RESUMEN
This work demonstrates that sesame (Sesamum indicum L.) hull, an unexploited food industrial waste, can be used as an efficient source for the extraction of hemicellulose and/or pectin polysaccharides to further obtain functional oligosaccharides. Different polysaccharides extraction methods were surveyed including alkaline and several enzymatic treatments. Based on the enzymatic release of xylose, arabinose, glucose, and galacturonic acid from sesame hull by using different enzymes, Celluclast®1.5 L, Pectinex®Ultra SP-L, and a combination of them were selected for the enzymatic extraction of polysaccharides at 50 °C, pH 5 up to 24 h. Once the polysaccharides were extracted, Ultraflo®L was selected to produce arabinoxylo-oligosaccharides (AXOS) at 40 °C up to 24 h. Apart from oligosaccharides production from extracted polysaccharides, alternative approaches for obtaining oligosaccharides were also explored. These were based on the analysis of the supernatants resulting from the polysaccharide extraction, alongside a sequential hydrolysis performed with Celluclast®1.5 L and Ultraflo®L of the starting raw sesame hull. The different fractions obtained were comprehensively characterized by determining low molecular weight carbohydrates and monomeric compositions, average Mw and dispersity, and oligosaccharide structure by MALDI-TOF-MS. The results indicated that sesame hull can be a useful source for polysaccharides extraction (pectin and hemicellulose) and derived oligosaccharides, especially AXOS.
Asunto(s)
Oligosacáridos , Sesamum , Sesamum/química , Oligosacáridos/química , Hidrólisis , Polisacáridos/química , Xilanos/química , Xilanos/aislamiento & purificación , Pectinas/química , Pectinas/aislamiento & purificación , Residuos Industriales , Arabinosa/química , Xilosa/químicaRESUMEN
Agro-industrial wastes such as sugarcane bagasse, wheat bran, rice bran, corn cob and wheat straw are cheapest and abundantly available natural carbon sources. The present study was aimed to production of amylase and xylanase simultaneously using agro-industrial waste as the sole carbon source. Seven thermophilic strains of actinomycete were isolated from the mushroom compost. Among of these, strain designated MSC702 having high potential to utilize agro-industrial wastes for the production of amylase and xylanase. Strain MSC702 was identified as novel species of Streptomyces through morphological characterization and 16S rRNA gene sequence. Enzyme production was determined using 1% (w/v) of various agro-industrial waste in production medium containing (g/100mL): K2HPO4(0.1), (NH4)2SO4(0.1), NaCl (0.1), MgSO4(0.1) at pH 7.0 after incubation of 48 h at 50°C. The amylase activity (373.89 IU/mL) and xylanase activity (30.15 IU/mL) was maximum in rice bran. The decreasing order of amylase and xylanase activity in different type of agro-industrial wastes were found rice bran (RB) > corn cob (CC) > wheat bran (WB) > wheat straw (WS) > sugarcane bagasse (SB) and rice bran (RB) > wheat bran (WB) > wheat straw (WS) > sugarcane bagasse (SB) > corn cob (CC), respectively. Mixed effect of different agro-industrial wastes was examined in different ratios. Enzyme yield of amylase and xylanase was ~1.3 and ~2.0 fold higher with RB: WB in 1:2 ratio.
Asunto(s)
Actinobacteria/aislamiento & purificación , Amilasas/análisis , Amilasas/aislamiento & purificación , Secuencia de Bases , Activación Enzimática , Residuos Industriales/análisis , Streptomyces/aislamiento & purificación , Xilanos/análisis , Xilanos/aislamiento & purificación , Microbiología Industrial , MétodosRESUMEN
Aspergillus niger F7 isolated from soil was found to be the potent producer of cellulase and xylanase. The residue of forest species Toona ciliata, Celtris australis, Cedrus deodara and Pinus roxburghii was selected as substrate for biodegradation study due to its easy availability and wide use in industry. It was subjected to alkali (sodium hydroxide) treatment for enhancing its degradation. Biodegradation of forest waste by hydrolytic enzymes (cellulase and xylanase) secreted by A. niger under solid state fermentation (SSF) was explored. SSF of pretreated forest biomass was found to be superior over untreated forest biomass. Highest extracellular enzyme activity of 2201±23.91 U/g by A. niger was shown in pretreated C. australis wood resulting in 6.72±0.20 percent hydrolysis and 6.99±0.23 biodegradation index (BI). The lowest BI of 1.40±0.08 was observed in untreated saw dust of C. deodara having the least enzyme activity of 238±1.36 U/g of dry matter. Biodegradation of forest biomass under SSF was increased many folds when moistening agent i.e. tap water had been replaced with modified basal salt media (BSM). In BSM mediated degradation of forest waste with A. niger, extracellular enzyme activity was increased up to 4089±67.11 U/g of dry matter in turn resulting in higher BI of 15.4±0.41 and percent hydrolysis of 19.38±0.81 in pretreated C. australis wood. A. niger exhibited higher enzyme activity on pretreated biomass when moistened with modified BSM in this study. Statistically a positive correlation has been drawn between these three factors i.e. enzyme activity, BI and percent hydrolysis of forest biomass thus proving their direct relationship with each other.
Asunto(s)
Zona Árida , Aspergillus niger/enzimología , Aspergillus niger/aislamiento & purificación , Biomasa , Celulasas/análisis , Celulasas/aislamiento & purificación , Xilanos/análisis , Xilanos/aislamiento & purificación , Biodegradación Ambiental , Activación Enzimática , Hidrólisis , Métodos , SueloRESUMEN
Holocellulose structures from agro-industrial residues rely on main and side chain attacking enzymes with different specificities for complete hydrolysis. Combinations of crude enzymatic extracts from different fungal species, including Aspergillus terreus, Aspergillus oryzae, Aspergillus niger and Trichoderma longibrachiatum, were applied to sugar cane bagasse, banana stem and dirty cotton residue to investigate the hydrolysis of holocellulose structures. A. terreus and A. oryzae were the best producers of FPase and xylanase activities. A combination of A. terreus and A. oryzae extracts in a 50% proportion provided optimal hydrolysis of dirty cotton residue and banana stem. For the hydrolysis of sugar cane bagasse, the best results were obtained with samples only containing A. terreus crude extract.
Asunto(s)
Agroindustria , Aspergillus niger/enzimología , Aspergillus niger/aislamiento & purificación , Aspergillus oryzae/enzimología , Aspergillus oryzae/aislamiento & purificación , Trichoderma/enzimología , Trichoderma/aislamiento & purificación , Xilanos/análisis , Xilanos/aislamiento & purificación , Biodegradación Ambiental , Activación Enzimática , Hidrólisis , Métodos , ResiduosRESUMEN
Xylanolytic enzymes produced by Lentinula edodes UFV70, cultivated in eucalyptus sawdust/rice bran medium, were stable at 50, 60 and 65ºC for 21 hours, losing only 15-25% activity. Fungus incubation at 50ºC for 12 hours and at 65ºC for 24 hours increased the amount of xylose produced.
Asunto(s)
Biomasa , Hongos Shiitake/aislamiento & purificación , Micelio/enzimología , Xilanos/aislamiento & purificación , Xilosa/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Pruebas Enzimáticas Clínicas , Activación Enzimática , MétodosRESUMEN
The analysis of individual gene product should enable to clarify the role of a particular enzyme in a complex xylanase system of A. niger. The two genes encoding precursors of co-produced endo-1,4-¥â-D-xylanases, xynA1 and xynB, were isolated from Aspergillus niger SCTCC 400264 (SCTCC, China) by using RT-PCR technique and then successfully expressed in Escherichia coli BL21. The nucleotide sequences of the xynA1 and xynB genes revealed that they were only 52.5 percent homology to each other. Characterization of the recombinant enzymes revealed the different properties: the specific activity of recombinant XYNA1 was 16.58 U/mg compared to 1201.7 U/mg for recombinant XYNB; The optimum temperature and pH of the recombinant XYNA1 were 35 ¨¬C and 3.0, respectively, whereas the corresponding values for the recombinant XYNB were 55 ¨¬C and 5.0, respectively; The recombinant XYNB showed much more thermostability than recombinant XYNA1; The recombinant XYNB showed 94 percent of maximal activity after incubating in water for 60 min at 60 ¨¬C compared to no activity for recombinant XYNA1. Various metal ions had different effects on activity between the two recombinant xylanases.
Asunto(s)
Aspergillus niger/aislamiento & purificación , Aspergillus niger/patogenicidad , Secuencia de Bases , Expresión Génica , Técnicas In Vitro , Reacción en Cadena de la Polimerasa , Xilanos/aislamiento & purificación , Activación Enzimática , Métodos , Métodos , VirulenciaRESUMEN
A purificação de uma etapa e caracterização de uma xilanase livre de celulase de uma linhagem recentemente isolada alcalofílicos e moderadamente termofílico de Bacillus subtilis ASH. Xilanase foi purificada à homogeneidade de 10,5 vezes, com ~ por cento de recuperação 43 através de cromatografia de troca iônica através de CM- Sephadex C -50. A enzima purificada revelou uma única banda no gel SDS-PAGE com uma massa molecular de 23 kDa. Ele mostrou um pH ótimo de 7,0 e manteve-se estável na faixa de pH 6,0-9,0 . A temperatura ótima para atividade da enzima foi 55 º C. A xilanase purificada não perder nenhuma atividade até 45 º C , no entanto, manteve 80 por cento e 51 por cento de sua atividade após pré-incubação a 55 º C e 60 º C , respectivamente. A enzima obedecido Michaelis- Menton cinética para xilano de madeira de bétula com aparente km 3,33 mg / ml e Vmax 100 UI / ml. A enzima foi fortemente inibida por Hg2 +, Cu2 + , enquanto reforçada por Co2 + e Mn2 +. A enzima purificada pode ser armazenado a 4 º C por seis semanas sem nenhuma perda de atividade catalítica. A purificação mais rápido e econômico da xilanase livre de celulase de B. subtilis ASH por um passo-a processo juntamente com a sua estabilidade sensível a alta temperatura e pH alcalino torna potencialmente eficazes para aplicações industriais.