Expression of low endotoxin 3-o-sulfotransferase in Bacillus subtilis and Bacillus megaterium

Wenya Wang; Jacob A. Englaender; Peng Xu; Krunal K. Mehta; Jiraporn Suwan; Jonathan S. Dordick; Fuming Zhang; Qipeng Yuan; Robert J. Linhardt; Mattheos Koffas

doi:10.1007/s12010-013-0415-8

Expression of low endotoxin 3-o-sulfotransferase in Bacillus subtilis and Bacillus megaterium

Wenya Wang, Jacob A. Englaender, Peng Xu, Krunal K. Mehta, Jiraporn Suwan, Jonathan S. Dordick, Fuming Zhang, Qipeng Yuan, Robert J. Linhardt^*, Mattheos Koffas

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

A key enzyme for the biosynthesis and bioengineering of heparin, 3-O-sulfotransferase-1 (3-OST-1), was expressed and purified in Gram-positive Bacillus subtilis and Bacillus megaterium. Western blotting, protein sequence analysis, and enzyme activity measurement confirmed the expression. The enzymatic activity of 3-OST-1 expressed in Bacillus species were found to be similar to those found when expressed in Escherichia coli. The endotoxin level in 3-OST-1 from B. subtilis and B. megaterium were 10⁴-10 ⁵-fold lower than that of the E. coli-expressed 3-OST-1, which makes the Bacillus expression system of particular interest for the generation of pharmaceutical grade raw heparin from nonanimal sources.

Original language	English
Pages (from-to)	954-962
Number of pages	9
Journal	Applied Biochemistry and Biotechnology
Volume	171
Issue number	4
DOIs	https://doi.org/10.1007/s12010-013-0415-8
State	Published - Oct 2013
Externally published	Yes

Keywords

3-O-Sulfotransferase
Bacillus megaterium
Bacillus subtilis
Bioengineered heparin
Endotoxin free
Nonanimal sources

Access to Document

10.1007/s12010-013-0415-8

Cite this

@article{9f06bb4727934657bb9c3e8ae6af658c,

title = "Expression of low endotoxin 3-o-sulfotransferase in Bacillus subtilis and Bacillus megaterium",

abstract = "A key enzyme for the biosynthesis and bioengineering of heparin, 3-O-sulfotransferase-1 (3-OST-1), was expressed and purified in Gram-positive Bacillus subtilis and Bacillus megaterium. Western blotting, protein sequence analysis, and enzyme activity measurement confirmed the expression. The enzymatic activity of 3-OST-1 expressed in Bacillus species were found to be similar to those found when expressed in Escherichia coli. The endotoxin level in 3-OST-1 from B. subtilis and B. megaterium were 104-10 5-fold lower than that of the E. coli-expressed 3-OST-1, which makes the Bacillus expression system of particular interest for the generation of pharmaceutical grade raw heparin from nonanimal sources.",

keywords = "3-O-Sulfotransferase, Bacillus megaterium, Bacillus subtilis, Bioengineered heparin, Endotoxin free, Nonanimal sources",

author = "Wenya Wang and Englaender, {Jacob A.} and Peng Xu and Mehta, {Krunal K.} and Jiraporn Suwan and Dordick, {Jonathan S.} and Fuming Zhang and Qipeng Yuan and Linhardt, {Robert J.} and Mattheos Koffas",

note = "Funding Information: Acknowledgments This work was supported by grants funded by the National Institutes of Health HL101721 and HL096972 (RJL), the Bioengineered Heparin Consortium, and 863 Hi-Tech Research and Development Program of the People{\textquoteright}s Republic of China (project no. 2012AA022300). The authors would also like to thank Dr. Sui-Lam Wong (University of Calgary), Dr. Xiaozhou Zhang (Virginia Tech), and Dr. Cynthia Collins (Rensselaer Polytechnic Institute) for supplying bacterial strains and plasmids.",

year = "2013",

month = oct,

doi = "10.1007/s12010-013-0415-8",

language = "英语",

volume = "171",

pages = "954--962",

journal = "Applied Biochemistry and Biotechnology",

issn = "0273-2289",

publisher = "Humana Press",

number = "4",

}

TY - JOUR

T1 - Expression of low endotoxin 3-o-sulfotransferase in Bacillus subtilis and Bacillus megaterium

AU - Wang, Wenya

AU - Englaender, Jacob A.

AU - Xu, Peng

AU - Mehta, Krunal K.

AU - Suwan, Jiraporn

AU - Dordick, Jonathan S.

AU - Zhang, Fuming

AU - Yuan, Qipeng

AU - Linhardt, Robert J.

AU - Koffas, Mattheos

N1 - Funding Information: Acknowledgments This work was supported by grants funded by the National Institutes of Health HL101721 and HL096972 (RJL), the Bioengineered Heparin Consortium, and 863 Hi-Tech Research and Development Program of the People’s Republic of China (project no. 2012AA022300). The authors would also like to thank Dr. Sui-Lam Wong (University of Calgary), Dr. Xiaozhou Zhang (Virginia Tech), and Dr. Cynthia Collins (Rensselaer Polytechnic Institute) for supplying bacterial strains and plasmids.

PY - 2013/10

Y1 - 2013/10

N2 - A key enzyme for the biosynthesis and bioengineering of heparin, 3-O-sulfotransferase-1 (3-OST-1), was expressed and purified in Gram-positive Bacillus subtilis and Bacillus megaterium. Western blotting, protein sequence analysis, and enzyme activity measurement confirmed the expression. The enzymatic activity of 3-OST-1 expressed in Bacillus species were found to be similar to those found when expressed in Escherichia coli. The endotoxin level in 3-OST-1 from B. subtilis and B. megaterium were 104-10 5-fold lower than that of the E. coli-expressed 3-OST-1, which makes the Bacillus expression system of particular interest for the generation of pharmaceutical grade raw heparin from nonanimal sources.

AB - A key enzyme for the biosynthesis and bioengineering of heparin, 3-O-sulfotransferase-1 (3-OST-1), was expressed and purified in Gram-positive Bacillus subtilis and Bacillus megaterium. Western blotting, protein sequence analysis, and enzyme activity measurement confirmed the expression. The enzymatic activity of 3-OST-1 expressed in Bacillus species were found to be similar to those found when expressed in Escherichia coli. The endotoxin level in 3-OST-1 from B. subtilis and B. megaterium were 104-10 5-fold lower than that of the E. coli-expressed 3-OST-1, which makes the Bacillus expression system of particular interest for the generation of pharmaceutical grade raw heparin from nonanimal sources.

KW - 3-O-Sulfotransferase

KW - Bacillus megaterium

KW - Bacillus subtilis

KW - Bioengineered heparin

KW - Endotoxin free

KW - Nonanimal sources

UR - http://www.scopus.com/inward/record.url?scp=84885190040&partnerID=8YFLogxK

U2 - 10.1007/s12010-013-0415-8

DO - 10.1007/s12010-013-0415-8

M3 - 文章

C2 - 23912211

AN - SCOPUS:84885190040

SN - 0273-2289

VL - 171

SP - 954

EP - 962

JO - Applied Biochemistry and Biotechnology

JF - Applied Biochemistry and Biotechnology

IS - 4

ER -

Expression of low endotoxin 3-o-sulfotransferase in Bacillus subtilis and Bacillus megaterium

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this