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Tandem repeated expression of lunasin gene in Pichia pastoris and its anti-inflammatory activity in vitro

文献类型： 外文期刊

作者： Zhu, Yingying 1 ; Nadia, Everaert 2 ; Yao, Yang 1 ; Shi, Zhenxing 1 ; Ren, Guixing 1 ;

作者机构： 1.CAAS, Inst Crop Sci, 80 South Xueyuan Rd, Beijing 100081, Peoples R China

2.Univ Liege, TERRA Teaching & Res Ctr, Gembloux Agrobio Tech, Precis Livestock & Nutr Unit, Passage Deportes 2, B-5030 Gembloux, Belgium

3.Univ Liege, Gembloux Agrobio Tech, Lab Biomass & Green Technol, Passage Deportes 2, B-5030 Gembloux, Belgium

关键词： Lunasin; Pichia pastoris; Fermentation conditions; Anti-inflammatory; Akt/mTOR/p70s6k pathway

期刊名称：JOURNAL OF BIOSCIENCE AND BIOENGINEERING （ 影响因子：2.894； 五年影响因子：2.746 ）

ISSN： 1389-1723

年卷期： 2018 年 126 卷 1 期

页码：

收录情况： SCI

摘要： Lunasin is a novel promising health-beneficial peptide derived from soybean. However, the application of lunasin is limited by its high cost. In this study, we developed a successful protocol for expression of a dimer formation protein containing 4 tandem repeated lunasin analogs (lunasin-4) in Pichia pastoris. The expression level at the optimal condition (initial pH 7.0,1.0% final methanol concentration and induction for 72 h at 26 degrees C) was 0.24 mg/mL cell-free broth. Lunasin analog, obtained from purified lunasin-4 protein through enterokinase digestion and ultrafiltration, significantly decreased (p < 0.05) the release of nitric oxide (NO), tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages in a dose-dependent manner. In addition, intracellular signaling array analysis demonstrated down-regulated levels of phosphorylated Akt, mechanistic target of rapamycin (mTOR) and p70 s6 kinase (p70s6k) and an up-regulated level of glycogen synthase kinase-3 beta (GSK-3 beta) after lunasin analog treatment. These results suggest that lunasin analog exerted anti-inflammatory activities in LPS-stimulated RAW264.7 cells partly via inhibiting the activation of Ala/mTOR/p70s6k signaling pathway. In conclusion, this study provides a potential strategy for recombinant production of bioactive lunasin in industry. (C) 2018, The Society for Biotechnology, Japan. All rights reserved.

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