Chitosanase signal peptide from Bacillus subtilis its lead target protein to periplasmic space in Escherichia coli

Title
Chitosanase signal peptide from Bacillus subtilis its lead target protein to periplasmic space in Escherichia coli
Author(s)
오철홍; 문송; 조은영; 이수진; 이영득; 박건후; 권영경; Hettiarachchi Sachithra Amarin; 강도형
KIOST Author(s)
Oh, Chulhong(오철홍)Jo, Eunyoung(조은영)Lee, Youngdeuk(이영득)Kang, Do-Hyung(강도형)
Publication Year
2016-11-10
Abstract
We isolated chitosanase secreting B. subtilis CH2 and identified the chitosanase sequence. Analyzed the sequence showed that it consisted of 813 bp, including 87 bp signal sequence. The signal sequence leads the target protein to the cell-membrane of the B. subtilis CH2 and then secret the chitosanase out of the cell. The signal peptide showed 6 amino acid deletion compared to other B. subtilis chitosnase signal peptides. The chitosanase sequence including signal peptide was cloned into pET11a vector without fusion and expressed in E. coli BL21(DE3). The expressed chitosanase in E. coli showed two distinct bands which represent the pro-chitosanase in cytoplasm and mature chitosanase in periplasm. Time frame induction and results showed that muture chitosanase was increased. Subsequently, we linked this chitosanase signal sequence in front of B. subtilis CH2 mature lichenase and xylanase sequences, and expressed it in E. coli BL21(DE3). The recombinant xylanase moved to periplasmic space. However, the recombinant lichenase did not move to periplasm.embrane of the B. subtilis CH2 and then secret the chitosanase out of the cell. The signal peptide showed 6 amino acid deletion compared to other B. subtilis chitosnase signal peptides. The chitosanase sequence including signal peptide was cloned into pET11a vector without fusion and expressed in E. coli BL21(DE3). The expressed chitosanase in E. coli showed two distinct bands which represent the pro-chitosanase in cytoplasm and mature chitosanase in periplasm. Time frame induction and results showed that muture chitosanase was increased. Subsequently, we linked this chitosanase signal sequence in front of B. subtilis CH2 mature lichenase and xylanase sequences, and expressed it in E. coli BL21(DE3). The recombinant xylanase moved to periplasmic space. However, the recombinant lichenase did not move to periplasm.
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/24345
Bibliographic Citation
International Conference of the Genetics Society of Korea 2016, pp.182, 2016
Publisher
The Genetics Society of Korea
Type
Conference
Language
English
Publisher
The Genetics Society of Korea
Related Researcher
Files in This Item:
There are no files associated with this item.

qrcode

Items in ScienceWatch@KIOST are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse