北京大学学报(自然科学版)

斜卧青霉P6木素过氧化物酶的褐煤降解活性及cDNA克隆

杨金水1,2,袁红莉1,3,陈文新1   

  • 收稿日期:2004-11-29 出版日期:2005-11-20 发布日期:2005-11-20

cDNA Clone and Lignite Degradation Ability of Lignin Peroxidase from Penicillium decumbens P6

YANG Jinshui1, 2, YUAN Hongli1, 3, CHEN Wenxin1   

  1. 1College of Biological Science, China Agricultural University, Beijing, 100094;
  • Received:2004-11-29 Online:2005-11-20 Published:2005-11-20

摘要: 对纯化得到的斜卧青霉P6分泌的木素过氧化物酶的褐煤降解活性进行了研究,发现纯酶的褐煤降解活性依赖于 H2O2 的激活。降解后产物中小分子黄腐酸含量显著提高,褐煤及降解产物的经验分子式分别为 C100H86.5O36.5N2.09 和 C100H103.9O48.6N11.4,降解产物比原褐煤具有更高的 H、O、N 含量,而 C 含量明显下降。并且降解产物可提高豌豆种子的发芽率并明显缩短发芽时间。根据 N 末端氨基酸序列设计特异性引物,利用 RT-PCR 方法扩增得到一条约 2000 bp 的 cDNA 片段,序列测定的大小为 1956 bp,通过序列比对,与已报道的 Piloderma croceum 菌的 mnp2 基因具有 45% 的同源性,具有过氧化氢酶的保守氨基酸残基。

关键词: 褐煤, 斜卧青霉, 木素过氧化物酶, 克隆

Abstract: The present study was undertaken to investigate the degradation ability and biological activity of purification lignin peroxidase (LiP) from Penicillium decumbens P6. P6 LiP had very good degradation ability on lignite, but its activity must depend on H2O2 activiation. After degradation, the contents of humic acid and fulvic acid increased distinctly, especially the content of fulvic acid. Results of elemental analysis indicated that the biodegraded products had higher H, O and N levels than the original lignite, although the carbon content was distinctly lower. The experimental molecular formula for lignite and its biodegraded products were C100H86.5O36.5N2.09 and C100H103.9O48.6N11.4 separately. The products of degradation could increase the germination rate and the speed of germination of pea. Furthermore, the authors designed two special primers according to N terminal amino acid sequence and got a 1956 bp cDNA fragment. Sequence comparison demonstrated that it had 45% homology with Piloderma croceum mnp2 gene and had conservative amino acids of catalase.

Key words: Penicillium decumbens, lignin peroxidase, clone

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