Comparative of Recombinant Vespa affinis Hyaluronidase Expressed in Different Cloning Vectors and their Biological Properties

Piyapon Janpan, Yutthakan Saengkun, Prapenpuksiri Rungsa, Mongkol Vesaratchavest, Tewa Upathanpreecha, Patthana Tastub and Sakda Daduang
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Piyapon Janpan: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
Yutthakan Saengkun: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
Prapenpuksiri Rungsa: Faculty of Pharmaceutical Sciences, Protein and Proteomics Research Center for Commercial and Industrial Purposes Khon Kaen University, Khon Kaen, Thailand
Mongkol Vesaratchavest: Research and Development Center Betagro Public Company Limited Klong Nueng, Klong Luang, Pathumthani
Tewa Upathanpreecha: Research and Development Center, Betagro Public Company Limited, Klong Nueng, Klong Luang, Pathumthani
Patthana Tastub: Research and Development Center, Betagro Public Company Limited, Klong Nueng, Klong Luang, Pathumthani
Sakda Daduang: Faculty of Pharmaceutical Sciences Protein and Proteomics Research Center for Commercial and Industrial Purposes Khon Kaen University, Khon Kaen, Thailand

International Journal of Applied and Physical Sciences, 2018, vol. 4, issue 2, 38-44

Abstract: Cloning and expression of recombinant Vespa affinis hyaluronidase (rVesA2) were successfully expressed in Escherichia coli system. The VesA2 gene was cloned into pET-17b and pET-32a cloning vectors which had molecular weight 41.71 and 59.0 kDa, respectively. The recombinant plasmid of pET-17b was composed 1.08 kDa his-tag at the N-terminal. The 17.14 kDa of fusion tag; thioredoxin tag, histidine tag, and S-tag, was found in pET-32a. The verified expression conditions of rVesA2 induced under the conditions of 0.1 mM IPTG at 37â—¦C for 4 hrs gave the highest quantity of protein expression. The colony PCR and sequencing analysis were used to verify the rVesA2. The positive clones were detected the hyaluronidase activity by a zymographic gel. Recombinant proteins from both cloning vectors were insoluble. However, the recombinant from pET-32a showed higher solubility than that form pET-17b, after dissolving in 4 M urea solution. This result suggests that the fusion tag increase protein solubility.

Keywords: Hyaluronidase enzyme; hyaluronic acid; banded tiger wasp (search for similar items in EconPapers)
Date: 2018
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Persistent link: https://EconPapers.repec.org/RePEc:apa:ijapss:2018:p:38-44

DOI: 10.20469/ijaps.4.50001-2.pdf

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