Learning from marine wood borers; enzymes and mechanisms of lignocellulose digestion

向海洋蛀木虫学习；

• 批准号: BB/L001926/1
• 负责人: Simon McQueen-Mason
• 金额: $ 359.35万
• 依托单位: University of York
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FL001926%2F1
• 关键词: Learning; marine; wood; borers; enzymes

The development of sustainable biofuels based on woody (lignocellulosic) plant biomass is generally seen as an essential element in the move to a low carbon economy within the constraints of food security. The key to unlock this potential lies in developing technologies for the cost effective deconstruction of lignocellulose into sugars for fermentation. A number of animals have evolved to live on the large amounts of woody biomass that enter the marine environment from river estuaries. Such animals may provide useful understanding and enzymes for lignocellulose deconstruction, as they thrive on a diet of wood, and digest it under ambient temperatures and pressures. The project builds on an existing programme funded as part of the BBSRC Sustainable Bioenergy Centre that has focused on Limnoria quadripunctata, a wood borer, which is unusual in having a digestive tract free of microbial life. Our studies indicate that this animal has a gut with an inert chitinous lining in which free radical chemistry is deployed as a pretreatment for enzymatic hydrolysis, and in which enzymes are produced in a separate organ and transposed to the gut, which functions as an enzyme reactor. In the programme we have established a wide range of specialist capabilities in the area of producing and characterising recombinant secreted Eukaryotic enzymes, as well as methods for studying digestive processes in vivo.Building from this basis of knowledge, skills and resources, we wish to expand the work to take in a wider range of wood-eating marine invertebrates. Our initial understanding was that the microbe-free gut was peculiar to the Limnoriids, but recent studies at Portsmouth show a similar situation in a marine amphipod, Chelura telebrans. We have now obtained substantial EST databases from the digestive tracts of three arthropods, two Limnoriids (L. quadripunctata and L. lignorum), and C. telebrans. Initial scrutiny reveals some broad similarities in the suite of obvious digestive genes expressed in the three, but also considerable divergence in the range and relative expression levels of these genes. We will undertake detailed comparative gut and hepatopancreas transcriptomic and proteomic studies to identify the range of proteins involved in digestion and also study the biochemical processes in the gut. C. terebrans has the advantage of being considerably larger than the Limnoriid species, which will make these studies easier to undertake.In addition to crustacean wood borers, we also propose to initiate studies in Lyrodus pedicellatus, a representative of the shipworms. Shipworms are large bivalve molluscs with little relationship to the arthropods. The bivalve shells of shipworms have evolved into a highly effective rasp at the anterior of the animal, with which they bore through wood, consuming particles as they go. These animals have a complex and poorly understood digestive that includes a large chamber where wood particles are stored and a region where wood particles are phagocytosed indicating intracellular digestion. In addition, shipworms harbour endo-symbiotic bacteria in their gills that have been suggested to aid in digestion, and have intestinal microflora that could also be involved. We will investigate genes and enzymes expressed, and biochemical processes in different regions of the shipworm digestive system in order to identify new enzymes for study. We will produce recombinant forms of hydrolytic and oxidative enzymes involved in wood digestion and characterise these in detail in terms of structure and activity. We will work closely with industry to examine the commercial potential of new enzymes and pretreatment approaches developed in the project.

基于木质（木质纤维素）植物生物量的可持续生物燃料的发展通常被视为在粮食安全限制内转移到低碳经济的基本要素。释放这种潜力的关键在于开发木质纤维素成本有效解构的技术，以进行发酵。许多动物已经演变为从河流河口进入海洋环境的大量木质生物质上。这样的动物可以为木质纤维素解构提供有用的理解和酶，因为它们在木材的饮食中壮成长，并在环境温度和压力下消化。该项目建立在一个现有的计划基础上，该计划是BBSRC可持续生物能源中心的一部分，该计划的重点是木毛毛毛龙骨Quadripunctata，这对于拥有没有微生物寿命的消化道是不寻常的。我们的研究表明，该动物的肠道具有惰性齿状衬里，其中将自由基化学作用作为酶水解的预处理，并且在单独的器官中生产酶并将其转移到肠道上，该酶充当酶反应器。在该计划中，我们在生产和表征重组分泌的真核生物酶的领域建立了广​​泛的专业能力，以及研究体内消化过程的方法。从这一知识，技能和资源的基础上构建消化过程，我们希望将工作扩展到较宽的木质范围内，以扩大固定范围内的固有属于固有的固有属于固有的海上。我们最初的理解是，不含微生物的肠道是Limnoriids的特殊肠道，但是朴次茅斯的最近研究表明，海洋两领域的Chelura Telebrans也有类似的情况。现在，我们从三个节肢动物，两个limnoriids（L. Quadripunctata和L. lignorum）和C. telebrans的消化道中获得了大量数据库。最初的审查揭示了在这三个中表达的明显消化基因套件中的一些广泛相似性，但在这些基因的范围和相对表达水平上也有很大的差异。我们将进行详细的比较肠道和肝癌转录组和蛋白质组学研究，以鉴定肠道中涉及的蛋白质范围，并研究肠道中的生化过程。 C. terebrans具有比Limnoriid物种大得多的优势，这将使这些研究更容易进行。除了甲壳类动物的鲍尔（Crustacean Wood Borers）外，我们还建议在船虫的代表中启动Pedicellatus的研究。虫子是大的双壳类软体动物，与节肢动物几乎没有关系。船虫的双壳类壳已演变成动物前部的高效rasp，它们通过木材散发出来，随着木材而散发出颗粒。这些动物具有复杂且知之甚少的消化率，其中包括一个大室，其中存储了木颗粒和一个木颗粒被吞噬的区域，表明细胞内消化。此外，虫子携带的g中含有有助于消化的g，并具有也可能涉及的肠道微生物。我们将研究所表达的基因和酶，以及在虫虫消化系统不同地区的生化过程，以鉴定新的酶进行研究。我们将产生参与木材消化的水解和氧化酶的重组形式，并在结构和活性方面详细介绍这些酶。我们将与行业紧密合作，研究项目中开发的新酶和预处理方法的商业潜力。

项目成果

MOESM6 of Discovery, activity and characterisation of an AA10 lytic polysaccharide oxygenase from the shipworm symbiont Teredinibacter turnerae

MOESM6 来自船虫共生体 Teredinibacter Turnerae 的 AA10 裂解多糖加氧酶的发现、活性和表征

• DOI: 10.6084/m9.figshare.9923507
• 发表时间: 2019
• 作者: Claire. Fowler

Supercritical extraction as an effective first-step in a maize stover biorefinery

• DOI: 10.1039/c5ra07485a
• 发表时间: 2015-01-01
• 期刊: RSC ADVANCES
• 影响因子: 3.9
• 作者: Attard, Thomas M.;Theeuwes, Elke;Hunt, Andrew J.
• 通讯作者: Hunt, Andrew J.

Bracing copper for the catalytic oxidation of C-H bonds

• DOI: 10.1038/s41929-018-0110-9
• 发表时间: 2018-08-01
• 期刊: NATURE CATALYSIS
• 影响因子: 37.8
• 作者: Ciano, Luisa;Davies, Gideon J.;Walton, Paul H.
• 通讯作者: Walton, Paul H.

Hemocyanin facilitates lignocellulose digestion by wood-boring marine crustaceans.

• DOI: 10.1038/s41467-018-07575-2
• 发表时间: 2018-12-03
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Besser K;Malyon GP;Eborall WS;Paro da Cunha G;Filgueiras JG;Dowle A;Cruz Garcia L;Page SJ;Dupree R;Kern M;Gomez LD;Li Y;Elias L;Sabbadin F;Mohamad SE;Pesante G;Steele-King C;Ribeiro de Azevedo E;Polikarpov I;Dupree P;Cragg SM;Bruce NC;McQueen-Mason SJ
• 通讯作者: McQueen-Mason SJ

Defining functional diversity for lignocellulose degradation in a microbial community using multi-omics studies.

• DOI: 10.1186/s13068-018-1164-2
• 发表时间: 2018
• 期刊: Biotechnology for biofuels
• 影响因子: 6.3
• 作者: Alessi AM;Bird SM;Oates NC;Li Y;Dowle AA;Novotny EH;deAzevedo ER;Bennett JP;Polikarpov I;Young JPW;McQueen-Mason SJ;Bruce NC
• 通讯作者: Bruce NC