생성능의 개량)

  • 이영록;고상균;김봉수
    • Korean Journal of Microbiology
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    • v.18 no.4
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    • pp.161-171
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    • 1980
  • A mutant strain having increased productivity of both enzymes, protease and amylase, was obtained from A. flavus KU 153, isolatd from South Korea for its high protease production by successive ultra-violet light irradiation, Two glucoamylases from the mutant strain selected were purified from wheat branculture by successive salting out, followed by dialysis and column chromatography, and their characteristics were compared with those of the wild strain. Glucoamylase production of the mutant selected was increased about 3.3 times compared with the wild strain, and 2.1 times compared with the parental strain, ${\alpha}-amylase$ activity of the mutant selected was about 2 times hugher than that of the wild strain or the parental strain. Protease and cellulase productivities of the muant selected were all alike compared with those of the highly proteolytic mutant, the parental strain. Therefore, it was considered that the back mutation on the protease production did not occurred in the formation process of the glucoamylase producing mutant. Total activities of glucoamylase I and II from the mutant selected were 2.86 and 3.65 times higher compared with those from the wild strain, respectively. Considering the optimal pH-thermal stability and Km-Vmax value of glucoamylase I and II from both strains, wild and mutant, it was deduced that the characteristics of glucoamylase I and II from the wild strain did not altered during the mutation process. Therefore, it was concluded that the selected mutant did not induce the formation of another glucoamylase isozyme, or the changes in the characteristics of the glucoamylase, but induce the productivity of the same glucoamylase I and II by the action of regulatory gene.

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Potentials for Biological Control of Blue Stain on Woods Caused by Ophiostomatoid Fungi

  • Lee, Jong-Kyu;Oh, Eun-Sung
    • The Plant Pathology Journal
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    • v.16 no.4
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    • pp.200-205
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    • 2000
  • Biological control of blue stain fungi, such as Ophiostoma and Leptographium spp., that reduce the quality of logs and cause economic losses in wood product industry, was carried out in laboratory and field trials by a colorless strain of Ophiostoma quercus, BSFcs-1. Inoculation of pine wood chips with the colorless strain 1 wk before inoculating wild-type strain demonstrated that BSFcs-1 colonized wood chips and excluded blue stain fungi from being established. Efficacy of BSFcs-1 was compared with colorless strain of O. piliferum, which is commercially available under the trade name of Cartapip. Inoculation of pine wood logs with the colorless strain 1 wk before inoculating wild-type strain of blue stain in isolated wood chips, while O. quercus and O. floccosum colonized 0% and 17%, respectively. Simultaneous inoculation of logs with the colorless and wild-type strains resulted in decreased colonization (28%) by BSFcs-1, but increased colonization by O. quercus (185) and O. floccosum (29%). On the other hand, BSFcs-1 and wild-type strain alone colonized 75% and 71%, respectively. Treatment of the surface of log ends with mycelial suspension of BSFcs-1 after cutting also showed good control of blue stain fungi in a pine forest stands.

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Increased Production of Recombinant Protein by Escherichia coli Deficient in Acetic Acid Formation

  • Koo, Tae-Young;Park, Tai-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.789-793
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    • 1999
  • The effect of acetic acid formation deficiency on recombinant E. coli fermentation was investigated using a mutant strain deficient in acetic acid formation. A mutant strain which does not grow under anaerobic conditions was isolated. The acetic acid production in this strain was negligible in aerobic batch fermentation. The cloned-gene expression in the mutant strain was higher than the wild-type strain. Fed-batch fermentations with controlled specific growth rates were carried out in order to compare the cloned-gene expression between the wild-type and the mutant strains. The expression decreased along with the specific growth rate in both strains. The cloned-gene expression in the mutant strain was 60% higher than in the wild-type strain at the same specific growth rate.

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Sigma S Involved in Bacterial Survival of Ralstonia pseudosolanacearum (Ralstonia pseudosolanacearum 생존에 관여하는 Sigma S 역할)

  • Hye Kyung Choi;Eun Jeong Jo;Jee Eun Heo;Hyun Gi Kong;Seon-Woo Lee
    • Research in Plant Disease
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    • v.30 no.2
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    • pp.148-156
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    • 2024
  • Ralstonia pseudosolanacearum, a plant pathogenic bacterium that can survive for a long time in soil and water, causes lethal wilt in the Solanaceae family. Sigma S is a part of the RNA polymerase complex, which regulates gene expression during bacterial stress response or stationary phase. In this study, we investigated the role of sigma S in R. pseudosolanacearum under stress conditions using a rpoS-defective mutant strain of R. pseudosolanacearum and its wild-type strain. The phenotypes of rpoS-defective mutant were complemented by introducing the original rpoS gene. There were no differences observed in bacterial growth rate and exopolysaccharide production between the wild-type strain and the rpoS mutant. However, the wild-type strain responded more sensitively to nutrient deficiency compared to the mutant strain. Under the nutrient deficiency, the rpoS mutant maintained a high bacterial viability for a longer period, while the viability of the wild-type strain declined rapidly. Furthermore, a significant difference in pH was observed between the culture supernatant of the wild-type strain and the mutant strain. The pH of the culture supernatant for the wild-type strain decreased rapidly during bacterial growth, leading to medium acidification. The rapid decline in the wild-type strain's viability may be associated with medium acidification and bacterial sensitivity to acidity during transition to the stationary phase. Interestingly, the rpoS mutant strain cannot utilize acetic acid, D-alanine, D-trehalose, and L-histidine. These results suggest that sigma S of R. pseudosolanacearum regulates the production or utilization of organic acids and controls cell death during stationary phase under nutrient deficiency.

Comparisons of growth and biofilm production with Vibrio fluvialis and mutants deficient in oligopeptide permease gene (Vibrio fluvialis의 Oligopeptide Permease Gene 결손에 의한 생육과 Biofilm 생산의 비교)

  • Lee, Eun Mi;Ahn, Sun Hee;Kong, In Soo
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.2
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    • pp.84-90
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    • 2006
  • Various ${\Delta}opp$ mutants of Vibrio fluvialis were constructed by allelic exchange method. The mutants occurred in target genes were confirmed by PCR and Southern hybridization analyses. After the exact mutants were identified, cell growth and biofilm production were examined using the respective mutants. The growth of wild strain was more rapid than mutants within 4hr incubation. Thereafter, the growth of wild strain and mutants reached to same level. When the productivities of wild strain and mutants were examined, ${\Delta}oppA$ mutant showed the highest productivity. Though ${\Delta}oppC,D$ and F mutants produced the lower production than that of ${\Delta}oppA$ mutant, the productivities of those mutants were much higher than that of wild strain.

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Selective Isolation and Characterization of Schwanniomyces castellii Mutants with Increased Production of a-Amylase and Glucoamylase

  • Ryu, Yeon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.3 no.2
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    • pp.95-98
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    • 1993
  • This study was carried out to isolate and characterize the mutant strains of Schwanniomyces castellii NRRL Y-2477. Mutants were prepared with the treatment of ethyl methane sulfonate. 2-deoxy-D-glucose resistant mutants were isolated and two mutants were selected based on their high production of amylolytic enzymes and their ability to ferment starch. The mutants selected had higher a-amylase and glucoamylase activities than the wild type strain from several other carbon sources. Especially, it was revealed that mutant strain M-9, when cultured in the presence of glucose as a sole carbon source, shows relatively high activities of a-amylase and glucoamylase compared to those of the wild type strain. In result, this mutant strain can be considered as a constitutive producer of amylolytic enzymes. To compare the ethanol production ability of wild type strain and of mutant strains selected, an alcohol fermentation was carried out using 100 g/l soluble starch. Mutant strain M-9 did not improve the direct alcohol fermentation of starch, despite its excellent amylolytic activities performance. On the other hand, mutant strain M-6 produced 37.9 g/l (4.8%, v/v) ethanol by utilizing about 82% of substrate.

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Comparison of Bacterial Cellulose Production in a Jar Fermentor Between Acetobacter xylinum BPR2001 and its Mutant, Acetan-Nonproducing Strain EP1

  • BAE SANG OK;SUGANO YASUSHI;SHODA MAKOTO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.247-253
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    • 2005
  • The bacterial cellulose (BC) production by a wild­strain Acetobacter xylinum BPR2001 and that by its acetan­nonproducing mutant, EPI, were compared in a jar fermentor. EPI produced about $28\%$ less BC than the wild-strain. The apparent difference in the cultivation of the two strains was the viscosity increase in the culture broth that was closely associated with acetan production. Increasing the viscosity of the culture broth of EPI by adding agar led to the formation of relatively small and uniform BC pellets, and BC production consequently became two-fold higher than that in the absence of agar and was almost equal to that by BPR2001. Therefore, acetan has an important role in BC production by inducing physical changes in the culture broth of the wild-type strain.