International Journal of Biological Macromolecules 99 (2017) 721–730 Contents lists available at ScienceDirect International Journal of Biological Macromolecules j ourna l h o mepa ge: www.elsevier.com/locate/ijbiomac Review Heparin depolymerization by immobilized heparinase: A review Indu Bhushan a,b,c,∗ , Alhumaidi Alabbas c,d,e , Jyothi C. Sistla c , Rashmi Saraswat a , Umesh R. Desai c,d , Ram B. Gupta b a Department of Biotechnology, Shri Mata Vaishno Devi University, Katra, J&K 182320, India b Department of Chemical and Life Science Engineering, Virginia Commonwealth University, Richmond, VA 23284-3068, USA c Institute for Structural Biology, Drug Discovery and Development Virginia Commonwealth University, Richmond, VA 23298-0540, USA d Department of Medicinal Chemistry, Virginia Commonwealth University, Richmond, VA 23298-0540, USA e Department of Pharmaceutical Chemistry, Prince Sattam bin Abdulaziz University, Alkharj 11942, Saudi Arabia a r t i c l e i n f o Article history: Received 17 November 2016 Received in revised form 19 February 2017 Accepted 6 March 2017 Available online 11 March 2017 Keywords: Immobilization Enzymes Heparin Low molecular weight heparins Heparinase and glycosaminoglycans a b s t r a c t Heparin is a member of the glycosaminoglycan (GAG) family composed of glucosamine and uronic acid units containing O-sulfo, N-acetyl and N-sulfo groups, which are alternating in the chain and linked by 1→4 manner. It is a naturally occurring anticoagulant that prevents the formation of clots and their growth within blood. Certain low molecular weight heparins (LMWHs) are considered as better ther- apeutic agents than natural heparin because of the reduced side effects and smaller risk of bleeding. LMWHs can be produced from heparin by chemical or enzymatic depolymerizations. Heparinases cat- alyze the cleavage of glycosidic linkage between amino sugars and uronic acids in heparin. There are three kinds of heparinases which are frequently used for depolymerization of heparin. Despite wide range of applications of heparinases in health care, their use still has been hampered due to poor stability and high cost. To overcome this problem heparinases are recommended for immobilization to reduce the cost of product and enhance stability. Heparinases have been successfully immobilized using various methods and supports, mostly for deheparinization of blood through extracorporeal devices. The focus of this review is to present the current status of heparinase immobilization including various supports and methods used, stability and applications. © 2017 Elsevier B.V. All rights reserved. Contents 1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 721 2. Heparin and low molecular weight heparins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 722 3. Enzyme immobilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 722 4. Heparinases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 724 5. Immobilization of heparinases: applications to health care . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 725 6. Conclusion and future prospects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 727 Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .727 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 727 1. Introduction Enzymes are the biological macromolecules which biocatalyze specific biochemical reactions. Similar to chemical catalysts, these also enhance the rate of reaction without any loss and remain unchanged at the end of reaction. To address the challenges of diffi- culty in availability, stability and specificity, many scientists started ∗ Corresponding author at: Department of Biotechnology, Shri Mata Vaishno Devi University, Katra, J&K 182320, India. E-mail address: sharmasmvdu92@gmail.com (I. Bhushan). application of genetic engineering and immobilization techniques. In turn this helped toward the innovation of new technologies to improve the production of enzymes and also to alter the cer- tain properties of enzymes by protein engineering. In addition, the development of fermentation processes using selective strains helped to produce high quality, high purity, well-characterized enzymes on a large scale for the industrial use. Recently, for large scale production, microbial enzymes have replaced the enzymes from animal and plant sources. Furthermore, the recombinant DNA http://dx.doi.org/10.1016/j.ijbiomac.2017.03.036 0141-8130/© 2017 Elsevier B.V. All rights reserved.