| 引用本文: | 陈芳芳, 曹曦跃, 刘 颖, 刘梦佳, 颜佳超, 杨富强, 曹 毅, 乔代蓉.Bacillus pumilus阿拉伯呋喃糖苷酶的重组表达及水解木聚糖的研究[J].广西植物,2018,38(4):444-450.[点击复制] |
| CHEN Fangfang, CAO Xiyue, LIU Ying, LIU Mengjia, YAN Jiachao,
YANG Fuqiang, CAO Yi, QIAO Dairong.Recombinant expression of α-L-Arabinofuranosidase from Bacillus pumilus and hydrolysis of xylan[J].Guihaia,2018,38(4):444-450.[点击复制] |
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| Bacillus pumilus阿拉伯呋喃糖苷酶的重组表达及水解木聚糖的研究 |
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陈芳芳1, 曹曦跃2, 刘 颖1, 刘梦佳1, 颜佳超1, 杨富强1, 曹 毅1, 乔代蓉1*
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1. 四川大学 生命科学学院 微生物与代谢工程四川省重点实验室, 成都 610065;2. 东北农业大学 食品学院, 哈尔滨 150036
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| 摘要: |
| 从短小芽孢杆菌中克隆阿拉伯呋喃糖苷酶基因xyn43并重组表达,有利于将该酶分离纯化后应用于其他半纤维素多糖的水解。该研究利用E. coli BL21表达系统对实验室克隆到的短小芽孢杆菌的α-L-阿拉伯呋喃糖苷酶基因xyn43进行重组表达并分析其酶学性质,将重组α-L-阿拉伯呋喃糖苷酶Xyn43和来源于棒曲霉突变菌株的商业木聚糖酶联合作用于燕麦木聚糖。结果表明:以燕麦木聚糖为底物,重组α-L-阿拉伯呋喃糖苷酶Xyn43的最适温度为50 ℃,最适pH为6.0。该酶在pH 5.0～10.0和45～55 ℃下较稳定。与木聚糖酶单独作用相比,重组Xyn43酶与商业木聚糖酶同时加入以及先用木聚糖酶水解后加入Xyn43酶,水解产物中的还原糖含量分别增加了16%和20%,木糖含量增加了35%和48%。该结果研究结果表明重组Xyn43酶能够和商业木聚糖酶协同降解燕麦木聚糖,提高水解效率,产生更多的木寡糖,阿拉伯糖和木糖。 |
| 关键词: α-L-阿拉伯呋喃糖苷酶, 木聚糖酶, 燕麦木聚糖, 重组表达, 协同作用 |
| DOI：10.11931/guihaia.gxzw201705006 |
| 分类号:Q946 |
| 文章编号:1000-3142(2018)04-0444-07 |
| 基金项目:国家资源平台项目(NIMR 2017-8-1); 四川省科技厅项目(18ZDYF1802, 2014GZX0005, 18PTDJ0034)[Supported by the National Infrastructure of Natural Resources for Science and Technology Program of China(NIMR 2017-8-1); Sichuan Science and Technology Bureau(18ZDYF1802,2014GZX0005,18PTDJ0034)]。 |
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| Recombinant expression of α-L-Arabinofuranosidase from Bacillus pumilus and hydrolysis of xylan |
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CHEN Fangfang1, CAO Xiyue2, LIU Ying1, LIU Mengjia1, YAN Jiachao1,
YANG Fuqiang1, CAO Yi1, QIAO Dairong1*
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1. Microbiology and Metabolic Engineering of Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University,
Chengdu 610065, China;2. College of Food Sciences, Northeast Agricultural University, Harbin 150036, China
1. Microbiology and Metabolic Engineering of Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University,
Chengdu 610065, China; 2. College of Food Sciences, Northeast Agricultural University, Harbin 150036, China
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| Abstract: |
| Hemicellulose is a rich renewable resource, which is one of the most promising raw materials for the production of biofuels. α-L-arabinofuranosidase(EC 3.2.1.55)is an auxiliary enzyme in hemicellulose hydrolase system which catalyzes the α-L-1,2, α-L-1,3 and α-L-1.5-arabinofuranoside residues in non-reducing terminal of various oligosaccharides and polysaccharides. Bacillus pumilus is a widely used, bio-friendly feed microbial strain that can degrades mannan, xylan, cellulose and so on. Therefore, cloning the α-L-arabinofuranosidase gene xyn43 from B. pumilus and recombinant expression is beneficial to the separation and purification of the enzyme and it can be applied to the hydrolysis of other hemicellulose polysaccharide. In this study, we used the E. coli BL21 expression system to express the α-L-arabinofuranosidase gene xyn43 isolated from the B. pumilus and then analyzed the enzymatic properties of recombinant enzyme Xyn43. And we used Xyn43 and commercial Xylanase derived from the mutant strain of Aspergillve clavatus to degrade the oat spelt xylan. The results showed that the optimal temperature of Xyn43 was 50 ℃ and the optimum pH was 6.0. It was stable over a pH range of 5.0 -10.0 and temperatures of 45-55 ℃. Compared with Xylanase alone, the reducing sugar content in the hydrolyzate of Xyn43 and Xylanase added simultaneously and Xyn43 added after Xylanase increased 16% and 20% respectively, and the xylose content increased of 35% and 48%. The studies indicate that Xyn43 is able to synergistically with commercial Xylanase for oat spelt xylan degradation and can improve the hydrolysis efficiency, produced more xylosaccharides, arabinose and xylose. |
| Key words: α-L-arabinofuranosidase, xylanase, oat spelt xylan, recombinant expression, synergistic effect |
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