Plasma Bioscience Research Center, Department of Plasma-Bio Display, Kwangwoon University, Seoul 01897, Republic of Korea.
Department of Electrical and Biological Physics, Kwangwoon University, Seoul 01897, Republic of Korea.
Int J Mol Sci. 2023 Feb 24;24(5):4503. doi: 10.3390/ijms24054503.
Although molecular regulation of cellulolytic enzyme production in filamentous fungi has been actively explored, the underlying signaling processes in fungal cells are still not clearly understood. In this study, the molecular signaling mechanism regulating cellulase production in was investigated. We found that the transcription and extracellular cellulolytic activity of four cellulolytic enzymes (, , , and ) increased in Avicel (microcrystalline cellulose) medium. Intracellular nitric oxide (NO) and reactive oxygen species (ROS) detected by fluorescent dyes were observed in larger areas of fungal hyphae grown in Avicel medium compared to those grown in glucose medium. The transcription of the four cellulolytic enzyme genes in fungal hyphae grown in Avicel medium was significantly decreased and increased after NO was intracellularly removed and extracellularly added, respectively. Furthermore, we found that the cyclic AMP (cAMP) level in fungal cells was significantly decreased after intracellular NO removal, and the addition of cAMP could enhance cellulolytic enzyme activity. Taken together, our data suggest that the increase in intracellular NO in response to cellulose in media may have promoted the transcription of cellulolytic enzymes and participated in the elevation of intracellular cAMP, eventually leading to improved extracellular cellulolytic enzyme activity.
虽然丝状真菌中纤维素酶产生的分子调控已经得到了积极的探索,但真菌细胞中潜在的信号转导过程仍不清楚。在本研究中,研究了调节纤维素酶产生的分子信号机制。我们发现,在微晶纤维素(Avicel)培养基中,四种纤维素酶(、、、和)的转录和细胞外纤维素酶活性增加。用荧光染料检测到,与在葡萄糖培养基中生长的真菌菌丝相比,在 Avicel 培养基中生长的真菌菌丝中细胞内的一氧化氮(NO)和活性氧(ROS)在更大的区域内被检测到。在 Avicel 培养基中生长的真菌菌丝中,这四种纤维素酶基因的转录在细胞内去除 NO 和细胞外添加 NO 后分别显著降低和增加。此外,我们发现,细胞内去除 NO 后,真菌细胞中环腺苷酸(cAMP)水平显著降低,添加 cAMP 可以增强纤维素酶活性。综上所述,我们的数据表明,培养基中纤维素诱导的细胞内 NO 增加可能促进了纤维素酶的转录,并参与了细胞内 cAMP 的升高,最终导致细胞外纤维素酶活性的提高。