Studies on the structure and functions of cellulase

纤维素酶的结构与功能研究

• 批准号: 07306005
• 负责人: ARAI Motoo
• 金额: $ 12.03万
• 依托单位: Osaka Prefecture University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07306005/
• 关键词: Cellulase; Glucanase; Glucosidase; Gene; Cloning; DNA; hydrogen gas

The results were as follows :(1) Avicelase gene of A.aculeatus was cloned and elucidated its nucleotide sequence. The gene was expressed in yeast. (ARAI)(2) Subsite of beta-glucosidase of A.niger recognized glucose strictly and this may reflect substrate specificity. (OKADA)(3) Two endoglucanase genes of A.oryzae were cloned and its nucleotide sequences were elucidated.(4) Cellulase gene (CelE) of Clostridium thermocellum was cloned and elucidated its nucleotide sequence. The gene was expressed in Escherichia coli. (OHMIYA)(5) Endoglucanase gene of T.reesei was cloned and elucidated its nucleotide sequence. The gene was expressed. (MORIKAWA)(6) Substrate specificities of 3 endoglucanases from fungi. Enzymatic hydrolysis mechanism depended on the structure of substrate. (KANDA)(7) Highly expressed cDNA at stationary phase of tabacco culture cell was searched and that DNA was isolated. (SHINMYO)(8) Celloanhydrooligosaccharides were useful for elucidation of reaction mechanism of cellulases because of marking effect of anhydroring. (ADUMA)(9) Rhodobacter capsulatus produced effectively hydrogen gas from cellobiose. (OHSHIMA)(10) Three cellulases from A.niger were purified and there characteristics were elucidated. (OGAWA)(11) Three type of chimeric enzymes were prepared from glucosidases from Celluvibrio gilvus (CG) and Agrobacterium tumefaciens (AT). There properties were studied. (HAYASHI)

结果如下：（1）克隆了棘孢蘑菇Avicelase基因，并对其核苷酸序列进行了分析。该基因在酵母中表达。（2）尼日尔曲霉β-葡萄糖苷酶的亚位点严格识别葡萄糖，这可能反映了底物的特异性。（3）克隆了两个葡聚糖内切酶基因，并对其核苷酸序列进行了分析。(4)克隆了热纤梭菌纤维素酶基因（CelE），并对其核苷酸序列进行了分析。该基因在大肠杆菌中表达。（5）克隆了里氏木霉内切葡聚糖酶基因，并对其核苷酸序列进行了分析。基因被表达了。（6）3种真菌内切葡聚糖酶的底物特异性。酶解机理取决于底物的结构。（7）寻找烟草培养细胞稳定期高表达的cDNA，并提取其DNA。（8）纤维素脱水低聚糖具有脱水环标记作用，可用于阐明纤维素酶的反应机理。（9）荚膜红细菌（Rhodobacter capsulatus）能有效地利用纤维二糖产生氢气。（OHSHIMA）（10）从黑曲霉（A.尼日尔）中分离纯化了3种纤维素酶，并对其酶学性质进行了研究。（11）从来自黄色纤维弧菌（CG）和根癌土壤杆菌（AT）的葡糖苷酶制备三种类型的嵌合酶。研究了它们的性质。（林）

项目成果

T.Kawaguchi et al: "Cloning and sequencing of the cDNA encoding β-gluosidase 1 frum Aspergillus aculeatus" Gene. 173. 287-288 (1996)

T. Kawaguchi 等人：“来自棘孢曲霉的 β-葡萄糖苷酶 1 编码 cDNA 的克隆和测序”基因 173. 287-288 (1996)。

岡田 厳太郎: "セルラーゼ研究の進歩と将来への展望" キナンキトサン研究. 2. 16-27 (1996)

冈田源太郎：“纤维素酶研究进展及未来展望”Quinane-Chitosan Research，2. 16-27 (1996)。

T.Kawaguchi et al: "Cloning and seguencing of the cDNA encoding β-dlucosidase1 from Aspergtllus aculeatus" Gene. 173. 287-288 (1996)

T. Kawaguchi 等人：“来自棘孢曲霉的编码 β-葡萄糖苷酶 1 的 cDNA 的克隆和测序”基因 173. 287-288 (1996)

T.Kawaguchi, et al.: "Cloning and sequencing of the cDNA the cDNA encoding beta-glucosidase 1 from Asp. aculeatus" Gene. 173. 287-288 (1996)

T.Kawaguchi 等人：“对棘孢曲霉中编码 β-葡萄糖苷酶 1 的 cDNA 进行克隆和测序”基因。

N.Kitamoto, et al.: "Molecular cloning, purification and characterization of two endoglucanase from Asp. oryzae" Appl.Microbiol.Biotechnol.(in press). (1997)

N.Kitamoto 等人：“米曲霉中两种内切葡聚糖酶的分子克隆、纯化和表征”Appl.Microbiol.Biotechnol.（出版中）。