里氏木霉纤维生物水解酶2的组成型过表达揭示了其引发纤维素降解的能力

Yubo Wang , Meibin Ren , Yifan Wang , Lu Wang , Hong Liu , Mei Shi , Yaohua Zhong
{"title":"里氏木霉纤维生物水解酶2的组成型过表达揭示了其引发纤维素降解的能力","authors":"Yubo Wang ,&nbsp;Meibin Ren ,&nbsp;Yifan Wang ,&nbsp;Lu Wang ,&nbsp;Hong Liu ,&nbsp;Mei Shi ,&nbsp;Yaohua Zhong","doi":"10.1016/j.engmic.2022.100059","DOIUrl":null,"url":null,"abstract":"<div><p>Cellulose degradation results from the synergistic effect of different enzymes, but which enzyme is involved in the initial stage of cellulose degradation is still not well understood. Cellobiohydrolase 2 (CBH2) attached to the conidial surface is possibly associated with the initial stage. However, its specific mechanism is still incompletely known. This study explored the potential role of CBH2 in initiating cellulose degradation using a constitutive overexpression strategy. First, the CBH2-overexpression <em>Trichoderma reesei</em> strains Qgc2–5 and Qrc2–40 were constructed using the constitutive promoters P<em>gpd1</em> and P<em>rpS30</em>, respectively. It was found that <em>cbh2</em> was expressed at a high level under the glucose conditions and was significantly higher than that of the parental strain QM9414 at the early stage of 29 h when cellulose was used as the carbon source. Particularly, the constitutive overexpression of <em>cbh2</em> caused the strong expression of major cellulase-encoding genes (<em>cbh1, eg1</em>, and <em>eg2</em>) and the rapid decomposition of cellulosic material. Meanwhile, the scanning electron microscope showed that the groove-like structure of the cellulose surface was eroded seriously owing to CBH2 overexpression, which caused the cellulose surface to be smooth. These results showed that the overexpression of CHB2 caused the major cellulase enzymes to be expressed and contributed to cellulose degradation, showing the potential role of CBH2 in the initial stage of the cellulose hydrolytic process.</p></div>","PeriodicalId":100478,"journal":{"name":"Engineering Microbiology","volume":"3 1","pages":"Article 100059"},"PeriodicalIF":0.0000,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"3","resultStr":"{\"title\":\"Constitutive overexpression of cellobiohydrolase 2 in Trichoderma reesei reveals its ability to initiate cellulose degradation\",\"authors\":\"Yubo Wang ,&nbsp;Meibin Ren ,&nbsp;Yifan Wang ,&nbsp;Lu Wang ,&nbsp;Hong Liu ,&nbsp;Mei Shi ,&nbsp;Yaohua Zhong\",\"doi\":\"10.1016/j.engmic.2022.100059\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Cellulose degradation results from the synergistic effect of different enzymes, but which enzyme is involved in the initial stage of cellulose degradation is still not well understood. Cellobiohydrolase 2 (CBH2) attached to the conidial surface is possibly associated with the initial stage. However, its specific mechanism is still incompletely known. This study explored the potential role of CBH2 in initiating cellulose degradation using a constitutive overexpression strategy. First, the CBH2-overexpression <em>Trichoderma reesei</em> strains Qgc2–5 and Qrc2–40 were constructed using the constitutive promoters P<em>gpd1</em> and P<em>rpS30</em>, respectively. It was found that <em>cbh2</em> was expressed at a high level under the glucose conditions and was significantly higher than that of the parental strain QM9414 at the early stage of 29 h when cellulose was used as the carbon source. Particularly, the constitutive overexpression of <em>cbh2</em> caused the strong expression of major cellulase-encoding genes (<em>cbh1, eg1</em>, and <em>eg2</em>) and the rapid decomposition of cellulosic material. Meanwhile, the scanning electron microscope showed that the groove-like structure of the cellulose surface was eroded seriously owing to CBH2 overexpression, which caused the cellulose surface to be smooth. These results showed that the overexpression of CHB2 caused the major cellulase enzymes to be expressed and contributed to cellulose degradation, showing the potential role of CBH2 in the initial stage of the cellulose hydrolytic process.</p></div>\",\"PeriodicalId\":100478,\"journal\":{\"name\":\"Engineering Microbiology\",\"volume\":\"3 1\",\"pages\":\"Article 100059\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-03-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"3\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Engineering Microbiology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2667370322000509\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Engineering Microbiology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2667370322000509","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 3

摘要

纤维素降解是不同酶协同作用的结果,但哪种酶参与纤维素降解的初始阶段还不清楚。附着在分生孢子表面的纤维素酶2(CBH2)可能与初始阶段有关。然而,其具体机制尚不完全清楚。本研究利用组成型过表达策略探讨了CBH2在引发纤维素降解中的潜在作用。首先,分别使用组成型启动子Pgpd1和PrpS30构建CBH2过表达的里氏木霉菌株Qgc2–5和Qrc2–40。发现cbh2在葡萄糖条件下以高水平表达,并且在使用纤维素作为碳源的29小时的早期阶段显著高于亲本菌株QM9414。特别地,cbh2的组成型过表达导致主要纤维素酶编码基因(cbh1、eg1和eg2)的强烈表达和纤维素材料的快速分解。同时,扫描电子显微镜显示,由于CBH2过表达,纤维素表面的凹槽状结构被严重侵蚀,导致纤维素表面光滑。这些结果表明,CHB2的过表达导致主要纤维素酶的表达并有助于纤维素降解,表明CBH2在纤维素水解过程的初始阶段具有潜在作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Constitutive overexpression of cellobiohydrolase 2 in Trichoderma reesei reveals its ability to initiate cellulose degradation

Constitutive overexpression of cellobiohydrolase 2 in Trichoderma reesei reveals its ability to initiate cellulose degradation

Cellulose degradation results from the synergistic effect of different enzymes, but which enzyme is involved in the initial stage of cellulose degradation is still not well understood. Cellobiohydrolase 2 (CBH2) attached to the conidial surface is possibly associated with the initial stage. However, its specific mechanism is still incompletely known. This study explored the potential role of CBH2 in initiating cellulose degradation using a constitutive overexpression strategy. First, the CBH2-overexpression Trichoderma reesei strains Qgc2–5 and Qrc2–40 were constructed using the constitutive promoters Pgpd1 and PrpS30, respectively. It was found that cbh2 was expressed at a high level under the glucose conditions and was significantly higher than that of the parental strain QM9414 at the early stage of 29 h when cellulose was used as the carbon source. Particularly, the constitutive overexpression of cbh2 caused the strong expression of major cellulase-encoding genes (cbh1, eg1, and eg2) and the rapid decomposition of cellulosic material. Meanwhile, the scanning electron microscope showed that the groove-like structure of the cellulose surface was eroded seriously owing to CBH2 overexpression, which caused the cellulose surface to be smooth. These results showed that the overexpression of CHB2 caused the major cellulase enzymes to be expressed and contributed to cellulose degradation, showing the potential role of CBH2 in the initial stage of the cellulose hydrolytic process.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
CiteScore
3.90
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信