Isolation and partial characterization of an α-amylase inhibitor present in Phaseolus lunatus seeds and its relationship with the induction of α-amylases in larval Zabrotes subfasciatus

IF 2.7 2区 农林科学 Q1 ENTOMOLOGY Journal of Stored Products Research Pub Date : 2024-07-24 DOI:10.1016/j.jspr.2024.102385
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Abstract

The Mexican bean weevil, Zabrotes subfasciatus, causes significant economic losses by infesting the seeds of three major legume species cultivated in Brazil, namely Phaseolus lunatus, Phaseolus vulgaris, and Vigna unguiculata. α-Amylase inhibitors found in P. vulgaris seeds, known as α-AI-1, are efficient against Old World bruchids but are ineffective against Z. subfasciatus. Exposure to α-AI-1 induces the expression and secretion of two α-amylase isoforms, both insensitive to this inhibitor when compared to the constitutive isoform. Interestingly, this induction was also observed when these insects were cultivated on P. lunatus seeds. In this study, we demonstrated the induction of digestive amylases in Z. subfasciatus on P. lunatus seeds through feeding trials. Separation of P. lunatus seed extracts via ion-exchange chromatography led to the identification of fractions containing inhibitory activity against porcine pancreatic amylase. Enzymatic kinetic parameters revealed the presence of a mixed non-competitive inhibitor in P. lunatus fractions, akin to the pattern observed in P. vulgaris. Furthermore, the protein profiles of both types of seeds were similar, with the enrichment of a 30–32 kD protein, possibly representing a precursor form of the α-amylase inhibitor. Thus, our findings show the presence of an α-AI-1-like inhibitor in P. lunatus seeds, which may play a pivotal role in modulating digestive amylases in Z. subfasciatus larvae.

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月见草种子中一种α-淀粉酶抑制剂的分离和部分鉴定及其与诱导亚法氏囊虫幼虫体内α-淀粉酶的关系
墨西哥豆象甲虫(Zabrotes subfasciatus)侵染巴西栽培的三种主要豆科植物(Phaseolus lunatus、Phaseolus vulgaris 和 Vigna unguiculata)的种子,造成重大经济损失。在 P. vulgaris 种子中发现的 α-淀粉酶抑制剂(称为 α-AI-1)对旧世界青虫有效,但对 Z. subfasciatus 无效。暴露于 α-AI-1 会诱导两种 α 淀粉酶异构体的表达和分泌,与组成型异构体相比,这两种异构体对这种抑制剂都不敏感。有趣的是,当这些昆虫在 P. lunatus 种子上培养时,也观察到了这种诱导。在本研究中,我们通过喂食试验证明了亚法西亚蝇对月季种子消化淀粉酶的诱导作用。通过离子交换色谱法分离月桂树种子提取物,鉴定出了对猪胰淀粉酶具有抑制活性的馏分。酶动力学参数显示,月牙藻籽萃取物中存在一种混合的非竞争性抑制剂,这与在粗壮月牙藻中观察到的模式相似。此外,两类种子的蛋白质特征相似,都富含 30-32 kD 蛋白质,可能代表 α 淀粉酶抑制剂的前体形式。因此,我们的研究结果表明,P. lunatus 种子中存在一种类似于 α-AI-1 的抑制剂,它可能在调节 Z. subfasciatus 幼虫的消化淀粉酶方面起着关键作用。
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来源期刊
CiteScore
5.70
自引率
18.50%
发文量
112
审稿时长
45 days
期刊介绍: The Journal of Stored Products Research provides an international medium for the publication of both reviews and original results from laboratory and field studies on the preservation and safety of stored products, notably food stocks, covering storage-related problems from the producer through the supply chain to the consumer. Stored products are characterised by having relatively low moisture content and include raw and semi-processed foods, animal feedstuffs, and a range of other durable items, including materials such as clothing or museum artefacts.
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