Thermostable Kunitz trypsin inhibitor with cytokine inducing, antitumor and HIV-1 reverse transcriptase inhibitory activities from Korean large black soybeans

• Published in: Journal of bioscience and bioengineering 2010-03, Vol.109 (3), p.211-217
• Main Authors: Fang, Evandro Fei, Wong, Jack Ho, Ng, Tzi Bun
• Format: Article
• Language: English
• Subjects: Animals; Antifungal activity; ARN MENSAJERO; ARN MESSAGER; Biological and medical sciences; Biological properties; Biotechnology; Cell Line, Tumor; CITOQUINAS; CYTOKINE; CYTOKINES; Cytokines - metabolism; Drug Stability; Fundamental and applied biological sciences. Psychology; GLYCINE MAX; Glycine max - chemistry; GRAINE; HIV Reverse Transcriptase - antagonists & inhibitors; http://www.fao.org/aos/agrovoc#c_27598; http://www.fao.org/aos/agrovoc#c_3301; http://www.fao.org/aos/agrovoc#c_35078; http://www.fao.org/aos/agrovoc#c_35873; http://www.fao.org/aos/agrovoc#c_37855000037859; http://www.fao.org/aos/agrovoc#c_3914; http://www.fao.org/aos/agrovoc#c_4765; http://www.fao.org/aos/agrovoc#c_6377; http://www.fao.org/aos/agrovoc#c_6946; HUMAN IMMUNODEFICIENCY VIRUS; Human immunodeficiency virus 1; Humans; INHIBIDORES DE TRIPSINA; INHIBITEUR DE TRYPSINE; INTERFERON; INTERFERONAS; INTERFERONS; Korean large black soybean; MESSENGER RNA; Neoplasms - drug therapy; Neoplasms - metabolism; Neoplasms - pathology; Peptides - administration & dosage; Peptides - chemistry; Phytotherapy - methods; Plant Extracts - administration & dosage; Plant Extracts - chemistry; Plant Proteins - administration & dosage; Plant Proteins - chemistry; PURIFICACION; PURIFICATION; REVERSE TRANSCRIPTASE; SEEDS; SEMILLA; Temperature; TRANSCRIPTASA INVERSA; TRANSCRIPTASE INVERSE; Treatment Outcome; Trypsin inhibitor; TRYPSIN INHIBITORS; VIRUS DE L'IMMUNODEFICIENCE HUMAINE; VIRUS DE LA INMUNODEFICIENCIA HUMANA
• ISSN: 1389-1723; 1347-4421
• DOI: 10.1016/j.jbiosc.2009.08.483

A large number of trypsin inhibitors belonging to various types have been purified from different kinds of legumes. In this study, by using liquid chromatography, a Kunitz type trypsin inhibitor (KBTI) with a molecular weight of 20107.645 Da was purified from Korean large black soybeans. KBTI reduced the proteolytic activities of trypsin and α-chymotrypsin with the activity of ∼ 8520 BAEE units/mg and ∼ 24 BTEE units/mg, respectively. It showed high thermal stability (0–100 °C) as well as stability over a large range of pH values (pH 3–11). Furthermore, KBTI inhibited HIV-1 reverse transcriptase activity with an IC 50 value of 0.71 μM and induced the release of pro-inflammatory cytokines such as TNF-α, IL-1β, IL-2 and interferon-γ at the mRNA level. KBTI exerted weak antiproliferative activity toward CNE-2 and HNE-2 nasopharyngeal cancer cells, MCF-7 breast cancer cells, and Hep G2 hepatoma cells. KBTI was destitute of mitogenic, ribonuclease and antifungal activities.