Kracher Daniel, Oros Damir, Yao Wanying, Preims Marita, Rezic Iva, Haltrich Dietmar, Rezic Tonci, Ludwig Roland
Department of Food Sciences and Technology, Food Biotechnology Laboratory, BOKU - University of Natural Resources and Life Sciences, Vienna, Austria.
Biotechnol J. 2014 Apr;9(4):483-92. doi: 10.1002/biot.201300214. Epub 2014 Mar 7.
The recalcitrance of lignocellulose makes enzymatic hydrolysis of plant biomass for the production of second generation biofuels a major challenge. This work investigates an efficient and economic approach for the enzymatic hydrolysis of sugar beet pulp (SBP), which is a difficult to degrade, hemicellulose-rich by-product of the table sugar industry. Three fungal strains were grown on different substrates and the production of various extracellular hydrolytic and oxidative enzymes involved in pectin, hemicellulose, and cellulose breakdown were monitored. In a second step, the ability of the culture supernatants to hydrolyze thermally pretreated SBP was tested in batch experiments. The supernatant of Sclerotium rolfsii, a soil-borne facultative plant pathogen, was found to have the highest hydrolytic activity on SBP and was selected for further hydrolyzation experiments. A low enzyme load of 0.2 mg g(-1) protein from the culture supernatant was sufficient to hydrolyze a large fraction of the pectin and hemicelluloses present in SBP. The addition of Trichoderma reesei cellulase (1-17.5 mg g(-1) SBP) resulted in almost complete hydrolyzation of cellulose. It was found that the combination of pectinolytic, hemicellulolytic, and cellulolytic activities works synergistically on the complex SBP composite, and a combination of these hydrolytic enzymes is required to achieve a high degree of enzymatic SBP hydrolysis with a low enzyme load.
木质纤维素的顽固性使得通过酶促水解植物生物质来生产第二代生物燃料成为一项重大挑战。这项工作研究了一种高效且经济的方法用于甜菜粕(SBP)的酶促水解,甜菜粕是制糖工业中一种难以降解且富含半纤维素的副产品。三种真菌菌株在不同底物上生长,并监测参与果胶、半纤维素和纤维素分解的各种细胞外水解酶和氧化酶的产生。在第二步中,在分批实验中测试了培养上清液水解热预处理SBP的能力。发现一种土壤传播的兼性植物病原菌齐整小核菌的上清液对SBP具有最高的水解活性,并被选用于进一步的水解实验。来自培养上清液的低酶负载量为0.2 mg g(-1)蛋白质就足以水解SBP中大部分的果胶和半纤维素。添加里氏木霉纤维素酶(1 - 17.5 mg g(-1) SBP)导致纤维素几乎完全水解。研究发现,果胶分解、半纤维素分解和纤维素分解活性在复杂的SBP复合物上协同作用,并且需要这些水解酶的组合才能以低酶负载实现高度的SBP酶促水解。