Abstract
Microbial fibrinolytic enzyme is a promising candidate for thrombolytic therapy. Non-sterile production of fibrinolytic enzyme by marine Bacillus subtilis D21-8 under submerged fermentation was realized at a mild temperature of 34°C, using a unique combination of starvation strategy and self-production of antibacterial agents. A medium composed of 18.5 g/L glucose, 6.3 g/L yeast extract, 7.9 g/L tryptone, and 5 g/L NaCl was achieved by conventional and statistical methods. Results showed efficient synthesis of fibrinolytic enzyme and antibacterial compounds required the presence of both yeast extract and tryptone in the medium. At shake-flask level, the non-sterile optimized medium resulted in higher productivity of fibrinolytic enzyme than the sterile one, with an enhanced yield of 3,129 U/mL and a production cost reduced by 24%. This is the first report dealing with non-sterile submerged fermentation of fibrinolytic enzyme, which may facilitate the development of feasible techniques for non-sterile production of raw materials for the preparation of potential drugs with low operation cost.
Highlights
Formation of endogenous thrombi in blood vessels leads to cardiovascular diseases which have become the leading cause of morbidity and mortality worldwide (Mozaffarian et al, 2015; Seo et al, 2018)
Our preliminary experiments showed that glucose present in the basal medium was rapidly consumed after inoculation of the strain D21-8, which was responsible for the establishment of a carbon-deficient environment
The basal medium was employed to evaluate the effect of starvation strategy aimed at creating a nutrient-deficient condition for biological control (Figure 1)
Summary
Formation of endogenous thrombi in blood vessels leads to cardiovascular diseases which have become the leading cause of morbidity and mortality worldwide (Mozaffarian et al, 2015; Seo et al, 2018). Fibrinolytic enzyme catalyzes the breakdown of the fibrin mesh of thrombi, thereby playing an important role in thrombolytic therapy (Kotb, 2013). The high expenses and undesired side effects caused by clinical use of traditional thrombolytic agents have prompted researchers toward isolating fibrinolytic enzymes from various sources such as snakes, earthworms, insects, plants, algae, and microorganisms (Peng et al, 2005; Kotb, 2014; Silva et al, 2018). Fibrinolytic enzymes originated from microbes have attracted much more medical interest because of relatively high fibrin specificity and low production cost (Cai et al, 2017; Silva et al, 2018). There is no research concerning fibrinolytic enzyme production by non-sterile submerged fermentation
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