title: Controllable production of low molecular weight heparins by combinations of heparinase I/II/III.
abstract: Enzymatic depolymerization of heparin by heparinases is promising for production of low molecular weight heparins (LMWHs) as anticoagulants, due to its mild reaction conditions and high selectivity. Here, different heparinase combinations were used to depolymerize heparin. Heparinase I and heparinase II can depolymerize heparin more efficiently than heparinase III, respectively, but heparinase III was the best able to protect the anticoagulant activities of LMWHs. Heparinase III and heparinase I/II combinations were able to efficiently depolymerize heparin to LMWHs with higher anticoagulant activity than the LMWHs produced by the respective heparinase I and heparinase II. HepIII and HepI is the best combination for maintaining high anti-IIa activity (75.7 ± 4.21 IU/mg) at the same Mw value. Furthermore, considering both the changes in molecular weight and anticoagulant activity, the action patterns of heparinase I and heparinase II were found not to follow the exolytic and processive depolymerizing mechanism from the reducing end of heparin.
authors.affiliation: Key Laboratory for Industrial Biocatalysis of Ministry of Education, Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Haidian, Beijing 100084, People's Republic of China. Electronic address: wujingjun2009@foxmail.com.
authors.forename: Jingjun
authors.initials: J
authors.lastname: Wu
authors.forename: Chong
authors.initials: C
authors.lastname: Zhang
authors.forename: Xiang
authors.initials: X
authors.lastname: Mei
authors.forename: Ye
authors.initials: Y
authors.lastname: Li
authors.forename: Xin-Hui
authors.initials: XH
authors.lastname: Xing
mesh_terms: D006493:Heparin / Q000737:chemistry*; D019763:Heparin Lyase / Q000378:metabolism; D008970:Molecular Weight; D058105:Polymerization; D011133:Polysaccharide-Lyases / Q000378:metabolism*
chemical_list: D006493:Heparin; D011133:Polysaccharide-Lyases; C114663:heparinase II; D019763:Heparin Lyase; C021349:heparitinsulfate lyase
keywords: 2-O-sulfo-α-l-iduronic acid; 3,6-di-O-sulfo-N-sulfo-α-d-glucosamine; 6-O-sulfo-N-sulfo-α-d-glucosamine; 6-O-sulfo-α-d-N-acetylglucosamine; AT-III; Enzymatic depolymerization; GPC; GlcA; GlcNAc; GlcNAc6S; GlcNS; GlcNS3S6S; GlcNS6S; H-d-phenylalanyl-l-pipecolyl-arginine-p-nitroaniline dihydrochloride; HIT; HPLC; HepI; HepII; HepIII; Heparin; Heparinases; IPTG; IdoA; IdoA2S; LC–MS; LMWHs; Low molecular weight heparin; M(n); M(w); MBP; N-sulfo-α-d-glucosamine; N-α-benzyloxycarbonyl-d-arginyl-l-glycyl-l-arginine-p-nitroaniline-dihydrochloride; PA; PAGE; RI; S-2238; S-2765; UV; antithrombin III; degree of polymerization; dp; gel permeation chromatography; heparin-induced thrombocytopenia; heparinase I; heparinase II; heparinase III; high performance chromatography; isopropyl-β-d-1-thiogalactopyranoside; liquid chromatography–mass spectrometry; low molecular weight heparins; maltose binding protein; number average molecular weight; photodiode array; polyacrylamide gel electrophoresis; refractive index; ultraviolet; weight average molecular weight; α-d-N-acetylglucosamine; α-l-iduronic acid; β-d-glucuronic acid