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首页> 外文期刊>Bioprocess and Biosystems Engineering >Cloning and characterization of a thermostable endo-arabinanase from Phanerochaete chrysosporium and its synergistic action with endo-xylanase
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Cloning and characterization of a thermostable endo-arabinanase from Phanerochaete chrysosporium and its synergistic action with endo-xylanase

机译:拟南芥(Phanerochaete chrysosporium)热稳定内阿拉伯聚糖酶的克隆,鉴定及其与木聚糖内切酶的协同作用

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摘要

Putative arabinanase (PcARA) was cloned from cDNA of Phanerochaete chrysosporium. The gene sequencing indicated that PcARA consisted of 939 nucle-otides that encodes for 312 amino acid arabinanase-poly-peptide chain, including a signal peptide of 19 amino acids. Three-dimensional homology indicated that this enzyme is a five-bladed β-propeller, belonging to glycosidase family 43 and its secondary structure is consisted of 24 β-sheets. The PcARA-cDNA was expressed in Pichia pastoris using pPICZαC. SDS-PAGE of purified arabinanase showed a single band of 33 kDa that is very close to theoretical molecular mass of 33.9 kDa calculated by its amino acid content. Recombinant arabinanase (rPcARA) exhibited maximum activity at pH and temperature of 5.0 and 60 ℃, respectively. End-product analysis of debranched arabinan hydrolysis by thin-layer chromatography indicated that rPcARA acted as endo-type. The synergistic action of rPcARA with recombinant xylanase resulted in 72 and 9.3 % release of total soluble sugar of arabinoxylan and NaOH-pretreated barley straw, respectively.
机译:从拟南芥Chrysosporium的cDNA克隆推定的阿拉伯聚糖酶(PcARA)。基因测序表明,PcARA由939个核苷酸组成,编码312个氨基酸的阿拉伯聚糖酶-多肽链,包括19个氨基酸的信号肽。三维同源性表明该酶是一种五叶β-螺旋桨,属于糖苷酶家族43,其二级结构由24个β-折叠组成。使用pPICZαC在巴斯德毕赤酵母中表达PcARA-cDNA。纯化的阿拉伯聚糖酶的SDS-PAGE显示一条33 kDa的条带,非常接近其氨基酸含量计算得出的理论分子量33.9 kDa。重组阿拉伯聚糖酶(rPcARA)分别在pH和5.0和60℃下表现出最大的活性。通过薄层色谱法对脱支的阿拉伯聚糖水解的最终产物进行分析表明,rPcARA充当内源型。 rPcARA与重组木聚糖酶的协同作用分别导致阿拉伯木聚糖和NaOH预处理的大麦秸秆的总可溶性糖释放分别为72%和9.3%。

著录项

  • 来源
    《Bioprocess and Biosystems Engineering》 |2013年第6期|677-685|共9页
  • 作者单位

    Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, Jeonbuk 570-752, Korea;

    Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, Jeonbuk 570-752, Korea;

    Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, Jeonbuk 570-752, Korea,LED-Agribio Fusion Technology Research Center,Chonbuk National University, Iksan, Jeonbuk 570-752, Korea;

    Institute of Resources, Environment and Biotechnology,Hue University, Hue, Vietnam,Department of Molecular Biology, College of Natural Sciences, Chonbuk National University, Jeonju, Jeonbuk 561-756, Korea;

    Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, Jeonbuk 570-752, Korea,LED-Agribio Fusion Technology Research Center,Chonbuk National University, Iksan, Jeonbuk 570-752, Korea;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Arabinanase; Phanerochaete chrysosporium; Pichia pastoris; Hemicellulose; Barley straw;

    机译:阿拉伯聚糖酶;Phanerochaete chrysosporium;巴斯德毕赤酵母;半纤维素;大麦秸秆;

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