通过结合不同的 LPMO 活性,提高高固体负荷下纤维素的酶法糖化产量。

IF 6.1 1区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology for Biofuels Pub Date : 2024-03-09 DOI:10.1186/s13068-024-02485-6
Camilla F. Angeltveit, Anikó Várnai, Vincent G. H. Eijsink, Svein J. Horn
{"title":"通过结合不同的 LPMO 活性,提高高固体负荷下纤维素的酶法糖化产量。","authors":"Camilla F. Angeltveit,&nbsp;Anikó Várnai,&nbsp;Vincent G. H. Eijsink,&nbsp;Svein J. Horn","doi":"10.1186/s13068-024-02485-6","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><p>The polysaccharides in lignocellulosic biomass hold potential for production of biofuels and biochemicals. However, achieving efficient conversion of this resource into fermentable sugars faces challenges, especially when operating at industrially relevant high solid loadings. While it is clear that combining classical hydrolytic enzymes and lytic polysaccharide monooxygenases (LPMOs) is necessary to achieve high saccharification yields, exactly how these enzymes synergize at high solid loadings remains unclear.</p><h3>Results</h3><p>An LPMO-poor cellulase cocktail, Celluclast 1.5 L, was spiked with one or both of two fungal LPMOs from <i>Thermothielavioides terrestris</i> and <i>Thermoascus aurantiacus, Tt</i>AA9E and <i>Ta</i>AA9A, respectively, to assess their impact on cellulose saccharification efficiency at high dry matter loading, using Avicel and steam-exploded wheat straw as substrates. The results demonstrate that LPMOs can mitigate the reduction in saccharification efficiency associated with high dry matter contents. The positive effect of LPMO inclusion depends on the type of feedstock and the type of LPMO and increases with the increasing dry matter content and reaction time. Furthermore, our results show that chelating free copper, which may leak out of the active site of inactivated LPMOs during saccharification, with EDTA prevents side reactions with in situ generated H<sub>2</sub>O<sub>2</sub> and the reductant (ascorbic acid).</p><h3>Conclusions</h3><p>This study shows that sustaining LPMO activity is vital for efficient cellulose solubilization at high substrate loadings. LPMO cleavage of cellulose at high dry matter loadings results in new chain ends and thus increased water accessibility leading to decrystallization of the substrate, all factors making the substrate more accessible to cellulase action. Additionally, this work highlights the importance of preventing LPMO inactivation and its potential detrimental impact on all enzymes in the reaction.</p></div>","PeriodicalId":494,"journal":{"name":"Biotechnology for Biofuels","volume":null,"pages":null},"PeriodicalIF":6.1000,"publicationDate":"2024-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://biotechnologyforbiofuels.biomedcentral.com/counter/pdf/10.1186/s13068-024-02485-6","citationCount":"0","resultStr":"{\"title\":\"Enhancing enzymatic saccharification yields of cellulose at high solid loadings by combining different LPMO activities\",\"authors\":\"Camilla F. Angeltveit,&nbsp;Anikó Várnai,&nbsp;Vincent G. H. Eijsink,&nbsp;Svein J. Horn\",\"doi\":\"10.1186/s13068-024-02485-6\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><p>The polysaccharides in lignocellulosic biomass hold potential for production of biofuels and biochemicals. However, achieving efficient conversion of this resource into fermentable sugars faces challenges, especially when operating at industrially relevant high solid loadings. While it is clear that combining classical hydrolytic enzymes and lytic polysaccharide monooxygenases (LPMOs) is necessary to achieve high saccharification yields, exactly how these enzymes synergize at high solid loadings remains unclear.</p><h3>Results</h3><p>An LPMO-poor cellulase cocktail, Celluclast 1.5 L, was spiked with one or both of two fungal LPMOs from <i>Thermothielavioides terrestris</i> and <i>Thermoascus aurantiacus, Tt</i>AA9E and <i>Ta</i>AA9A, respectively, to assess their impact on cellulose saccharification efficiency at high dry matter loading, using Avicel and steam-exploded wheat straw as substrates. The results demonstrate that LPMOs can mitigate the reduction in saccharification efficiency associated with high dry matter contents. The positive effect of LPMO inclusion depends on the type of feedstock and the type of LPMO and increases with the increasing dry matter content and reaction time. Furthermore, our results show that chelating free copper, which may leak out of the active site of inactivated LPMOs during saccharification, with EDTA prevents side reactions with in situ generated H<sub>2</sub>O<sub>2</sub> and the reductant (ascorbic acid).</p><h3>Conclusions</h3><p>This study shows that sustaining LPMO activity is vital for efficient cellulose solubilization at high substrate loadings. LPMO cleavage of cellulose at high dry matter loadings results in new chain ends and thus increased water accessibility leading to decrystallization of the substrate, all factors making the substrate more accessible to cellulase action. Additionally, this work highlights the importance of preventing LPMO inactivation and its potential detrimental impact on all enzymes in the reaction.</p></div>\",\"PeriodicalId\":494,\"journal\":{\"name\":\"Biotechnology for Biofuels\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":6.1000,\"publicationDate\":\"2024-03-09\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://biotechnologyforbiofuels.biomedcentral.com/counter/pdf/10.1186/s13068-024-02485-6\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biotechnology for Biofuels\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://link.springer.com/article/10.1186/s13068-024-02485-6\",\"RegionNum\":1,\"RegionCategory\":\"工程技术\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biotechnology for Biofuels","FirstCategoryId":"5","ListUrlMain":"https://link.springer.com/article/10.1186/s13068-024-02485-6","RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

背景:木质纤维素生物质中的多糖具有生产生物燃料和生物化学品的潜力。然而,将这种资源高效转化为可发酵糖类面临着挑战,尤其是在工业相关的高固体负荷下运行时。显然,要获得高糖化产量,必须将传统水解酶和溶解多糖单氧酶(LPMOs)结合起来,但这些酶在高固体负荷下究竟如何协同作用仍不清楚:结果:以 Avicel 和蒸汽爆破的小麦秸秆为底物,在低 LPMO 贫纤维素酶鸡尾酒 Celluclast 1.5 L 中添加了两种真菌 LPMO(Thermothielavioides terrestris 和 Thermoascus aurantiacus,分别为 TtAA9E 和 TaAA9A)中的一种或两种,以评估它们在高干物质负荷下对纤维素糖化效率的影响。结果表明,LPMOs 可以缓解高干物质含量导致的糖化效率降低。加入 LPMO 的积极效果取决于原料类型和 LPMO 类型,并且随着干物质含量和反应时间的增加而增加。此外,我们的研究结果表明,在糖化过程中,游离铜可能会从失活的 LPMO 的活性位点漏出,用 EDTA 与游离铜螯合可防止与原位生成的 H2O2 和还原剂(抗坏血酸)发生副反应:这项研究表明,维持 LPMO 的活性对于在高底物负荷下高效溶解纤维素至关重要。在干物质负荷较高时,LPMO 裂解纤维素会产生新的链端,从而增加水的可及性,导致底物脱晶,所有这些因素都使底物更容易被纤维素酶作用。此外,这项工作还强调了防止 LPMO 失活及其对反应中所有酶的潜在不利影响的重要性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Enhancing enzymatic saccharification yields of cellulose at high solid loadings by combining different LPMO activities

Background

The polysaccharides in lignocellulosic biomass hold potential for production of biofuels and biochemicals. However, achieving efficient conversion of this resource into fermentable sugars faces challenges, especially when operating at industrially relevant high solid loadings. While it is clear that combining classical hydrolytic enzymes and lytic polysaccharide monooxygenases (LPMOs) is necessary to achieve high saccharification yields, exactly how these enzymes synergize at high solid loadings remains unclear.

Results

An LPMO-poor cellulase cocktail, Celluclast 1.5 L, was spiked with one or both of two fungal LPMOs from Thermothielavioides terrestris and Thermoascus aurantiacus, TtAA9E and TaAA9A, respectively, to assess their impact on cellulose saccharification efficiency at high dry matter loading, using Avicel and steam-exploded wheat straw as substrates. The results demonstrate that LPMOs can mitigate the reduction in saccharification efficiency associated with high dry matter contents. The positive effect of LPMO inclusion depends on the type of feedstock and the type of LPMO and increases with the increasing dry matter content and reaction time. Furthermore, our results show that chelating free copper, which may leak out of the active site of inactivated LPMOs during saccharification, with EDTA prevents side reactions with in situ generated H2O2 and the reductant (ascorbic acid).

Conclusions

This study shows that sustaining LPMO activity is vital for efficient cellulose solubilization at high substrate loadings. LPMO cleavage of cellulose at high dry matter loadings results in new chain ends and thus increased water accessibility leading to decrystallization of the substrate, all factors making the substrate more accessible to cellulase action. Additionally, this work highlights the importance of preventing LPMO inactivation and its potential detrimental impact on all enzymes in the reaction.

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
Biotechnology for Biofuels
Biotechnology for Biofuels 工程技术-生物工程与应用微生物
自引率
0.00%
发文量
0
审稿时长
2.7 months
期刊介绍: Biotechnology for Biofuels is an open access peer-reviewed journal featuring high-quality studies describing technological and operational advances in the production of biofuels, chemicals and other bioproducts. The journal emphasizes understanding and advancing the application of biotechnology and synergistic operations to improve plants and biological conversion systems for the biological production of these products from biomass, intermediates derived from biomass, or CO2, as well as upstream or downstream operations that are integral to biological conversion of biomass. Biotechnology for Biofuels focuses on the following areas: • Development of terrestrial plant feedstocks • Development of algal feedstocks • Biomass pretreatment, fractionation and extraction for biological conversion • Enzyme engineering, production and analysis • Bacterial genetics, physiology and metabolic engineering • Fungal/yeast genetics, physiology and metabolic engineering • Fermentation, biocatalytic conversion and reaction dynamics • Biological production of chemicals and bioproducts from biomass • Anaerobic digestion, biohydrogen and bioelectricity • Bioprocess integration, techno-economic analysis, modelling and policy • Life cycle assessment and environmental impact analysis
期刊最新文献
Metabolic and tolerance engineering of Komagataella phaffii for 2-phenylethanol production through genome-wide scanning Transcriptomic and metabolic changes in Trichoderma reesei caused by mutation in xylanase regulator 1 (xyr1) An integrated QTL and RNA-seq analysis revealed new petal morphology loci in Brassica napus L. Enzymatic hydrolysis of waste streams originating from wastewater treatment plants Balancing pH and yield: exploring itaconic acid production in Ustilago cynodontis from an economic perspective
×
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
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 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 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1