Production, purification and biochemical characterization of the microbial protease produced by Lactobacillus fermentum R6 isolated from Harbin dry sausages

Published on Feb 1, 2020in Process Biochemistry2.952
· DOI :10.1016/J.PROCBIO.2019.10.029
Fangda Sun10
Estimated H-index: 10
(NEAU: Northeast Agricultural University),
Yingying Hu6
Estimated H-index: 6
(NEAU: Northeast Agricultural University)
+ 2 AuthorsLigang Qin3
Estimated H-index: 3
(NEAU: Northeast Agricultural University)
Abstract This study investigated the purification and biochemical characterization of the protease produced by Lactobacillus fermentum R6 isolated from Harbin dry sausages. The optimized fermentation conditions were as follows: a fermentation time of 48 h, an initial pH of 6 and a fermentation temperature of 37 °C. The 37.7 kDa extracellular protease was purified using ammonium sulphate deposition, an ion exchange layer system and gel filtration. The protease produced by L. fermentum R6 had the highest initial velocity and kcat/Km at pH 6, 40 °C. The microbial protease activity could be inhibited by ethylene diamine tetraacetic acid disodium salt (EDTA). The Vmax and Km of the protease were 58.2 ± 1.42 mg/min and 17.3 ± 0.85 mg/mL, respectively. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) reflected the ability of the protease to hydrolyse myofibrillar and sarcoplasmic proteins, in particular, myosin heavy chain, paramyosin, phosphorylase and creatine kinase-M type. In conclusion, L. fermentum R6 can be used as a starter culture or an enzyme-producing strain for the inoculation of Harbin dry sausages.
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