Bohlmann Lisa, Chang Chih-Wei, Beacham Ifor, von Itzstein Mark
Institute for Glycomics, Gold Coast Campus, Griffith University, Queensland, 4222 (Australia).
Chembiochem. 2015 May 26;16(8):1205-11. doi: 10.1002/cbic.201500081. Epub 2015 Apr 23.
Bacterial heparinases that cleave heparan sulfate (HS) and heparin are widely used to generate low-molecular-weight heparins (LMWHs) and to structurally and functionally characterise heparin and HS biomolecules. We provide novel insights into the substrate specificity of heparinase II from two different bacteria: Pedobacter heparinus (formerly Flavobacterium heparinum) and Bacteroides eggerthii. The activity towards various well-defined HS oligosaccharides was investigated by (1) H NMR spectroscopy; this revealed distinct specificities for the two heparinases. Heparinase II from P. heparinus appears to be more active and displays a broader substrate specificity than B. eggerthii heparinase II. Furthermore, HS di- and tetrasaccharides inhibited B. eggerthii heparinase II activity. A better understanding of heparinase substrate specificity will contribute to the production of homogenous LMWHs, provide better characterisation of heparin and HS and assist therapeutic applications.
能够切割硫酸乙酰肝素（HS）和肝素的细菌肝素酶被广泛用于生成低分子量肝素（LMWHs），以及对肝素和HS生物分子进行结构和功能表征。我们对来自两种不同细菌——食碱杆菌（以前的肝素黄杆菌）和埃氏拟杆菌的肝素酶II的底物特异性提供了新的见解。通过¹H NMR光谱研究了对各种明确的HS寡糖的活性；这揭示了两种肝素酶不同的特异性。来自食碱杆菌的肝素酶II似乎比埃氏拟杆菌肝素酶II更具活性，并且表现出更广泛的底物特异性。此外，HS二糖和四糖抑制了埃氏拟杆菌肝素酶II的活性。对肝素酶底物特异性的更好理解将有助于生产同质的LMWHs，更好地表征肝素和HS，并辅助治疗应用。
