Biossurfactants of endophytic fungi isolated from Gustavia cf. hexapetala (Alb.) Sm. (Lecythidaceae) in the Amazon region, against oral cavity pathogens

Autor: Ferreira, Francisca da Silva
Přispěvatelé: Pereira, Jos?? Odair, Souza, Antonia Queiroz Lima de, Albuquerque, Patricia Melchinna, Conde, Nikeila Chacon de Oliveira, Souza, Jo??o Vicente Braga de, Matsuura, Ani Beatriz Jackisch
Jazyk: portugalština
Rok vydání: 2020
Předmět:
Zdroj: Biblioteca Digital de Teses e Dissertações da UFAM
Universidade Federal do Amazonas (UFAM)
instacron:UFAM
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Modelo dispon??vel em https://biblioteca.ufam.edu.br/deposito-de-teses-e-dissertacoes.html on 2020-12-16T14:55:42Z (GMT) Submitted by Francisca Ferreira (fransferreirauea@gmail.com) on 2020-12-21T15:23:14Z No. of bitstreams: 3 Tese - Francisca da Silva Ferreira.pdf: 5552009 bytes, checksum: 469f3f1fc074f7cd9dda3e433b37438c (MD5) 330 Ata de Defesa da Tese FRANCISCA FERREIRA.pdf: 388077 bytes, checksum: 96e91a6eb3c02ccc0c3d1795a1999c79 (MD5) CartaEncaminhamentoAutodepo??sito (Assinada).pdf: 42096 bytes, checksum: f1f5e804790806449ca27868edefbe61 (MD5) Approved for entry into archive by PPG BIONORTE (secestadualppgbionorte@uea.edu.br) on 2020-12-28T14:08:20Z (GMT) No. of bitstreams: 3 Tese - Francisca da Silva Ferreira.pdf: 5552009 bytes, checksum: 469f3f1fc074f7cd9dda3e433b37438c (MD5) 330 Ata de Defesa da Tese FRANCISCA FERREIRA.pdf: 388077 bytes, checksum: 96e91a6eb3c02ccc0c3d1795a1999c79 (MD5) CartaEncaminhamentoAutodepo??sito (Assinada).pdf: 42096 bytes, checksum: f1f5e804790806449ca27868edefbe61 (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2020-12-28T17:05:48Z (GMT) No. of bitstreams: 3 Tese - Francisca da Silva Ferreira.pdf: 5552009 bytes, checksum: 469f3f1fc074f7cd9dda3e433b37438c (MD5) 330 Ata de Defesa da Tese FRANCISCA FERREIRA.pdf: 388077 bytes, checksum: 96e91a6eb3c02ccc0c3d1795a1999c79 (MD5) CartaEncaminhamentoAutodepo??sito (Assinada).pdf: 42096 bytes, checksum: f1f5e804790806449ca27868edefbe61 (MD5) Made available in DSpace on 2020-12-28T17:05:48Z (GMT). No. of bitstreams: 3 Tese - Francisca da Silva Ferreira.pdf: 5552009 bytes, checksum: 469f3f1fc074f7cd9dda3e433b37438c (MD5) 330 Ata de Defesa da Tese FRANCISCA FERREIRA.pdf: 388077 bytes, checksum: 96e91a6eb3c02ccc0c3d1795a1999c79 (MD5) CartaEncaminhamentoAutodepo??sito (Assinada).pdf: 42096 bytes, checksum: f1f5e804790806449ca27868edefbe61 (MD5) Previous issue date: 2020-02-13 FAPEAM - Funda????o de Amparo ?? Pesquisa do Estado do Amazonas CNPq - Conselho Nacional de Desenvolvimento Cient??fico e Tecnol??gico Biosurfactants (BS) are amphiphilic molecules which, due to their chemical characteristics, have been of great industrial interest. There is evidence of application in dentistry due to their antimicrobial action against pathogens of interest. Thus, this study evaluated the production of BS in strains of endophytic fungi isolated from Gustavia cf. hexapetala, in the city of Manaus, with antimicrobial action against pathogens in the oral cavity. The isolation of endophytic fungi from branches and leaves, purification, conservation and morphological characterization of the isolates were carried out. Screening was performed to select BS-producing fungi grown in liquid (BDL and mineral) supplemented with soybean oil. The culture conditions were: pH 7.0, temperature of 28 ??C at 170 rpm, for 10 days. BS production was evaluated using the emulsification index (E24) and the Surface Tension (ST) of the culture medium. A new culture was made on an expanded scale for the extraction of BS. With the raw BS, it was possible to determine the CMC, antimicrobial tests and chemical profile. The selected fungi were identified using molecular techniques. Three hundred ninety-nine (399) fungi and a black yeast of G. cf. hexap??tala were isolated. The isolation frequency was 74.6% from branches and 49.6% from leaves. They were grouped into 20 morphogroups with eight identified genera: Colletotrichum, Fusarium, Pestalotiopsis, Penicillium, Guigniardia, Phomopsis, Xylaria, Diaphorte and the others, unknown (Micelia sterilia). One hundred eighty-seven (187) endophytic fungi were conserved, of which 24 were evaluated, and of these, nine stood out as BS producers, and five were selected: three branch isolates were identified as Fusarium decemcellulare (FF17), Diaporthe sp. (FF36) and Fusarium solani (FF45); two leaf isolates were identified as Fusarium solani (FF51) and Coletotrichum vietnamensi (FF33). The best E24 results were: C. vietnamensi (FF33) with 33.99%, Diaporthe sp. (FF36) with 31.97%, and F. solani (FF51) with 26.70%. There was ST reduction below 35.0 mN/m in 09 strains, with emphasis on F. solani (FF51) with 28.7 and 30.1 mN/m; F. solani (FF45) with 31.5 and 30.1 mN / m; C. vietnamensi (FF33) with 32.4 and 31.8 mN/m; Diaporthe sp. (FF36) with 32.7 and 30.9 mN/m grown in BDL and mineral medium, respectively. The ST of the mineral medium decreased from 61.0 mN/m to 30.9 mN/m (49.3% reduction) for fungus Diaporthe sp. (FF36), and for fungi F. solani (FF45 and FF51) with 30.1 mN/m (50.6% reduction). The highest yield of BS production was 0.66 g/L, obtained from the extract of fungus C. vietnamensi (FF33). This endophyte produced a BS with the lowest CMC (1.08 mg/mL) reducing the ST of ultrapure water from 70.6 to 31.94 mN/m (54.8% reduction). In the antimicrobial tests, the MIC of F. decemcellulare (FF17) extract against Streptococcus mutans (ATCC25175) was 0.4 mg/mL and the MBC was 0.5 mg/mL; for Lactobacillus casei (ATCC7469), the MIC was 1.5 mg/ml, and the MBC was 1.75 mg/ml. The bioactives produced by Diaporthe sp. (FF36) showed an MIC of 2.4 mg/mL against S. mutans. The MIC of F. solani mycelium extract (FF51) for L. casei was 1.8 mg/mL. The mycelium extract of F. solani (FF45) showed bacteriostatic action for L. casei and fungistatic for C. albicans (MIC of 2.1 mg/mL and 4.2 mg/mL, respectively). The results indicate the importance of research aimed at the prospection of endophytic fungi with a potential to produce BS and other bioactives of biotechnological interest, against pathogens in the oral cavity, allowing the use of natural molecules to control oral diseases. Os biossurfactantes (BS) s??o mol??culas anfif??licas que por suas caracter??sticas qu??micas t??m apresentado grande interesse industrial. H?? evid??ncias de aplica????o na ??rea odontol??gica por apresentarem a????o antimicrobiana contra pat??genos de interesse. Nesse aspecto o presente estudo avaliou a produ????o de BS em linhagens de fungos endof??ticos isolados de Gustavia cf. hexapetala, na cidade de Manaus, com a????o antimicrobiana contra pat??genos da cavidade oral. Foi realizado isolamento de fungos endof??ticos de galhos e folhas, purifica????o, conserva????o e caracteriza????o morfol??gica dos isolados. Realizou-se uma triagem para selecionar fungos produtores de BS cultivados em meio l??quido (BDL e mineral) suplementados com ??leo de soja. As condi????es de cultivo foram: pH 7.0, temperatura de 28??C ?? 170rpm, por 10 dias. A produ????o de BS foi avaliada por meio do ??ndice de emulsifica????o (E24) e da Tens??o Superficial (TS) do meio de cultivo. Foi realizado um novo cultivo em escala ampliada para a extra????o dos BS. Com os BS brutos foi poss??vel determinar a CMC, testes antimicrobianos e perfil qu??mico. Os fungos selecionados foram identificados por meio de t??cnicas moleculares. Foram isolados 399 fungos e uma levedura negra de G. Cf. hexap??tala. A frequ??ncia de isolamento foi 74,6% a partir de galhos e 49,6% a partir da folha. Reagrupou-se em 20 morfogrupos com oito g??neros identificados: Colletotrichum, Fusarium, Pestalotiopsis, Penicillium, Guigniardia, Phomopsis, Xylaria, Diaphorte e os demais desconhecidos (Micelia sterilia). Foram conservados 187 fungos endof??ticos,, avaliados 24 e destes, nove se destacaram como produtores de BS, sendo cinco selecionados, tais quais: tr??s isolados de galho identificados como Fusarium decemcellulare (FF17), Diaporthe sp. (FF36) e Fusarium solani (FF45); dois isolados de folha, identificados como Fusarium solani (FF51) ) e Coletotrichum vietnamensi (FF33). Os melhores resultados do E24 foram: C. vietnamensi (FF33) com 33,99%, Diaporthe sp. (FF36) com 31,97%, e F. solani (FF51) com 26,70%. Houve redu????o da TS abaixo de 35,0 mN/m em 09 cepas com destaque para F. solani (FF51) com 28,7 e 30,1 mN/m; F. solani (FF45) com 31,5 e 30,1 mN/m; C. vietnamensi (FF33) com 32,4 e 31,8 mN/m; Diaporthe sp. (FF36) com 32,7 e 30,9 mN/m cultivados em meio BDL e mineral respectivamente. A TS do meio mineral reduziu de 61,0 mN/m para 30,9 mN/m (49,3% de redu????o) para o fungo Diaporthe sp. (FF36), e para os fungos F. solani (FF45 e FF51) com 30,1 mN/m (50,6% de redu????o). O maior rendimento da produ????o de BS foi de 0,66 g/L, obtido com o extrato do fungo C. vietnamensi (FF33). Este endof??fito produziu um BS com a menor CMC (1,08 mg/mL) reduzindo a TS da ??gua ultrapura de 70,6 para 31,94 mN/m (54,8% de redu????o). Nos testes antimicrobianos a CIM do extrato de F. decemcellulare (FF17) contra Streptococcus mutans (ATCC25175) foi de 0,4 mg/mL e a CBM foi 0,5 mg/mL; para Lactobacillus casei (ATCC7469), a CIM foi 1,5 mg/mL, e a CBM foi 1,75 mg/mL. Os bioativos produzidos por Diaporthe sp. (FF36) apresentaram CIM de 2,4 mg/mL contra S. mutans. A CIM do extrato de mic??lio de F. solani (FF51) para o L. casei foi 1,8 mg/mL. O extrato de mic??lio de F. solani (FF45) apresentou a????o bacteriost??tica para L. casei e fungist??tica para C. albicans (CIM de 2,1 mg/mL e 4,2 mg/mL, respectivamente). Os resultados indicam a import??ncia de pesquisas que visam a prospec????o de fungos endof??ticos com potencial para a produ????o de BS e outros bioativos de interesse biotecnol??gicos, contra pat??genos da cavidade oral, permitindo inserir o uso de mol??culas naturais para o controle de doen??as bucais. A PESQUISA ABORDA A PRODU????O DE BIOSSURFACTANTES POR FUNGOS ENDOF??TICOS COM A????O CONTRA PAT??GENOS DA CAVIDADE ORAL.
Databáze: OpenAIRE
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