Secretion of laccase and manganese peroxidase by Pleurotus strains cultivate in solid-state using Pinus spp. sawdust

Pleurotus species secrete phenol oxidase enzymes: laccase (Lcc) and manganese peroxidase (MnP). New genotypes of these species show potential to be used in processes aiming at the degradation of phenolic compounds, polycyclic aromatic hydrocarbons and dyes. Hence, a screening of some strains of Pleurotus towards Lcc and MnP production was performed in this work. Ten strains were grown through solid-state fermentation on a medium based on Pinus spp. sawdust, wheat bran and calcium carbonate. High Lcc and MnP activities were found with these strains. Highest Lcc activity, 741 ± 245 U gdm-1 of solid state-cultivation medium, was detected on strain IB11 after 32 days, while the highest MnP activity occurred with strains IB05, IB09, and IB11 (5,333 ± 357; 4,701 ± 652; 5,999 ± 1,078 U gdm-1, respectively). The results obtained here highlight the importance of further experiments with lignocellulolytic enzymes present in different strains of Pleurotus species. Such results also indicate the possibility of selecting more valuable strains for future biotechnological applications, in soil bioremediation and biological biomass pre-treatment in biofuels production, for instance, as well as obtaining value-added products from mushrooms, like phenol oxidase enzymes.(AU)

Bacillus thuringiensis isolates entomopathogenic for Culex quinquefasciatus (Diptera: Culicidae) and Anticarsia gemmatalis (Lepidoptera: Noctuidae) / Isolados de Bacillus thuringiensis entomopatogênicos para Culex quinquefasciatus (Diptera: Culicidae) e Anticarsia gemmatalis (Lepidoptera: Noctuidae)

Samples of the Bacillus thuringiensis (Bt) were collected from soil and insects. Eight isolates were selected from rural soil, 15 from urban soil and 11 from insects. These were evaluated for entomopathogenicity against larvae of Anticarsia gemmatalis and Culex quinquefasciatus. The pathogenicity tests showed that a higher percentage of isolates were active against A. gemmatalis (60 percent) compared to C. quinquefasciatus (31 percent). Probit analysis (LC50) indicated that against A. gemmatalis four of the isolates presented values similar to the reference strain against A. gemmatalis, while against C. quinquefasciatus one isolate showed an LC50 similar to the reference strain (IPS-82). SDS-PAGE characterisation of two isolates showed a 27 kDa protein fraction related to the Bt subspecies israelensis cytolytic toxin (cyt) gene. One 130 kDa protein, possibly related to the Bt crystal inclusions (cry1) gene, was identified in the other two isolates, which were more toxic for lepidoptera; another isolate presented a protein of 100 kDa. Some new local Bt isolates had similar LC50 probit values to the reference strains.

Amostras de Bacillus thuringiensis (Bt) foram coletadas do solo e de insetos. Oito isolados foram coletados de solo rural, 15 de solo urbano e 11 de insetos, os quais foram avaliados quanto a sua entomopatogenicidade contra larvas de Anticarsia gemmatalis e Culex quinquefasciatus. Os testes de patogenicidade mostraram uma alta porcentagem de isolados ativos contra A. gemmatalis (60 por cento), comparado a C. quinquefasciatus (31 por cento). Por análise de probit (CL50), verificou-se que quatro isolados apresentaram valores similares aos da estirpe de referência contra A. gemmatalis, enquanto somente um isolado mostrou CL50 similar à estirpe de referência (IPS 82) contra C.quinquefasciatus. A caracterização por SDS-PAGE de dois isolados mostrou uma proteína de 27 kDa relativa à toxina citolítica (Cyt) de B. thuringiensis subespécie israelensis. Uma proteína de 130 kDa, possivelmente relacionada à família do gene cry 1, foi identificada em outros dois isolados, os quais foram mais tóxicos para lepidópteros, enquanto que os outros dois isolados apresentaram uma proteína de 100 kDa. Alguns novos isolados locais de B. thuringiensis apresentaram valores de CL50 similares às estirpes de referência.

Bacillus thuringiensis isolates entomopathogenic for Culex quinquefasciatus (Diptera: Culicidae) and Anticarsia gemmatalis (Lepidoptera: Noctuidae)

Samples of the Bacillus thuringiensis (Bt) were collected from soil and insects. Eight isolates were selected from rural soil, 15 from urban soil and 11 from insects. These were evaluated for entomopathogenicity against larvae of Anticarsia gemmatalis and Culex quinquefasciatus. The pathogenicity tests showed that a higher percentage of isolates were active against A. gemmatalis (60%) compared to C. quinquefasciatus (31%). Probit analysis (LC50) indicated that against A. gemmatalis four of the isolates presented values similar to the reference strain against A. gemmatalis, while against C. quinquefasciatus one isolate showed an LC50 similar to the reference strain (IPS-82). SDS-PAGE characterisation of two isolates showed a 27 kDa protein fraction related to the Bt subspecies israelensis cytolytic toxin (cyt) gene. One 130 kDa protein, possibly related to the Bt crystal inclusions (cry1) gene, was identified in the other two isolates, which were more toxic for lepidoptera; another isolate presented a protein of 100 kDa. Some new local Bt isolates had similar LC50 probit values to the reference strains.

Amostras de Bacillus thuringiensis (Bt) foram coletadas do solo e de insetos. Oito isolados foram coletados de solo rural, 15 de solo urbano e 11 de insetos, os quais foram avaliados quanto a sua entomopatogenicidade contra larvas de Anticarsia gemmatalis e Culex quinquefasciatus. Os testes de patogenicidade mostraram uma alta porcentagem de isolados ativos contra A. gemmatalis (60%), comparado a C. quinquefasciatus (31%). Por análise de probit (CL50), verificou-se que quatro isolados apresentaram valores similares aos da estirpe de referência contra A. gemmatalis, enquanto somente um isolado mostrou CL50 similar à estirpe de referência (IPS 82) contra C.quinquefasciatus. A caracterização por SDS-PAGE de dois isolados mostrou uma proteína de 27 kDa relativa à toxina citolítica (Cyt) de B. thuringiensis subespécie israelensis. Uma proteína de 130 kDa, possivelmente relacionada à família do gene cry 1, foi identificada em outros dois isolados, os quais foram mais tóxicos para lepidópteros, enquanto que os outros dois isolados apresentaram uma proteína de 100 kDa. Alguns novos isolados locais de B. thuringiensis apresentaram valores de CL50 similares às estirpes de referência.