• Title/Summary/Keyword: Mutant strain

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Immunological Roles of Pasteurella multocida Toxin (PMT) Using a PMT Mutant Strain

  • Kim, Tae-Jung;Toan, Nguyen Tat;Jang, Eun-Jin;Jung, Bock-Gie;Lee, Jae-Il;Lee, Bong-Joo
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.364-366
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    • 2007
  • The immunological role of the Pasteurella multocida toxin (PMT) in mice was examined using a PMT mutant strain. After a nasal inoculation, the mutant strain failed to induce interstitial pneumonia. Moreover, PMT had no significant effect on the populations of CD4+, CD8+, CD3+, and CD19+ immunocytes in blood or on the populations of CD4+ and CD8+ splenocytes (P<0.01). However, there was a significant increase in the total number of cells in the BAL samples obtained from the wild-type P. multocida-inoculated mice. On the other hand, the level of IL-l expression decreased when the macrophages from the bronchio-alveolar lavage were stimulated with PMT. Overall, PMT appears to play some role (stimulating and/or inhibiting) in the immunological responses but further studies will be required to confirm this.

High-Level Production of Astaxanthin by Fed-Batch Culture of Mutant Strain Phaffia rhodozyma AJ-6-1

  • KIM, SU-JIN;GEUN-JOONG KIM;DON-HEE PARK;YEON-WOO RYU
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.175-181
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    • 2003
  • The production of a carotenoid astaxanthin, a growth-associated principal pigment, is limited in a batch cultivation, because a high glucose concentration severely inhibits the cell growth and also influences the carotenoid production. Therefore, a fermentation strategy including effective chemicals for the high-level production of cells and astaxanthin by a mutant strain Phaffia rhodozyma AJ-6-1 was developed in a fed-batch culture. First, a production medium for maximizing the cell and astaxanthin yields was formulated and optimized. Using this optimized medium, the highest cell and astaxanthin concentrations obtained were about 38.25 g/1 and 34.77 mg/1, respectively. In addition, an attempt was made to increase the amount of astaxanthin using effective chemicals such as ethanol and acetic acid, which are known at an inducer and/or precursor of carotenoid synthesis. When either 10g/1 ethanol or 5 g/1 acetic acid was added to investigate the resulting astaxanthin content, a relatively high astaxanthin concentration or 45.62 mg/l and 43.87 mg/1, respectively, was obtained, and the cell concentrations also increased slightly under these conditions. Therefore, these results imply that a fed-batch culture of the mutant strain P. rhodozyma AJ-6-1 could be effectively employed in the commercial production of astaxanthin, although the factors affecting the productivity remain to be elucidated.

Overexpression of Mutant Galactose Permease (ScGal2_N376F) Effective for Utilization of Glucose/Xylose or Glucose/Galactose Mixture by Engineered Kluyveromyces marxianus

  • Kwon, Deok-Ho;Kim, Saet-Byeol;Park, Jae-Bum;Ha, Suk-Jin
    • Journal of Microbiology and Biotechnology
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    • v.30 no.12
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    • pp.1944-1949
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    • 2020
  • Mutant sugar transporter ScGAL2-N376F was overexpressed in Kluyveromyces marxianus for efficient utilization of xylose, which is one of the main components of cellulosic biomass. K. marxianus ScGal2_N376F, the ScGAL2-N376F-overexpressing strain, exhibited 47.04 g/l of xylose consumption and 26.55 g/l of xylitol production, as compared to the parental strain (24.68 g/l and 7.03 g/l, respectively) when xylose was used as the sole carbon source. When a mixture of glucose and xylose was used as the carbon source, xylose consumption and xylitol production rates were improved by 195% and 360%, respectively, by K. marxianus ScGal2_N376F. Moreover, the glucose consumption rate was improved by 27% as compared to that in the parental strain. Overexpression of both wild-type ScGAL2 and mutant ScGAL2-N376F showed 48% and 52% enhanced sugar consumption and ethanol production rates, respectively, when a mixture of glucose and galactose was used as the carbon source, which is the main component of marine biomass. As shown in this study, ScGAL2-N376F overexpression can be applied for the efficient production of biofuels or biochemicals from cellulosic or marine biomass.

노랑초파리의 날개성체원기의 결정성에 관한 연구: I. 정상종과 흔적시의 성체원기에서의 단백질 합성

  • 이양림;박성순
    • The Korean Journal of Zoology
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    • v.25 no.2
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    • pp.93-106
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    • 1982
  • Proteins, which may be closely related to differentiation of a cell group into a predestined fate, were investigated using imaginal discs of a wing mutant, vestigial of Drosophila melanogaster. The wing discs of the mutant fail to differentiate into normal wings, even though the third instar larvae form wing discs, which are very similar to those of the normal strain in size and shape. Patterns of proteins of accumulated or synthesized in the discs of the third instar larvae of the normal and mutant strains were analyzed by acrylamide gel electrophoresis. The patterns of accumulated proteins were found to be slightly different between two strains in quantity rather than in quality. The patterns of proteins synthesized at various times of the third instar were found to be very similar to each other, even though there were a few proteins specific to the normal or to the mutant strain.

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Hyper-CMCase-Producing Mutants of Bacillus sp. 79-23 Induced by Gamma- Radiation

  • Yoon, Ki-Hong;Shin, In-Kyung;Jung, Kyung-Hwa;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.518-521
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    • 1999
  • Bacillus sp. 79-23 spores were irradiated with $^{60}Co$ gamma-rays at doses ranging from 0.5 to 5 kGy. Following gamma-irradiation, seven mutant strains were isolated by scoring the halo sizes formed around the colonies grown on LB agar plates containing 4% carboxymethylcellulose (CMC) and trypan blue. The mutant strains showed a 1.5 to 2-fold increase in carboxymethylcellulase (CMCase) activity over the parent strain. Wheat bran acted as an effective inducer for CMCase production in the parent and mutant strains. Mutant strains 68 and 70 were identified as exhibiting higher CMCase activities than those of other mutants in LB media both with and without 3% wheat bran. In addition, these strains seem to produce substantially lower amounts of capsular materials, whereas the parent strain produced large amounts of them in both liquid and solid LB media. In flask cultures, the CMCase production by mutants 68 and 70 reached maximum levels of 17.5 unit/ml and 15.7 unit/ml, respectively, in an LB medium containing 3% wheat bran.

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Isolation of Glucose Isomerase Hyperproducing Strain, Streptomyces sp. SM 805 and Its Enzymatic Properties

  • Kim, Hong-Rip
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.78-84
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    • 1992
  • Streptomyces sp. No.8, which produced glucose isomerase was isolated from soil samples. The isolated strain, No.8, was identified as belonging to the Genus Streptomyces. A mutant strain, SM 805, showed the greatest ability to produce glucose isomerase. It was developed from the strain, No.8, by mutagenesis induced by NTG and UV treatment. The mutant strain, SM 805, produced about 7 times more glucose isomerase than the parental strain, No.8. This enzyme catalyzed the isomerization of D-xylose, D-glucose and D-ribose. It was inactive in the absence of metal ions, but was activated by the addition of $Mg^{2+}$ or $Co^{2+}$. The optimum temperature and pH for enzyme activity were $80^\circ{C}$ and pH 8.5, respectively. The enzyme was stable in a pH range of 6.0 to 10.0, and it was highly thermostable. There was no activity loss below $80^\circ{C}$, and even above $90^\circ{C}$ about 45% of its activity was retained. The reaction equilibrium was reached when about 53% fructose was present in the reaction mixture. Whole cells containing glucose isomerase from Streptomyces sp. SM 805 were immobilized by glutaraldehyde treatment. The resultant immobilized enzyme pellets showed a relatively long stability during the isomerizing reaction. The half-life of the immobilized enzyme during the operating was 45 days in the presence of 10mM $Mg^{2+}$.

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Strain Improvement of Penicillium verruculosum for High Cellulase Production by Induced Mutation (섬유소분해효소 생산증진을 위한 Penicillium verruculosum의 균주개량)

  • 정기철
    • Microbiology and Biotechnology Letters
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    • v.15 no.6
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    • pp.388-395
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    • 1987
  • In order to obtain a regulatory mutant strain with high cellulase activity, a newly isolated Penicillium verrculosum, strain F-3 was used as parental strain since it was proved to be an efficient cellulase producer. A number of experiments were conducted to determine the optimum conditions to in-duce mutagenesis and isolate the desirable mutant strains. Out of several restriction compounds tested, 1.5% oxgall was found to be most effective to restrict the colony size by suppressing overgrowth. Derepression of catabolites was employed as a criterion in selecting mutant strains with high cellulase productivity. Production of cellulase by Penicillium venculosum F-3 was suppressed when cultured on the media with more than 1% of glucose or glycerol. It was found that either irradiation with UV light for 19 mins or treatment with nitrosoguanidine at 200$\mu\textrm{g}$/m1 for 60 mins, induced mutagenesis at desired level, when the survival rate of the spore was 0.2% and 48%, respectively. Three mutant strains of F-3, UV-9, UV-10, and NTG-3 that had the highest cellulase productivity were finally selected, based on filter paper degradation rate, size of clearing zone on the screening plate and cellulase activity in the medium containing cellulose powder. When the mutant strains were compared with parental strain F-3, on the KC-M-W medium containing cellulose powder, the filter paper activities of UV-9, UV-10, and NTG-3 were increased by 34%, 55%, and 41%, respectively. However, the assimilation of cellobiose octaacetate by UV-9 or NTG-3 was markedly reduced. When the mutant UV-10 was grown on cellobiose octaacetate medium (CCA-4) in shaking flasks, the cellulase activities of the mutant increased by 20 to 50% compared to the parental strain. Excreation of soluble protein from the mutant also elevated up to 30%. The mutant also constitutively produced both CMCase and $\beta$-glucosidase, though at relatively low level, in the presence of glucose or cellobiose as carbon sources.

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Isolation of Mutant Strains from Keratinase Producing Bacillus subtilis SMMJ-2 and Comparision of Their Enzymatic Properties (Keratinase 생산균 Bacillus subtilis SMMJ-2의 변이주 분리와 효소학적 특성 비교)

  • Ko, Hee-Sun;Kim, Hyun-Soo
    • KSBB Journal
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    • v.25 no.5
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    • pp.429-436
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    • 2010
  • Keratinase is widely used in certain industrial applications. The present study sought to improve the culture conditions of Bacillus subtilis SMMJ-2 to facilitate mass production of keratinase. Strain SMMJ-2 was irradiated by ultraviolet light and the resulting isolates were tested for keratinase activity. Isolates displaying elevated keratinase activity were selected and used to determine the optimum temperature (24, 30, 37, 45, $55^{\circ}C$) for bacterial keratinase production during a 4 day incubation period. The highest enzyme activity (55 units/mL/min), from a Bacillus subtilis SMMJ-2 mutant (mutant No. 2) was demonstrated following incubation at $30^{\circ}C$. The effects of carbon and nitrogen sources on keratinase production were confirmed by measuring the enzyme activity from the culture broth of the mutant strain cultured in various media containing different carbon source and nitrogen sources during a 4 day period. The optimal medium composition for producing keratinase consisted of 1% glucose, 0.7% $K_2HPO_4$, 0.2% $K_2HPO_4$, and 1.2% soybean meal. Optimal initial pH and temperature for producing keratinase were 7.0 and $30^{\circ}C$, respectively. Keratinases produced by B. subtilis SMMJ-2 and the mutant No. 2 were purified from the culture broth which used soybean meal as a nitrogen source. Membrane ultrafiltration, DEAE-sephacel ion exchange and Sephadex G-100 gel chromatography were used to purify the enzymes. The purified keratinases from both B. subtilis SMMJ-2 and the mutant No. 2 showed single bands and their molecular weights were estimated as 28 kDa and 42 kDa, respectively on SDS-polyacrylamide gel electrophoresis.

Selection of a Mutant Strain with High Yield of Cellulose Production Derived from $Acetobacter$ sp. A9 ($Acetobacter$ sp. A9에서 셀룰로오스 생산량이 높은 변이주 선별)

  • Lee, O-Mi;Son, Hong-Joo;Lee, Sang-Joon
    • Korean Journal of Environmental Biology
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    • v.29 no.4
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    • pp.321-325
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    • 2011
  • The mutant strain M6 derived from Acetobacter sp. A9, which produces high levels of the bacterial cellulose derived by random mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine or UV treatment, was selected by a Hestrin and Schramm medium (HSB) plate assay. The characterization of the cellulose production was studied in flask culture to improve the productivity of bacterial cellulose by $Acetobacter$ sp. A9 and mutant strain M6. The yield of cellulose production was superior to mutant M6 than $Acetobacter$ sp. A9. Cellulose was produced 0.12 g $L^{-1}$ by $Acetobacter$ sp. A9 at HS medium and the mutant M6 produced the cellulose 6.95 g $L^{-1}$at HS medium. Strain M6 produced less amount of gluconic acid than A9, thus showing that cellulose production is negatively relted with the gluconic acid production.

Construction of a Pure Cryparin-null Mutant for the Promoter Analysis of Cryparin Gene (Cryparin 유전자의 promoter 분석을 위한 cryparin 유전자 치환체의 순수 제조)

  • Kim, Myoung-Ju;Yang, Moon-Sik;Kim, Dae-Hyuk
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.450-457
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    • 1998
  • The cryparin of Cryphonectria parasitica belongs to a cell wall associated fungal hydrophobin. The cryparin, though it is encoded by a single copy gene, is known for the high expression during the liquid culture of C. parasitica, and it turns out that 22% of total mRNA was transcribed for cryparin at 48hr after the liquid culture. In addition, it is also known as one of down-regulated fungal proteins by the presence of double stranded RNA virus, Cryphonectria hypovirus 1. In previous studies (Kim et al., 1999), we have constructed a cryparin-null mutant by replacing the cryparin gene with hygromycin B resistance gene due to site directed homologous recombination. In order for the promoter analysis of cryparin which seems to be very strong as well as mycoviral specific, it is preferable to have a strain with only a target promoter replaced and a discernable target site for incoming vectors. However, the cryparin-null mutant revealed the presence of an additional copy of transforming vector except the one which replaced the cryparin gene. In addition, the cryparin-null mutant did not contain any markers for targeted integration of incoming vectors. This prompts us to design an experiment to obtain a strain for promoter analysis of cryparin gene. A different mating type strain EP6(Mata, $met^-$) was mated with the cryparin-null mutant ${\triangle}$Crp194-7(MatA, Crp${\triangle}$::hph) to make the progenies with only a single replacement vector and $met^-$ characteristic remained. Nutritional assay as well as Southern blot analysis revealed that the progeny, ${\triangle}$Crp194-a6, was the methionine auxotroph with a single replacing vector in genome. Northern blot analysis and PAGE showed that there was no cryparin produced in this bred strain either.

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