Control of the heparosan N-deacetylation leads to an improved bioengineered heparin

• Published in: Applied microbiology and biotechnology Vol. 91; no. 1; pp. 91 - 99
• Main Authors: Wang, Zhenyu, Yang, Bo, Zhang, Zhenqing, Ly, Mellisa, Takieddin, Majde, Mousa, Shaker, Liu, Jian, Dordick, Jonathan S., Linhardt, Robert J.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-07-2011; Springer; Springer Nature B.V
• Subjects: Acetylglucosamine - metabolism; Anticoagulants; Anticoagulants (Medicine); Anticoagulants - chemistry; Anticoagulants - metabolism; Anticoagulants - pharmacology; Binding sites; Bioengineering; Biological and medical sciences; Biotechnological Products and Process Engineering; Biotechnology; Caustic soda; Chondroitin sulfate; Contamination; Disaccharides - chemistry; Disaccharides - metabolism; E coli; Enzymes; Escherichia coli - genetics; Escherichia coli - metabolism; Fermentation; Fundamental and applied biological sciences. Psychology; Glycosaminoglycans; Heparin - chemistry; Heparin - metabolism; Heparin - pharmacology; Humans; Industrial Microbiology - methods; Life Sciences; Microbial Genetics and Genomics; Microbiology; Molecular Structure; Molecular Weight; Pharmaceutical industry; Pharmaceuticals; Pharmacy; Quality control; Sodium; Studies; Heparin; Heparosan; Deacetylation; Porcine intestine; Vertebrata; Mammalia; Intestine; Artiodactyla; Ungulata; Pig
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-011-3231-5

The production of the anticoagulant drug heparin from non-animal sources has a number of advantages over the current commercial production of heparin. These advantages include better source material availability, improved quality control, and reduced concerns about animal virus or prion impurities. A bioengineered heparin would have to be chemically and biologically equivalent to be substituted for animal-sourced heparin as a pharmaceutical. In an effort to produce bioengineered heparin that more closely resembles pharmaceutical heparin, we have investigated a key step in the process involving the
N-
deacetylation of heparosan. The extent of
N-
deacetylation directly affects the
N-
acetyl/
N-
sulfo ratio in bioengineered heparin and also impacts its molecular weight. Previous studies have demonstrated that the presence and quantity of
N-
acetylglucosamine in the nascent glycosaminoglycan chain, serving as the substrate for the subsequent enzymatic modifications (C5 epimerization and
O-
sulfonation), can impact the action of these enzymes and, thus, the content and distribution of iduronic acid and
O-
sulfo groups. In this study, we control the
N-
deacetylation of heparosan to produce a bioengineered heparin with an
N-
acetyl/
N-
sulfo ratio and molecular weight that is similar to animal-sourced pharmaceutical heparin. The structural composition and anticoagulant activity of the resultant bioengineered heparin was extensively characterized and compared to pharmaceutical heparin obtained from porcine intestinal mucosa.