双水相木聚糖酶水解体系制备木二糖及其纯化

doi:10.3969/j.issn.0253-2417.2015.05.015

林产化学与工业 ›› 2015, Vol. 35 ›› Issue (5): 93-97.doi: 10.3969/j.issn.0253-2417.2015.05.015

双水相木聚糖酶水解体系制备木二糖及其纯化

李鑫, 顾夕梅, 绳敖楠, 张丽, 陈晓佩, 勇强

南京林业大学化学工程学院, 江苏南京 210037

收稿日期:2014-10-27 出版日期:2015-10-25 发布日期:2015-10-24
作者简介:李鑫(1975—),男,辽宁大连人,副教授,博士,从事木质纤维原料生物催化与生物转化的研究;E-mail:xli@njfu.edu.cn
基金资助:
江苏省高校自然科学研究重大项目(14KJA220003);江苏省自然科学基金项目(BK20131426);江苏高校优势学科建设工程项目资助项目(无编号)

Preparation of Xylobiose by Xylanase Hydrolysis in Aqueous Two-phase System and Its Purification

LI Xin, GU Xi-mei, SHENG Ao-nan, ZHANG Li, CHEN Xiao-pei, YONG Qiang

College of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, China

Received:2014-10-27 Online:2015-10-25 Published:2015-10-24

摘要/Abstract

摘要： 采用聚乙二醇6000/柠檬酸钠的双水相木聚糖酶水解体系制备木二糖,以10 g/L桦木木聚糖为底物,木聚糖酶的酶用量为20 IU/g(以木聚糖质量计),在50℃、pH值5.6条件下,水解8 h制备木二糖,木二糖的酶解得率为13.8%。双水相水解体系对木聚糖酶回收率为66.4%,对木二糖回收率为87.2%;采用凝胶层析纯化方法,以葡聚糖凝胶LH-20为分离介质,脱气超纯水为洗脱剂,在柱温35 ℃、洗脱溶剂流速0.1 mL/min、上样量0.3 mL条件下,分离纯化木二糖,木二糖纯度为98.5%。

关键词: 木聚糖酶, 双水相体系, 酶水解, 凝胶层析, 木二糖

Abstract: Xylobiose was prepared by xylanase hydrolysis of birchwood xylan in the PEG6000/sodium citrate aqueous two-phase system (ATPS). The optimized conditions for enzymatic hydrolysis in ATPS were birchwood xylan 10 g/L, xylanase 20 IU/g (calculated by the mass of xylan), 50 ℃ and pH 5.6. The enzymatic hydrolysis yield of xylobiose was 13.8% for 8h. The recoveries of xylanase and xylobiose were 66.4% and 87.2%, respectively. Then, purification of xylobiose was carried out on Sephadex LH-20 with using degassed high purified water as eluent. The optimized conditions for purification of xylobiose were column temperature 35 ℃, 0.1 mL/min and sample volume 0.3 mL. After purification, the purity of xylobiose was improved to 98.5% under the optimized conditions.

Key words: xylanase, aqueous two-phase system, enzymatic hydrolysis, gel chromatography, xylobiose

中图分类号:

TQ35
Q556

李鑫, 顾夕梅, 绳敖楠, 张丽, 陈晓佩, 勇强. 双水相木聚糖酶水解体系制备木二糖及其纯化[J]. 林产化学与工业, 2015, 35(5): 93-97.

LI Xin, GU Xi-mei, SHENG Ao-nan, ZHANG Li, CHEN Xiao-pei, YONG Qiang. Preparation of Xylobiose by Xylanase Hydrolysis in Aqueous Two-phase System and Its Purification[J]. Chemistry and Industry of Forest Products, 2015, 35(5): 93-97.

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双水相木聚糖酶水解体系制备木二糖及其纯化

Preparation of Xylobiose by Xylanase Hydrolysis in Aqueous Two-phase System and Its Purification

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