Marine-derived Fungi: Diversity Of Enzymes And Biotechnological Applications

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)The ocean is considered to be a great reservoir of biodiversity. Microbial communities in marine environments are ecologically relevant as intermediaries of energy, and play an important role in nutrient regeneration cycles as decomposers of dead and decaying organic matter. In this sense, marine-derived fungi can be considered as a source of enzymes of industrial and/or environmental interest. Fungal strains isolated from different substrates, such as invertebrates, decaying wood, seawater, sediments, and mangrove detritus, have been reported to be producers of hydrolytic and/or oxidative enzymes, with alginate lyase, amylase, cellulase, chitinase, glucosidase, inulinase, keratinase, ligninase, lipase, nuclease, phytase, protease, and xylanase being among the enzymes produced by fungi of marine origin. These enzymes present temperature and pH optima ranging from 35 to 70 degrees C, and 3.0 to 11.0, respectively. High-level production in bioreactors is mainly performed using submerged-state fermentation. Certain marine-derived fungal strains present enzymes with alkaline and cold-activity characteristics, and salinity is considered an important condition in screening and production processes. The adaptability of marine-derived fungi to oceanic conditions can be considered an attractive point in the field of fungal marine biotechnology. In this review, we focus on the advances in discovering enzymes from marine-derived fungi and their biotechnological relevance.6Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)FAPESP [2013/19486-0, 2013/08617-7]CNPq [304103/2013-6, 301248/2010-9]FAPESP [FAPESP 2009/18399-1, FAPESP 2011/18769-3, FAPESP 2008/06720-7, FAPESP 2012/12622-3, FAPESP 2013/12505-0, FAPESP 2014/12430-2, CNPq 159488/2014, FAPESP 2013/00286-1

The Production of Ligninolytic Enzymes by Marine-Derived Basidiomycetes and Their Biotechnological Potential in the Biodegradation of Recalcitrant Pollutants and the Treatment of Textile Effluents

Filamentous fungi derived from marine environments are well known as a potential genetic resource for various biotechnological applications. Although terrestrial fungi have been reported to be highly efficient in the remediation of xenobiotic pollutants, fungi isolated from the marine environment may possess biological advantages over terrestrial fungi because of their adaptations to high salinity and pH extremes. The present study describes the production of ligninolytic enzymes under saline and non-saline conditions and the decolorization of Remazol Brilliant Blue R (RBBR) dye by three basidiomycetes recovered from marine sponges (Tinctoporellus sp. CBMAI 1061, Marasmiellus sp. CBMAI 1062, and Peniophora sp. CBMAI 1063). Ligninolytic enzymes were primarily produced by these fungi in a salt-free malt extract and malt extract formulated with artificial seawater (saline condition). CuSO4 and wheat bran were the best inducers of lignin peroxidase and manganese peroxidase activity. RBBR was decolorized up to 100% by the three fungi, and Tinctoporellus sp. CBMAI 1061 was the most efficient. Our results revealed the biotechnological potential of marine-derived basidiomycetes for dye decolorization and the treatment of colored effluent as well as for the degradation of other organopollutants by ligninolytic enzymes in non-saline and saline conditions that resemble the marine environment.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Enhanced Textile Dye Decolorization By Marine-derived Basidiomycete Peniophora Sp Cbmai 1063 Using Integrated Statistical Design

In the present study, the biotechnological potential of the marine-derived fungus Peniophora sp. CBMAI 1063 was investigated in relation to Reactive Black 5 (RB5) dye decolorization and degradation using an integrated statistical design composed of Plackett-Burman design (P&B), central composite design (CCD), and response surface methodology (RSM). RB5 dye was effectively decolorized (94 %) in saline conditions, without any detection of mutagenic compounds, and simultaneously, 57 % of total organic carbon (TOC) was removed in 7 days. The activity of lignin peroxidase (LiP) was not detected during the process. The gene expression of laccase (Lac) and manganese peroxidase (MnP) enzymes produced during the process was evaluated, and results from this experiment coupled with LC-MS analyses revealed that in the early stage of dye decolorization, a higher MnP gene expression and significant enzymatic activity was detected in Peniophora sp. CBMAI 1063 with the formation of p-Base and TAHNDS compounds. This paper reports innovative data related to the textile dye decolorization by the marine-derived basidiomycete Peniophora sp. CBMAI 1063, showing the metabolites formed and enzymatic action throughout the process in saline condition. The strategy used showed to be an efficient statistical approach that provides an attractive solution for the screening and simultaneous optimization of the degradation process.239865986681st International Conference on Chemical Biological Radiological and Nuclear, Research and Innovation (CBRN-RI)MAR 16-18, 2015Antibes Juan les Pins, FRANC

Biotechnological potential of microorganisms from landfill leachate: isolation, antibiotic resistance and leachate discoloration

Abstract Disposal of municipal solid waste (MSW) can be considered a risk to human health representing a great environmental problem in several countries. MSW landfills are a significant source of toxic elements in the environment. Microorganisms able to thriving in leachate wastewater may exhibit metabolic machinery to synthesize a wide range of enzymes able to degrade and/or discolor toxic compounds from leachate. The use of non-pathogenic microbial cells for human health, recovered from leachate for biotechnological application, can be considered a promising approach in bioremediation processes of toxic compounds found in these environments. The present work aimed to the isolation, antibiotic resistance evaluation and leachate discoloration by microorganisms isolated from landfill leachate of Foz do Iguaçu. Forty bacteria and fifteen filamentous fungi were isolated. From these, six bacterial showed resistance at least one tested antibiotic, while six fungal isolates showed resistance to the antimycotic nystatin. CCMIBA_4L (unidentified bacteria) and Paecilomyces sp. CCMIBA_5N, were able to discolor 19.15% and 25.26% of the leachate, respectively. The results of the present work encourage future studies to characterize the enzymes involved in the discoloration and degradation of the leachate. The findings demonstrated the potential for the use of microorganisms from landfill leachate as bioremediation tools

Improvement in extracellular protease production by the marine antarctic yeast Rhodotorula mucilaginosa L7

Microorganisms from extreme and restrictive eco systems, such as the Antarctic continent, are of great interest due to their ability to synthesize products of commercial value. Among these, enzymes from psychrotolerant and psychrophilic microorganisms offer potential economical benefits due to their high activity at low and moderate temperatures. The cold adapted yeast Rhodotorula mucilaginosa L7 was selected out of 97 yeasts isolated from Antarctica as having the highest extracellular proteolytic activity in preliminary tests. The present study was aimed at evaluating the effects of nutrient composition (peptone, rice bran extract, ammonium sulfate, sodium chloride) and physicochemical parameters (temperature and pH) on its proteolytic activity. A 26−2 fractional factorial design experiment followed by a central composite design (CCD 23) was performed to optimize the culture conditions and improve the extracellular proteolytic activity. The results indicated that the presence of peptone in the medium was the most influential factor in protease production. Enzymatic activity was enhanced by the interaction between low glucose and peptone concentrations. The optimization of culture conditions with the aid of mathematical modeling enabled a c. 45% increase in proteolytic activity and at the same time reduced the amount of glucose and peptone required for the culture. Thus culture conditions established in this work may be employed in the biotechnological production of this protease.Fil: Chaud, Luciana C. S.. Universidade de Sao Paulo; BrasilFil: Lario, Luciana Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidade de Sao Paulo; BrasilFil: Bonugli Santos, Rafaella C.. Federal University of Latin American Integration; BrasilFil: Sette, Lara D.. Universidade de Sao Paulo; BrasilFil: Pessoa Junior, Adalberto. Universidade de Sao Paulo; BrasilFil: Felipe, Maria das Graças de A.. Universidade de Sao Paulo; Brasi

Enhanced Textile Dye Decolorization By Marine-derived Basidiomycete Peniophora Sp. Cbmai 1063 Using Integrated Statistical Design.

In the present study, the biotechnological potential of the marine-derived fungus Peniophora sp. CBMAI 1063 was investigated in relation to Reactive Black 5 (RB5) dye decolorization and degradation using an integrated statistical design composed of Plackett-Burman design (P&B), central composite design (CCD), and response surface methodology (RSM). RB5 dye was effectively decolorized (94 %) in saline conditions, without any detection of mutagenic compounds, and simultaneously, 57 % of total organic carbon (TOC) was removed in 7 days. The activity of lignin peroxidase (LiP) was not detected during the process. The gene expression of laccase (Lac) and manganese peroxidase (MnP) enzymes produced during the process was evaluated, and results from this experiment coupled with LC-MS analyses revealed that in the early stage of dye decolorization, a higher MnP gene expression and significant enzymatic activity was detected in Peniophora sp. CBMAI 1063 with the formation of p-Base and TAHNDS compounds. This paper reports innovative data related to the textile dye decolorization by the marine-derived basidiomycete Peniophora sp. CBMAI 1063, showing the metabolites formed and enzymatic action throughout the process in saline condition. The strategy used showed to be an efficient statistical approach that provides an attractive solution for the screening and simultaneous optimization of the degradation process.2

Enhanced textile dye decolorization by marine-derived basidiomycete Peniophora Sp CBMAI 1063 using integrated statistical design

In the present study, the biotechnological potential of the marine-derived fungus Peniophora sp. CBMAI 1063 was investigated in relation to Reactive Black 5 (RB5) dye decolorization and degradation using an integrated statistical design composed of Plackett-Burman design (P&B), central composite design (CCD), and response surface methodology (RSM). RB5 dye was effectively decolorized (94 %) in saline conditions, without any detection of mutagenic compounds, and simultaneously, 57 % of total organic carbon (TOC) was removed in 7 days. The activity of lignin peroxidase (LiP) was not detected during the process. The gene expression of laccase (Lac) and manganese peroxidase (MnP) enzymes produced during the process was evaluated, and results from this experiment coupled with LC-MS analyses revealed that in the early stage of dye decolorization, a higher MnP gene expression and significant enzymatic activity was detected in Peniophora sp. CBMAI 1063 with the formation of p-Base and TAHNDS compounds. This paper reports innovative data related to the textile dye decolorization by the marine-derived basidiomycete Peniophora sp. CBMAI 1063, showing the metabolites formed and enzymatic action throughout the process in saline condition. The strategy used showed to be an efficient statistical approach that provides an attractive solution for the screening and simultaneous optimization of the degradation process2398659866

Pyrene degradation by marine-derived ascomycete: process optimization, toxicity, and metabolic analyses

Marine-derived fungi are relevant genetic resources for bioremediation of saline environments/processes. Among the five fungi recovered from marine sponges able to degrade pyrene (Py) and benzo[a]pyrene (BaP), Tolypocladium sp. strainCBMAI 1346 and Xylaria sp. CBMAI 1464 presented the best removal rates of Py and BaP, respectively. Since the decrease in BaP was related to mycelial adsorption, a combined strategy was applied for the investigation of Py degradation by the fungus Tolypocladium sp. CBMAI 1346. The selected fungus was able to degrade about 95% of Py after 7days of incubation (optimized conditions), generating metabolites different from the ones found before optimization. Metabolites and transcriptomic data revealed that the degradation occurred mainly by the cytochrome P450 pathway. Putative monooxygenases and dioxygenases found in the transcriptome may play an important role. After 21days of degradation, no toxicity was found in the optimized culture conditions. The findings from the present study highlight the potential of marine-derived fungi to degrade environmental pollutants and convey innovative information related to the metabolism of pyrene26121241212424FAPESP – Fundação de Amparo à Pesquisa Do Estado De São Paulo2013/19486-0; 2016/07957-

Microbial Diversity Associated With Algae, Ascidians And Sponges From The North Coast Of São Paulo State, Brazil.

Little is known about the microbial diversity associated with marine macroorganisms, despite the vital role microorganisms may play in marine ecosystems. The aim of the present study was to investigate the diversity of bacteria and fungi isolated from eight marine invertebrate and one algae samples. Data derived from ARDRA and sequencing analyses allowed the identification of marine-derived microorganisms isolated from those samples. Microbial strains identified up to the genus level revealed 144 distinct ribotypes out of 256 fungal strains and 158 distinct ribotypes out of 181 bacterial strains. Filamentous fungi were distributed among 24 different genera belonging to Ascomycota, Zygomycota and Basidiomycota, some of which had never been reported in the literature as marine invertebrate-inhabiting fungi (Pestalotiopsis, Xylaria, Botrysphaeria and Cunnninghamella). Bacterial isolates were affiliated to 41 different genera, being Bacillus, Ruegeria, Micrococcus, Pseudovibrio and Staphylococcus the most abundant ones. Results revealed an unexpected high microbial diversity associated to the macroorganisms which have been collected and suggested the selection of certain microbial taxonomic groups according to the host. The combined data gathered from this investigation contribute to broaden the knowledge of microbial diversity associated to marine macroorganisms, including as a promising source for the discovery of new natural products.165466-8