Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot, Israel.
Appl Environ Microbiol. 2013 Sep;79(17):5242-9. doi: 10.1128/AEM.01211-13. Epub 2013 Jun 28.
Lactobacillus plantarum is an attractive candidate for bioprocessing of lignocellulosic biomass due to its high metabolic variability, including its ability to ferment both pentoses and hexoses, as well as its high acid tolerance, a quality often utilized in industrial processes. This bacterium grows naturally on biomass; however, it lacks the inherent ability to deconstruct lignocellulosic substrates. As a first step toward engineering lignocellulose-converting lactobacilli, we have introduced genes coding for a GH6 cellulase and a GH11 xylanase from a highly active cellulolytic bacterium into L. plantarum. For this purpose, we employed the recently developed pSIP vectors for efficient secretion of heterologous proteins. Both enzymes were secreted by L. plantarum at levels estimated at 0.33 nM and 3.3 nM, for the cellulase and xylanase, respectively, in culture at an optical density at 600 nm (OD600) of 1. Transformed cells demonstrated the ability to degrade individually either cellulose or xylan and wheat straw. When mixed together to form a two-strain cell-based consortium secreting both cellulase and xylanase, they exhibited synergistic activity in the overall release of soluble sugar from wheat straw. This result paves the way toward metabolic harnessing of L. plantarum for novel biorefining applications, such as production of ethanol and polylactic acid directly from plant biomass.
植物乳杆菌因其高度的代谢多样性而成为木质纤维素生物加工的有吸引力的候选者,包括其能够发酵戊糖和己糖的能力,以及其耐酸能力,这一特性通常在工业过程中得到利用。这种细菌自然生长在生物质上;然而,它缺乏分解木质纤维素底物的固有能力。作为工程木质纤维素转化乳杆菌的第一步,我们已经将来自高度活跃的纤维素分解菌的 GH6 纤维素酶和 GH11 木聚糖酶的基因引入到植物乳杆菌中。为此,我们采用了最近开发的 pSIP 载体,用于高效分泌异源蛋白。在 OD600 为 1 的培养物中,这两种酶分别以估计为 0.33 nM 和 3.3 nM 的水平分泌,纤维素酶和木聚糖酶的水平分别为 0.33 nM 和 3.3 nM。转化细胞表现出单独降解纤维素或木聚糖和小麦秸秆的能力。当混合在一起形成分泌纤维素酶和木聚糖酶的两菌株基于细胞的联合体时,它们在从小麦秸秆中整体释放可溶性糖方面表现出协同活性。这一结果为利用植物乳杆菌进行新型生物精炼应用铺平了道路,例如直接从植物生物质生产乙醇和聚乳酸。