Purification and characterization of a serine protease from the fruit of Ficus carica cultivar Masui Dauphine

被引：0

作者：

Tokonami, Mizuki ^{[1
]}

Horiguchi, Akari ^{[1
]}

Ueshima, Naoki ^{[1
]}

Takita, Teisuke ^{[1
]}

Takahashi, Toru ^{[2
]}

Nishimura, Kosaku ^{[2
]}

Yasukawa, Kiyoshi ^{[1
]}

机构：

[1] Kyoto Univ, Grad Sch Agr, Div Food Sci & Biotechnol, Sakyo Ku, Kyoto, Japan

[2] Toyo Inst Food Technol, 4-23-2 Minami Hanayashiki, Kawanishi, Hyogo, Japan

来源：

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 2023年 / 87卷 / 05期

基金：

日本学术振兴会;

关键词：

Ficus carica; serine protease; subtilisin-like protease; SUBTILASE; CUCUMISIN; SARCOCARP; FAMILIES; FICIN;

D O I：

10.1093/bbb/zbad028

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Ficus carica produces, in addition to the cysteine protease ficin, a serine protease (FSP). Here, we purified FSP to homogeneity from the fruit of F. carica cultivar Masui Dauphine. An 81-fold enrichment in specific activity of FSP with 2.1% recovery was attained. Three protein bands (70, 62, and 60 kDa) were identified on SDS-PAGE. Each band was identified as a subtilisin-like protease (661 amino acids) by trypsin digestion, LC-MS/MS analysis, and the partial N-terminal amino acid sequence analysis. Gelatin zymography revealed that the active FSP exists as a dimer. The optimum hydrolysis pH of FSP was 7.5, and the pHs at which the enzyme retained its initial activity by 70% in 24 h were 8.0-11.0. The optimum hydrolysis temperature of FSP was 50-60 degrees C, and the temperature required to reduce the initial activity by 50% in 15 min was 70 degrees C. These results will inform the industrial use of FSP.

引用

页码：532 / 540

页数：9

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