In vitro synthesis of high molecular weight rubber by Hevea, small rubber particles(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
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概要
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Hevea brasiliensis is one of few higher plants producing the commercial natural rubber used in many significant applications. The biosynthesis of high molecular weight rubber molecules by the higher plants has not been clarified yet. Here, the in vitro rubber biosynthesis was performed by using enzymatically active small rubber particles (SRP) from Hevea. The mechanism of the in vitro rubber synthesis was investigated by the molecular weight distribution (MWD). The highly purified SRP prepared by gel filtration and centrifugation in the presence of Triton^[○!R] X-100 showed the low isopentenyl diphosphate (IPP) incorporation for the chain extension mechanism of pre-existing rubber. The MWD of in vitro rubber elongated from the pre^-existing rubber chains in SRP was analyzed for the first time in the case of H. brasiliensis by incubating without the addition of any initiator. The rubber transferase activity of 70% incorporation of the added IPP (w/w) was obtained when farnesyl diphosphate was present as the allylic diphosphate initiator. The in vitro synthesized rubber showed a typicalbimodal MWD of high and low molecular weight fractions in GPC analysis, which was similar to that of the in vivo rubber with peaks at around 10^6 and 10^5 Da or lower. The reaction time independence and dependence of molecular weight of high and low molecular weight fractions, respectively, indicated that the high molecular weight rubber was synthesized from the chain extension of pre-existing rubber molecules whereas the lower one was from the chain elongation of rubber molecules newly synthesized from the added allylic substrates.
著者
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Rojruthai Porntip
Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ma
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Sakdapipanich Jitladda
Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ma
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Takahashi Seiji
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University
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Hyegin Lee
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University
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Noike Motoyoshi
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University
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Koyama Tanetoshi
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University
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Tanaka Yasuyuki
Institute of Molecular Bioscience, Mahidol University
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Hyegin Lee
Institute Of Multidisciplinary Research For Advanced Materials Tohoku University
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Takahashi Seiji
Department Of Biomolecular Engineering Graduate School Of Engineering Tohoku University
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Koyama Tanetoshi
Institute For Chemical Reaction Science Tohoku University
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Rojruthai Porntip
Department Of Chemistry And Center Of Excellence For Innovation In Chemistry Faculty Of Science Mahi
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Sakdapipanich Jitladda
Department Of Chemistry And Center Of Excellence For Innovation In Chemistry Faculty Of Science Mahi
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Takahashi Seiji
Institute Of Multidisciplinary Research For Advanced Materials Tohoku University
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Noike Motoyoshi
Institute Of Multidisciplinary Research For Advanced Materials Tohoku University
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Tanaka Yasuyuki
Institute Of Molecular Bioscience Mahidol University
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