Food-grade expression of nattokinase in Lactobacillus delbrueckii subsp. bulgaricus and its thrombolytic activity in vit
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ORIGINAL RESEARCH PAPER
Food-grade expression of nattokinase in Lactobacillus delbrueckii subsp. bulgaricus and its thrombolytic activity in vitro Chen Li . Zaihui Du . Shaoqing Qi . Xin Zhang . Miaoshu Wang . Yangping Zhou . Haiqiang Lu . Xinxi Gu . Hongtao Tian
Received: 20 October 2019 / Accepted: 16 July 2020 Ó Springer Nature B.V. 2020
Abstract Objectives To produce nattokinase in a food-grade expression system and evaluate its thrombolytic activity in vitro. Results No nattokinase activity from reconstituted strains was observed in simulated gastric juice, but the enzyme was stable in intestinal fluid, the relative activity of which was found to be 60% after 4 h. Due to the nattokinase being produced intracellularly by recombinant bacterial strains, the persistence of the bacteria in gastric juice ensured transmission of the nattokinase into intestinal juice. Because of subsequent disintegration of the bacteria, the highest nattokinase activity was observed after 3 h at
approximately 32%, following its carriage within the recombinant strains to the intestinal fluid. Conclusions This study demonstrated that nattokinase from recombinant strains exhibited good thrombolytic activity in vitro and may be used by the dairy fermentation industry for the development of novel thrombolytic functional foods. Keywords Lactobacillus delbrueckii subsp. bulgaricus thyA-deficient strain Nattokinase Food-grade expression Thrombolytic In vitro
Introduction
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-02974-2) contains supplementary material, which is available to authorized users. Chen Li, Zaihui Du, and Shaoqing Qi have contributed equally to this study. C. Li Z. Du S. Qi X. Zhang Y. Zhou H. Lu X. Gu H. Tian (&) College of Food Science and Technology, Agricultural University of Hebei, Baoding, China e-mail: [email protected] M. Wang New Hope Tensun (Hebei) Dairy Co., Ltd, Baoding, China
Lactic acid bacteria (LAB) play an important role in starter cultures and have a history of safety in dairy products and the development of functional foods (Leroy et al. 2004). At present, a large variety of production systems, including inducible gene expression systems, food-grade genetic modification systems, and chromosomal integration systems, have been developed and used in the production of heterologous proteins in LAB (Sietske et al. 1991; Leite et al. 2013; Yin et al. 2016). Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus) has been among the most commonly-used LAB in the industrial fermentation of dairy products for several centuries.
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Biotechnol Lett
Nattokinase has excellent thrombolytic properties and has beneficial therapeutic effects in serious cardiovascular diseases, a major cause of morbidity, which has attracted considerable attention over recent years (Mathers et al. 2006). At present, traditional drugs used in thrombolytic therapy are generally fibrinolytic enzymes, includ
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