College of Pharmaceutical Science & Collaborative Innovation Center for Yangtze River Delta Region Green Pharmaceuticals, Key Laboratory of Marine Fishery Resources Exploitment & Utilization of Zhejiang Province, Zhejiang University of Technology, 310014 Hangzhou, People's Republic of China.
Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA; Department of Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
Carbohydr Polym. 2022 Nov 1;295:119825. doi: 10.1016/j.carbpol.2022.119825. Epub 2022 Jul 5.
Low-molecular-weight heparin (LMWH) is prepared from the controlled chemical or enzymatic depolymerization of animal sourced heparins. It has been widely used as an anticoagulant. Concerns about the shortcomings of animal-derived heparin and the contamination of supply chain demand biochemical approaches for synthesizing LMWH. In the present study, two LMWHs were enzymatically synthesized from low molecular weight N-sulfated heparosan (LMW-NSH) cleaved by recombinant hydrolase, endo-β-glucuronidase, (HepBp) or heparin lyase III (HepIII), followed by subsequent sulfotransferase modifications. Structural characterization shows that LMWH chains prepared using HepBp had a saturated uronic acid residue at their reducing ends, while chains of LMWH prepared using HepIII had an unsaturated uronic acid residue at their non-reducing end. Both LMWHs had anti-factor Xa and anti-factor IIa activities comparable to enoxaparin. This approach demonstrates that the hydrolase, HepBp, can be used to prepare a new type of LMWH that has no unsaturated uronic acid at its non-reducing end.
低分子量肝素（LMWH）是通过动物源肝素的受控化学或酶解聚制备的。它已被广泛用作抗凝剂。由于担心动物源性肝素的缺点和供应链污染，因此需要生化方法来合成 LMWH。在本研究中，使用重组水解酶内切-β-葡糖醛酸酶（HepBp）或肝素 lyase III（HepIII）从低分子量 N-硫酸化肝素聚糖（LMW-NSH）切割后，通过随后的磺基转移酶修饰，酶法合成了两种 LMWH。结构表征表明，使用 HepBp 制备的 LMWH 链在其还原端具有饱和的糖醛酸残基，而使用 HepIII 制备的 LMWH 链在其非还原端具有不饱和的糖醛酸残基。两种 LMWH 均具有与依诺肝素相当的抗 Xa 因子和抗 IIa 因子活性。该方法表明，水解酶 HepBp 可用于制备新型 LMWH，其非还原端没有不饱和糖醛酸。
