Sato Shin, Feltus F Alex, Iyer Prashanti, Tien Ming
Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, 16802, USA.
Curr Genet. 2009 Jun;55(3):273-86. doi: 10.1007/s00294-009-0243-0. Epub 2009 Apr 26.
As part of an effort to determine all the gene products involved in wood degradation, we have performed massively parallel pyrosequencing on an expression library from the white rot fungus Phanerochaete chrysosporium grown in shallow stationary cultures with red oak as the carbon source. Approximately 48,000 high quality sequence tags (246 bp average length) were generated. 53% of the sequence tags aligned to 4,262 P. chrysosporium gene models, and an additional 18.5% of the tags reliably aligned to the P. chrysosporium genome providing evidence for 961 putative novel fragmented gene models. Due to their role in lignocellulose degradation, the secreted proteins were focused upon. Our results show that the four enzymes required for cellulose degradation: endocellulase, exocellulase CBHI, exocellulase CBHII, and beta-glucosidase are all produced. For hemicellulose degradation, not all known enzymes were produced, but endoxylanases, acetyl xylan esterases and mannosidases were detected. For lignin degradation, the role of peroxidases has been questioned; however, our results show that lignin peroxidase is highly expressed along with the H(2)O(2) generating enzyme, alcohol oxidase. The transcriptome snapshot reveals that H(2)O(2) generation and utilization are central in wood degradation. Our results also reveal new transcripts that encode extracellular proteins with no known function.
作为确定所有参与木材降解的基因产物的努力的一部分,我们对来自以红橡木为碳源在浅层静止培养中生长的白腐真菌黄孢原毛平革菌的表达文库进行了大规模平行焦磷酸测序。产生了约48,000个高质量序列标签(平均长度246 bp)。53%的序列标签与4,262个黄孢原毛平革菌基因模型匹配,另外18.5%的标签可靠地与黄孢原毛平革菌基因组匹配,为961个推定的新片段化基因模型提供了证据。由于其在木质纤维素降解中的作用,重点研究了分泌蛋白。我们的结果表明,纤维素降解所需的四种酶:内切纤维素酶、外切纤维素酶CBHI、外切纤维素酶CBHII和β-葡萄糖苷酶均有产生。对于半纤维素降解,并非所有已知酶都有产生,但检测到了内切木聚糖酶、乙酰木聚糖酯酶和甘露糖苷酶。对于木质素降解,过氧化物酶的作用一直受到质疑;然而,我们的结果表明,木质素过氧化物酶与产生H(2)O(2)的酶醇氧化酶一起高度表达。转录组快照显示,H(2)O(2)的产生和利用在木材降解中至关重要。我们的结果还揭示了编码无已知功能的细胞外蛋白的新转录本。