反相高效液相色谱法分离麻风树种子糜蛋白酶水解液中的抗菌肽

Q3 Chemistry
Zehan Andriana, T. Wahyuningsih, T. Raharjo
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引用次数: 0

摘要

抗菌肽(Antimicrobial peptide, AMP)作为一种新型抗生素,在克服耐药性方面具有广阔的应用前景。AMP的生产可以通过蛋白水解酶进行。蓖麻豆(R. communis)中的蛋白质是有毒的,因此它可能是AMP的潜在来源。本研究旨在通过几种提取方法,包括SDS透析,SDS-凝胶过滤,三氟乙酸(TFA)和丙酮沉淀,从蓖麻豆(R. communis)中获得蛋白质水解物。采用胰凝乳酶对蛋白进行水解,肽解液采用反相高效液相色谱进行分馏。采用琼脂扩散法和微量稀释法对肽段进行抑菌活性测定,采用LC-HRMS法对活性部位的氨基酸序列进行鉴定。结果表明,丙酮沉淀法是最佳的水解法,水解度为83.9%。有效组分6和10对大肠杆菌的IC50分别为14.1和14.5µg/mL,对金黄色葡萄球菌的IC50分别为13.3和14.4µg/mL。在馏分10中发现了NVLRGKGMASL肽,在馏分6中发现了GIILLSSK、NMIAKR和LLDILTKK肽,它们具有α -螺旋二级结构,可引起膜损伤。肽NVLRGKGMASL, GIILLSSK, NMIAKR和LLDILTKK被认为具有潜在的抗菌化合物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Antibacterial Peptide from Chymotrypsin Hydrolysate of Jatropha Seeds with RP-HPLC Fractionation
Antimicrobial peptide (AMP), as a new antibiotic agent, has promising prospects in overcoming the problem of resistance. AMP production can be carried out by proteolytic enzymes. Protein from castor bean (R. communis) is toxic, so it can potentially be a source of AMP. This study aims to obtain protein hydrolysate from the castor bean (R. communis) using several extraction methods, including SDS dialysis, SDS-gel filtration, trifluoroacetic acid (TFA), and acetone precipitation. Chymotrypsin enzyme was used to hydrolyze the protein, and the peptide hydrolysate was fractionated using RP-HPLC. The peptide fraction was tested for its antibacterial activity by agar diffusion and microdilution methods, and the most active fraction was identified for its amino acid sequence by LC-HRMS. The results showed that the acetone precipitation extraction method was the best method, with a degree of hydrolysis of 83.9%. The active fractions 6 and 10 of RP-HPLC had IC50 values of 14.1 and 14.5 µg/mL for E. coli and 13.3 and 14.4 µg/mL for S. aureus, respectively. NVLRGKGMASL peptides were found in fraction 10, and GIILLSSK, NMIAKR, and LLDILTKK peptides were found in fraction 6 with an alpha helix secondary structure that can cause membrane damage. The peptides NVLRGKGMASL, GIILLSSK, NMIAKR, and LLDILTKK are thought to have potential as antibacterial compounds.
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来源期刊
Molekul
Molekul Chemistry-Chemistry (all)
CiteScore
1.30
自引率
0.00%
发文量
31
审稿时长
4 weeks
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