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Characterization of the latex clearing protein of the poly(cis-1,4-isoprene) and poly(trans-1,4-isoprene) degrading bacterium Nocardia nova SH22a
| dc.contributor.author | Vivod, Robin | |
| dc.contributor.author | Andler-Osorio, Rodrigo | |
| dc.contributor.author | Oetermann, Sylvia | |
| dc.contributor.author | Altenhoff, A. L. | |
| dc.contributor.author | Seipel, Nele | |
| dc.contributor.author | Holtkamp, Michael | |
| dc.contributor.author | Hogeback, Jens | |
| dc.contributor.author | Karst, Uwe | |
| dc.contributor.author | Steinbüchel, A. | |
| dc.date.accessioned | 2020-08-20T13:13:56Z | |
| dc.date.available | 2020-08-20T13:13:56Z | |
| dc.date.issued | 2019 | |
| dc.identifier.uri | http://repositorio.ucm.cl/handle/ucm/3016 | |
| dc.description.abstract | Nocardia nova SH22a is an actinobacterium capable of degrading the polyisoprenes poly(cis-1,4-isoprene) and poly(trans-1,4-isoprene). Sequencing and annotating the genome of this strain led to the identification of a single gene coding for the key enzyme for the degradation of rubber: the latex clearing protein (Lcp). In this study, we showed that LcpSH22a-contrary to other already characterized rubber cleaving enzymes-is responsible for the initial cleavage of both polyisoprene isomers. For this purpose, lcpSH22a was heterologously expressed in an Escherichia coli strain and purified with a functional His6- or Strep-tag. Applying liquid chromatography electrospray ionization time-of-flight mass spectrometry (LC/ESI-ToF-MS) and a spectrophotometric pyridine hemochrome assay, heme b was identified as a cofactor. Furthermore, heme-associated iron was identified using total reflection X-ray fluorescence (TXRF) analysis and inhibition tests. The enzyme's temperature and pH optima at 30°C and 7, respectively, were determined using an oxygen consumption assay. Cleavage of poly(cis-1,4-isoprene) and poly(trans-1,4-isoprene) by the oxygenase was confirmed via detection of carbonyl functional groups containing cleavage products, using Schiff's reagent and electrospray ionization mass spectrometry (ESI-MS). | es_CL |
| dc.language.iso | en | es_CL |
| dc.rights | Atribución-NoComercial-SinDerivadas 3.0 Chile | * |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/3.0/cl/ | * |
| dc.source | The Journal of General and Applied Microbiology, 65(6), 293-300 | es_CL |
| dc.subject | Nocardia nova SH22a | es_CL |
| dc.subject | Gutta-percha | es_CL |
| dc.subject | Microbial rubber degradation | es_CL |
| dc.subject | Poly(cis-1,4-isoprene) rubber | es_CL |
| dc.subject | Poly(trans-1,4-isoprene) rubber | es_CL |
| dc.title | Characterization of the latex clearing protein of the poly(cis-1,4-isoprene) and poly(trans-1,4-isoprene) degrading bacterium Nocardia nova SH22a | es_CL |
| dc.type | Article | es_CL |
| dc.ucm.facultad | Facultad de Ciencias Agrarias y Forestales | es_CL |
| dc.ucm.indexacion | Scopus | es_CL |
| dc.ucm.indexacion | Isi | es_CL |
| dc.ucm.uri | www.jstage.jst.go.jp/article/jgam/65/6/65_2019.01.003/_article | es_CL |
| dc.ucm.doi | doi.org/10.2323/jgam.2019.01.003 | es_CL |
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Excepto si se señala otra cosa, la licencia de la publicación se describe como Atribución-NoComercial-SinDerivadas 3.0 Chile