Brasseur Catherine, Bauwens Julien, Tarayre Cédric, Mattéotti Christel, Thonart Philippe, Destain Jacqueline, Francis Frédéric, Haubruge Eric, Portetelle Daniel, Vandenbol Micheline, Focant Jean-François, De Pauw Edwin
Mass Spectrometry Laboratory, Department of Chemistry, University of Liège, Allée du Six Aout, B6c, Sart-Tilman, B-4000 Liège, Belgium.
Department of Functional and Evolutionary Entomology, Gembloux Agro-Bio Tech, University of Liège, 2 Passage des Déportés, B-5030 Gembloux, Belgium.
Molecules. 2014 Apr 11;19(4):4578-94. doi: 10.3390/molecules19044578.
Hindgut homogenates of the termite Reticulitermes santonensis were incubated with carboxymethyl cellulose (CMC), crystalline celluloses or xylan substrates. Hydrolysates were analyzed with matrix-assisted laser desorption/ionization coupled to time-of-flight mass spectrometry (MALDI-TOF MS). The method was first set up using acid hydrolysis analysis to characterize non-enzymatic profiles. Commercial enzymes of Trichoderma reesei or T. longibrachiatum were also tested to validate the enzymatic hydrolysis analysis. For CMC hydrolysis, data processing and visual display were optimized to obtain comprehensive profiles and allow rapid comparison and evaluation of enzymatic selectivity, according to the number of substituents of each hydrolysis product. Oligosaccharides with degrees of polymerization (DPs) ranging from three to 12 were measured from CMC and the enzymatic selectivity was demonstrated. Neutral and acidic xylo-oligosaccharides with DPs ranging from three to 11 were measured from xylan substrate. These results are of interest for lignocellulose biomass valorization and demonstrated the potential of termites and their symbiotic microbiota as a source of interesting enzymes for oligosaccharides production.
将白蚁桑氏散白蚁(Reticulitermes santonensis)的后肠匀浆与羧甲基纤维素(CMC)、结晶纤维素或木聚糖底物一起孵育。水解产物用基质辅助激光解吸/电离与飞行时间质谱联用(MALDI-TOF MS)进行分析。该方法首先通过酸水解分析来表征非酶解图谱进行建立。还测试了里氏木霉(Trichoderma reesei)或长枝木霉(T. longibrachiatum)的商业酶,以验证酶水解分析。对于CMC水解,根据每个水解产物的取代基数,对数据处理和可视化显示进行了优化,以获得全面的图谱,并能够快速比较和评估酶的选择性。从CMC中测量了聚合度(DPs)为3至12的寡糖,并证明了酶的选择性。从木聚糖底物中测量了DPs为3至11的中性和酸性木寡糖。这些结果对于木质纤维素生物质的增值具有重要意义,并证明了白蚁及其共生微生物群作为生产寡糖的有趣酶来源的潜力。