Gene analysis and application of RuBisCO from nitiifying bacteria

硝化菌RuBisCO基因分析及应用

• 批准号: 09660103
• 负责人: TAKAHASHI Reiji
• 金额: $ 1.28万
• 依托单位: NIHON UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09660103/
• 关键词: nitrifying bacteiia; RuBisCO; gene analysis; pure isolation; high-concentration-ammonium-rquring ammonia-oxidizing bacterium; 炭酸固定; cbbL; cbbS

1) Gene analysis of RuBisCO : DNA sequencing indicated cbbL horn Nitrobacter agilis ATCC14123 and Nitrobacter winogradskyi IFO14297 to consist of 1,422bp coding 473 amino acids aid cbbS from these strains, of 330bp coding 108 amino acids. Two genes were found to line tandemly with 74bp of an intervening region. 24 residues from N-terminal of the L subunit and 19 residues from the N-terminal of the S subunit on proposed amino acid sequences completely corresponded to those of previously purified protein. cbbR would thus appear situated 232bp upstream of cbbL with opposite orientation of trascription against cbbL/S and some decades of bp from 5'terminal of cbbR were determined. On the intergenic region within cbbR and cbbL/S, the promoter of cbbL/S was found and CbbR binding sites were present in the same region. A comparison with the gene structure and amine acid sequence of RubisCO may be useful for one of the indexes which subdivided classification with 16S rRNA sequence in nitrifying bacteria.2) Pure isolation A high -concentraion- ammonium sulfate-requiring ammonia-oxidizing bacterium, strain K1 was newly isolated from packed tower biological deodorization plants of chicken farmas. This bacterium has a higher optimal growth temperature (35℃) than is usually the case and tolerance up to 40℃. The optimum concentration of ammonia sulfate in the medium is 303 mM, which should make it applicable for use in deodorization plants for enhancing the efficiency of deodorization.

1)RuBisCO基因序列分析表明：敏捷硝化杆菌ATCC 14123和温氏硝化杆菌IFO 14297的cbbL基因全长1,422 bp，编码473个氨基酸;发现两个基因串联在一起，中间有一个74 bp的插入区。L亚基N端24个氨基酸残基和S亚基N端19个氨基酸残基的氨基酸序列与已纯化蛋白完全一致。cbbR位于cbbL上游232 bp处，与cbbL/S的转录方向相反，并确定了cbbR 5 '端的几十个bp。在cbbR和cbbL/S的基因间隔区，发现cbbL/S的启动子和CbbR结合位点。与RubisCO的基因结构和氨基酸序列的比较，可作为硝化细菌16 SrRNA序列细分分类的指标之一。2）纯分离从鸡场填料塔生物反应器中新分离到一株高浓度硫酸铵氨氧化菌K1。该菌的最适生长温度（35℃）比通常的情况高，耐受性可达40℃。培养基中硫酸铵的最佳浓度为303 mM，这应使其适用于在植物中用于提高发酵效率。

项目成果

Hatayama,R., et al.: "Characteristics of high-concentration-ammonium sulfate-requiring ammonia oxidizing bacterium isolated from deodorization plants of chicken farms."Journal of Bioscience and Bioengineering. 87(2). 245-248 (1999)

Hatayama，R.，等人：“从养鸡场除臭装置中分离出的高浓度硫酸铵需求氨氧化细菌的特性。”生物科学与生物工程杂志。

Ryouta Hatayama, Kumiko Chiba, Kazue Noda, Reiji Takahashi, Tsutomu Kanehira, Kazuhiro Setara, Masayuki Shinohara, and Tatsuaki Tokuyama: "Characteristics of high-concentration-ammonium sulfate-requiring ammonia-oxidizing bacterium isolated from deodoriza

Ryouta Hatayama、Kumiko Chiba、Kazue Noda、Reiji Takahashi、Tsutomu Kanehira、Kazuhiro Setara、Masayuki Shinohara 和 Tatsuaki Tokuyama：“从除臭剂中分离出的高浓度硫酸铵需求氨氧化细菌的特性