Recycle of woody wastes with useful microorganisms

研究了从日本柳杉（Cryptomeria japonica D.Don）健康叶片上分离到的拟盘多毛孢菌YK-3的培养特性。该菌在葡萄糖液体培养基中生长良好，与黑曲霉（Aspergillusniger）货车Tieghem相似。该菌株对单糖、双糖和多糖的利用范围较广，并产生聚糖降解酶。它甚至生长在杉木屑或冷冻干燥的杉叶培养基上，并且木聚糖酶在每种杉培养基中产生。与尼日尔曲霉货车Tieghem相比，该菌株对杉叶提取物具有耐受性或利用能力。拟盘多毛孢属还利用了不溶性多糖如木聚糖，并产生了木聚糖水解酶。在含有木聚糖和邻苯二酚的溶液中，木聚糖水解酶也催化木聚糖的降解和木糖苷的产生。具有木糖基转移活性的木聚糖水解酶已被应用于各种糖苷的酶促合成。从秋田杉叶分离的拟盘多毛孢属ST菌株的木聚糖酶在波罗斯50 HA（Perseptive Biosystems）、波罗斯50 HS和Sephadex G-100上连续分级分离纯化。经SDS-PAGE电泳证实，纯化的酶分子量为25 kDa，具有电泳均一性。该酶的最适pH为6.0，最适温度为50℃。该酶不能水解木二糖和木三糖，从绿色木霉（Trichoderma viride）中分离纯化了一种新的木聚糖降解酶。经凝胶过滤和SDS-PAGE分析，纯化的酶的表观分子量为73 kDa。该酶在以木聚糖为底物的溶液中几乎不水解木聚糖，但在含有木聚糖和邻苯二酚的溶液中催化降解木聚糖并产生木糖苷，即邻苯二酚-β-木糖苷。该酶对间苯二酚、邻苯三酚、没食子酸等酚类物质也有转木糖活性，但对甲醇等醇类物质无转木糖活性。

The cultural properties of Pestalotiopsis sp.YK-3 isolated from healthy sugi (Cryptomeria japonica D.Don) leaves were investigated. The fungus grew well as the useful fungus, Aspergillus niger Van Tieghem, in the glucose liqiud medium. It exhibited wider ranges of utilizing abilities for mono-, di-and polysaccharides tested and produced glycan-degrading enzymes. It was grown even on the sugi sawdust or the freeze-dryed sugi leaf medium, and xylanases were produced in each sugi medium. It was also suggested that it had tolerances or utilizing abilities for extractives of sugi leaves when compared with Aspergillus niger Van Tieghem. Pestalotiopsis spp. also utilized insoluble polysaccharides such as xylan, and produced xylan-hydrolyzing enzymes. The xylan-hydrolyzing enzymes also catalyzed degradation of xylan and production of xylosides in solution containing xylan and catechol. It has suggested that xylan-hydrolyzing enzymes with the xylosyl transfer activity has been applied for enzymatic synthesis of various glycosides. A xylanase of Pestalotiopsis sp.ST, a strain isolated from Akita-sugi leaf, was purified successive fractionation on POROS 50HA (Perseptive Biosystems), POROS 50HS and Sephadex G-100. The purified enzyme, 25kDa, was confirmed to be electophretically homogenious on SDS-PAGE (Fig.2). The optimum pH and temperature for this enzyme were 6.0 and 50℃, respectively. This enzyme could not hydrolyze xylobiose and xylotriose.A novel xylan-degrading enzyme was purified from Trichoderma viride. The purified enzyme had the apparent molecular mass of 73 kDa by gel filtration and SDS-PAGE. The enzyme hardly hydrolyzed xylan in solution containing xylan as a substrate, but catalyzed degradation of xylan and production of xylosides as catechol β-xylobioside in solution containing xylan and catechol. The enzyme had also transxylosyl activity for various phenols as resorcinol, pylogallol, gallic acid, but not for alcohols as methanol.