A studv on the structure and function of a fungal β-1, 6-glucanase

真菌β-1,6-葡聚糖酶结构与功能的研究

• 批准号: 13660077
• 负责人: NAKAJIMA Tasuku
• 金额: $ 1.98万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13660077/
• 关键词: β-glucanase; β-1,6-glucanase gene; Neurospora crassa; fungal cell wall gene; 1β-1,6-glucan; 1β-1,6-glucanase gene disruptant; アカパンカビ; β-1,6-グルカナーゼ; 細胞壁β-グルカン; β-1,6-グルカナーゼ破壊株; Neurosopora rassa; β-グルカン

A gene (neg1) encoding endo-β-1,6-glucanase from Neurospora crassa was cloned. The putative neg1 was 1443-bp long and encoded a mature endo-β-1,6-glucanpse protein of 463 amino acids and signal peptide of 17 amino.acids. The purified recombinant protein (Neg1) obtained from Escherichia coli showed β-1,6-D-glucanase activity. No homologous genes are found in yeasts. and fungi.The β-1,6-glucanase gene (neg1) was disrupted by using repar-induced point mutation(RIP). The gene disruptant strain (R12-1) had no β-1,6-glucanase activity. The phenotype analyses showed thet the sisuptant had no apparent morphological changes, but protein denaturing agents, such as SDS or CTAB inhibited the mycerial growth.

从粗糙脉孢菌（Neurospora crassa）中克隆了一个编码内切β-1，6-葡聚糖酶的基因（pGE 1）。该基因全长1443 bp，编码463个氨基酸的成熟β-1，6-葡聚糖酶蛋白和17个氨基酸的信号肽。从大肠杆菌中纯化的重组蛋白（Neg 1）具有β-1，6-D-葡聚糖酶活性。在酵母中没有发现同源基因。利用repar-induced point mutation（RIP）技术，对β-1，6-葡聚糖酶基因（β-1，6-glucanase，β-1，6-glucanase）进行了突变。该基因突变株（R12-1）不具有β-1，6-葡聚糖酶活性。表型分析表明，菌丝体形态无明显变化，但SDS、CTAB等蛋白质变性剂对菌丝生长有抑制作用。

项目成果

K.Furukawa, Y.Katsuro, T.Urao, T.Yabe, T.Yarnada-Okabe, H.Yamada-Okabe, Y.Yamagata, I.Abe and T.Nakajima: "Isolahon and functional analysis of a gene, tcsB, encoding a transmembrane hybrid-type histidine kinase from Aspergillus nidulans"Appl.Envioln.Micro

K.Furukawa、Y.Katsuro、T.Urao、T.Yabe、T.Yarnada-Okabe、H.Yamada-Okabe、Y.Yamagata、I.Abe 和 T.Nakajima：“Isolahon 和基因 tcsB 的功能分析

千木良裕子 他: "chsZ, a gene for a novel class fo chitin synthase from Aspergillus oryzae"Cuff Genet. 41. 261-267 (2002)

Yuko Chigira 等人：“chsZ，来自米曲霉的新型几丁质合酶基因”Cuff Genet. 41. 261-267 (2002)

小山誠司 他: "Cloning and expression of an endo-1,6-β-D-glucanase gene (neg1) from Nerospora crassa"Bisci. Biotechnol. Biochem.. 66・6. 1387-1381 (2002)

Seiji Koyama 等：“粗糙脉孢菌内切 1,6-β-D-葡聚糖酶基因 (neg1) 的克隆和表达” Bisci Biochem. 66・6 (2002)。

古川健太郎 他: "Isolation and functional analysis of a gene, tcsB, encoding a transrnembrane dybrid-type histidine kinase from Aspergillus nidulans"Appl.Envioln.Microbiol.. 68. 5304-5310 (2002)

Kentaro Furukawa 等人：“编码来自构巢曲霉的跨膜杂合型组氨酸激酶的基因 tcsB 的分离和功能分析”Appl.Envioln.Microbiol.. 68. 5304-5310 (2002)

S.Oyama, Y.Yamagata, K.Abe, and T.Nakajima: "Cloning and expresion of an endo-1,6-β-D-glucanase gene (negl) from Neurospora crassa"Biosi.Biotechnol.Biochem.. 66. 1378-1381 (2002)

S.Oyama、Y.Yamagata、K.Abe 和 T.Nakajima：“粗糙脉孢菌内切 1,6-β-D-葡聚糖酶基因 (negl) 的克隆和表达”Biosi.Biotechnol.Biochem.. 66 .1378-1381 (2002)