• 제목/요약/키워드: Substrate culture

검색결과 655건 처리시간 0.026초

High-Yield Production of Xylitol from Xylose by a Xylitol Dehydrogenase Defective Mutant of Pichia stipitis

  • Kim, Min-Soo;Chung, Yun-Seung;Seo, Jin-Ho;Jo, Do-Hyun;Park, Yun-Hee;Ryu, Yeon-Woo
    • Journal of Microbiology and Biotechnology
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    • 제11권4호
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    • pp.564-569
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    • 2001
  • This study was carried out in order to investigate the characteristics of xylitol fermentation by a xylitol dehydrogenase defective mutant PXM-4 of P stipitis CBS 5776 and to determime optimum conditions for the high yield ofxylitol production from xylose. Gluconic acid was selected as a co substrate for the xylitol fermentation, since gluconic acid neither blocked xylose transport nor repressed xylose reductase expression. An increase of gluconic acid concentration reduced the rates of xylitol production and cell growth by decreasing medium pH, and the optimal concentration of gluconic acid was determined to be 20 gll with approximately 100% xylitol conversion yield. A fed-batch cell culture resulted in a 44.8 g/l xylitol concentration with 100% yield, based on the amount of xylose consumed.

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Molecular and Morphological Characterization of Green Mold, Trichoderma spp. isolated from Oyster Mushrooms

  • Choi, In-Young;Hong, Seung-Beom;Yadav, Mahesh C.
    • Mycobiology
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    • 제31권2호
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    • pp.74-80
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    • 2003
  • Isolates of Trichoderma spp. collected from Pleurotus ostreatus and P. eryngii beds, which included loosened substrate compactness and development of green colour, were grouped into three species. The occurrence of different species of Trichoderma was as T. cf. virens(70.8%), T. longibrachiatum(16.7%) and T. harzianum(12.5%). The conidia of Trichoderma spp. were ellipsoidal, obovoid and phialides were bowling pins, lageniform and the length of phialides was $3.5{\sim}10.0{\times}1.3{\sim}3.3{\mu}m$. Phialides of T. cf. virens and T. harzianum were tending clustered, but it was solitary disposition in T. longibrachiatum. T. cf. virens was characterized by predominantly effuse conidiation, sparingly branched, and fertile to the apex and it was penicillate type. RAPD analysis could detect variability amongst three different species of Trichoderma using two newly designed URP-primers. However, intra-specific variation could not be detected in all the isolates except for rDNA sequence data classified Trichoderma isolates into three distinct groups representing three species. The profiles of rDNA sequences of isolates representing a species showed high similarity in T. cf. virens and T. harzianum. However, there was a variation in rDNA sequences of isolates representing T. longibrachiatum. The results of present study reveals that molecular techniques of RAPD and rDNA sequencing can greatly aid in classification based on morphology and precise identification of fast evolving species of Trichoderma.

Evaluation of a Chromogenic Medium Supplemented with Glucose for Detecting Enterobacter sakazakii

  • Song, Kwang-Young;Hyeon, Ji-Yeon;Shin, Ho-Chul;Park, Chan-Kyu;Choi, In-Soo;Seo, Kun-Ho
    • Journal of Microbiology and Biotechnology
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    • 제18권3호
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    • pp.579-584
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    • 2008
  • A commercial chromogenic agar medium (DFI) was supplemented with glucose (mDFI) to enhance the specificity of Enterobacter sakazakii (E. sakazakit) detection. Escherichia vulneris (E. vulneris), a putative false-positive strain on the DFI medium, produces ${\alpha}$-glucosidase. The enzyme ${\alpha}$-glucosidase hydrolyzes a substrate, 5-bromo-4-chloro-3-indolyl-${\alpha}$, D-glucopyranoside $(X{\alpha}Glc)$, producing green colonies. E. sakazakii strains produced green colonies on both DFI and mDFI agar, whereas E. vulneris produced green colonies on DFI agar but small white colonies on mDFI agar. E. sakazakii and E. vulneris were also readily differentiated by colony color when the mixed culture of the two strains was plated on mDFI agar and incubated for 24 h at $37^{\circ}C$. The results indicate that the selectivity of the commercial chromogenic agar medium could be improved by a simple supplementation with glucose.

번데기동충하초 균사 중의 Branched-Chain Amino Acid Aminotransferase의 분리정제 및 그 특성에 관한 연구 (A Study on the Purification and Characteristics of Branched-Chain Amino Acid Aminotransferase in Cultural Mycelia of Cordyceps militaris)

  • 김성태;박정오
    • 대한임상검사과학회지
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    • 제37권2호
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    • pp.78-83
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    • 2005
  • The optimum conditions of Cordyceps militaris mycelial growth, purification and characteristics of branched-chain amino acid aminotransferase [BCAT(EC 2.6.1.42)] in this mycelium were studied. Optimum pH, temperature and medium of culture of mycelia were 5.5, $22.5^{\circ}C$ and Hamada medium (HM), respectively. BCAT in homogenate of this mycelia was precipitated by 20-40% saturated solution of ammonium sulfate and then purified by DEAE (diethylaminoethyl)-Sephadex A-50 column chromatography with linear concentration gradient and Sephadex G-200 gel filtration. A single band of purified enzyme was detected on SDS-PAGE (sodium dodecylsulfate-polyacrylamide gel electrophoresis). Optimum pH and temperature of BCAT were found to be 7.8 and $29^{\circ}C$, respectively. It showed activity toward L-leucine, L-isoleucine and L-valine as a substrate. The Km values of this enzyme for L-leucine were determined to be 5.88 mM for L-leucine.

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Leuconostoc mesenteroides KCTC3505를 이용한 발효자몽 추출물 생산 조건의 최적화 (Optimum Conditions for Production of Fermented Grapefruit Extract using Leuconostoc mesenteroides KCTC3505)

  • 홍경표
    • 동아시아식생활학회지
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    • 제21권5호
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    • pp.661-668
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    • 2011
  • This study was conducted in order to determine the optimum conditions for the production of fermented grapefruit extract showing high cell growth, antioxidant activity and total flavonoid content. Five lactic acid bacterial strains - Lactobacillus plantarum KCTC3104, Lactobacillus brevis KCTC3102, Weisella cibaria KCTC3746, Leuconostoc citreum KCTC3526 and Leuconostoc mesenteroides KCTC3505 - were evaluated first in order to determine the optimum strain able to grow with high efficiency on grapefruit as a substrate and possesses higher antioxidant activity and flavonoids content. Among these strains, L. mesenteroides KCTC3505 was selected as a starter culture. To estimate the available or effective content of grapefruit in basal medium, the effects of 30%, 50%, and 70% grapefruit contents on the performance of fermentation were tested, and it was found that grapefruit can be added at 70% levels to medium. In this study, three factors of fermentation conditions - incubation time, sucrose, and glucose contents - were evaluated for their effects on fermentation performance. Taguchi experiment design was employed and the responses of experiments were calculated using signal and noise ratio calculation with larger-the-best characteristics. Finally, the optimum conditions for the manufacture of fermented grapefruit extract were as follows: grapefruit 70%, sucrose 10 g/L, glucose 10 g/L, sodium acetate 1 g/L, NaCl 1 g/L, dipotassium phosphate 0.1 g/L, magnesium sulfate 0.01 g/L and 16 hr of incubation.

Enzymatic Conjugation of RGD Peptides on the Surface of Fibroin Microspheres

  • Jeon, Hyun Sang;Lee, Jin Sil;Hur, Won
    • 공업화학
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    • 제31권1호
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    • pp.67-72
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    • 2020
  • Biomaterials are frequently functionalized with Arg-Gly-Asp (RGD) peptides to provide cell adhesion sites. In this study, RGD peptides were enzymatically coupled on to the surface of fibroin microspheres. Papain exhibited a strong preference for dansyl phenylalanine for the peptide formation with fibroin microspheres. Thus, RGD1 peptide was designed to carry cysteine to both sides of the sequence, glycine as a spacer and two residues of phenylalanine at the C-terminal (CRGDCGFF). The enzymatic modification facilitated by an increasing amount of substrate and by the presence of organic solvent, dimethylsulfoxide at 25% (v/v). Microspheres coupled with RGD1, showed a significantly different precipitation property and an increased apparent volume, possibly due to the steric hindrance of RGD peptides on the surface. Transmission electron microscopy also confirmed the presence of cysteine residues in RGD1 coupled on the surface of microspheres stained with gold nanoparticles. RGD1-microspheres significantly facilitated the growth of murine fibroblast 3T3 cells even under non-adhesion culture conditions.

Plasma Surface Modification of Patterned Polyurethane Acrylate (PUA) Film for Biomedical Applications

  • Yun, Young-Shik;Kang, Eun-Hye;Yun, In-Sik;Kim, Yong-Oock;Yeo, Jong-Souk
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2015년도 제49회 하계 정기학술대회 초록집
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    • pp.223.2-223.2
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    • 2015
  • Polyurethane acrylate (PUA) has been introduced to utilize as a mold material for sub-100 nm lithography as it provides advantages of stiffness for nanostructure formation, short curing time, flexibility for large area replication and transparency for relevant biomedical applications. Due to the ability to fabricate nanostructures on PUA, there have been many efforts to mimic extracellular matrix (ECM) using PUA especially in a field of tissue engineering. It has been demonstrated that PUA is useful for investigating the nanoscale-topographical effects on cell behavior in vitro such as cell attachment, spreading on a substrate, proliferation, and stem cell fate with various types of nanostructures. In this study, we have conducted surface modification of PUA films with micro/nanostructures on their surfaces using plasma treatment. In general, it is widely known that the plasma treated surface increases cell attachment as well as adsorption of ECM materials such as fibronectin, collagen and gelatin. Effect of plasma treatment on PUA especially with surface of micro/nanostructures needs to be understood further for its biomedical applications. We have evaluated the modified PUA film as a culture platform using adipose derived stem cells. Then, the behavior of stem cells and the level of adsorbed protein have been analyzed.

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Pseudomonas aeruginosa BYK-2에 의한 생물유화제에 발효생산

  • 김학주;이경미;정혜성;김봉조;강양순;공재열
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 춘계학술발표대회
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    • pp.263-266
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    • 2000
  • The purified biosurfactant $3.16g/{\ell}$ was obtained after cultivation for 104hr at $25^{\circ}C$ with an optimal agitation speed of 200rpm, an aeration rate of 2vvm in a $14{\ell}$ fermenter containing $5.5{\ell}$ of LB medium and 1%(w/v) olive oil as a carbon source. For the kinetic studies, the optimal substrate concentration was analyzed on different olive oil concentrations(0.1, 0.5, 1.0, 1.5, 2.0%(w/v)) and optimal culture conditions(MLBM, 200rpm, 2vvm at $25^{\circ}C$) in a $14{\ell}$ jar fermenter. The results obtained indicate that $K_s$=0.0086 $g/{\ell}$, $q_s$= 0.664 $g/g{\cdot}h$, $q_p$= $4.2{\times}10^{-3}$ $g/g{\cdot}h$, and ${\mu}_{max}$ was determined as $0.1449h^{-1}$.

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Enzymatic saccarification of cellulosic wastes by pectinase

  • Lee, Ji-Eun;Kim, Sam-Gon;Kim, Seong-Jun
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
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    • pp.512-516
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    • 2003
  • 토양에서 분리된 pectinase 생산균 strain KL34를 이용하여 폐섬유소의 효소학적 당화에 관한 연구를 수행하였다. 섬유소폐기물을 이용한 효소생산에서 폐야채보다는 과일껍질을 기질로 사용했을 때 높은 효소활성을 보여주었으며, 과일껍질 1.0%(w/v)를 기질로 사용했을 때 3.8U/ml의 pectinase활성을 얻을 수 있었다. 폐야채와 과일껍질을 기질로 사용하고, 5U/ml의 pectinase 활성을 가지는 배양 상등액을 사용하여 효소학적 당화를 수행한 결과 사과껍질 10%를 기질로 사용했을 때 7.1%의 당화율을 얻을 수 있었으며, 이때 생산된 환원당은 9.5g/L였다. 또한 생산된 효소액을 음식물 쓰레기의 당화에 적용하여 pectinase의 사용 가능성을 검토한 결과 KL34의 배양 상등액과 cellulase를 함께 사용한 당화액에서 12.7g/L의 환원당을 얻어, cellulase만을 사용했을 때 보다 1.6배의 당화효율을 얻을 수 있었다.

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Selection and Characterization of Pseudomonas aeruginosa EMS1 Mutant strain Showing Enhanced Biosurfactant Production

  • Cha, Mi-Sun;Lee, Kuen-Hee;Lee, Na-Eun;Lee, Sang-Joon
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
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    • pp.434-437
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    • 2003
  • A new bacterial strain, was isolated from activated sludge, identified and named P. aeruginosa EMS1. The new strain produced surface-active rhamnolipids by batch cultivation in mineral salts medium with waste flying oils. The mutant strain KH7, designated P. aeruginosa EMS1, derived by random mutagenesis with N-methyl-N-nitro-N-nitrosogoanidine treatment producing high levels of the biosurfactants was selected by an ion-pair plate assay. The mutant strain KH7 showed 4-5 times more hydrocarbon emulsification as compared to the parent when grown on waste frying oils and various hydrocarbons. Furthermore, P. aeruginosa EMS1 and mutant strain KH7 was also able to use whey as a co-substrate for growth and biosurfactant production. As results of this study, mutant strain KH7 is a very efficient biosurfactant producer, and its culture conditions are relatively inexpensive and economical. Rhamnolipid is synthesized by the rhlAB-encoded rhamnosyltransferase. To be convinced of these genes, we performed PCR based on P. aeruginosa PAO1 whole-genome database. rhl gene cluster nucleotide and amino acid sequences were compared for both parent and mutant. Comparison of nucleotide sequence of rhlAB, there were usually terminal's codons exchange.

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