Antibacterial Activity of Recombinant Porcine β-Defensin 2

IF 3.8 3区农林科学 Q1 VETERINARY SCIENCES Pakistan Veterinary Journal Pub Date : 2019-07-01 DOI:10.29261/pakvetj/2019.030

K. Fan

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引用次数: 3

Abstract

Received: Revised: Accepted: Published online: September 03, 2018 December 03, 2018 December 12, 2018 March 07, 2019 To express and purify the recombinant porcine β-defensin 2 (rpBD-2) using yeast Pichia Pastoris expression system and in vitro characterization of it’s antibacterial activity. rpBD-2 was expressed and then purified using methylotrophic yeast Pichia Pastoris expression system. To assess its antimicrobial activity against Escherichia coli, Salmonella paratyphi A, Pseudomonas aeruginosa, Staphylococcus epidermidis, Staphylococcus aureus, Bacillus pumilus, and Bacillus subtilis, liquid growth inhibition assay was used. The antibacterial activity was further kinetically evaluated against representative bacterial strains E. coli and S. aureus. The hemolysis activity of rpBD-2 was also checked against porcine erythrocytes. rpBD2 was expressed and purified successfully using yeast Pichia Pastoris expression system. The yield of final product was 9 mg per liter of the harvested supernatant. rpBD-2 possessed antimicrobial activity against E. coli, S. aureus, S. epidermidis, B. subtilis, and B. pumilus at 33.3-66.6 μM and killed all E. coli and S. aureus within 30 min. Negligible hemolytic activity was observed against porcine erythrocytes. We have successfully developed rpBD-2 as an effective antibacterial peptide against both gram-positive and negative bacteria. rpBD-2 will be a potential novel therapeutic agent against bacterial infection. ©2018 PVJ. All rights reserved

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重组猪β-防御素2的抗菌活性

接收：修订：接受：在线发布：2018年9月3日2018年12月3日2019年12月12日2019年3月7日使用酵母毕赤酵母表达系统表达和纯化重组猪β-防御素2（rpBD-2），并对其抗菌活性进行体外表征。表达rpBD-2，然后使用甲基营养酵母毕赤酵母表达系统进行纯化。为了评估其对大肠杆菌、甲型副伤寒沙门氏菌、铜绿假单胞菌、表皮葡萄球菌、金黄色葡萄球菌、短小芽孢杆菌和枯草芽孢杆菌的抗菌活性，使用液体生长抑制试验。对代表性菌株大肠杆菌和金黄色葡萄球菌的抗菌活性进行了进一步的动力学评价。还检测了rpBD-2对猪红细胞的溶血活性。利用毕赤酵母表达系统成功表达并纯化了rpBD2。最终产物的产率为每升收获的上清液9mg。rpBD-2对大肠杆菌、金黄色葡萄球菌、表皮葡萄球菌、枯草芽孢杆菌和短小芽孢杆菌具有33.3-66.6μM的抗菌活性，并在30分钟内杀死所有大肠杆菌和金黄色葡萄杆菌。对猪红细胞的溶血活性可忽略不计。我们已经成功开发出rpBD-2，作为一种有效的抗菌肽，可以对抗革兰氏阳性菌和阴性菌。rpBD-2将是一种潜在的新型细菌感染治疗剂。©2018 PVJ。保留所有权利

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来源期刊

Pakistan Veterinary Journal 兽医-兽医学

CiteScore

4.20

自引率

13.00%

发文量

审稿时长

4-8 weeks

期刊介绍： The Pakistan Veterinary Journal (Pak Vet J), a quarterly publication, is being published regularly since 1981 by the Faculty of Veterinary Science, University of Agriculture, Faisalabad, Pakistan. It publishes original research manuscripts and review articles on health and diseases of animals including its various aspects like pathology, microbiology, pharmacology, parasitology and its treatment. The “Pak Vet J” (www.pvj.com.pk) is included in Science Citation Index Expended and has got 1.217 impact factor in JCR 2017. Among Veterinary Science Journals of the world (136), “Pak Vet J” has been i) ranked at 75th position and ii) placed Q2 in Quartile in Category. The journal is read, abstracted and indexed internationally.