• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to project page

1990 Fiscal Year Final Research Report Summary

Cellular Localization and Processing of Artificially Constructed Proteins.

Research Project

Project/Area Number 01470120
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 応用生物化学・栄養化学
Research InstitutionUniversity of Tsukuba

Principal Investigator

YAMANE Kunio  University of Tsukuba, Institute of Biological Sciences. Professor, 生物科学系, 教授 (20013336)

Project Period (FY) 1989 – 1990
KeywordsProtein processing / extracellular enzymes / fused proteins / Signal peptide / thermostable alpha-amylase / Bacillus subtilis / alkaline protease
Research Abstract

A large number of proteins and hormones are synthesized as precursors and are converted into the mature forms by selective enzymatic cleavage of peptide bonds. As extracellular alpha-amylase, neutral protease and alkaline protease in Bacillus subtilis are synthesized as prepro enzymes, their signal peptides for secretion are removed at first, and then the prosequences in the secreted proenzymes are processed to form the mature enzymes. In order to analyze the processing system of the prosequences in. subtilis, we constructed artificial hybrid genes, amyE^1 - ^1pBR322^1 - ^<>amyT.
In these hybrid genes, small peptides encoded in the fragments of pBR322 DNA were inserted between the signal peptide of B. subtilis alpha-amylase (amyE^1) and the mature thermostable alpha-amylase (^1amyT) of. stearothermophilus A631 using dlll linker DNA. Among the constructed hybrid genes, the gene in the plasmid pTUBE638 induced a 1.7-fold higher production of extracellular thermostable alpha-amylase in. Subtilis than by the parental amyE^1 - ^1amyT gene. In B. Coli (pTUBE638), an N-terminally extended thermostable alpha-amylase accumilates in the periplasm of the cells, where it retains the 21-amino-acid-extra-peptide. In contrast, another N-terminally extended thermostable alpha-amylase, in which an 18-amino-acid-extra-peptide, is secreted from B. Subtilis (pTUBE638). We purified the N-terminally extended thermostable alpha-amylase from. Coli (pTUBE638). Then we developed an in vitro processing system for the analysis of the protein processing in B. Subtilis. Then we demonstrated that the extracellular alkaline protease acts as the major processing enzyme for the selective cleavage of the ide.

  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] Nakamura. K., Fujita, T., Itoh, Y. and Yamano, K.: "Moclification of length,hydrophobic propenties and electroc change of Bacillus sultles αーamylaseaignel peptide and their different effects on the production of aecrefory proteins in B・suletilis and Eschenichia cob" Mol. Gen. Genet. 216. 1-9 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kimura, K., Kataoku, S., Nakamura, A., Takano, T., Kobayashi, S., and Yamane, K.: "Function of the COOH.terminal region of cycloclextrin ghrcano trausfenase of alkalophilor Bauillus sp. #104 : Relation to cotalizing acticity and pH stability" Biochem. Biophys. Res, Commun.161. 1273-1279 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sasamoto, H., Nakasawa, K., Tsutsumi, K., Tahase, K., and Yamane, K.: "Signal pepticle of Bacillus subtilis αーamylase" J. Biochem.106. 376-382 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Itoh, Y., Kanoh, K., Nakamura, K., Takasa, K., and Yamada, K.: "Artificial insertion of peptides between signalpeptide and mcture protein : effect on secretion and proassing of hyprid therid thenmostable αーamylases in Bacillus subtilis and Escherichia cob ulls" J. Gen. Micrbiol. 136. 1551-1558 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Itoh, Y., Sumi, Nakamura, K., and Yamane, K.: "processing of an NH_2 terminally extended thernoshable αーamylase by Baciltuo subtilis protease" J. Biochem.108. 954-959 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sadaie, Y., Takamatsu, H., Nakamura, K., ans Yamaue, K.: "Seuercing reoeals Somilarity of the wildーtype diot gene of Bacillus subtilis to the Escherichia coli seeA gere" Gene. (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamane, K.: "Bacilluo subtilin : Molecalar Biology and industrial application(ed. Maruo, B., and Yoshikawa, H.)Extraulluler engyass of B. subfilis" Elsevler Scionce Pulishens., 162-190 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yanane, K., Itah, Y., Nakamura, K., Tsutsumi, K., and Takase, K.: "Genet. s and Biotechnology of Bacilli(ed. J. A. Hoch)Signal poptide cleauage and proussing of artificial extracellala hybrid αーamylaces in Bacillus subtilis" Academic Press Inc.,

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakamura, K., Fujita, K., Itoh, Y., and Yamane, K. :"Modification of length, hydrophobic properties, and electric chage of acillus subtilis alpha-amylase signal peptide and their different effects on the production of secretoty proteins in B. subtilis and Escherichia coli." Mol. Gen. Genet.216 ;. 1-9 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kimura, K., Kataoka, S., Nakamura, A., Takano, T., Kobayashi, S., and Yamane, K.: "Function of the COOH-terminal region of cyclodextrin glucanotransferase of alkalophilic Bacillus sp. #1011 : relation to catalyzing actevity a-d pH stability" Biochem. Biophys. Res. Commun.161 ;. 1273-1279 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sasamoto, H., Nakazawa, K., Tsutsumi, K., Takase, K., and Tamane K.: "Signal peptide of Bacillus subtilis alpha-amylade." J. Biochem.,. 106 ;. 376-382 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakamura, J., Yamane, K., Yoda, K., and Yamasaki, M.: "Amplification of an Integration plasmid into the tmr amplification unit of a Bacillus subtilis tunicamycin-resistant mutant." Agric. Biol. Chem.53 ;. 19-24 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamoto, J., Shimizu, M., and Yamane, K. :"Nucleotide sequence of the diaminopimelate-de carboxylase gene from Bacillus subtilis" Nucleic Acids Res.17 ;. 10105 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamane, K.: "Extracellular enzymes of B. subtilis." Bacillus subtilis : Molecular biology and industrial application. ed. by Maruo, B., and Yoshikawa, H.162-189 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Itoh, Y., Kanoh, K., Nakamura, K., Takase, K., and Yamane, K.: "Artificial insertion of peptides between signal pepetide and mature protein : effect on secretion and processing of hybrid thermostable alpha-amylases in Bacillus subtilis and Escherichia coli cells." J. Gen. Microbiol.,. 136 ;. 1551-1558 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Itoh, Y., Sumi, M., Nakamura, K., and Yamane, K.: "Processing of an NH_2-terminally extended thermostable alpha-amylase by Bacilus subtilis alkaline protease." J. Biochim.,. 108 ;. 954-959 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kimura, K., Ishii, Y., Kataoka, S., Takano, T., and Yamane, K.: "Expression of the beta-cyclodextrin glucanotransferase gene of an alkalophilic Bacillus sp. #1011 in Escherichia coli cells and characterization of the synthesized enzyme." Agric. Biol. Chem.54. 641-648 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakamura, K., Takamatsu, H., Nakamura, A., Yashikawa, H., and Yamane, K.: "Cloning and characterization of a Bacillus subtilis gene homologous to E. coli secY." J. Biochem.107 ;. 603-607 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakamura, K., Takamatsu, H., Akiyama, Y., Ito, K., and Yamane, K.: "Complementation of the protein transport defect of an Escherichia coli secY mutant (secY24) by Bacillus subtilis secY homologue." FEBS Letters. 273 ;. 75-78 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sadaie, Y., Takamatsu, H., Nakamura, K., and Yamane, K.: "Sequencing reveals similarity of the wild type div gene of Bacillus subtilis to the Escherichia coli secA gene." Gene.

    • Description
      「研究成果報告書概要(欧文)」より

URL: 

Published: 1993-08-12  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi