Cleavage of poly(cis-1,4-isoprene) by the latex clearing protein (Lcp) and involvement of a molybdenum-dependent oxidoreductase (OxiBA) in early steps of rubber degradation

橡胶降解早期步骤中乳胶清除蛋白 (Lcp) 裂解聚（顺式 1,4-异戊二烯）以及钼依赖性氧化还原酶 (OxiBA) 的参与

• 批准号: 42348726
• 负责人: Professor Dr. Alexander Steinbüchel
• 金额: --
• 依托单位: Institut für Molekulare Mikrobiologie und Biotechnologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/42348726?language=en
• 关键词: Cleavage; poly; cis; isoprene; latex

This project investigates initial cleavage of poly(cis-1,4-isoprene) by Lcp (Latex clearing protein) from Streptomyces sp. K30. Lcp homologues occur in all other Gram-positive rubber and gutta percha degrading bacteria; this protein seems to be generally responsible for cleavage of polyisoprenoides by Actinobacteria. The aims are as follows: (i) Secretion of Lcp via the Tat pathway will be investigated. (ii) Random mutagenesis employing a mutator strain and an E.coli-Streptomyces shuttle vector will unravel essential amino acid residues of Lcp, (iii) which will be verified by site specific mutagenesis. (iv) An enzyme activity assay for Lcp will be developed to characterize wild type and mutated Lcp proteins and to investigate regulation of Lcp. (v) During cleavage of poly(cis-1,4-isoprene), oxygen is incorporated into the degradation products. An enzyme activity assay and knowledge of amino acids involved in cleavage will enable to determine the molecular mechanism of rubber cleavage by Lcp. In this context, putative co-factors and the 3D structure of Lcp will be identified. (vi) A putative aldehyde dehydrogenase encoded by oxiA and oxiB located downstream of lcp obviously oxidizes the cleavage products produced by Lcp. Function and regulation of oxiAB will be investigated. (vii) 2D gel electrophoresis employing Nocardia farcinica IFM10152, the only rubber degrading bacterium with known genome sequence, will unravel further proteins involved in poly(cis-1,4-isoprene) degradation.

本项目研究了链霉菌K30的乳胶透明蛋白（LCP）对聚（顺式-1，4-异戊二烯）的初始裂解。LCP同源物存在于所有其他革兰氏阳性橡胶和古塔胶降解细菌中;这种蛋白质似乎通常负责放线菌对聚异戊二烯类的裂解。目的如下：（i）将研究LCP经由达特途径的分泌。(ii)随机诱变采用的诱变菌株和大肠杆菌-链霉菌穿梭载体将解开必需的氨基酸残基的LCP，（iii）这将通过位点特异性诱变验证。(iv)将开发LCP的酶活性测定以表征野生型和突变型LCP蛋白并研究LCP的调节。(v)在聚（顺式-1，4-异戊二烯）的裂解过程中，氧被掺入降解产物中。酶活性测定和裂解中涉及的氨基酸的知识将能够确定LCP裂解橡胶的分子机制。在这种情况下，推定的辅因子和LCP的三维结构将被确定。(vi)由oxiA和oxiB编码的位于LCP下游的推定的醛脱氢酶明显地氧化由LCP产生的裂解产物。将研究oxiAB的功能和调节。(vii)采用Nocardia farcinica IFM 10152（唯一具有已知基因组序列的橡胶降解细菌）的2D凝胶电泳将进一步揭示参与聚（顺式-1，4-异戊二烯）降解的蛋白质。

项目成果

Secretion and Transcriptional Regulation of the Latex-Clearing Protein, Lcp, by the Rubber-Degrading Bacterium Streptomyces sp. Strain K30

• DOI: 10.1128/aem.01001-08
• 发表时间: 2008-07
• 期刊: Applied and Environmental Microbiology
• 影响因子: 4.4
• 作者: Meral Yikmis;M. Arenskötter;K. Rose;N. Lange;Henrike Wernsmann;Lars Wiefel;A. Steinbüchel

Transfer of megaplasmid pKB1 from the rubber-degrading bacterium Gordonia westfalica strain Kb1 to related bacteria and its modification

橡胶降解菌 Gordonia westfalica Kb1 菌株巨质粒 pKB1 向相关菌的转移及其修饰

• DOI: 10.1007/s00253-007-1262-8
• 发表时间: 2008
• 期刊: Applied Microbiology and Biotechnology
• 影响因子: 5
• 作者: Bröker;Arenskötter;A. Steinbüchel
• 通讯作者: A. Steinbüchel

Historical and Recent Achievements in the Field of Microbial Degradation of Natural and Synthetic Rubber

• DOI: 10.1128/aem.00001-12
• 发表时间: 2012-04
• 期刊: Applied and Environmental Microbiology
• 影响因子: 4.4
• 作者: Meral Yikmis;A. Steinbüchel

Megaplasmid pKB1 of the Rubber-Degrading Bacterium Gordonia westfalica Strain Kb1

橡胶降解细菌 Gordonia westfalica 菌株 Kb1 的巨质粒 pKB1

• DOI: 10.1007/978-3-540-85467-8_14
• 发表时间: 2009
• 作者: Bröker;A. Steinbüchel
• 通讯作者: A. Steinbüchel

Microbial Gutta-Percha Degradation Shares Common Steps with Rubber Degradation by Nocardia nova SH22a

• DOI: 10.1128/aem.03016-12
• 发表时间: 2013-02-01
• 期刊: APPLIED AND ENVIRONMENTAL MICROBIOLOGY
• 影响因子: 4.4
• 作者: Luo, Quan;Hiessl, Sebastian;Steinbuechel, Alexander
• 通讯作者: Steinbuechel, Alexander