HPLC结合MALDI-TOF-MS分析木聚糖水解产物

doi:10.3969/j.issn.0253-2417.2015.05.003

林产化学与工业 ›› 2015, Vol. 35 ›› Issue (5): 10-14.doi: 10.3969/j.issn.0253-2417.2015.05.003

HPLC结合MALDI-TOF-MS分析木聚糖水解产物

黎海龙¹, 李成国¹, 秦光辉¹, 刘健¹, 甘礼惠¹, 谢茹胜², 龙敏南¹

1. 厦门大学能源学院, 福建厦门 361102;
2. 福建生物工程职业技术学院药学系, 福建福州 350002

收稿日期:2014-09-25 出版日期:2015-10-25 发布日期:2015-10-24
通讯作者: 龙敏南(1965—),男,教授,博士,博士生导师,从事生物质转化制生物基燃料与生物基化学品的研究;E-mail:longmn@xmu.edu.cn E-mail:longmn@xmu.edu.cn
作者简介:黎海龙 (1988—),男,湖南耒阳人,博士生,主要从事生物质转化制生物基燃料与生物基化学品的研究;E-mail:longtimegogo@126.com
基金资助:
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)

HPLC and MALDI-TOF-MS Analysis of Hydrolysates of Xylan

LI Hai-long¹, LI Cheng-guo¹, QIN Guang-hui¹, LIU Jian¹, GAN Li-hui¹, XIE Ru-sheng², LONG Min-nan¹

1. College of Energy, Xiamen University, Xiamen 361102, China;
2. Department of Pharmacy, Fujian Vocational College of Bio-engineering, Fuzhou 350002, China

Received:2014-09-25 Online:2015-10-25 Published:2015-10-24

摘要/Abstract

摘要： 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AnXyn10C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HoXyn11A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。

关键词: 1-苯基-3-甲基-5-吡唑啉酮, 低聚木糖, 4-O-甲基葡萄糖醛酸, MALDI-TOF-MS, HPLC

Abstract: 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 MALDI-TOF-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-O-methylglucuronic acid (B₁) in the hydrolysates of beechwood xylan by sulfuric acid. Recombinant endo-β-1,4-xylanase AnXyn10C 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 HoXyn11A released xylose, xylobiose, xylotriose, 4-O-methyl-glucuronic acid-xylotetrose (B₄) and -xylopentaose (B₅), and aldohexaouronic acid. These results revealed that HPLC assisted with MALDI-TOF-MS based on PMP derivatization was a very useful and robust method for the determination of products in hydrolysis of xylan.

Key words: 1-phenyl-3-methyl-5-pyrazolone, xylo-oligosaccharides, 4-O-methylglucuronic acid, MALDI-TOF-MS, HPLC

中图分类号:

TQ35

黎海龙, 李成国, 秦光辉, 刘健, 甘礼惠, 谢茹胜, 龙敏南. HPLC结合MALDI-TOF-MS分析木聚糖水解产物[J]. 林产化学与工业, 2015, 35(5): 10-14.

LI Hai-long, LI Cheng-guo, QIN Guang-hui, LIU Jian, GAN Li-hui, XIE Ru-sheng, LONG Min-nan. HPLC and MALDI-TOF-MS Analysis of Hydrolysates of Xylan[J]. Chemistry and Industry of Forest Products, 2015, 35(5): 10-14.

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HPLC结合MALDI-TOF-MS分析木聚糖水解产物

HPLC and MALDI-TOF-MS Analysis of Hydrolysates of Xylan

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