Protein Engineering of Novel Proteinase Inhibitors and Their Effects on the Growth of Spodoptera exigua Larvae(Biochemistry & Molecular Biology)

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概要

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• Novel types of proteinase inhibitors with multi-inhibitory activities were generated by replacement of phytocystatin domains in sunflower multi-cystatin (SMC) by the serine proteinase inhibitor BGIT from bitter gourd seeds. Two chimeric inhibitors SMC-T3 and SMC-T23, in which the third domain in SMC and the second and third domains in SMC were replaced by BGIT, acquired trypsin inhibitory activity (Ki : 1.46×10^<-7>м and 1.75×10^<-7>м), retaining inhibitory activity toward papain (Ki : 4.5×10^<-8>м and 1.52×10^<-7>м), respectively. We compared the chimeric inhibitors and the recombinant SMC (r-SMC) in relation to their effects on the growth of larval Spodoptera exigua. When the second instar larvae were reared on a diet containing rSMC, SMC-T3, or SMC-T23 for ten days, a significant reduction in weight gain was observed. Mean weights for rSMC, SMC-T3, and SMC-T23 were 43 mg, 32 mg, and 43 mg, respectively, as compared with that (60 mg) for the absence of the inhibitor. In contrast, BGIT had little effect on the growth of the S. exigua larvae. This result indicated that the chimeric inhibitor SMC-T3 with two phytocystatin domains and one serine proteinase inhibitor domain is an efficient inhibitor of proteinases in the S. exigua larvae. Therefore, this novel type of proteinase inhibitor with multi-inhibitory activities may represent a promising protein for successful application to a transgenic plant with insect resistance.

• 社団法人日本農芸化学会の論文
• 2001-10-23

著者

• KIMURA Makoto

  Laboratory of Structural Biology, Graduate School of Systems Life Sciences, Kyushu University

• KOUZUMA Yoshiaki

  Laboratory of Biochemistry, Faculty of Agriculture, Graduate School, Kyushu University

• Kimura M

  Laboratory Of Biochemistry Faculty Of Agriculture Graduate School Kyushu University

• INANAGA Hideko

  Laboratory of Biochemisty, Graduate School, Kyushu University

• KOBAYASI Daiki

  Laboratory of Biochemisty, Graduate School, Kyushu University

• AOKI-YASUNAGA Chisa

  Institute of Biological Control, Faculty of Agriculture, Graduate School, Kyushu University

• IIYAMA Kazuhiro

  Institute of Biological Control, Faculty of Agriculture, Graduate School, Kyushu University

• Inanaga Hideko

  Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat

• Aoki-yasunaga Chisa

  Institute Of Biological Control Faculty Of Agriculture Graduate School Kyushu University

• Kouzuma Yoshiaki

  Laboratory Of Food Molecular Functionality College Of Agriculture Ibaraki University

• Kouzuma Yoshiaki

  Laboratory Of Food Functionality School Of Agriculture Ibaraki University

• Kawabe Masato

  United Graduate School Of Agricultural Science Tokyo University Of Agriculture And Technology (tuat)

• Iiyama Kazuhiro

  Institute Of Biological Control Faculty Of Agriculture Graduate School Kyushu University

• Kobayasi Daiki

  Laboratory Of Biochemisty Graduate School Kyushu University

• Kimura Makoto

  Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Graduate School Faculty Of Agr

• Kimura Makoto

  Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat

• Kimura Makoto

  Laboratory Of Biochemistry Department Of Agricultural Chemistry Faculty And Graduate School Of Agric

• KOUZUMA Yoshiaki

  Laboratory of Biochemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Graduate School, Kyushu University

• KIMURA Makoto

  Laboratory for Remediation Research, Plant Science Center, RIKEN

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