Munir Riffat I, Spicer Victor, Krokhin Oleg V, Shamshurin Dmitry, Zhang XiangLi, Taillefer Marcel, Blunt Warren, Cicek Nazim, Sparling Richard, Levin David B
Department of Biosystems Engineering, University of Manitoba, R3T 5N6, Winnipeg, MB, Canada.
Department of Physics and Astronomy, University of Manitoba, R3T 5N6, Winnipeg, MB, Canada.
BMC Microbiol. 2016 May 23;16:91. doi: 10.1186/s12866-016-0711-x.
Clostridium termitidis CT1112 is an anaerobic, Gram-positive, mesophilic, spore-forming, cellulolytic bacterium, originally isolated from the gut of a wood feeding termite Nasusitermes lujae. It has the ability to hydrolyze both cellulose and hemicellulose, and ferment the degradation products to acetate, formate, ethanol, lactate, H2, and CO2. It is therefore ges in gene and gene product expression during growth of C. termitidis on cellobiose, xylose, xylan, and α-cellulose.
Correlation of transcriptome and proteome data with growth and fermentation profiles identified putative carbon-catabolism pathways in C. termitidis. The majority of the proteins associated with central metabolism were detected in high abundance. While major differences were not observed in gene and gene-product expression for enzymes associated with metabolic pathways under the different substrate conditions, xylulokinase and xylose isomerase of the pentose phosphate pathway were found to be highly up-regulated on five carbon sugars compared to hexoses. In addition, genes and gene-products associated with a variety of cellulosome and non-cellulosome associated CAZymes were found to be differentially expressed. Specifically, genes for cellulosomal enzymes and components were highly expressed on α-cellulose, while xylanases and glucosidases were up-regulated on 5 carbon sugars with respect to cellobiose. Chitinase and cellobiophosphorylases were the predominant CAZymes expressed on cellobiose. In addition to growth on xylan, the simultaneous consumption of two important lignocellulose constituents, cellobiose and xylose was also demonstrated.
There are little changes in core-metabolic pathways under the different carbon sources compared. The most significant differences were found to be associated with the CAZymes, as well as specific up regulation of some key components of the pentose phosphate pathway in the presence of xylose and xylan. This study has enhanced our understanding of the physiology and metabolism of C. termitidis, and provides a foundation for future studies on metabolic engineering to optimize biofuel production from natural biomass.
白蚁梭菌CT1112是一种厌氧、革兰氏阳性、嗜温、产芽孢、可分解纤维素的细菌,最初从以木材为食的白蚁卢氏鼻白蚁的肠道中分离得到。它具有水解纤维素和半纤维素的能力,并将降解产物发酵为乙酸、甲酸、乙醇、乳酸、氢气和二氧化碳。因此,研究了白蚁梭菌在纤维二糖、木糖、木聚糖和α-纤维素上生长过程中的基因和基因产物表达情况。
转录组和蛋白质组数据与生长及发酵谱的相关性确定了白蚁梭菌中假定的碳分解代谢途径。与中心代谢相关的大多数蛋白质被检测到高丰度表达。虽然在不同底物条件下,与代谢途径相关的酶的基因和基因产物表达未观察到主要差异,但戊糖磷酸途径的木酮糖激酶和木糖异构酶在五碳糖上的表达相比六碳糖被发现高度上调。此外,发现与多种纤维素体和非纤维素体相关的碳水化合物活性酶(CAZyme)的基因和基因产物存在差异表达。具体而言,纤维素体酶和组分的基因在α-纤维素上高度表达,而木聚糖酶和葡糖苷酶在五碳糖上相对于纤维二糖上调表达。几丁质酶和纤维二糖磷酸化酶是在纤维二糖上表达的主要CAZyme。除了在木聚糖上生长外,还证明了能同时消耗两种重要的木质纤维素成分,即纤维二糖和木糖。
相比之下,不同碳源下核心代谢途径变化不大。发现最显著的差异与CAZyme有关,以及在木糖和木聚糖存在时戊糖磷酸途径的一些关键组分的特异性上调。本研究增进了我们对白蚁梭菌生理学和代谢的理解,并为未来代谢工程研究优化从天然生物质生产生物燃料奠定了基础。