Mayura Veerana, Nan-Nan Yu, Si-Jin Bae, Ikhwan Kim, Eun-Seong Kim, Wirinthip Ketya, Hak-Yong Lee, Nam-Young Kim, Gyungsoon Park
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引用次数: 3
Abstract
Enzyme production by microorganisms on an industrial scale has demonstrated technical bottlenecks, such as low efficiency in enzyme expression and extracellular secretion. In this study, as a potential tool for overcoming these technical limits, radio-frequency electromagnetic field (RF-EMF) exposure was examined for its possibility to enhance production of an enzyme, α-amylase, in a filamentous fungus, Aspergillus oryzae. The RF-EMF perfectly resonated at 2 GHz with directivity radiation pattern and peak gain of 0.5 dB (0.01 Watt). Total protein concentration and activity of α-amylase measured in media were about 1.5-3-fold higher in the RF-EMF exposed (10 min) sample than control (no RF-EMF) during incubation (the highest increase after 16 h). The level of α-amylase mRNA in cells was approximately 2-8-fold increased 16 and 24 h after RF-EMF exposure for 10 min. An increase in vesicle accumulation within fungal hyphae and the transcription of some genes involved in protein cellular trafficking was observed in RF-EMF-exposed samples. Membrane potential was not changed, but the intracellular Ca2+ level was elevated after RF-EMF exposure. Our results suggest that RF-EMF can increase the extracellular level of fungal total proteins and α-amylase activity and the intracellular level of Ca2+.
工业规模的微生物酶生产存在技术瓶颈,如酶表达和细胞外分泌效率低。在这项研究中,作为克服这些技术限制的潜在工具,研究了射频电磁场(RF-EMF)暴露在丝状真菌米曲霉(Aspergillus oryzae)中提高α-淀粉酶生产的可能性。RF-EMF在2ghz下完美谐振,具有指向性辐射方向图,峰值增益为0.5 dB (0.01 w)。在培养过程中,RF-EMF暴露(10分钟)的样品中α-淀粉酶的总蛋白浓度和活性比对照组(无RF-EMF)高1.5-3倍(16小时后增幅最大)。在RF-EMF暴露10分钟后的16和24小时,细胞中α-淀粉酶mRNA的水平增加了约2-8倍RF-EMF-exposed样本。膜电位没有改变,但细胞内Ca2+水平升高后RF-EMF暴露。研究结果表明,RF-EMF可以提高真菌胞外总蛋白水平和α-淀粉酶活性以及细胞内Ca2+水平。