利用PEF辅助自溶法无干扰提取酵母胞内分子

IF 0.4 4区工程技术 Q4 ENGINEERING, ELECTRICAL & ELECTRONIC

Electrical Engineering in Japan Pub Date : 2022-02-22 DOI:10.1002/eej.23372

Shuji Okamoto, Yoji Murakami, Gen Urabe, S. Katsuki

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引用次数: 0

摘要

本文探讨了脉冲电场法结合自溶法无损提取酵母胞内蛋白的可能性。首先，通过荧光染色形态学观察，研究了20 kV/cm, 600 ns的PEFs对酵母的影响以及随后的细胞壁塌缩现象。形态学观察表明，PEFs增加了细胞膜的通透性，也破坏了细胞内的液泡，而细胞壁形态保持不变。然而，在pef后的培养过程中，细胞壁塌陷。接下来，用BCA法评估提取蛋白的量。PEF处理后的悬浮液中提取的蛋白质量很少，而额外的孵育促进了提取。最后，用Native - PAGE和SDS - PAGE对提取蛋白的大小进行了研究。经PEF处理和孵育的悬浮液含有大于250 kDa的大蛋白。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Nondistractive extraction of intracellular molecules from yeast using PEF‐assisted autolysis

This paper discusses the possibility of nondestructive extraction of intracellular proteins of yeast (Saccharomyces cerevisiae) by means of pulsed electric fields (PEFs) combined with autolysis. First, the effects of PEFs (20 kV/cm, 600 ns) on yeast and the subsequent cell wall collapse phenomenon were studied by morphological observation of fluorescence staining. Morphological observation shows that the PEFs increase the permeability of the cell membrane and also disrupt the intracellular vacuoles, whereas the cell wall morphology remains unchanged. However, the cell wall was collapse during the postPEF incubation. Next, the amount of extracted protein was evaluated by using the BCA assay. The amount of extracted protein in the PEF treated suspension was only a little, whereas the additional incubation promotes the extraction. Lastly, the size of the extracted proteins was investigated by using both Native‐PAGE and SDS‐PAGE. The PEF‐treated and incubated suspension contains the large protein over 250 kDa.

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来源期刊

Electrical Engineering in Japan 工程技术-工程：电子与电气

CiteScore

0.80

自引率

0.00%

发文量

审稿时长

4-8 weeks

期刊介绍： Electrical Engineering in Japan (EEJ) is an official journal of the Institute of Electrical Engineers of Japan (IEEJ). This authoritative journal is a translation of the Transactions of the Institute of Electrical Engineers of Japan. It publishes 16 issues a year on original research findings in Electrical Engineering with special focus on the science, technology and applications of electric power, such as power generation, transmission and conversion, electric railways (including magnetic levitation devices), motors, switching, power economics.