• Title/Summary/Keyword: recombinant fermentation

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Transcription Analysis of Recombinant Trichoderma reesei HJ-48 to Compare the Molecular Basis for Fermentation of Glucose and Xylose

  • Huang, Jun;Lin, Mei;Liang, Shijie;Qin, Qiurong;Liao, Siming;Lu, Bo;Wang, Qingyan
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1467-1479
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    • 2020
  • Profiling the transcriptome changes involved in xylose metabolism by the fungus Trichoderma reesei allows for the identification of potential targets for ethanol production processing. In the present study, the transcriptome of T. reesei HJ-48 grown on xylose versus glucose was analyzed using next-generation sequencing technology. During xylose fermentation, numerous genes related to central metabolic pathways, including xylose reductase (XR) and xylitol dehydrogenase (XDH), were expressed at higher levels in T. reesei HJ-48. Notably, growth on xylose did not fully repress the genes encoding enzymes of the tricarboxylic acid and respiratory pathways. In addition, increased expression of several sugar transporters was observed during xylose fermentation. This study provides a valuable dataset for further investigation of xylose fermentation and provides a deeper insight into the various genes involved in this process.

Prospects for Recombinant Protein Production in Dairy Cattle (유우로부터 재조합단백질 생산에 대한 전망)

  • Bremel, Robert D.
    • Korean Journal of Animal Reproduction
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    • v.20 no.4
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    • pp.365-370
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    • 1997
  • Historical programs for genetic improvement of dairy cattle are discussed in the context of new genetic technologies resulting in a hetero zygous gain of function. Concepts are outlined pointing to the importance of breaking the well established genetic correlation between fat content and protein content of milk to provide flexibility in the dairy industry. The concept of value added genetics is introduced and the econ omic mpetitiveness of the mammary glands of livestock are considered in relationship to mammalian cell culture bacterial fermentation technology.

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Improved Technologies to Produce Heterologous Proteins in Recombinant Escherichia coli. (재조합 대장균에서 외래단백질 발현을 위한 기술개발)

  • 박용철;권대혁;이대희;서진호
    • KSBB Journal
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    • v.16 no.1
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    • pp.1-10
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    • 2001
  • Escherichia coli has been used as an expression work horse for foreign genes. This article summarized recent development in genetic engineering techniques for overproduction of medical proteins and industrial enzymes. Special emphasis was placed upon research activities concerning folding and refolding of inclusion bodies at genetic and fermentation levels. Plasmid and mRNA stabilization, development of strong inducible promoters, modification of translational elements and reduction of rpoteolytic degradation were carried out to elevate an expression level of a target protein. Optimization of culture conditions, improvement of denaturation and renaturation steps and coexpression of molecular chaperones or foldase were accomplished to produce active proteins in soluble form. Fusion protein systems with selective separation and surface display technology were also performed in an effort to make the E. coli expression system more effective and versatile.

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PCA를 이용한 유전자 재조합 대장균의 ALA 생산공정의 해석

  • Gang, Tae-Hyeong;Jeong, Sang-Yun;Im, Yong-Sik;Kim, Chun-Gwang;Jeong, Sang-Uk;Lee, Jong-Il
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.157-160
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    • 2003
  • ALA is an intermediate in the tetrapyrrole biosynthesis pathway and has extensive applications as a biodegradable herbicide and insecticide as well as medical applications including photodynamic therapy of cancers. For the development of mass production process of ALA it is necessary to on-line monitor some metabolites such as glycine, succinate, LA and ALA. In this study, medium compositions and fermentation conditions were investigated for enhancement of ALA production by recombinant E. coli. A 2-dimensional fluorescence sensor was employed to monitor the bioprocess of ALA production. The monitored data is analyzed using principal component analysis, a powerful tool for multivariate statistical analysis.

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On-Line Induction of Fermentation with recombinant cells: Optimization and Data Acquision

  • 이철균;최차용
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.514.3-515
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    • 1986
  • λP$_{L}$ promoter와 Infiuenza virus의 NS1 Structural gene이 있는 pASl EH801 plasmid를 E. coli host N5' 과 AR120에 각각 transformation하여 온도와 nalidixic acid로 각각 induction 하여 보았다 N5151의 경우, O.D.600 1.2에서 42$^{\circ}C$로 induction하였을 때 maximum productivity를 보였으며 AR120의 경우는 O. D. 600 1.2, 37$^{\circ}C$, 40$\mu\textrm{g}$ nalidixic acid/$m\ell$ induction 하였을 때 maximum yield를 보여주었다. 이때 pH, DO, temperature, $O_2$%, $CO_2$%를 A/D converter통해 computer에 연결시켜 data acquision을 한 결과, 접종 후 ON-line induction이 가능함을 알 수 있었다.

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Production System for Biodegradable Polyester Polyhydroxybutyrate by Corynebacterium glutamicum

  • Jo, Sung-Jin;Ooi, Toshihiko;Taguchi, Seiichi
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.352-352
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    • 2006
  • Corynebacterium glutamicum, which is well known as an amino acid fermentation bacterium, has been used as a producer of poly(3-hydroxybutyrate) [P(3HB)]. P(3HB) was synthesized in recombinant C. glutamicum harboring the expression plasmid vector with a strong promoter for cell surface protein gene derived from C. glutamicum and P(3HB) biosynthetic gene operon derived from Ralstonia eutropha. The expression of P(3HB) synthase gene was detected by enzyme activity assay. Intracellular P(3HB) was microscopically observed as inclusion granules and its content was calculated to be 22.5 % (w/w) with molecular weight of $2.1{\times}10^{5}$ and polydispersity of 1.63.

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Purification of Single Chain Human Insulin Precursors Using Various Fusion Proteins

  • Park, Seon-Ho;Jo, Jeong-U;Nam, Du-Hyeon
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.619-622
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    • 2000
  • For the production of $B^{30}-homoserine$ human insulin precursor, four types of fusion peptides LacZ, MBP, GST, and His-tagged sequence were studied in this work. Recombinant E. coli JM 103 and E. coli JM 109 containing fusion peptides were cultivated at $37^{\circ}C$ for 1hr, and gene expression was occurred when 0.5mM of isopropyl-D-thiogalactoside(IPTG) was added to the culture broth, and followed by longer than 4hr fermentation respectively. DEAE-Sphacel and gel filtration chromatography, amylose and glutathione-Sepharose 4B affinity chromatography, and nickel-affinity chromatography system were employed as purification of $B^{30}-homoserine$ human insulin precursor. Recovery yields of His-tagged, LacZ, GST, and MBP fused $B^{30}-homoserine$ human insulin precursor resulted in 47%, 20%, 20%, and 18%, respectively.

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Optimization and Adaptive Control for Fed-Batch Culture of Yeast (효모 배양을 위한 발효공정의 최적화 및 적응제어)

  • 백승윤;유영제이광순
    • KSBB Journal
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    • v.6 no.1
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    • pp.15-25
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    • 1991
  • The optimal glucose concentration for the high-density culture of recombinant yeasts was obtained using dynamic simulation. An adaptive and predictive algoritilm complimented by the rule base was proposed for the control of the fed-batch fermentation process. The measurement of process variables has relatively long sampling period and relatively long time delay characteristics. As one of the solution on these problems, prediction techniques and rule bases were added to a classical recursive identification and control algorithm. Rule bases were used in the determination of control input considering the difference between the predicted value and the measured value. A mathelnatical model was used in the estimation and interpretation of the changes of state variables and parameters. Better performances were obtained by employing the control algorithm proposed in the present study compared to the conventional adaptive control method.

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Influence of Gluconeogenic Phosphoenolpyruvate Carboxykinase (PCK) Expression on Succinic Acid Fermentation in Escherichia coli Under High Bicarbonate Condition

  • Kwon Yeong-Deok;Lee Sang-Yup;Kim Pil
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1448-1452
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    • 2006
  • The effects of amplifying the gluconeogenic phosphoenolpyruvate carboxykinase of Escherichia coli ($pck_{Ec}$) on succinic acid production in E. coli were examined under anaerobic condition. No significant increase in succinic acid production was observed in E. coli overexpressing the $pck_{Ec}$ gene without supplementing $NaHCO_{3}$ or $MgCO_{3}$. On the other hand, succinic acid production was enhanced as the $NaHCO_{3}$ concentration was increased. When 20 g/l of $NaHCO_{3}$ was added, succinic acid production in recombinant E. coli overexpressing PCK was 2.2-fold higher than that observed in the wild-type strain. It was concluded that the gluconeogenic $pck_{Ec}$ overexpression enabled E. coli to enhance succinic acid production only under the high bicarbonate supplementation condition.

Optimizing the Production of 5-Aminolevulinic Acid by Recombinant Escherichia coli Containing the Rhodobacter capsulatus hemA Gene (Rhodobacter capsulatus hemA 유전자 발현 대장균에 의한 5-Aminolevulinic Acid 생산의 최적화)

  • Yang, Dong-Soo;Park, Moon-Won;Lim, Soo-Jin;Kim, Min-Jeong;Shin, Yu-Ri;Park, Chan-Soo;Hyun, Young;Kang, Dae-Kyung
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.153-159
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    • 2009
  • Recombinant Escherichia coli BLR(DE3) harboring the hemA gene from Rhodobacter capsulatus under the control of a constitutive promoter, which we constructed previously, was used for the extracellular production of 5-aminolevulinic acid (ALA). The effects of several factors on ALA production were investigated in flask culture. ALA production by the recombinant E. coli was more efficient at $30^{\circ}C$ than $37^{\circ}C$. The glycine concentration had an important effect on cell growth. Glycine and succinic acid concentration of 5-10 and 10-20 g/L, respectively, resulted in high ALA production. In addition, the partial replacement of succinic acid by sodium glutamate increased the ALA production. The ALA production was inhibited by the presence of glucose in the medium. Using the optimal conditions, an ALA concentration of 8.2 g/L was achieved in jar fermentation without an added inducer or ALA dehydratase inhibitor; this is the highest reported concentration.