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  • Title: Gene cloning and expression of the l-asparaginase from Bacillus cereus BDRD-ST26 in Bacillus subtilis WB600.
    Author: Feng Y, Liu S, Jiao Y, Wang Y, Wang M, Du G.
    Journal: J Biosci Bioeng; 2019 Apr; 127(4):418-424. PubMed ID: 30344018.
    Abstract:
    l-Asparaginase (ASN; EC 3.5.1.1) shows great commercial value because of its ability to reduce toxic levels of acrylamide in foods. To achieve high-efficiency production of l-asparaginase, an open reading frame of 978 bp encoding asparaginase (BcA) was amplified from Bacillus cereus BDRD-ST26, followed by its expression in Bacillus subtilis WB600, with the highest yield of 374.9 U/ml obtained using an amyE-signal peptide. A four-step purification protocol was used to purify BcA, resulting in a 15.1-fold increase in purification yield, with a specific activity of purified BcA at 550.8 U/mg and accompanied by detection of minimal l-glutaminase activity. Maximum BcA activity was detected at 50°C and pH 9.0 in 20 mM Tris-HCl buffer, with a half-life at 50°C of 17.35 min and a Km and kcat of 9.38 mM and 63.6 s-1, respectively. Compared with untreated potato strips, 72% acrylamide (2.35 mg/kg) was removed from potato strips pretreated with BcA. These results indicated that this novel BcA variant represents a potential candidate for application in the food-processing industry.
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