Prospection and Evaluation of (Hemi) Cellulolytic Enzymes Using Untreated and Pretreated Biomasses in Two Argentinean Native Termites

Saccharum officinarum bagasse (common name: sugarcane bagasse) and Pennisetum purpureum (also known as Napier grass) are among the most promising feedstocks for bioethanol production in Argentina and Brazil. In this study, both biomasses were assessed before and after acid pretreatment and following hydrolysis with Nasutitermes aquilinus andCortaritermes fulviceps termite gut digestome. The chemical composition analysis of the biomasses after diluted acid pretreatment showed that the hemicellulose fraction was partially removed. The (hemi) cellulolytic activities were evaluated in bacterial culture supernatantsof termite gut homogenates grown in treated and untreated biomasses. In all cases, we detected significantly higher endoglucanase and xylanase activities using pretreated biomasses compared to untreated biomasses, carboxymethylcellulose and xylan. Several protein bands with (hemi) cellulolytic activity were detected in zymograms and two-dimensionalgel electrophoresis. Some proteins of these bands or spots were identified as xylanolytic peptides by mass spectrometry. Finally, the diversity of cultured cellulolytic bacterial endosymbionts associated to both Argentinean native termite species was analyzed. This study describes, for the first time, bacterial endosymbionts and endogenous (hemi) cellulases of two Argentinean native termites as well as their potential application in degradation of lignocellulosic biomass for bioethanol production.Fil: Ben Guerrero, Emiliano. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Arneodo Larochette, Joel Demián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; ArgentinaFil: Bombarda Campanha, Raquel. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; BrasilFil: Oliveira, Patrícia Abrão de. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; BrasilFil: Labate, Mônica T. Veneziano. Universidade de Sao Paulo; BrasilFil: Cataldi, Thaís Regiani. Universidade de Sao Paulo; BrasilFil: Campos, Eleonora. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cataldi, Ángel Adrián. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Labate, Carlos A.. Universidade de Sao Paulo; BrasilFil: Rodrigues, Clenilson Martins. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; BrasilFil: Talia, Paola Monica. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Características físico-químicas e estruturais de amidos nativos e suas dextrinas Naegeli

Amido é um polímero semicristalino constituído basicamente por amilose e amilopectina. Seus grânulos são formados por regiões amorfas intercaladas com áreas mais densas compostas por lamelas alternadas de material cristalino e amorfo. O empacotamento das moléculas no grânulo é dependente da fonte botânica. Raízes e tubérculos são ricos em amido e seu plantio não requer muito investimento. Assim, o interesse industrial vem aumentando. O conhecimento da estrutura granular é fundamental para melhor entendimento das propriedades funcionais e melhor utilização do amido. A hidrólise ácida é uma técnica muito utilizada no estudo da estrutura do amido. O tratamento do amido com H2SO4 15% produz uma fração ácido-resistente, com baixa massa molar, conhecida como dextrina Naegeli (DN). O objetivo deste trabalho foi determinar a estrutura granular dos amidos de mandioca, batata, batata-doce e mandioquinha-salsa através da hidrólise ácida. Os amidos foram incubados com H2SO4 15,3% a 38ºC, por até 30 dias, e suas DN foram analisadas. A hidrólise ácida ocorreu em duas etapas: a primeira foi atribuída à rápida degradação das áreas amorfas dos grânulos, enquanto a segunda fase correspondeu à lenta hidrólise das regiões cristalinas. O amido de mandioquinha-salsa foi o mais suscetível à hidrólise, enquanto que os amidos de batata e batata-doce foram os mais resistentes. Os amidos de batata e mandioquinha-salsa apresentaram padrões cristalinos tipo B e mostraram maiores proporções de cadeias laterais longas da amilopectina (GP 37) do que os amidos de mandioca (tipo CA) e batata-doce (tipo C). Além de grande proporção de cadeias longas, o amido de mandioquinha-salsa também apresentou grande proporção de cadeias curtas (32,0%). O amido de mandioca também apresentou grande proporção de cadeias curtas (30,5%). Esses amidos mostraram um ombro em GP 17-21...Starch is a semi-crystalline polymer composed mainly of amylose and amylopectin. The starch granules are formed by successive layers that include amorphous areas interspersed with more dense areas composed of alternated lamellae of crystalline and amorphous material. Packing of amylose and amylopectin into granules is dependent of botanical source. Tubers and roots are rich in starch and their cultivation does not require much investment. Thus, the industrial interest has increased. Knowledge of the starch structure is fundamental for better understanding the functional properties and better utilization of the starch. Acid hydrolysis is widely used to study starch structure. Treatment of starch with 15% H2SO4 solution produces an acid-resistant fraction with low molecular weight known as Naegeli dextrin (ND). Objective of this study was to determine the structural characteristics of cassava, potato, sweet potato and Peruvian carrot starches by using acid hydrolysis. The starches were submitted to 15.3% H2SO4 at 38ºC for up to 30 days and their ND were analyzed. Acid hydrolysis displayed two phases: the first one was attributed to faster degradation of amorphous areas of granules, whereas the second phase corresponded to slower hydrolysis of crystalline regions. Peruvian carrot starch was the most susceptible to hydrolysis, whereas potato and sweet potato starches were the most resistant. Potato and Peruvian carrot starches displayed B-type X-ray pattern and had larger proportions of long branch-chains of amylopectin (DP 37) than cassava (CA-type) and sweet potato (C-type) starches. Beyond the great proportion of long branch-chains, the Peruvian carrot starch also showed great proportion of short branch-chains (32.0%). Cassava starch also had great proportion of short branch-chains (30.5%). These starches displayed a shoulder at DP 17-21 on branch chain-length distribution... (Complete abstract click electronic access below)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Gelatinization properties of native starches and their Naegeli dextrins

Cassava, potato, sweet potato, and Peruvian carrot starches were hydrolyzed with 15% v/v sulfuric acid solution for up to 30 days. Naegeli dextrins obtained from 1, 3, 6, 12, and 30 days were evaluated using differential scanning calorimeter (DSC) and scanning electron microscopy (SEM). Two phases of hydrolysis were found. The first phase was attributed to faster degradation of amorphous areas of granules, whereas the second phase corresponded to slower degradation of crystalline regions. Peruvian carrot starch was the most susceptible to acid, whereas potato and sweet potato starches were the most resistant. From DSC, it was observed a progressive reduction in peak height and a broadening of peaks with increasing hydrolysis time. The peaks shifted to higher temperatures. Onset temperature decreased on first day of hydrolysis for cassava and Peruvian carrot starches, and on third day for potato and sweet potato. Enthalpy decreased during first stage of hydrolysis in cassava and Peruvian carrot starches, and during second phase, it reduced in all starches. SEM showed that the granule surfaces were degraded by erosion on the first day of treatment, followed by degradation of amorphous areas. on third day, potato and sweet potato starches still displayed some granules almost intact, whereas cassava and Peruvian carrot starch granules were totally degraded, confirming their high susceptibility to acid attack. on sixth day of hydrolysis, starch granules had faceted structures, characteristic of crystalline material. The effect that acid hydrolysis had on thermal properties of starches depended on both hydrolysis stage and starch source.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Extração e caracterização do amido de diferentes genótipos de bananeira

A banana, uma das principais herbáceas cultivadas em grande quantidade no mundo e importante fonte de energia para as pessoas, tem obtido interesse como matéria-prima para a produção de farinha e fécula para uso alimentício e outros fins industriais, devido ao elevado conteúdo de amido nos frutos verdes. O trabalho teve como objetivo caracterizar os frutos verdes de sete genótipos de bananeira quanto à composição química e avaliar o conteúdo de amilose, propriedades de pasta e térmicas dos amidos destes genótipos. Os resultados obtidos mostraram diferenças significativas na composição química dos frutos, com maior teor de amido na cultivar Figo Cinza. As análises dos amidos mostraram valores de amilose de 26,68 a 33,95% e propriedades de pasta com elevado pico de viscosidade, baixa resistência à temperatura e agitação (quebra de viscosidade) e tendência à retrogradação para todos os amidos. Nas propriedades térmicas, os amidos mostraram um endoderma, temperatura de gelatinização na faixa de 60 a 73ºC com DH variando de 11,6 a 16,99 J g-1 na gelatinização e 4,11 a 7,97 J g-1 na retrogradação

Quantitative enzymatic activities in cell-free culture supernatants.

<p>Endoglucanase. (B) Xylanase. Data were calculated as the means ± SD of triplicates assay ****<i>P<</i>0.0001; **<i>P</i><0.01.</p

Rarefaction analysis of 16S rRNA gene sequences.

<p>(A) CMC—cultures from <i>N</i>. <i>aquilinus</i>. (B) CMC—cultures from <i>C</i>. <i>fulviceps</i>.</p

Percentage of biomass composition of the untreated or pretreated (-P) <i>Saccharum officinarum</i> bagasse (SOB) and <i>Pennisetum purpureum</i> (PP)^*.

<p>*Values with the same lower case in the same column are not significantly different at <i>p</i> < 0.05; values with the same capital letter in the same column are not significantly different at <i>p</i> < 0.05.</p><p>Percentage of biomass composition of the untreated or pretreated (-P) <i>Saccharum officinarum</i> bagasse (SOB) and <i>Pennisetum purpureum</i> (PP)^{<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0136573#t001fn001" target="_blank">*</a>}.</p