Zwickl P, Fabry S, Bogedain C, Haas A, Hensel R
Max-Planck-Institut für Biochemie, Martinsried, Federal Republic of Germany.
J Bacteriol. 1990 Aug;172(8):4329-38. doi: 10.1128/jb.172.8.4329-4338.1990.
The glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic archaebacterium Pyrococcus woesei (optimal growth temperature, 100 to 103 degrees C) was purified to homogeneity. This enzyme was strictly phosphate dependent, utilized either NAD+ or NADP+, and was insensitive to pentalenolactone like the enzyme from the methanogenic archaebacterium Methanothermus fervidus. The enzyme exhibited a considerable thermostability, with a 44-min half-life at 100 degrees C. The amino acid sequence of the glyceraldehyde-3-phosphate dehydrogenase from P. woesei was deduced from the nucleotide sequence of the coding gene. Compared with the enzyme homologs from mesophilic archaebacteria (Methanobacterium bryantii, Methanobacterium formicicum) and an extremely thermophilic archaebacterium (Methanothermus fervidus), the primary structure of the P. woesei enzyme exhibited a strikingly high proportion of aromatic amino acid residues and a low proportion of sulfur-containing residues. The coding gene of P. woesei was expressed at a high level in Escherichia coli, thus providing an ideal basis for detailed structural and functional studies of that enzyme.
对嗜热古细菌沃氏嗜热栖热菌（最佳生长温度为100至103摄氏度）的甘油醛-3-磷酸脱氢酶进行了纯化，使其达到同质。该酶严格依赖磷酸盐，可利用NAD⁺或NADP⁺，并且与产甲烷古细菌嗜热栖热菌的酶一样，对戊烯醇内酯不敏感。该酶表现出相当高的热稳定性，在100摄氏度下的半衰期为44分钟。沃氏嗜热栖热菌甘油醛-3-磷酸脱氢酶的氨基酸序列是根据编码基因的核苷酸序列推导出来的。与嗜温古细菌（布氏甲烷杆菌、甲酸甲烷杆菌）和极端嗜热古细菌（嗜热栖热菌）的酶同源物相比，沃氏嗜热栖热菌酶的一级结构显示出芳香族氨基酸残基的比例极高，而含硫残基的比例极低。沃氏嗜热栖热菌的编码基因在大肠杆菌中高水平表达，从而为该酶的详细结构和功能研究提供了理想的基础。
