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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2011.tde-09022012-105548
Document
Author
Full name
Daniela Carolina Pinto e Licio
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Gomez, José Gregorio Cabrera (President)
Maldonado, Gabriel Padilla
Spira, Beny
Title in Portuguese
Isolamento de bactérias produtoras de polihidroxialcanoatos e caracterização molecular de sua PHA sintase.
Keywords in Portuguese
Pseudomonas
Esgotos sanitários
Genética bacteriana
Linhagens bacterianas
Polihidroxialcanoatos
Abstract in Portuguese
Um total de 1.520 bactérias isoladas a partir de amostras de lodo de esgoto doméstico e de solo de área preservada foi avaliado para a produção de polihidroxialcanoatos (PHA) com os corantes lipofílicos Nile Red A e Sudan Black B, permitindo a detecção de 261 isolados. Ensaios de produção de PHA revelaram isolados produtores de P3HB apresentando valores superiores à linhagem Burkholderia sacchari LFM101 e valores de produção de PHAMCL superiores à linhagem Pseudomonas sp. LFM046, além de 6 isolados produtores da mistura de P3HB e PHAMCL. Os primers RECF1/RECR permitiram a detecção específica do gene phaC do tipo I de três bactérias produtoras de P3HB, e os primers P613F/P613R detectaram genes phaC do tipo I em bactérias produtoras da mistura de polímeros. Um fragmento de DNA de 4 kpb de uma biblioteca genômica do isolado RMP1058BII, produtor da mistura de P3HB e PHAMCL, foi detectado utilizando os primers P613F/P613R, e a expressão desse fragmento de DNA uma linhagem de Pseudomonas mutada no gene phaC levou à produção de pequena quantidade de P3HB.
Title in English
Isolation of polyhydroxyalkanoates producing bacteria and molecular characterization of their PHA synthase.
Keywords in English
Pseudomonas
Bacterial genetics
Bacterial strains
Polyhydroxyalkanoates
Sewage
Abstract in English
A total of 1520 isolates from domestic sewage sludge or soil of preserved area were evaluated in regards of PHA production using the lipophilic Nile Red A or Sudan Black B dyes, allowing the detection of 261 isolates. PHA production experiments revealed P3HB producing isolates at higher amounts than the strain Burkholderia sacchari LFM101 and PHAMCL production rates higher than those presented by the strain Pseudomonas sp. LFM046, besides 6 isolates producing P3HB and PHAMCL mixtures. The RECF1/RECR primers allowed the specific type I phaC genes detection in three P3HB producing bacteria and the P613F/P613R primers detected type I phaC genes in the polymer mixture producing isolates. A 4 kbp DNA fragment from a genomic library of the isolate RMP1058BII was detected using the P613F/P613R primers. This DNA fragment expression in a Pseudomonas strain mutated in phaC gene leaded to the production of small amounts of P3HB.
 
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Publishing Date
2012-03-01
 
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