Purification and Partial Characterization of an X-prolyl-dipeptidyl Aminopeptidase from <i>Lactobacillus gasseri</i> ME-284

• Miyabe Ryosuke

  Graduate School of Bioresource Sciences, Nihon University

• Takahashi Yuto

  Graduate School of Bioresource Sciences, Nihon University

• Matsufuji Hiroshi

  Graduate School of Bioresource Sciences, Nihon University

• Ogihara Jun

  Graduate School of Bioresource Sciences, Nihon University

• Itou Kentarou

  Graduate School of Bioresource Sciences, Nihon University Division of Research and Development, Meiji Co.

• Kawai Yasushi

  Graduate School of Bioresource Sciences, Nihon University

• Masuda Tetsuya

  Graduate School of Bioresource Sciences, Nihon University

• Suzuki Koichi

  Graduate School of Bioresource Sciences, Nihon University

• Oda Munehiro

  Graduate School of Bioresource Sciences, Nihon University

抄録

An X-prolyl-dipeptidyl aminopeptidase has been purified from Lactobacillus gasseri by lysozyme treatment and seven chromatographic steps. The yield was 1.4% and the specific activity increased 265-fold over the crude enzyme extract. SDS-PAGE provided a single band with a MW of approximately 82 kDa. Gel filtration chromatography showed that the native enzyme was approximately 173 kDa, suggesting that the protein is a dimer. The amino acid sequences of the peptide fragments obtained after tryptic treatment of the purified enzyme were determined and found to be consistent with that of an X-prolyl-dipeptidyl aminopeptidase from L. gasseri ATCC 33323 (LGAS_0712, coverage: 31.74%). Optimal activity was observed at pH 7.0 and 55°C. The activity was inhibited by diisopropylfluorophosphate and phenylmethylsulfonylfluoride, and by divalent cations (Cu²⁺, Hg²⁺, and Zn²⁺). From these inhibition characteristics, the enzyme is likely a serine proteinase. Beta-casomorphin 7 (BCM-7, Tyr-Pro-Phe-Pro-Gly-Pro-Ile) was hydrolyzed into X-proline fragments from the N-terminus of the peptide. The characteristics of the enzyme are very similar to those of the enzyme from Lactobacillus sakei. However, as free amino acids were detected among the degradation products of BCM-7, some aspects of substrate specificity require further clarification.

収録刊行物

• Food Science and Technology Research

  Food Science and Technology Research 21 (3), 445-451, 2015

  公益社団法人 日本食品科学工学会

キーワード

• <i>L. gasseri</i>
• X-prolyl-dipeptidyl aminopeptidase
• purification
• beta-casomorphin

詳細情報

• CRID: 1390282679435092352
• NII論文ID: 130005098074
• DOI: 10.3136/fstr.21.445
• ISSN: 18813984
• ISSN: 13446606
• Web Site: https://www.jstage.jst.go.jp/article/fstr/21/3/21_445/_pdf
• 本文言語コード: en
• データソース種別:
  • JaLC
  • Crossref
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可