PMID- 29292153 OWN - NLM STAT- MEDLINE DCOM- 20180829 LR - 20180829 IS - 1879-0003 (Electronic) IS - 0141-8130 (Linking) VI - 111 DP - 2018 May TI - The N-terminal-truncated recombinant fibrin(ogen)olytic serine protease improves its functional property, demonstrates in vivo anticoagulant and plasma defibrinogenation activity as well as pre-clinical safety in rodent model. PG - 462-474 LID - S0141-8130(17)34702-5 [pii] LID - 10.1016/j.ijbiomac.2017.12.140 [doi] AB - An N-terminal truncated fibrino(geno)lytic serine protease gene encoding a ~42kDa protein from Bacillus cereus strain AB01 was produced by error prone PCR, cloned into pET19b vector, and expressed in E5 coli BL21 DE3 cells. The deletion of 24 amino acid residues from N-terminal of wild-type Bacifrinase improves the catalytic activity of [Bacifrinase (DeltaN24)]. The anticoagulant potency of [Bacifrinase (DeltaN24)] was comparable to Nattokinase and Warfarin and results showed that its anticoagulant action is contributed by progressive defibrinogenation and antiplatelet activities. Nonetheless, at the tested concentration of 2.0muM [Bacifrinase (DeltaN24)] did not show in vitro cytotoxicity or chromosomal aberrations on human embryonic kidney cells-293 (HEK-293) and human peripheral blood lymphocytes (HPBL) cells. [Bacifrinase (DeltaN24)], at a dose of 2mg/kg, did not show toxicity, adverse pharmacological effects, tissue necrosis or hemorrhagic effect after 72h of its administration in Swiss albino mice. However, at the tested doses of 0.125 to 0.5mg/kg, it demonstrated significant in anticoagulant effect as well as defibrinogenation after 6h of administration in mice. We propose that [Bacifrinase (DeltaN24)] may serve as prototype for the development of potent drug to prevent hyperfibrinogenemia related disorders. CI - Copyright (c) 2018 Elsevier B.V. All rights reserved. FAU - Bora, Bandana AU - Bora B AD - Microbial Biotechnology and Protein Research Laboratory, Department of Molecular Biology and Biotechnology, School of Sciences, Tezpur University, Tezpur 784028, Assam, India. FAU - Gogoi, Debananda AU - Gogoi D AD - Microbial Biotechnology and Protein Research Laboratory, Department of Molecular Biology and Biotechnology, School of Sciences, Tezpur University, Tezpur 784028, Assam, India. FAU - Tripathy, Debabrata AU - Tripathy D AD - Department of Biotechnology & Bioinformatics, North Eastern Hill University, Shillong 793022, India. FAU - Kurkalang, Sillarine AU - Kurkalang S AD - Department of Biotechnology & Bioinformatics, North Eastern Hill University, Shillong 793022, India. FAU - Ramani, Sheetal AU - Ramani S AD - Microbial Biotechnology and Protein Research Laboratory, Department of Molecular Biology and Biotechnology, School of Sciences, Tezpur University, Tezpur 784028, Assam, India. FAU - Chatterjee, Anupam AU - Chatterjee A AD - Department of Biotechnology & Bioinformatics, North Eastern Hill University, Shillong 793022, India. FAU - Mukherjee, Ashis K AU - Mukherjee AK AD - Microbial Biotechnology and Protein Research Laboratory, Department of Molecular Biology and Biotechnology, School of Sciences, Tezpur University, Tezpur 784028, Assam, India. Electronic address: akm@tezu.ernet.in. LA - eng PT - Journal Article DEP - 20171229 PL - Netherlands TA - Int J Biol Macromol JT - International journal of biological macromolecules JID - 7909578 RN - 0 (Anticoagulants) RN - 0 (Recombinant Proteins) RN - 5Q7ZVV76EI (Warfarin) RN - EC 3.4.- (Serine Proteases) RN - EC 3.4.21.- (Subtilisins) RN - EC 3.4.21.5 (Thrombin) RN - H81695M5OP (nattokinase) SB - IM MH - Animals MH - Anticoagulants/*chemistry MH - Bacillus cereus/enzymology MH - Fibrinolysis/drug effects MH - HEK293 Cells MH - Humans MH - Mice MH - Recombinant Proteins/*chemistry/genetics/pharmacology MH - Serine Proteases/*chemistry/genetics/pharmacology MH - Substrate Specificity MH - Subtilisins/pharmacology MH - Thrombin/chemistry MH - Warfarin/pharmacology OTO - NOTNLM OT - Anticoagulant OT - Antiplatelet OT - Cell cycle OT - Fibrinogen degradation EDAT- 2018/01/03 06:00 MHDA- 2018/08/30 06:00 CRDT- 2018/01/03 06:00 PHST- 2017/11/27 00:00 [received] PHST- 2017/12/26 00:00 [accepted] PHST- 2018/01/03 06:00 [pubmed] PHST- 2018/08/30 06:00 [medline] PHST- 2018/01/03 06:00 [entrez] AID - S0141-8130(17)34702-5 [pii] AID - 10.1016/j.ijbiomac.2017.12.140 [doi] PST - ppublish SO - Int J Biol Macromol. 2018 May;111:462-474. doi: 10.1016/j.ijbiomac.2017.12.140. Epub 2017 Dec 29.