利用HPlC结合MAldI-TOf-MS对1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化后山毛榉木聚糖水解产物进行分析,检测到了难以获得标准品对照的木聚糖水解产物。结果表明,稀硫酸水解山毛榉木聚糖的主要水解产物有木糖和4-O-甲基葡萄糖醛酸-木糖(b2),以及少量4-O-甲基葡萄糖醛酸(b1)。内切重组木聚糖酶An Xyn10C水解山毛榉木聚糖产生木糖、木二糖和4-O-甲基葡萄糖醛酸-木三糖(b3),而内切重组木聚糖酶HO Xyn11A水解山毛榉木聚糖主要产生木糖、木二糖、木三糖、4-O-甲基葡萄糖醛酸-木四糖(b4)和4-O-甲基葡萄糖醛酸-木五糖(b5)。基于PMP柱前衍生化的HPlC结合MAldI-TOf-MS方法能高效地分析复杂的木聚糖水解产物。The hydrolysis end products of beechwood xylan,which were released by sulfuric acid or enzymes and then labeled at their reducing ends with 1-phenyl-3-methyl-5-pyrazolone( PMP) derivatization,were analyzed by HPLC assisted with MALDITof-MS.Some of the hydrolysates,which were lack of related commercial available standard substances,were determined.It was found that the xylose and 4-O-methyl-glucuronic acid-xylose( B2) were the main products with minor amounts of 4-Omethylglucuronic acid( B1) in the hydrolysates of beechwood xylan by sulfuric acid.Recombinant endo-β-1,4-xylanase An Xyn10 C released xylose,xylobiose,and 4-O-methyl-glucuronic acid-xylotriose( B3) as the main hydrolysates from beechwood xylan,whereas recombinant endo-β-1,4-xylanase Ho Xyn11 A released xylose,xylobiose,xylotriose,4-O-methyl-glucuronic acidxylotetrose( B4) and-xylopentaose( B5),and aldohexaouronic acid.These results revealed that HPLC assisted with MALDITof-MS based on PMP derivatization was a very useful and robust method for the determination of products in hydrolysis of xylan.国家自然科学基金资助项目(31170067); 厦门市海洋经济发展专项资金项目(14GZP59HJ29); 福建省海洋高新产业发展专项项目(闽海洋高新[2014]25号
