Functioning of a tripartite lignocellulolytic microbial consortium cultivated under two shaking conditions: a metatranscriptomic study.

Yanfang Wang, Diego Javier Jiménez, Zhenhua Zhang, Jan Dirk van Elsas
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Abstract

Background: In a previous study, shaking speed was found to be an important factor affecting the population dynamics and lignocellulose-degrading activities of a synthetic lignocellulolytic microbial consortium composed of the bacteria Sphingobacterium paramultivorum w15, Citrobacter freundii so4, and the fungus Coniochaeta sp. 2T2.1. Here, the gene expression profiles of each strain in this consortium were examined after growth at two shaking speeds (180 and 60 rpm) at three time points (1, 5 and 13 days).

Results: The results indicated that, at 60 rpm, C. freundii so4 switched, to a large extent, from aerobic to flexible (aerobic/microaerophilic/anaerobic) metabolism, resulting in continued slow growth till late stage. In addition, Coniochaeta sp. 2T2.1 tended to occur to a larger extent in the hyphal form, with genes encoding adhesion proteins being highly expressed. Much like at 180 rpm, at 60 rpm, S. paramultivorum w15 and Coniochaeta sp. 2T2.1 were key players in hemicellulose degradation processes, as evidenced from the respective CAZy-specific transcripts. Coniochaeta sp. 2T2.1 exhibited expression of genes encoding arabinoxylan-degrading enzymes (i.e., of CAZy groups GH10, GH11, CE1, CE5 and GH43), whereas, at 180 rpm, some of these genes were suppressed at early stages of growth. Moreover, C. freundii so4 stably expressed genes that were predicted to encode proteins with (1) β-xylosidase/β-glucosidase and (2) peptidoglycan/chitinase activities, (3) stress response- and detoxification-related proteins. Finally, S. paramultivorum w15 showed involvement in vitamin B2 generation in the early stages across the two shaking speeds, while this role was taken over by C. freundii so4 at late stage at 60 rpm.

Conclusions: We provide evidence that S. paramultivorum w15 is involved in the degradation of mainly hemicellulose and in vitamin B2 production, and C. freundii so4 in the degradation of oligosaccharides or sugar dimers, next to detoxification processes. Coniochaeta sp. 2T2.1 was held to be strongly involved in cellulose and xylan (at early stages), next to lignin modification processes (at later stages). The synergism and alternative functional roles presented in this study enhance the eco-enzymological understanding of the degradation of lignocellulose in this tripartite microbial consortium.

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在两种摇动条件下培养的三方木质纤维素分解微生物联合体的功能:一项转写组学研究。
背景:在之前的一项研究中发现,摇动速度是影响由副多动鞘杆菌w15、弗氏柠檬酸杆菌so4和真菌Coniochaeta sp. 2T2.1组成的合成木质纤维素降解微生物群落种群动态和降解木质纤维素活性的重要因素。在这里,我们检测了该联盟中每个菌株在三个时间点(1、5和13天)以两种摇动速度(180和60 rpm)生长后的基因表达谱。结果:结果表明,在60 rpm时,弗氏梭菌so4在很大程度上由好氧代谢切换到柔性(好氧/微氧/厌氧)代谢,导致生长持续缓慢直至后期。此外,Coniochaeta sp. 2T2.1更倾向于以菌丝形式出现,编码粘附蛋白的基因高表达。从cazy特异性转录本中可以发现,在180 rpm和60 rpm时,S. paramultivorum w15和Coniochaeta sp. 2T2.1在半纤维素降解过程中起关键作用。Coniochaeta sp. 2T2.1表达了编码阿拉伯木聚糖降解酶的基因(即CAZy组GH10, GH11, CE1, CE5和GH43),而在180 rpm时,这些基因中的一些在生长早期被抑制。此外,弗氏线虫so4稳定表达的基因被预测编码具有(1)β-木糖苷酶/β-葡萄糖苷酶和(2)肽聚糖/几丁质酶活性的蛋白,(3)应激反应和解毒相关蛋白。最后,副多动弧菌w15在两种摇速下的早期阶段参与了维生素B2的生成,而这一作用在摇速为60 rpm的后期阶段被弗氏弧菌so4取代。结论:我们提供的证据表明,副多胞杆菌w15主要参与半纤维素的降解和维生素B2的产生,而C. freundii so4则参与低聚糖或糖二聚体的降解,仅次于解毒过程。Coniochaeta sp. 2T2.1被认为强烈参与纤维素和木聚糖(早期阶段),其次是木质素修饰过程(后期阶段)。本研究中提出的协同作用和替代功能作用增强了对这三方微生物联盟中木质纤维素降解的生态酶学理解。
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