Phylogenetically distinct fungal expansins show different binding preferences towards cellulosic materials and enhance cellulase activity

IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Taru Koitto , Deepika Dahiya , Martina Huusela , Merja Penttilä , Emma Master
{"title":"Phylogenetically distinct fungal expansins show different binding preferences towards cellulosic materials and enhance cellulase activity","authors":"Taru Koitto ,&nbsp;Deepika Dahiya ,&nbsp;Martina Huusela ,&nbsp;Merja Penttilä ,&nbsp;Emma Master","doi":"10.1016/j.crbiot.2025.100296","DOIUrl":null,"url":null,"abstract":"<div><div>Expansins and expansin-like proteins are found in plants and microbes, and can disrupt the cellulosic microfibril network of plant cell walls. While plant expansins play a role in cell wall formation, microbial expansin-like proteins reportedly enhance the activity of lignocellulolytic enzymes. Herein, two novel fungal expansin-like proteins, <em>Ama</em>EXLX1 from <em>Allomyces macrogynus</em> and <em>Apu</em>EXLX1 from <em>Aureobasidium pullulans</em>, were recombinantly produced in <em>Pichia pastoris</em>. While both <em>Ama</em>EXLX1 and <em>Apu</em>EXLX1 retain typical expansin structure, they share low sequence identity (22.5 %) and different predicted pI values (5.8 and 8.8, respectively), which was expected to impact their action on cellulosic substrates. Accordingly, adsorption of the proteins on cellulose nanofibrils (CNF) and the impact of the expansin-like proteins on the hydrolysis of CNF by an endoglucanase (Cel7B) was investigated using quartz crystal microbalance with dissipation (QCM-D). <em>Ama</em>EXLX1 showed higher affinity towards cellulose compared to <em>Apu</em>EXLX1, which was correlated to missing key aromatic residues in the polysaccharide binding surface of <em>Apu</em>EXLX1. The pretreatment of a CNF film with <em>Ama</em>EXLX1 and <em>Apu</em>EXLX1 increased the initial rate of Cel7B activity. This study underscores similarities between the impacts that bacterial and fungal expansin-like proteins can have on the enzymatic deconstruction of cellulose, and sequence properties that could impact expansin performance.</div></div>","PeriodicalId":52676,"journal":{"name":"Current Research in Biotechnology","volume":"9 ","pages":"Article 100296"},"PeriodicalIF":3.6000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Research in Biotechnology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2590262825000279","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

Expansins and expansin-like proteins are found in plants and microbes, and can disrupt the cellulosic microfibril network of plant cell walls. While plant expansins play a role in cell wall formation, microbial expansin-like proteins reportedly enhance the activity of lignocellulolytic enzymes. Herein, two novel fungal expansin-like proteins, AmaEXLX1 from Allomyces macrogynus and ApuEXLX1 from Aureobasidium pullulans, were recombinantly produced in Pichia pastoris. While both AmaEXLX1 and ApuEXLX1 retain typical expansin structure, they share low sequence identity (22.5 %) and different predicted pI values (5.8 and 8.8, respectively), which was expected to impact their action on cellulosic substrates. Accordingly, adsorption of the proteins on cellulose nanofibrils (CNF) and the impact of the expansin-like proteins on the hydrolysis of CNF by an endoglucanase (Cel7B) was investigated using quartz crystal microbalance with dissipation (QCM-D). AmaEXLX1 showed higher affinity towards cellulose compared to ApuEXLX1, which was correlated to missing key aromatic residues in the polysaccharide binding surface of ApuEXLX1. The pretreatment of a CNF film with AmaEXLX1 and ApuEXLX1 increased the initial rate of Cel7B activity. This study underscores similarities between the impacts that bacterial and fungal expansin-like proteins can have on the enzymatic deconstruction of cellulose, and sequence properties that could impact expansin performance.
系统发育上不同的真菌扩张蛋白对纤维素材料表现出不同的结合偏好,并增强纤维素酶的活性
扩张蛋白和扩张蛋白样蛋白存在于植物和微生物中,可以破坏植物细胞壁的纤维素微纤维网络。虽然植物膨胀素在细胞壁形成中起作用,但据报道微生物膨胀素样蛋白增强了木质纤维素水解酶的活性。本研究在毕赤酵母中重组产生了两个新的真菌扩张蛋白样蛋白,分别是来自巨绿异酵母菌(alloomyces macrogynus)的AmaEXLX1和来自pululans的auobasidium ApuEXLX1。虽然AmaEXLX1和ApuEXLX1都保留了典型的扩展蛋白结构,但它们具有较低的序列同一性(22.5%)和不同的预测pI值(分别为5.8和8.8),这预计会影响它们对纤维素底物的作用。因此,利用石英晶体耗散微天平(QCM-D)研究了蛋白质在纤维素纳米原纤维(CNF)上的吸附以及膨胀蛋白样蛋白质对内切葡聚糖酶(Cel7B)水解CNF的影响。与ApuEXLX1相比,AmaEXLX1对纤维素的亲和力更高,这与ApuEXLX1的多糖结合表面缺失关键芳香残基有关。用AmaEXLX1和ApuEXLX1预处理CNF膜提高了Cel7B的初始活性率。这项研究强调了细菌和真菌类扩张蛋白对纤维素酶解的影响之间的相似性,以及可能影响扩张蛋白性能的序列特性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Current Research in Biotechnology
Current Research in Biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
6.70
自引率
3.60%
发文量
50
审稿时长
38 days
期刊介绍: Current Research in Biotechnology (CRBIOT) is a new primary research, gold open access journal from Elsevier. CRBIOT publishes original papers, reviews, and short communications (including viewpoints and perspectives) resulting from research in biotechnology and biotech-associated disciplines. Current Research in Biotechnology is a peer-reviewed gold open access (OA) journal and upon acceptance all articles are permanently and freely available. It is a companion to the highly regarded review journal Current Opinion in Biotechnology (2018 CiteScore 8.450) and is part of the Current Opinion and Research (CO+RE) suite of journals. All CO+RE journals leverage the Current Opinion legacy-of editorial excellence, high-impact, and global reach-to ensure they are a widely read resource that is integral to scientists' workflow.
×
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