• Title/Summary/Keyword: Rhamnosyltransferase

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Biosynthesis of Rhamnosylated Anthraquinones in Escherichia coli

  • Nguyen, Trang Thi Huyen;Shin, Hee Jeong;Pandey, Ramesh Prasad;Jung, Hye Jin;Liou, Kwangkyoung;Sohng, Jae Kyung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.398-403
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    • 2020
  • Rhamnose is a naturally occurring deoxysugar present as a glycogenic component of plant and microbial natural products. A recombinant mutant Escherichia coli strain was developed by overexpressing genes involved in the TDP-ʟ-rhamnose biosynthesis pathway of different bacterial strains and Saccharothrix espanaensis rhamnosyl transferase to conjugate intrinsic cytosolic TDP-ʟ-rhamnose with anthraquinones supplemented exogenously. Among the five anthraquinones (alizarin, emodin, chrysazin, anthrarufin, and quinizarin) tested, quinizarin was biotransformed into a rhamoside derivative with the highest conversion ratio by whole cells of engineered E. coli. The quinizarin glycoside was identified by various chromatographic and spectroscopic analyses. The anti-proliferative property of the newly synthesized rhamnoside, quinizarin-4-O-α-ʟ-rhamnoside, was assayed in various cancer cells.

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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    • 2003.10a
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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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Characterization of Biosurfactant Produced by Pseudomonas fluorescens PD101 (Pseudomonos fluorescens PD101이 생산하는 생물유화제 특성)

  • YOON Hong Mook;MOON Sung Hoon;SONG Young Hwan
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.3
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    • pp.230-238
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    • 2003
  • Biosurfactant-producing bacteria, showing strong crude oil degrading activity, were isolated from the caverns of National Oil Storage Basement. From the results of biochemical and molecular biological tests, the isolate was identified as Pseudomonas fluorescens PD101. It grows well on liquid media at temperature range from $20^{\circ}C\;to\;37^{\circ}C,$ but it does not produce biosurfactant when grown at $37^{\circ}C$ or at higher temperature. The biosurfactant was stable at broad pH range from 5 to 11 and under heat treatment condition of $100^{\circ}C$ for 30 min. The biosurfactant produced dark blue halo around the colony when grown on SW agar plates, which could confirm the biosurfactant as one of rhamnolipid group. The 700 bp of PCR product could be amplified from DNA of P. flurorescens PD101 by using PCR primers designed from rh1A gene of P. aeruginosa, and it showed $99\%$ of sequence homology with rh1A gene of P. aeruginosa encoding rhamnosyltransferase 1.