The purpose of this study was to construct a recombinant Enterococcus faecalis with constitutive secretory expression of protease Ker and evaluate its application effect by solid state fermentation of soybean meal. In this study, the recombinant plasmid pSIP401-kerhds was constructed and used as the basic framework. The recombinant vector pSIP401Z-s6-kerhds was constructed by the introduction of the constitutive promoter p10 and efficient signal peptide S6. The recombinant E. faecalis was constructed by transferring the recombinant vector pSIP401Z-s6-kerhds into the excellent probiotics E. faecalis EXW27. The extracellular protease activity of the recombinant E. faecalis reached 125.37 U/mL. The optimal reaction temperature of recombinant protease Ker was 40 ℃ and the high relative activity and stability was obtained at pH 5.0~10.0. Meanwhile, protease Ker exhibited good tolerance to bile salt solution. The enzymatic properties of Ker made it applicable to the intestinal environment of animals. Compared with E. faecalis EXW27, the recombinant E. faecalis could degrade the protein of soybean meal more effectively. In this study, the recombinant E. faecalis with the characters of probiotic and high protease activity was constructed. This stain showed its potential as a novel probiotic.
GUO Jianjun
,
ZENG Jing
,
YUAN Lin
,
WEI Guowen
. Construction and application of the recombinant Enterococcus faecalis with constitutive secretion of protease[J]. Food and Fermentation Industries, 2020
, 46(3)
: 14
-21
.
DOI: 10.13995/j.cnki.11-1802/ts.022317
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