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SUCCINIC ACID PRODUCTION FROM PULP AND PAPER INDUSTRY WASTE - A TRANSCRIPTOMIC APPROACH

机译:浆料和造纸工业废弃物的琥珀酸生产 - 转录组方法

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The utilization of renewable resources for the production of bio-based products is essential in order to develop sustainable bioprocesses and biorefineries. Xylose-rich hydrolysates produced from hemicelluloses contained in lignocellulosic resources could be used for the production of succinic acid, one of the most important platform chemicals in the bio-economy era. Exploitation of spent sulphite liquor (SSL), a xylose rich by-product from pulp and paper industry has been previously investigated for succinic acid production by Actinobacillus succinogenes, one of the most efficient natural succinic acid producers. In this study, the metabolic potential of this strain was evaluated through the RNA expression of the metabolic pathways involved in succinic acid production utilizing glucose, xylose or SSL as carbon sources. A transcriptomic approach of the key enzymes of glucose and xylose catabolism, carboxylic acid production as well as oxidative phosphorylation led to an improved understanding on the energy consuming metabolic pathways. The transcriptomic analysis was carried out in batch cultures. A cDNA library was constructed at different phases of the fermentation where major metabolic changes in extracellular metabolites or biomass production were observed. Real time PCR was used to determine the expression levels of the genes of interest throughout the fermentation. The bottlenecks of the fermentative production of succinic acid by A. succinogenes were addressed with particular focus on the effect of glucose and xylose catabolism on pathways that involve ATP consumption and NADH oxidation. All subunits of ATP synthase were highly expressed in all substrates. In particular ATP synthase FO (ATP SYN FO) was higher expressed when glucose was the only carbon source. Phosphoenol-pyruvate carboxykinase (PEPCK) expression was delayed when xylose was present in the medium. Despite the fact that extracellular lactic acid was not detected, low expression levels of lactic acid dehydrogenase (L.
机译:利用可再生资源生产基于生物的产品,以开发可持续的生物处理和生物料理至关重要。由木质纤维素资源中包含的半纤维素产生的木糖富水解产物可用于生产琥珀酸,是生物经济时代最重要的平台化学品之一。对废硫酸盐液(SSL)的剥削,富含纸浆和造纸工业的木糖富含副产物,已经研究了Actinobacillus Suctinogenes的琥珀酸产生,其中一种最有效的天然琥珀酸生产商之一。在该研究中,通过利用葡萄糖,木糖或SSL作为碳源的琥珀酸产生的代谢途径的RNA表达评估该菌株的代谢潜力。葡萄糖和木糖分解代谢的关键酶的转录组种方法,羧酸产生以及氧化磷酸化导致对能量消耗代谢途径的改进了解。转录组分析在分批培养物中进行。在发酵的不同阶段构建cDNA文库,观察到细胞外代谢物或生物质产生的主要代谢变化。实时PCR用于确定整个发酵过程中利益的表达水平。通过A琥珀酸的发酵产生的琥珀酸产生的瓶颈特别关注葡萄糖和木糖分解代谢对涉及ATP消耗和NADH氧化的途径的影响。 ATP合酶的所有亚基在所有基材中高度表达。特别地,当葡萄糖是唯一的碳源时,特别表达ATP合酶FO(ATP SYN FO)。当在培养基中存在木糖时,磷烯醇 - 丙酮酸羧酮糖酶(PEPCK)表达延迟。尽管未检测到细胞外乳酸,但乳酸脱氢酶的表达水平低(L.

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