橡胶加氧酶活性检测方法
Assays for the Detection of Rubber Oxygenase Activities.
作者信息
Röther Wolf, Birke Jakob, Jendrossek Dieter
机构信息
Institute of Microbiology, University of Stuttgart, Stuttgart, Germany.
出版信息
Bio Protoc. 2017 Mar 20;7(6):e2188. doi: 10.21769/BioProtoc.2188.
Abstract
Microbial biodegradation of rubber relies on extracellular rubber oxygenases that catalyze the oxidative cleavage of the double bond of the polyisoprene backbone into oligo-isoprenoids. This protocol describes the determination of rubber oxygenase activities by an online measurement of molecular oxygen consumption via a non-invasive fluorescence-based assay. The produced oligo-isoprenoid cleavage products with terminal keto- and aldehyde-groups are identified qualitatively and quantitatively by HPLC. Our method allows for the characterization of homologue rubber oxygenases, and can likely be adapted to assay other oxygenases consuming dioxygen. Here we describe the determination of rubber oxygenase activities at the examples of the so far two known types of rubber oxygenases, namely rubber oxygenase A (RoxA) and latex clearing protein (Lcp).
摘要
橡胶的微生物生物降解依赖于细胞外橡胶加氧酶，该酶催化聚异戊二烯主链双键的氧化裂解生成低聚异戊二烯类化合物。本方案描述了通过基于非侵入性荧光测定法在线测量分子氧消耗来测定橡胶加氧酶活性的方法。通过高效液相色谱对生成的具有末端酮基和醛基的低聚异戊二烯裂解产物进行定性和定量鉴定。我们的方法可用于表征同源橡胶加氧酶，并且可能适用于检测其他消耗双氧的加氧酶。在此，我们以迄今为止已知的两种橡胶加氧酶，即橡胶加氧酶A（RoxA）和乳胶清除蛋白（Lcp）为例，描述橡胶加氧酶活性的测定方法。
相似文献
1
Assays for the Detection of Rubber Oxygenase Activities.
Bio Protoc. 2017 Mar 20;7(6):e2188. doi: 10.21769/BioProtoc.2188.
2
RoxB Is a Novel Type of Rubber Oxygenase That Combines Properties of Rubber Oxygenase RoxA and Latex Clearing Protein (Lcp).
Appl Environ Microbiol. 2017 Jun 30;83(14). doi: 10.1128/AEM.00721-17. Print 2017 Jul 15.
3
Rubber oxygenase and latex clearing protein cleave rubber to different products and use different cleavage mechanisms.
Appl Environ Microbiol. 2014 Aug;80(16):5012-20. doi: 10.1128/AEM.01271-14. Epub 2014 Jun 6.
4
Biochemical and spectroscopic characterization of purified Latex Clearing Protein (Lcp) from newly isolated rubber degrading Rhodococcus rhodochrous strain RPK1 reveals novel properties of Lcp.
BMC Microbiol. 2016 May 23;16:92. doi: 10.1186/s12866-016-0703-x.
5
Latex Clearing Protein (Lcp) of Streptomyces sp. Strain K30 Is a b-Type Cytochrome and Differs from Rubber Oxygenase A (RoxA) in Its Biophysical Properties.
Appl Environ Microbiol. 2015 Jun;81(11):3793-9. doi: 10.1128/AEM.00275-15. Epub 2015 Mar 27.
6
Production of functionalized oligo-isoprenoids by enzymatic cleavage of rubber.
Microb Biotechnol. 2017 Nov;10(6):1426-1433. doi: 10.1111/1751-7915.12748. Epub 2017 Jul 11.
7
Rhizobacter gummiphilus NS21 has two rubber oxygenases (RoxA and RoxB) acting synergistically in rubber utilisation.
Appl Microbiol Biotechnol. 2018 Dec;102(23):10245-10257. doi: 10.1007/s00253-018-9341-6. Epub 2018 Sep 13.
8
Solimonas fluminis has an active latex-clearing protein.
Appl Microbiol Biotechnol. 2019 Oct;103(19):8229-8239. doi: 10.1007/s00253-019-10085-w. Epub 2019 Sep 4.
9
Rubber oxygenases.
Appl Microbiol Biotechnol. 2019 Jan;103(1):125-142. doi: 10.1007/s00253-018-9453-z. Epub 2018 Oct 30.
10
Towards the understanding of the enzymatic cleavage of polyisoprene by the dihaem-dioxygenase RoxA.
AMB Express. 2019 Oct 17;9(1):166. doi: 10.1186/s13568-019-0888-0.
引用本文的文献
1
Computational investigation of cis-1,4-polyisoprene binding to the latex-clearing protein LcpK30.
PLoS One. 2024 May 15;19(5):e0302398. doi: 10.1371/journal.pone.0302398. eCollection 2024.
2
Biodegradation of rubber in cultures of Rhodococcus rhodochrous and by its enzyme latex clearing protein.
Biodegradation. 2022 Dec;33(6):609-620. doi: 10.1007/s10532-022-09998-7. Epub 2022 Oct 5.
3
Towards the understanding of the enzymatic cleavage of polyisoprene by the dihaem-dioxygenase RoxA.
AMB Express. 2019 Oct 17;9(1):166. doi: 10.1186/s13568-019-0888-0.
4
Rubber oxygenases.
Appl Microbiol Biotechnol. 2019 Jan;103(1):125-142. doi: 10.1007/s00253-018-9453-z. Epub 2018 Oct 30.
5
Structural and Functional Analysis of Latex Clearing Protein (Lcp) Provides Insight into the Enzymatic Cleavage of Rubber.
Sci Rep. 2017 Jul 21;7(1):6179. doi: 10.1038/s41598-017-05268-2.
6
Production of functionalized oligo-isoprenoids by enzymatic cleavage of rubber.
Microb Biotechnol. 2017 Nov;10(6):1426-1433. doi: 10.1111/1751-7915.12748. Epub 2017 Jul 11.
7
RoxB Is a Novel Type of Rubber Oxygenase That Combines Properties of Rubber Oxygenase RoxA and Latex Clearing Protein (Lcp).
Appl Environ Microbiol. 2017 Jun 30;83(14). doi: 10.1128/AEM.00721-17. Print 2017 Jul 15.
本文引用的文献
1
Cleavage of Rubber by the Latex Clearing Protein (Lcp) of Streptomyces sp. Strain K30: Molecular Insights.
Appl Environ Microbiol. 2016 Oct 27;82(22):6593-6602. doi: 10.1128/AEM.02176-16. Print 2016 Nov 15.
2
Biochemical and spectroscopic characterization of purified Latex Clearing Protein (Lcp) from newly isolated rubber degrading Rhodococcus rhodochrous strain RPK1 reveals novel properties of Lcp.
BMC Microbiol. 2016 May 23;16:92. doi: 10.1186/s12866-016-0703-x.
3
Latex Clearing Protein (Lcp) of Streptomyces sp. Strain K30 Is a b-Type Cytochrome and Differs from Rubber Oxygenase A (RoxA) in Its Biophysical Properties.
Appl Environ Microbiol. 2015 Jun;81(11):3793-9. doi: 10.1128/AEM.00275-15. Epub 2015 Mar 27.
4
Latex clearing protein-an oxygenase cleaving poly(cis-1,4-isoprene) rubber at the cis double bonds.
Appl Environ Microbiol. 2014 Sep;80(17):5231-40. doi: 10.1128/AEM.01502-14. Epub 2014 Jun 13.
5
Rubber oxygenase and latex clearing protein cleave rubber to different products and use different cleavage mechanisms.
Appl Environ Microbiol. 2014 Aug;80(16):5012-20. doi: 10.1128/AEM.01271-14. Epub 2014 Jun 6.
6
Functional identification of rubber oxygenase (RoxA) in soil and marine myxobacteria.
Appl Environ Microbiol. 2013 Oct;79(20):6391-9. doi: 10.1128/AEM.02194-13. Epub 2013 Aug 9.
7
Structure of the processive rubber oxygenase RoxA from Xanthomonas sp.
Proc Natl Acad Sci U S A. 2013 Aug 20;110(34):13833-8. doi: 10.1073/pnas.1305560110. Epub 2013 Aug 6.
8
Phe317 is essential for rubber oxygenase RoxA activity.
Appl Environ Microbiol. 2012 Nov;78(22):7876-83. doi: 10.1128/AEM.02385-12. Epub 2012 Aug 31.
9
Spectroscopic properties of rubber oxygenase RoxA from Xanthomonas sp., a new type of dihaem dioxygenase.
Microbiology (Reading). 2010 Aug;156(Pt 8):2537-2548. doi: 10.1099/mic.0.038992-0. Epub 2010 Apr 22.
10
Heme-dependent rubber oxygenase RoxA of Xanthomonas sp. cleaves the carbon backbone of poly(cis-1,4-Isoprene) by a dioxygenase mechanism.
Appl Environ Microbiol. 2005 May;71(5):2473-8. doi: 10.1128/AEM.71.5.2473-2478.2005.
Zotero 插件