Abstract:
OBJECTIVES:To identify a new member of serine proteases from Deinagkistrodon acutus via phage display technique and appraise its biocatalytic activities. RESULTS:A novel thrombin-like enzyme gene was cloned by screening the phage display library of D. acutus venom gland. The gene has a 783 bp ORF encoding 260 amino acids. A recombinant enzyme expression vector was constructed and the fused protein was expressed in Escherichia coli. The protein was purified showing a single band of approx. 49.4 kDa after SDS-PAGE. The recombinant enzyme was capable of congealing normal human plasma in vitro with the minimum coagulant dose of 6 µg in 57 s. It exhibited fibrinogenolytic activity by hydrolyzing the Aα-chain of human fibrinogen. It was most active at pH 7.5-8.0 and 35-40 °C with the highest clotting activity of 120 NIH units/mg. It was completely inhibited by PMSF but not by EDTA. Multiple sequence alignments demonstrate that this protein shares high identity with other thrombin-like enzymes from snake venoms. CONCLUSIONS:A novel thrombin-like protein from D. acutus venom was identified, expressed and biologically characterized in vitro. Its fibrinogenolytic properties make the enzyme applicable for biochemical research and drug development on thrombolytic therapy.
journal_name
Biotechnol Lettjournal_title
Biotechnology lettersauthors
Li A,Zhang C,Wang J,Wang J,Jiang H,Li J,Ma X,Zhang W,Lu Ydoi
10.1007/s10529-017-2441-zsubject
Has Abstractpub_date
2018-01-01 00:00:00pages
93-102issue
1eissn
0141-5492issn
1573-6776pii
10.1007/s10529-017-2441-zjournal_volume
40pub_type
杂志文章abstract:OBJECTIVES:To exploit cold-inducible biochemical processes beneficial for foreign mRNA transcription, translation and storage, as well as protein product stability, during Agrobacterium-mediated transient expression. RESULTS:The efficiency of three different 5'-regulatory sequences to achieve transient expression of t...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-017-2336-z
更新日期:2017-07-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-011-0635-3
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pub_type: 杂志文章
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-011-0824-0
更新日期:2012-04-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
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更新日期:2008-03-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-011-0745-y
更新日期:2012-01-01 00:00:00
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pub_type: 杂志文章
doi:10.1007/s10529-009-0067-5
更新日期:2009-11-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/b:bile.0000025890.97067.98
更新日期:2004-05-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-012-1046-9
更新日期:2013-01-01 00:00:00
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pub_type: 杂志文章
doi:10.1007/s10529-015-1816-2
更新日期:2015-07-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章,评审
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更新日期:2018-08-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-020-02850-z
更新日期:2020-06-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/b:bile.0000023029.81600.91
更新日期:2004-04-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-021-03077-2
更新日期:2021-01-12 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-005-8655-5
更新日期:2005-08-01 00:00:00
abstract::The penicillin G amidase (PGA) from Alcaligenes faecalis, which has interesting properties for use in combinatorial biochemistry, was produced by recombinant expression in Escherichia coli. The corresponding gene was cloned into a multicopy vector under the strict regulatory control of the rhamnose inducible promoter....
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/a:1022498314354
更新日期:2003-03-01 00:00:00
abstract::Potato (Solanum tuberosum) hairy root cultures, established by infecting potato tuber discs with Agrobacterium rhizogenes, were used as a model system for the production of antimicrobial sesquiterpenes and lipoxygenase (LOX) metabolites. Of the four sesquiterpene phytoalexins (rishitin, lubimin, phytuberin and phytube...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/a:1023038804556
更新日期:2003-04-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-005-5794-7
更新日期:2005-06-01 00:00:00
abstract:OBJECTIVE:To examine the role of miR-124a in LPS-induced septic cardiac insufficiency where underlying mechanism is unclear. RESULTS:Expression of miR-124a was decreased in myocardium of LPS-induced septic cardiac dysfunction model. miR-124a antagomiR or agomiR were injected via tail vein to induce miR-124a-dysregulat...
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pub_type: 杂志文章
doi:10.1007/s10529-017-2368-4
更新日期:2017-09-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-008-9747-9
更新日期:2008-10-01 00:00:00
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pub_type: 杂志文章
doi:10.1007/s10529-012-1078-1
更新日期:2013-02-01 00:00:00
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journal_title:Biotechnology letters
pub_type: 杂志文章,评审
doi:10.1007/s10529-020-02901-5
更新日期:2020-10-01 00:00:00
abstract::Growth and sporulation of Verticillium lecanii on inert and organic carriers (sugar-cane bagasse, corncob, rice straw, polyurethane foam and activated carbon) in a solid-state fermentation process was studied. Sugar-cane bagasse and polyurethane foam produced 10(10) spores g(-1) dry carrier whereas corncob, rice straw...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-010-0496-1
更新日期:2011-04-01 00:00:00
abstract::Rhizopus arrhizus, strain DAR 36017, produced L(+)-lactic acid in a simultaneous saccharification and fermentation process using starch waste effluents. Lactic acid at 19.5-44.3 g l(-1) with a yield of 0.85-0.96 g g(-1) was produced in 40 h using 20-60 g starch l(-1). Supplementation of nitrogen source may be unnecess...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1023/b:bile.0000004389.53388.d0
更新日期:2003-12-01 00:00:00
abstract::Since T cells express diverse sex steroid hormone receptors, they might be a good model to evaluate the effects of sex steroid hormones on immune modulation. Porcine testicular extract contains several sex steroid hormones and may be useful to study the effects of sex steroid hormones during T cell activation. We have...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-012-0907-6
更新日期:2012-07-01 00:00:00
abstract::Previously, we constructed a genetically modified luciferase of Photinus pyralis that generates more than 10-fold higher luminescence intensity than the wild-type enzyme. In this study, we demonstrate that this modified luciferase enables us to detect ATP at 10(-18) mol, which is almost equal to the quantity contained...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-008-9643-3
更新日期:2008-06-01 00:00:00
abstract::The impact of the two adaptation-induced mutations in an improved xylose-fermenting Zymomonas mobilis strain was investigated. The chromosomal mutation at the xylose reductase gene was critical to xylose metabolism by reducing xylitol formation. Together with the plasmid-borne mutation impacting xylose isomerase activ...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-012-0970-z
更新日期:2012-10-01 00:00:00
abstract::Lentiviral vectors have been used for gene transfer into the liver but their ability to efficiently transduce quiescent hepatocytes remains controversial. Lentivirus-mediated gene transfer is more efficient in cycling cells. We determine the effect of H-IL6 in the lentiviral transduction. The lentiviral vector was use...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-007-9528-x
更新日期:2008-02-01 00:00:00
abstract::Some pest management programs employ PCR to identify cry1 genes from Bacillus thuringiensis to predict bacterial toxicity towards different insect pests. However, due to changes on the mode of action of the Cry proteins, new primers had to be designed to detect the new genes. Therefore, an 'in-silico' study of genetic...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-009-0088-0
更新日期:2009-12-01 00:00:00
abstract::An endoxylanase gene, xynA, was cloned from Bacillus pumilus ARA and expressed in Escherichia coli. The open reading frame of the xynA gene was 687 bp encoding a signal peptide and a mature xylanase with a molecular mass of 23 kDa. The enzyme was categorized as a glycosyl hydrolase family 11 member based on the sequen...
journal_title:Biotechnology letters
pub_type: 杂志文章
doi:10.1007/s10529-011-0568-x
更新日期:2011-07-01 00:00:00