Functional characterization of putative tat signal peptide on heterologous expression of amylase (HL11Amy) isolated from Roseateles terrae HL11

Name: Pongsatorn Khunrach

Organization : Thammasat University. Faculty of Science and Technology

Name: Daran Prongjit

Organization : Thammasat University. Faculty of Science and Technology

Name: Benjarat Bunterngsook

Organization : National Science and Technology Development Agency. National Center for Genetic Engineering and Biotechnology

Name: Katesuda Aiewviriyasakul

Organization : National Science and Technology Development Agency. National Center for Genetic Engineering and Biotechnology

Name: Wipawee Sritusnee

Organization : National Science and Technology Development Agency. National Center for Genetic Engineering and Biotechnology

Name: Hataikarn Lekakarn

Organization : Thammasat University. Faculty of Science and Technology

Email : hataikarn.lek@sci.tu.ac.th

keyword: Twin-arginine translocation signal peptide

ThaSH: Escherichia coli

; Roseateles terrae

ThaSH: Amylases

ThaSH: Proteins -- Analysis

Abstract: The bacterial twin-arginine translocation (Tat) pathway serves an important role in the transportation of fully folded proteins across the bacterial cytoplasmic membrane. The proteins harboring Tat signal peptide are translocated into the periplasm compartment of gram-negative bacteria or extracellular fraction. Furthermore, proteins that co-operate with Tat pathway can decrease the incidence of inclusion body in the cytoplasm. In this research, N-terminal Tat signal peptide was identified from amylase gene (hl11Amy) of Gram-negative bacteria Roseateles terrae HL11. Based on sequence analysis, Tat signal peptide contains 32 amino acids with S/T-RR-X-F-L-K conserved motif in N-region and signal peptidase recognition site (AXA) in C-region. To evaluate the function of Tat signal peptide on recombinant protein expression level and translocation, Tat signal peptide was deleted from the amylase gene. The expression level of full-length gene and mature gene in different cell compartment of Escherichia coli BL21(DE3) was then analyzed. According to the results, the expression level of full-length hl11Amy containing Tat signal peptide (19.78 mg/L culture medium) is 4.43 folds higher than that of the mature gene (4.47 mg/L culture medium). Soluble fulllength HL11Amy was translocated to periplasmic space and culture media, whereas soluble mature HL11Amy was not found in periplasmic space. These results demonstrated the great potential application of the identified Tat signal peptide for the heterologous expression of recombinant protein in periplasmic space of E. coli.

King Mongkut's University of Technology North Bangkok. Central Library

Address: BANGKOK

Email: library@kmutnb.ac.th

Created: 2021

Modified: 2023-05-31

Issued: 2023-05-31

บทความ/Article

application/pdf

BibliograpyCitation : In Thai Society for Biotechnology. Thai Society for Biotechnology and International Conference Online (TSB Online 2021) (pp.435-440). Bangkok : Thai Society for Biotechnology

eng

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