• Title/Summary/Keyword: high-yield strains

Search Result 159, Processing Time 0.028 seconds

Development of Miniaturized Culture Systems for Large Screening of Mycelial Fungal Cells of Aspergillus terreus Producing Itaconic Acid

  • Shin, Woo-Shik;Lee, Dohoon;Kim, Sangyong;Jeong, Yong-Seob;Chun, Gie-Taek
    • Journal of Microbiology and Biotechnology
    • /
    • v.27 no.1
    • /
    • pp.101-111
    • /
    • 2017
  • The task of improving a fungal strain is highly time-consuming due to the requirement of a large number of flasks in order to obtain a library with enough diversity. In addition, fermentations (particularly those for fungal cells) are typically performed in high-volume (100-250 ml) shake-flasks. In this study, for large and rapid screening of itaconic acid (IA) high-yielding mutants of Aspergillus terreus, a miniaturized culture method was developed using 12-well and 24-well microtiter plates (MTPs, working volume = 1-2 ml). These miniaturized MTP fermentations were successful, only when highly filamentous forms were induced in the growth cultures. Under these conditions, loose-pelleted morphologies of optimum sizes (less than 0.5 mm in diameter) were casually induced in the MTP production cultures, which turned out to be the prerequisite for the active IA biosynthesis by the mutated strains in the miniaturized fermentations. Another crucial factor for successful MTP fermentation was to supply an optimal amount of dissolved oxygen into the fermentation broth through increasing the agitation speed (240 rpm) and reducing the working volume (1 ml) of each 24-well microtiter plate. Notably, almost identical fermentation physiologies resulted in the 250 ml shake-flasks, as well as in the 12-well and 24-well MTP cultures conducted under the respective optimum conditions, as expressed in terms of the distribution of IA productivity of each mutant. These results reveal that MTP cultures could be considered as viable alternatives for the labor-intensive shake-flask fermentations even for filamentous fungal cells, leading to the rapid development of IA high-yield mutant strains.

Genetic Variability of Flammulina velutipes Monosporous Isolates (팽나무버섯(Flammulina velutipes) 단포자 분리주의 유전적 변이성)

  • Kong, Won-Sik;Kim, Dong-Hyun;Kim, Young-Ho;Kim, Kyoung-Soo;You, Chang-Hyun;Byun, Myung-Ok;Kim, Kwang-Ho
    • The Korean Journal of Mycology
    • /
    • v.25 no.2 s.81
    • /
    • pp.111-120
    • /
    • 1997
  • The characteristics of monosporous isolates of winter mushroom [Flammulina velutipes (Curt. ex Fr.)] were investigated to obtain useful breeding materials. Within monokaryons from cultivars which have a white colored fruitbody showed narrow genetic variation, while domestic strains which have a brown colored fruitbody showed wide variation. The mating type of the white strains was A1A2B1B2 genotype, but that of the domestic brown strains were A3A4B3B4. In intra-crossing, the genetic stability of dikaryons mated by monokaryons from white strain was less than that of parents. While in brown strain, dikaryons with high yield and color variation were obtained.

  • PDF

Towards Methionine Overproduction in Corynebacterium glutamicum - Methanethiol and Dimethyldisulfide as Reduced Sulfur Sources

  • Bolten, Christoph J.;Schroder, Hartwig;Dickschat, Jeroen;Wittmann, Christoph
    • Journal of Microbiology and Biotechnology
    • /
    • v.20 no.8
    • /
    • pp.1196-1203
    • /
    • 2010
  • In the present work, methanethiol and dimethyldisulfide were investigated as sulfur sources for methionine synthesis in Corynebacterium glutamicum. In silico pathway analysis predicted a high methionine yield for these reduced compounds, provided that they could be utilized. Wild-type cells were able to grow on both methanethiol and dimethyldisulfide as sole sulfur sources. Isotope labeling studies with mutant strains, exhibiting targeted modification of methionine biosynthesis, gave detailed insight into the underlying pathways involved in the assimilation of methanethiol and dimethyldisulfide. Both sulfur compounds are incorporated as an entire molecule, adding the terminal S-$CH_3$ group to O-acetylhomoserine. In this reaction, methionine is directly formed. MetY (O-acetylhomoserine sulfhydrylase) was identified as the enzyme catalyzing the reaction. The deletion of metY resulted in methionine auxotrophic strains grown on methanethiol or dimethyldisulfide as sole sulfur sources. Plasmid-based overexpression of metY in the ${\Delta}$metY background restored the capacity to grow on methanethiol or dimethyldisulfide as sole sulfur sources. In vitro studies with the C. glutamicum wild type revealed a relatively low activity of MetY for methanethiol (63 mU/mg) and dimethyldisulfide (61 mU/mg). Overexpression of metY increased the in vitro activity to 1,780 mU/mg and was beneficial for methionine production, since the intracellular methionine pool was increased 2-fold in the engineered strain. This positive effect was limited by a depletion of the metY substrate O-acetylhomoserine, suggesting a need for further metabolic engineering targets towards competitive production strains.

Selection of good bioactive strains from collected Phellinus sp. and development of suitable grain media (상황버섯 수집균주로 기능성 우수균주 선발 및 적합 곡물배지 개발)

  • Kim, Jeong-Han;Baek, Il-Sun;Shin, Bok-Eum;Gwon, Hee-Min;Lee, Yong-Seon
    • Journal of Mushroom
    • /
    • v.17 no.3
    • /
    • pp.107-112
    • /
    • 2019
  • This study was conducted to develop a healthy functional food material using mycelial extract of Phellinus sp. Jangsoo and GMPL48007 were selected as suitable strains because of good mycelial growth and high functionality. The highest total polyphenols and reducing power were observed in the Jangsoo strain and the radical scavenging was shown in GMPL48007. Among grains media, whole-grain wheat had the highest extraction yield from mycelia of selected strains; especially, the GMPL48007 strain showed the highest content of beta-glucan.

Statistical Optimization of Culture Conditions for Lactobacillus Strains using Response Surface Methodology (반응표면분석법을 이용한 Lactobacillus 균주 배양조건의 통계적 최적화)

  • Young Min Hwang;Hee-Seok Lee
    • Journal of Food Hygiene and Safety
    • /
    • v.38 no.5
    • /
    • pp.338-346
    • /
    • 2023
  • The demand for probiotic products has been steadily increasing, and Lactobacillus strains are widely used and are currently the most popular probiotics. Optimizing culture conditions for Lactobacillus production for use as probiotics will enhance their profitability by reducing production costs and time. Statistical analysis using response surface methodology revealed the following optimal sets of independent variables: 22.55 h (cultivation time), 25℃ (cultivation temperature), and 3.41% (w/w, prebiotics concentration) for Lactobacillus acidophilus; 24 h, 30.86℃, and 2% (w/w) for Lactiplantibacillus plantarum; 66.67 h, 35℃, and 3.41% (w/w) for Lacticaseibacillus rhamnosus. Actual outcomes using predicted optimal conditions for Lactobacillus strains have been confirmed to closely match predicted results. This study will provide valuable guidelines for high yield Lactobacillus production.

The Characteriastics of Viscosity Behavior of EMC for Semi-conductor Encapsulant - Containing One Kind of Spherical Silica (반도체 봉지재용 EMC의 점도거동 특성 - 한 종류의 구형 실리카 포함)

  • Kim, In Beom;Lee, Myung Cheon;Lee, Euy Soo
    • Applied Chemistry for Engineering
    • /
    • v.10 no.8
    • /
    • pp.1175-1179
    • /
    • 1999
  • The rheological properties of highly filled epoxy molding compound(EMC) for semi-conductor encapsulants are greatly affected by the content of filler loaded. In this study, the change of viscosity of EMC for semi-conductor encapsulants with the filler content was investigated. Also, both of Cox-Merz and modified Cox-Merz equations were applied to convert the viscosity change as a function of frequency to that of shear rate. It was ovserved that shear thinning and yield stress occured at high filler contents and that the Cox-Merz equation could not be applied at high filler contents because of the difference of viscosity according to the various strains. When the modified Cox-Merz equation was applied, the all the curves having different strain tend to be represented by one master curve, even though some deviation was obseved at high filler content and strain.

  • PDF

Production and Regeneration of Lactobacillus bulgaricus Protoplasts

  • Jun, Hong-Ki;Park, Hyun-Jeong;Baik, Hyung-Suk;Song, Jae-Chul
    • Journal of Microbiology and Biotechnology
    • /
    • v.1 no.4
    • /
    • pp.246-250
    • /
    • 1991
  • Conditions for the production and regeneration in Lactobacillus bulgaricus protoplasts were investigated. Protoplasts of L bulgaricus strains were obtained by treatment with mutanolysin and lysozyme together in a protoplast forming buffer containing 0.02 M N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) (pH 7.0) and 0.5 M sucrose. High protoplast yield was obtained from cells cultured in the de Man, Rogosa and Sharpe(MRS) medium at the middle to late logarithmic growth phase. Regeneration was efficiently accomplished with a complex medium containing 1% sucrose, 20 mM $MgCl_2$, 5% gelatin, and 0.5% bovine serum albumin. The frequency of regeneration of protoplasts was 10~20% after 5 days of incubation at $30^{\circ}C$.

  • PDF

A Study on the strength improvement in weldment by the impact loading (충격하중에 의한 용접구조물의 강도 증가에 관한 연구)

  • 양영수
    • Journal of the Korean Society of Manufacturing Technology Engineers
    • /
    • v.9 no.3
    • /
    • pp.76-82
    • /
    • 2000
  • It is well known that during the oxygen cutting process residual thermal stresses are produced in weldment. The local non-uniform heating and subsequent cooling which takes place during any welding process causes complex thermal strains and stresses to finally lead to residual stresses exceed to the yield stress. High tensile stresses combined with applied structural load in the region near the welded joint can given rise to distortion brittle fracture change of the fatigue strength and stress corrosion cracking. The appropriate treatment of the welded component which reduces the peak of he welding residual stresses is believed to lower risk of the fracture during the service of the structure. In this study the impact loading in oxygen cutting frame was applied to reduce the residual stress. After applying the impact loading redistribution of resid-ual stress was measured by cutting method and the effect of fatigue was tested.

  • PDF

Acetic Acid Fermentation by Acetobacter sp. SK-7 using Maesil Juice (Acetobacter sp. SK-7에 의한 매실식초 발효)

  • Kim, Mi-Hyang;Choi, Ung-Kyu
    • Journal of the Korean Society of Food Culture
    • /
    • v.21 no.4
    • /
    • pp.420-425
    • /
    • 2006
  • This study was conducted to produce vinegar using maesil. Acetic acid bacteria was 20 strains isolated from several conventional vinegars. Among the isolates, a strain showed highest acetic acid productivity was selected and identified as Acetobacter sp. SK-7. The optimum medium of acetic acid production by Acetobacter sp. SK-7 was 30% of maesil juice, 4% of ethanol, and 2% of starting acidity and 0.2% of glucose. Optomum condition for the high yield of acetic acid was in the shaking culture at 30$^{\cire}$. The acidity of culture medium was reached to 7.1% after 12 days fermentation. Organic acid was identify 6 kinds containing acetic acid. The total content was 7,068.7 mg% after 12 days and malic acid slowly decreased and acetaic and citirc acid gradationally increased according to fermentation

Isolation of a Mutant with Thermotolerance and Ethanol Tolerance Using Proofreading-deficient DNA Polymerases in Saccharomyces cerevisiae (출아효모에서 proofreading-deficient DNA polymerase를 이용한 내열성 및 에탄올내성 변이 주의 분리)

  • Kim, Yeon-Hee
    • Journal of Life Science
    • /
    • v.29 no.8
    • /
    • pp.916-921
    • /
    • 2019
  • In this study, we constructed a biological system that exhibited thermotolerance, ethanol tolerance, and increased ethanol productivity using a random mutagenesis method. We attempted to isolate a thermotolerant mutant using proofreading-deficient DNA polymerase ${\delta}$ and ${\varepsilon}$ encoded by the pol3 and pol2 genes, respectively, in Saccharomyces cerevisiae. To obtain mutants that could grow at high temperatures ($38^{\circ}C$ and $40^{\circ}C$), random mutagenesis of AMY410 (pol2-4) and AMY126 (pol3-01) strains was induced. The parental strains (AMY410 and AMY126) grew poorly at temperatures higher than $38^{\circ}C$. By stepwise elevation of the incubation temperature, AMY410-Ht (heat tolerance) and AMY126-Ht strains that proliferated at $40^{\circ}C$ were obtained. These strains were further incubated in medium containing 6% and 8% ethanol and then AMY410-HEt (heat and ethanol tolerance) and AMY126-HEt strain with ethanol tolerance at an 8% ethanol concentration was obtained. The AMY126-HEt strain grew even at an ethanol concentration of 10%. Furthermore, following the addition of high concentrations of glucose (5% and 10%), an AMY126-HEt3 strain with increased ethanol productivity was isolated. This strain produced 24.7 g/l of ethanol (95% theoretical conversion yield) from 50 g/l of glucose. The findings demonstrate that a new biological system (yeast strain) showing various phenotypes can be easily and efficiently bred by random mutagenesis of a proofreading- deficient mutant.