• Title/Summary/Keyword: Biomolecule immobilization

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Production of dTDP-4-keto-6-deoxy-D-glucose by Immobilization of dTDP-D-glucose 4,6-dehydratase

  • Kharel, Mandan-Kumar;Liou, Kwang-Kyoung;Sohng, Jae-Kyung;Lee, Hei-Chan
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.297-301
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    • 2004
  • The dTDP-D-glucose 4,6-dehydratase from Salmonella enterica was immobilized using covalent binding to cyanogen bromide activated sepharose. The immobilized enzyme was used to produce dTDP-4-keto-6-deoxy-D-glucose, a key sugar intermediate that can be used economically to produce diverse classes of unusual sugars appended in various antibiotics. The enzyme was immobilized on the sepharose after activation with cyanogen bromide. The maximum immobilization (80.03%) was achieved after 14 h of coupling. The covalently immobilized enzyme was stable, and an average of 78.4 % conversion was achieved until 120 h of immobilization when it was repeatedly used. Similar conversion was noticed for the first batch using the enzyme entrapped-hydrogel but activity was gradually decreased in the following batches. The production of dTDP-4-keto-6-deoxy-D-glucose by using an immobilized enzyme has high potential for commercial application.

Sustainable Production of Dihydroxybenzene Glucosides Using Immobilized Amylosucrase from Deinococcus geothermalis

  • Lee, Hun Sang;Kim, Tae-Su;Parajuli, Prakash;Pandey, Ramesh Prasad;Sohng, Jae Kyung
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1447-1456
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    • 2018
  • The amylosucrase encoding gene from Deinococcus geothermalis DSM 11300 (DgAS) was codon-optimized and expressed in Escherichia coli. The enzyme was employed for biosynthesis of three different dihydroxybenzene glucosides using sucrose as the source of glucose moiety. The reaction parameters, including temperature, pH, and donor (sucrose) and acceptor substrate concentrations, were optimized to increase the production yield. This study demonstrates the highest ever reported molar yield of hydroquinone glucosides 325.6 mM (88.6 g/l), resorcinol glucosides 130.2 mM (35.4 g/l) and catechol glucosides 284.4 mM (77.4 g/l) when 400 mM hydroquinone, 200 mM resorcinol and 300 mM catechol, respectively, were used as an acceptor substrate. Furthermore, the use of commercially available amyloglucosidase at the end of the transglycosylation reaction minimized the gluco-oligosaccharides, thereby enhancing the target productivity of mono-glucosides. Moreover, the immobilized DgAS on Amicogen LKZ118 beads led to a 278.4 mM (75.8 g/l), 108.8 mM (29.6 g/l) and 211.2 mM (57.5 g/l) final concentration of mono-glycosylated product of hydroquinone, catechol and resorcinol at 35 cycles, respectively, when the same substrate concentration was used as mentioned above. The percent yield of the total glycosides of hydroquinone and catechol varied from 85% to 90% during 35 cycles of reactions in an immobilized system, however, in case of resorcinol the yield was in between 65% to 70%. The immobilized DgAS enhanced the efficiency of the glycosylation reaction and is therefore considered effective for industrial application.

A Study on the Immobilization of Biomolecules on Poly(acrylic acid)-grafted MWCNTs Prepared by Radiation-Induced Graft Polymerization (방사선 그래프트 중합에 의하여 제조된 폴리(아크릴 산)이 그래프트된 탄소나노튜브에 생체분자 고정화에 관한 연구)

  • Jung, Chan-Hee;Lee, Byoung-Min;Hwang, In-Tae;Choi, Jae-Hak;Nho, Young-Chang;Hong, Sung-Kwon
    • Polymer(Korea)
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    • v.34 no.2
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    • pp.150-153
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    • 2010
  • In this research, biomolecule-immobilized multi-walled carbon nanotubes (MWCNTs) were prepared by using radiation-induced graft polymerization. For the immobilization of biomolecules, the surface of MWNCTs was functionalized by radiation-induced graft polymerization of acrylic acid. Based on the results of TGA and Raman spectroscopy it was found that acrylic acid was effectively graft-polymerized on the MWCNTs. Biomolecules such as DNA and proteins were immobilized onto the resultant poly(acrylic acid)-grafted MWCNTs. The results of the X-ray photoelectron spectroscopy and fluorescence microscopy confirmed that the biomoelcules were successfully immobilized on the poly(acrylic acid)-grafted MWCNTs.