Overexpression and secretion of AgaA7 from Pseudoalteromonas hodoensis sp. nov in Bacillus subtilis for the depolymerization of agarose

College

College of Science

Department/Unit

Biology

Document Type

Article

Source Title

Enzyme and Microbial Technology

Volume

90

First Page

19

Last Page

25

Publication Date

2016

Abstract

Interest in agar or agarose-based pharmaceutical products has driven the search for potent agarolytic enzymes. An extracellular -agarase (AgaA7) recently isolated from Pseudoalteromonas hodoensis sp. nov was expressed in Bacillus subtilis, which was chosen due to its capability to overproduce and secrete functional enzymes. Phenotypic analysis showed that the engineered B. subtilis secreted a functional AgaA7 when fused with the aprE signal peptide (SP) at the amino-terminus. The maximum agarolytic activity was observed during the late logarithmic phase. To further improve the secretion of AgaA7, an expression library of AgaA7 fused to different naturally occurring B. subtilis SPs was created. The amount of AgaA7 secreted by the clones was compared through activity assay, immuno-blot, and purification via affinity chromatography. Although the aprE SP can readily facilitate the secretion of AgaA7, other SPs such as yqgA, pel, and lipA were relatively more efficient. Among these SPs, lipA was the most efficient in improving the secretion of AgaA7.The use of B. subtilis as host for the expression and secretion of agarolytic and other hydrolytic enzymes can be a useful tool in the field of white biotechnology.

html

Digitial Object Identifier (DOI)

http://dx.doi.org/10.1016/j.enzmictec.2016.04.009

Disciplines

Biology

Keywords

Agar; Signal peptidases

Upload File

wf_no

This document is currently not available here.

Share

COinS