title: OmpA signal peptide leads to heterogenous secretion of B. subtilis chitosanase enzyme from E. coli expression system.
abstract: The production of secreted recombinant proteins from E. coli is pivotal to the biotechnological industry because it reduces the cost of downstream processing. Proteins destined for secretion contain an N-terminal signal peptide that is cleaved by secretion machinery in the plasma membrane. The resulting protein is released in an active mature form. In this study, Bacillus subtilis chitosanase (Csn) was used as a model protein to compare the effect of two signal peptides on the secretion of heterologous recombinant protein. The results showed that the E. coli secretion machinery could recognize both native bacillus and E. coli signal peptides. However, only the native bacillus signal peptide could generate the same N-terminal sequence as in the wild type bacteria. When the recombinant Csn constructs contained the E. coli OmpA signal peptide, the secreted enzymes were heterogeneous, comprising a mixed population of secreted enzymes with different N-terminal sequences. Nevertheless, the E. coli OmpA signal peptide was found to be more efficient for high expression and secretion of bacillus Csn. These findings may be used to help engineer other recombinant proteins for secretory production in E. coli.
authors: Molecular Biotechnology Laboratory, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology (SUT), 111 University Avenue, Meung District, Nakhon Ratchasima, 30000 Thailand.
authors: Phornsiri
authors: P
authors: Pechsrichuang
authors: Synchrotron Light Research Institute (Public Organization), Nakhon Ratchasima, Thailand.
authors: Chomphunuch
authors: C
authors: Songsiriritthigul
authors: Food Biotechnology Laboratory, BOKU - University of National Resources and Life Sciences, Vienna, Austria.
authors: Dietmar
authors: D
authors: Haltrich
authors: Genome Institute, National Center for Genetic Engineering and Biotechnology, Pathumthani, Thailand.
authors: Sittiruk
authors: S
authors: Roytrakul
authors: Molecular Biotechnology Laboratory, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology (SUT), 111 University Avenue, Meung District, Nakhon Ratchasima, 30000 Thailand.
authors: Peenida
authors: P
authors: Namvijtr
authors: Molecular Biotechnology Laboratory, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology (SUT), 111 University Avenue, Meung District, Nakhon Ratchasima, 30000 Thailand.
authors: Napolean
authors: N
authors: Bonaparte
authors: Molecular Biotechnology Laboratory, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology (SUT), 111 University Avenue, Meung District, Nakhon Ratchasima, 30000 Thailand.
authors: Montarop
authors: 0000-0003-2674-2419
authors: M
authors: Yamabhai
keywords: Bacillus; Chitosanase; E. coli; Expression; OmpA; Recombinant; Secretion; Signal peptide