1999 Fiscal Year Annual Research Report

櫛状ポリエチレングリコール誘導体による酵素の安定化

Research Project

Project/Area Number	10680806
Research Institution	UNIVERSITY OF YOKOHAMA
Principal Investigator	廣戸三佐雄桐蔭横浜大学, 工学部, 助教授 (00110172)
Co-Investigator(Kenkyū-buntansha)	二見瑞子桐蔭横浜大学, 工学部, 助教授 (30209142) 西村裕之桐蔭横浜大学, 工学部, 教授 (60189313) 稲田祐二桐蔭横浜大学, 工学部, 教授 (40016035) 小寺洋桐蔭横浜大学, 工学部, 講師 (80205426)
Keywords	ポリエチレングリコール / 化学修飾 / 安定化 / β-フルクトフラノシダーゼ / フルクトオリゴ糖 / 耐熱性 / pH依存症
Research Abstract	β-Fructofuranosidase(以下β-Fructofuranosidase(+CHO)と表す)を37℃のクエン酸ナトリウム緩衝液(pH5.5)中でエンドグリコシダーゼHを48時間作用させ糖鎖を除去し(この時点の酵素をβ-Fructofuranosidase(-CHO)と表す)、リン酸緩衝液(pH8.0)で限外濾過後、修飾剤PM_<100>を加え4℃で1時間反応させた後、酢酸緩衝液(pH4.5)で透析したものをPM_<100>-β-Fructofuranosidase(-CHO)とした。他に糖鎖除去を行わずに修飾剤を作用させたPM_<100>-β-Fructofuranosidase(+CHO)を合わせ4種類の試料について、耐熱性、pH依存性を比較した。耐熱性については、糖鎖を除去しないもの(β-Fructofuranosidase(+CHO)とPM_<100>-β-Fructofuranosidase(+CHO)についてはPM_<100>修飾の有無に関わらず耐熱性に変化がみられず、ともに温度の上昇に従って同じ様な曲線を描いて失活していった。糖鎖を除去した場合、β-Fructofuranosidase(-CHO)よりPM_<100>-β-Fructofuranosidase(-CHO)のほうが、あらゆる測定ポイントで常に5%程上回るという若干の効果が得られた。 pH依存症はPM_<100>-β-Fructofuranosidase(-CHO)だけがその活性提示範囲を拡げ、PM_<100>修飾効果を上げた他の試料では活性を失う範囲でもその活性を維持し他の試料の30%程のプラス効果が認められた。有機溶媒中での合成能については否定的な現状である。エタノール中での配糖体の合成反応は行われているのだが(TLCによる確認)フルクトオリゴ糖の合成は未だしの状態である。

Research Products
(8 results)

All Other

All Publications (8 results)

[Publications] Hiroto M. et. al.: "Chemical modification of lipase with a comb-shaped synthetic copolymer of polyoxyethylene allyl methyl diether and maleic anhydride."Biotech. Lett.. 14. 559-564 (1992)
[Publications] Kodera Y. et. al.: "Chemical modification of L-asparaginase with a comb-shaped copolymer of polyethylene derivative and anhydride."Biochem. Biophys. Res. Commun.. 184. 144-148 (1992)
[Publications] Sasaki H. et. al.: "Reduction of immunoreactivity of bovine serum albumin cojugated with comb-shaped polyethylene glycol derivatives."Biochem. Biophys. Res. Commun.. 197. 287-291 (1993)
[Publications] Kodera Y. et. al.: "Stabilization of L-asparaginase modified with comb-shaped polyethylene glycol derivatives, in vibo and in vitro."Bioconjugate Chemistry. 5. 283-286 (1994)
[Publications] Hiroto M. et. al.: "Stabilization of trypsin by modification with comb-shaped copolymer of poly(ethyleneglycol) derivative and maleic anhydride."Biotech. Techniques.. 9. 105-110 (1995)
[Publications] Saito T. et. al.: "Tolelogenic caoacity of poly (ethyleneglycol)(PEG)-modified ovalbumins in relation to their immunoreactivity towards anti-overlbumin antibody."J. Biomaster. Sci. Polymer Edn.. 8. 311-321 (1996)
[Publications] Inada Y. et. al.: "Modificatuon of proteins with polyethyleneglycol derivatives"Methods in Enzymology. 25 (1994)
[Publications] Hiroto M. et. al.: "Advanced Biomaterials in Biomedical Engineering and Drug Delivery Systems."Stabilization of L-asparaginase by chemicalmodification with poly(ethyleneglycol) derivatives.. 2 (1996)

1999 Fiscal Year Annual Research Report

櫛状ポリエチレングリコール誘導体による酵素の安定化

Principal Investigator

廣戸 三佐雄 桐蔭横浜大学, 工学部, 助教授 (00110172)

Research Products

[Publications] Hiroto M. et. al.: "Chemical modification of lipase with a comb-shaped synthetic copolymer of polyoxyethylene allyl methyl diether and maleic anhydride."Biotech. Lett.. 14. 559-564 (1992)

[Publications] Kodera Y. et. al.: "Chemical modification of L-asparaginase with a comb-shaped copolymer of polyethylene derivative and anhydride."Biochem. Biophys. Res. Commun.. 184. 144-148 (1992)

[Publications] Sasaki H. et. al.: "Reduction of immunoreactivity of bovine serum albumin cojugated with comb-shaped polyethylene glycol derivatives."Biochem. Biophys. Res. Commun.. 197. 287-291 (1993)

[Publications] Kodera Y. et. al.: "Stabilization of L-asparaginase modified with comb-shaped polyethylene glycol derivatives, in vibo and in vitro."Bioconjugate Chemistry. 5. 283-286 (1994)

[Publications] Hiroto M. et. al.: "Stabilization of trypsin by modification with comb-shaped copolymer of poly(ethyleneglycol) derivative and maleic anhydride."Biotech. Techniques.. 9. 105-110 (1995)

[Publications] Saito T. et. al.: "Tolelogenic caoacity of poly (ethyleneglycol)(PEG)-modified ovalbumins in relation to their immunoreactivity towards anti-overlbumin antibody."J. Biomaster. Sci. Polymer Edn.. 8. 311-321 (1996)

[Publications] Inada Y. et. al.: "Modificatuon of proteins with polyethyleneglycol derivatives"Methods in Enzymology. 25 (1994)

[Publications] Hiroto M. et. al.: "Advanced Biomaterials in Biomedical Engineering and Drug Delivery Systems."Stabilization of L-asparaginase by chemicalmodification with poly(ethyleneglycol) derivatives.. 2 (1996)

廣戸三佐雄桐蔭横浜大学, 工学部, 助教授 (00110172)