Analysis of E. coli K5 capsular polysaccharide heparosan.
Heparosan is the key precursor for the preparation of bioengineered heparin, a potential replacement for porcine intestinal heparin, an important anticoagulant drug. The molecular weight (MW) distribution of heparosan produced by the fermentation of E. coli K5 was investigated. Large-slab isocratic and mini-slab gradient polyacrylamide gel electrophoresis (PAGE) were used to analyze the MW and polydispersity of heparosan. A preparative method that allowed fractionation by continuous-elution PAGE was used to obtain heparosan MW standards. The MWs of the heparosan standards were determined by electrospray ionization Fourier-transform mass spectrometry (ESI-FT-MS). A ladder of the standards was then used to determine the MW properties of polydisperse heparosan samples. Unbleached and bleached heparosan produced by fermentation of E. coli K5 had similar number-averaged MWs (M(N)), weight-averaged MWs (M(W)), and MW ranges of 3,000 to 150,000 Da.
Ly
Mellisa
M
Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, 110 Eight Street, Troy, NY 12180, USA.
Wang
Zhenyu
Z
Laremore
Tatiana N
TN
Zhang
Fuming
F
Zhong
Weihong
W
Pu
Dennis
D
Zagorevski
Dmitri V
DV
Dordick
Jonathan S
JS
Linhardt
Robert J
RJ
D000925:Anticoagulants / Q000737:chemistry* / Q000378:metabolism; D016667:Bacterial Capsules / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry* / Q000378:metabolism; D005285:Fermentation; D005583:Fourier Analysis; D008970:Molecular Weight; D021241:Spectrometry, Mass, Electrospray Ionization
D000925:Anticoagulants; C050673:capsular polysaccharide K5
J Med Chem. 2005 Jan 27;48(2):349-52
15658847
Anal Biochem. 1984 Feb;137(1):25-32
6203432
J Pharm Sci. 1992 Aug;81(8):823-7
1328601
Thromb Haemost. 2009 Nov;102(5):874-8
19888522
Eur J Biochem. 1981 May 15;116(2):359-64
7018909
Angew Chem Int Ed Engl. 2002 Feb 1;41(3):391-412
12491369
Carbohydr Res. 2010 Jan 26;345(2):250-6
19945695
Anal Biochem. 2010 Jun 15;401(2):236-41
20211145
Nat Prod Rep. 2009 May;26(5):610-27
19387498
Nat Biotechnol. 2008 Jun;26(6):669-75
18437154
J Mol Microbiol Biotechnol. 2009;17(2):71-82
19401610
Arterioscler Thromb Vasc Biol. 2004 Sep;24(9):1549-57
15231514
J Biol Chem. 2003 Dec 26;278(52):52613-21
14519763
Proc Natl Acad Sci U S A. 2009 Oct 6;106(40):16956-61
19805108
Electrophoresis. 2004 Oct;25(18-19):3307-12
15472951
Chem Biol. 2005 Mar;12(3):267-77
15797210
J Am Chem Soc. 2008 Oct 1;130(39):12998-3007
18767845
N Engl J Med. 2008 Jun 5;358(23):2457-67
18434646
J Pharm Sci. 2009 Nov;98(11):4017-26
19642166
Mini Rev Med Chem. 2007 Apr;7(4):383-7
17430224
J Med Chem. 2003 Jun 19;46(13):2551-64
12801218
Nat Prod Rep. 2009 Mar;26(3):313-21
19240943