Novel Chymotrypsin Purified and Biochemically Characterized from Digestive Organs of Bigfin Reef Squid (Sepioteuthis lessoniana)

IF 4.3 3区材料科学 Q1 ENGINEERING, ELECTRICAL & ELECTRONIC ACS Applied Electronic Materials Pub Date : 2024-05-14 DOI:10.3390/resources13050067

Jirapan Satjarak, S. Klomklao, Yi Zhang, K. Thongprajukaew

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Abstract

Chymotrypsin from the digestive organs of bigfin reef squid, a major commercial squid species in Thailand, was purified to 41-fold with 5.7% yield by a sequential purification process including ammonium sulfate precipitation, size exclusion, and ion exchange chromatography. According to SDS-PAGE, the molecular weight of purified chymotrypsin was 43 kDa. Native-PAGE analysis revealed a single band for this purified enzyme. The optimum pH and temperature for chymotrypsin activity of the purified enzyme were a pH of 7.0 with a temperature of 55 °C. The purified chymotrypsin remained stable throughout a wide range of pH levels (6–11) and at relatively high temperature (55 °C). It was significantly inhibited by PMSF and TPCK. The values of the kinetic constants Km and Kcat were found to be 1.33 mM and 31.46 s−1, respectively. The purified chymotrypsin has the N-terminal amino acid IVGGQEATPGEWPWQAALQV. This study provided new information about the biochemical properties of pure chymotrypsin from bigfin reef squid, which will be useful in the future investigation, aquaculture, and application of bigfin reef squid.

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从大鳍礁鱿鱼（Sepioteuthis lessoniana）消化器官中纯化出的新型糜蛋白酶及其生物化学特征

通过硫酸铵沉淀、尺寸排阻和离子交换色谱等连续纯化过程，从泰国一种主要商业鱿鱼品种--大鳍礁鱿鱼的消化器官中纯化了 41 倍的糜蛋白酶，产率为 5.7%。根据 SDS-PAGE 分析，纯化的糜蛋白酶分子量为 43 kDa。原生聚合酶链分析显示，纯化的糜蛋白酶只有一个条带。纯化酶的糜蛋白酶活性的最佳 pH 值和温度为 pH 值 7.0 和温度 55 ℃。纯化的糜蛋白酶在很宽的 pH 值范围（6-11）和相对较高的温度（55 °C）下都保持稳定。它受到 PMSF 和 TPCK 的明显抑制。动力学常数 Km 和 Kcat 的值分别为 1.33 mM 和 31.46 s-1。纯化的糜蛋白酶的 N 端氨基酸为 IVGGQEATPGEWPWQAALQV。该研究为纯化的大翅礁鱿鱼糜蛋白酶的生化特性提供了新的信息，将有助于今后大翅礁鱿鱼的研究、养殖和应用。

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来源期刊

ACS Applied Electronic Materials Multiple-

CiteScore

7.20

自引率

4.30%

发文量

567

期刊介绍： ACS Applied Electronic Materials is an interdisciplinary journal publishing original research covering all aspects of electronic materials. The journal is devoted to reports of new and original experimental and theoretical research of an applied nature that integrate knowledge in the areas of materials science, engineering, optics, physics, and chemistry into important applications of electronic materials. Sample research topics that span the journal's scope are inorganic, organic, ionic and polymeric materials with properties that include conducting, semiconducting, superconducting, insulating, dielectric, magnetic, optoelectronic, piezoelectric, ferroelectric and thermoelectric. Indexed/Abstracted： Web of Science SCIE Scopus CAS INSPEC Portico