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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제28권 제9호
- 발행연도
- 2018.1
- 수록면
- 1,482 - 1,492 (11page)
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초록· 키워드
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Fusarium oxysporum trypsin (FOT) is a fungal serine protease similar to mammal trypsin. The FOT could be successfully expressed in Pichiapastoris by engineering the natural propeptide APQEIPN. In this study, we constructed two recombinant enzymes with engineered amino acid sequences added to the N-terminus of FOT and expressed in P. pastoris. The N-terminal residues had various effects on the structural and functional properties of trypsin. The FOT, and the recombinants TE (with peptide YVEF) and TS (with peptide YV) displayed the same optimum temperature (40°C) and pH (8.0). However, the combinants TE and TS showed significantly increased thermal stability at 40°C and 50°C. Moreover, the combinants TE and TS also showed enhanced tolerance of alkaline pH conditions. Compared with those of wildtype FOT, the intramolecular hydrogen bonds and the cation π-interactions of the recombinants TE and TS were significantly increased. The recombinants TE and TS also had significantly increased catalytic efficiencies (referring to the specificity constant, kcat/Km), 1.75- fold and 1.23-fold than wild-type FOT. In silico modeling analysis uncovered that the introduction of the peptides YVEF and YV resulted in shorter distances between the substrate binding pocket (D174, G198, and G208) and catalytic triad (His42, Asp102, and Ser180), which would improve the electron transfer rate and catalytic efficiency. In addition, N-terminal residues modification described here may be a useful approach for improving the catalytic efficiencies and characteristics of other target enzymes.
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참고문헌 (34)
참고문헌 신청
Walsh K / 1970 / Trypsinogens and trypsins of various species / Methods Enzymol. 19:41~63
Ling Z / 2012 / Functional expression of trypsin from Streptomyces griseus by Pichia pastoris / J. Ind. Microbiol. Biotechnol. 39:1651~1662
Muhlia-Almazan A / 2008 / Invertebrate trypsins: a review / J. Comp. Physiol. 178:655~672
Klomklao S / 2008 / Digestive proteinases from marine organisms and their applications / Songklanakarin J. Sci. Technol. 30:37~46
Zugno LA / 1992 / The effect of trypsin on soaking of salt cured hides / J. Am. Leather Chem. Assoc. 78:207~220
Ling Z / 2012 / Functional expression of trypsin from Streptomyces griseus by Pichia pastoris / J. Ind. Microbiol. Biotechnol. 39:1651~1662
Muhlia-Almazan A / 2008 / Invertebrate trypsins: a review / J. Comp. Physiol. 178:655~672
Klomklao S / 2008 / Digestive proteinases from marine organisms and their applications / Songklanakarin J. Sci. Technol. 30:37~46
Zugno LA / 1992 / The effect of trypsin on soaking of salt cured hides / J. Am. Leather Chem. Assoc. 78:207~220
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