Yano Shigekazu, Yamamoto Sachiko, Toge Toshihiko, Wakayama Mamoru, Tachiki Takashi
Department of Bioscience and Biotechnology, Faculty of Science and Technology, Ritsumeikan University, Shiga, Japan.
Biosci Biotechnol Biochem. 2003 Sep;67(9):1976-82. doi: 10.1271/bbb.67.1976.
KA-prep, a culture filtrate of Bacillus circulans KA-304 grown on a cell-wall preparation (CWP) of Schizophyllum commune, has been reported to have an activity to form protoplasts from S. commune mycelia. The SDS-polyacrylamide gel electrophoreses described here demonstrated that a specific proteinous component (molecular weight: 150,000) occurred in KA-prep. The protein (P150T) was also formed in culture filtrates with CWP of several basidiomycetes, which could release the protoplasts, suggesting that the component was an indispensable factor for protoplast formation. P150T, isolated from an ammonium sulfate fraction of KA-prep (0-30% saturation), did not have any protoplast-forming activity. Results were obtained indicating that P150T participates in protoplast formation together with chitinase(s) and beta-glucanase(s) in KA-prep. The N-terminal amino acid sequence indicated an analogy of P150T to mutanase (alpha-1,3-glucanase) from Bacillus sp. RM1, and actually P150T hydrolyzed mutan as well as S-(alpha-1,3) glucan from S. commune.
KA制剂是环状芽孢杆菌KA - 304在裂褶菌细胞壁制剂(CWP)上生长得到的一种培养滤液,据报道它具有使裂褶菌菌丝体形成原生质体的活性。本文所述的SDS - 聚丙烯酰胺凝胶电泳表明,KA制剂中存在一种特定的蛋白质成分(分子量:150,000)。该蛋白质(P150T)在几种能释放原生质体的担子菌的含CWP培养滤液中也有形成,这表明该成分是原生质体形成所必需的因素。从KA制剂的硫酸铵分级分离物(0 - 30%饱和度)中分离得到的P150T没有任何原生质体形成活性。结果表明,P150T与KA制剂中的几丁质酶和β - 葡聚糖酶一起参与原生质体形成。N端氨基酸序列表明P150T与芽孢杆菌属RM1的变聚糖酶(α - 1,3 - 葡聚糖酶)有相似性,实际上P150T能水解变聚糖以及裂褶菌的S -(α - 1,3)葡聚糖。