Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo n. 542/2, 166 10 Prague 6, Czech Republic.
Molecules. 2021 Sep 16;26(18):5636. doi: 10.3390/molecules26185636.
The problem of a growing resistance of bacteria and other microorganisms to conventional antibiotics gave rise to a search for new potent antimicrobial agents. Insect antimicrobial peptides (AMPs) seem to be promising novel potential anti-infective therapeutics. The dipeptide β-alanyl-tyrosine (β-Ala-Tyr) is one of the endogenous insect toxins exhibiting antibacterial activity against both Gram-negative and Gram-positive bacteria. Prior to testing its other antimicrobial activities, it has to be prepared in a pure form. In this study, we have developed a capillary zone electrophoresis (CZE) method for analysis of β-Ala-Tyr isolated from the extract of the hemolymph of larvae of the fleshfly by reversed-phase high-performance liquid chromatography (RP-HPLC). Based on our previously described correlation between CZE and free-flow zone electrophoresis (FFZE), analytical CZE separation of β-Ala-Tyr and its admixtures have been converted into preparative purification of β-Ala-Tyr by FFZE with preparative capacity of 45.5 mg per hour. The high purity degree of the β-Ala-Tyr obtained by FFZE fractionation was confirmed by its subsequent CZE analysis.
细菌和其他微生物对传统抗生素的耐药性日益增强的问题，促使人们寻找新的强效抗菌药物。昆虫抗菌肽（AMPs）似乎是很有前途的新型潜在抗感染治疗药物。二肽β-丙氨酰-酪氨酸（β-Ala-Tyr）是一种内源性昆虫毒素，对革兰氏阴性和革兰氏阳性菌均具有抗菌活性。在测试其其他抗菌活性之前，必须将其制备成纯形式。在这项研究中，我们开发了一种毛细管区带电泳（CZE）方法，用于分析从肉蝇幼虫血淋巴提取物中分离出的β-Ala-Tyr，方法是通过反相高效液相色谱（RP-HPLC）进行分析。基于我们之前描述的 CZE 与自由流区电泳（FFZE）之间的相关性，通过 FFZE 将β-Ala-Tyr 及其混合物的分析性 CZE 分离转化为β-Ala-Tyr 的制备性纯化，其制备能力为每小时 45.5 毫克。通过 FFZE 分级获得的β-Ala-Tyr 的高纯度程度，通过其随后的 CZE 分析得到了证实。
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