Ohtoko K, Ohkuma M, Moriya S, Inoue T, Usami R, Kudo T
RIKEN, The Institute of Physical and Chemical Research, Wako, Saitama, Japan.
Extremophiles. 2000 Dec;4(6):343-9. doi: 10.1007/s007920070003.
Diverse genes encoding cellulase homologues belonging to glycosyl hydrolase family 45 were identified from the symbiotic protists in the hindgut of the termite Reticulitermes speratus through the use of consensus PCR and the screening of a cDNA library. Fifteen full-length cDNA clones were isolated and sequenced, which encoded polypeptides consisting of 218-221 amino acid residues showing up to 63% identity to known family 45 cellulases. The cellulase sequences of the termite symbiotic protists were phylogenetically monophyletic, showing more than 75% amino acid identity with each other. These enzymes consist of a single catalytic domain, lacking the ancillary domains found in most microbial cellulases. By whole-cell in situ hybridization using oligonucleotide probes specific for regions conserved in some of the sequences, the origin of the genes was identified as symbiotic hypermastigote protists. The presence of diverse cellulase homologues suggests that symbiotic protists of termites may be rich reservoirs of novel cellulase sequences.
通过使用简并PCR和筛选cDNA文库,从日本白蚁后肠共生原生生物中鉴定出了属于糖基水解酶家族45的多种编码纤维素酶同源物的基因。分离并测序了15个全长cDNA克隆,它们编码的多肽由218 - 221个氨基酸残基组成,与已知的45家族纤维素酶具有高达63%的同一性。白蚁共生原生生物的纤维素酶序列在系统发育上是单系的,彼此之间显示出超过75%的氨基酸同一性。这些酶由单个催化结构域组成,缺乏大多数微生物纤维素酶中发现的辅助结构域。通过使用对某些序列中保守区域特异的寡核苷酸探针进行全细胞原位杂交,确定这些基因的来源是共生超鞭毛虫原生生物。多种纤维素酶同源物的存在表明,白蚁的共生原生生物可能是新型纤维素酶序列的丰富来源。