Fungus-growing termites are eusocial insects that represent one of the most efficient and unique ... more Fungus-growing termites are eusocial insects that represent one of the most efficient and unique systems for lignocellulose bioconversion, evolved from a sophisticated symbiosis with lignocellulolytic fungi and gut bacterial communities. Despite a plethora of information generated during the last century, some essential information on gut bacterial profiles and their unique contributions to wood digestion in some fungus-growing termites is still inadequate. Hence, using the culture-dependent approach, the present study aims to assess and compare the diversity of lignocellulose-degrading bacterial symbionts within the gut systems of three fungus-growing termites: Ancistrotermes pakistanicus, Odontotermes longignathus, and Macrotermes sp. A total of 32 bacterial species, belonging to 18 genera and 10 different families, were successfully isolated and identified from three fungus-growing termites using Avicel or xylan as the sole source of carbon. Enterobacteriaceae was the most domina...
The recalcitrance of lignocellulosic biomass is a major constraint to its high-value use at indus... more The recalcitrance of lignocellulosic biomass is a major constraint to its high-value use at industrial scale. In nature, microbes play a crucial role in biomass degradation, nutrient recycling and ecosystem functioning. Therefore, the use of microbes is an attractive way to transform biomass to produce clean energy and high-value compounds. The microbial degradation of lignocelluloses is a complex process which is dependent upon multiple secreted enzymes and their synergistic activities. The availability of the cutting edge proteomics and highly sensitive mass spectrometry tools make possible for researchers to probe the secretome of microbes and microbial consortia grown on different lignocelluloses for the identification of hydrolytic enzymes of industrial interest and their substrate-dependent expression. This review summarizes the role of secretomics in identifying enzymes involved in lignocelluloses deconstruction, the development of enzyme cocktails and the construction of syn...
A salt-tolerant yeast named G1 which could decolorize various azo dyes was recently isolated and ... more A salt-tolerant yeast named G1 which could decolorize various azo dyes was recently isolated and identified as Pichia occidentalis. Systematic researches on characterization, degradation pathway, detoxification effects and enzymes analysis of this yeast were done. The results showed that the optimal metabolism and growth parameters for strain G1 were: 2.0gL(-1) glucose, 0.6gL(-1) ammonium sulfate, 0.08gL(-1) yeast extract, 30gL(-1) NaCl, 160rmin(-1), 30°C and pH 5.0. More than 98% of 50mgL(-1) Acid Red B (ARB) could be decolorized within 16h under the optimal conditions. Additionally, strain G1 degraded and obviously detoxified ARB through a possible pathway successively consisting of decolorization, deamination/desulfonation and TCA cycle processes. Moreover, NADH-DCIP reductase was estimated as the key reductase for decolorization and ligninases including lignin peroxidase, manganese peroxidase and laccase were important oxidoreductases for further degradation of decolorization in...
Sporulation characteristics and virulence of Metarhizium anisopliae and Beauveria bassiana were e... more Sporulation characteristics and virulence of Metarhizium anisopliae and Beauveria bassiana were examined in relation to laboratory transmission in Coptotermes formosanus. Fungal isolates significantly affected disease prevalence in termite populations. Sporulation of M. anisopliae played a more important role than virulence in producing epizootics within small groups of termites, but this was not the case for B. bassiana. Isolates characterized by quick sporulation (day 2 after death) did not exhibit better transmission in termites than those with high total sporulation (day 11 after death) in either fungal species. An isolate of M. anisopliae ranking highly in all three categories (virulence, quick sporulation, and total sporulation) produced better epizootics than an isolate that was inferior in all three characteristics. High temperatures (35 degrees C) significantly reduced fungal germination rates, leading to significant reduction of epizootics. M. anisopliae was better than B. bassiana in producing epizootics at 27 degrees C. Thus, fungal characteristics other than virulence should be considered for the seasonal colonization approach to termite microbial control.
Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae) was used as “bait” to isolate pathogen... more Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae) was used as “bait” to isolate pathogenic fungi from soil. Ninety soil samples were collected from woodlands and pastures in the vicinities of Baton Rouge, New Orleans, and Lake Charles, LA, from which six Metarhizium anisopliae (Metsch.) Sorokin and nine Beauveria bassiana (Balsamo) Vuillemin isolates were obtained. Numbers of fungal isolates from the three sampling locations did not differ, but more isolates were found in woodlands than in pastures. Median lethal doses (LD50s) of these fungal species to C. formosanus were interspersed, indicating that fungal isolates rather than species had the greatest effect on virulence. Among nine Louisiana and two USDA isolates of B. bassiana, LD50s ranged from 4.95 × 103 to 4.96 × 105 conidia/termite, a difference of 100×. LD50s of six Louisiana and four USDA isolates of M. anisopliae ranged from 7.89 × 103 to 1.22 × 105 conidia/termite. Survival time also was used to compare virulence; M. anisopliae infections caused significantly shorter host survival times than B. bassiana. In vitro growth characteristics were significantly correlated with virulence against termites, suggesting that the characteristics of a fungus growing on agar might contribute to estimating the fungal virulence in vivo.
Identification of novel enzymes from lignin degrading microorganisms will help to develop biotech... more Identification of novel enzymes from lignin degrading microorganisms will help to develop biotechnologies for biomass valorization and aromatic hydrocarbons degradation. Bacillus ligniniphilus L1 grows with alkaline lignin as the single carbon source and is a great candidate for ligninolytic enzyme identification. The first dioxygenase from strain L1 was heterologously expressed, purified, and characterized with an optimal temperature and pH of 32.5 °C and 7.4, respectively. It showed the highest activity with 3-ethylcatechol and significant activities with other substrates in the decreasing order of 3-ethylcatechol > 3-methylcatechol > 3-isopropyl catechol > 2, 3-dihydroxybiphenyl > 4-methylcatechol > catechol. It did not show activities against other tested substrates with similar structures. Most reported catechol 2,3-dioxygenases (C23Os) are Fe2+-dependent whereas Bacillus ligniniphilus catechol 2,3-dioxygenase (BLC23O) is more Mn2+- dependent. At 1 mM, Mn2+ led t...
Background Cotton bollworm, Helicoverpa armigera is a widely distributed, devastating pest of ove... more Background Cotton bollworm, Helicoverpa armigera is a widely distributed, devastating pest of over 200 crop plants that mainly consist of some cellulosic materials. Despite its economic importance as a pest, little is known about the diversity and community structure of gut symbiotic bacteria potentially functioned in cellulose digestion in different gut-sections of H. armigera. In view of this lacuna, we attempted to evaluate and characterize cellulose-degrading bacteria (CDB) from foregut, midgut, and hindgut -regions of H. armigera by using a culture-dependent approach. Methodology The symbiotic bacteria were isolated from different gut-systems of H. armigera by enrichment techniques using Carboxymethyl cellulose sodium salt (CMC) as carbon source. The isolated bacteria were purified and subsequently screened for cellulose-degradation by plate-based method to display the zones of CMC clearance around the colonies. The identification and phylogeny of the gut-bacteria were reconstr...
Fungus-growing termites are eusocial insects that represent one of the most efficient and unique ... more Fungus-growing termites are eusocial insects that represent one of the most efficient and unique systems for lignocellulose bioconversion, evolved from a sophisticated symbiosis with lignocellulolytic fungi and gut bacterial communities. Despite a plethora of information generated during the last century, some essential information on gut bacterial profiles and their unique contributions to wood digestion in some fungus-growing termites is still inadequate. Hence, using the culture-dependent approach, the present study aims to assess and compare the diversity of lignocellulose-degrading bacterial symbionts within the gut systems of three fungus-growing termites: Ancistrotermes pakistanicus, Odontotermes longignathus, and Macrotermes sp. A total of 32 bacterial species, belonging to 18 genera and 10 different families, were successfully isolated and identified from three fungus-growing termites using Avicel or xylan as the sole source of carbon. Enterobacteriaceae was the most domina...
The recalcitrance of lignocellulosic biomass is a major constraint to its high-value use at indus... more The recalcitrance of lignocellulosic biomass is a major constraint to its high-value use at industrial scale. In nature, microbes play a crucial role in biomass degradation, nutrient recycling and ecosystem functioning. Therefore, the use of microbes is an attractive way to transform biomass to produce clean energy and high-value compounds. The microbial degradation of lignocelluloses is a complex process which is dependent upon multiple secreted enzymes and their synergistic activities. The availability of the cutting edge proteomics and highly sensitive mass spectrometry tools make possible for researchers to probe the secretome of microbes and microbial consortia grown on different lignocelluloses for the identification of hydrolytic enzymes of industrial interest and their substrate-dependent expression. This review summarizes the role of secretomics in identifying enzymes involved in lignocelluloses deconstruction, the development of enzyme cocktails and the construction of syn...
A salt-tolerant yeast named G1 which could decolorize various azo dyes was recently isolated and ... more A salt-tolerant yeast named G1 which could decolorize various azo dyes was recently isolated and identified as Pichia occidentalis. Systematic researches on characterization, degradation pathway, detoxification effects and enzymes analysis of this yeast were done. The results showed that the optimal metabolism and growth parameters for strain G1 were: 2.0gL(-1) glucose, 0.6gL(-1) ammonium sulfate, 0.08gL(-1) yeast extract, 30gL(-1) NaCl, 160rmin(-1), 30°C and pH 5.0. More than 98% of 50mgL(-1) Acid Red B (ARB) could be decolorized within 16h under the optimal conditions. Additionally, strain G1 degraded and obviously detoxified ARB through a possible pathway successively consisting of decolorization, deamination/desulfonation and TCA cycle processes. Moreover, NADH-DCIP reductase was estimated as the key reductase for decolorization and ligninases including lignin peroxidase, manganese peroxidase and laccase were important oxidoreductases for further degradation of decolorization in...
Sporulation characteristics and virulence of Metarhizium anisopliae and Beauveria bassiana were e... more Sporulation characteristics and virulence of Metarhizium anisopliae and Beauveria bassiana were examined in relation to laboratory transmission in Coptotermes formosanus. Fungal isolates significantly affected disease prevalence in termite populations. Sporulation of M. anisopliae played a more important role than virulence in producing epizootics within small groups of termites, but this was not the case for B. bassiana. Isolates characterized by quick sporulation (day 2 after death) did not exhibit better transmission in termites than those with high total sporulation (day 11 after death) in either fungal species. An isolate of M. anisopliae ranking highly in all three categories (virulence, quick sporulation, and total sporulation) produced better epizootics than an isolate that was inferior in all three characteristics. High temperatures (35 degrees C) significantly reduced fungal germination rates, leading to significant reduction of epizootics. M. anisopliae was better than B. bassiana in producing epizootics at 27 degrees C. Thus, fungal characteristics other than virulence should be considered for the seasonal colonization approach to termite microbial control.
Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae) was used as “bait” to isolate pathogen... more Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae) was used as “bait” to isolate pathogenic fungi from soil. Ninety soil samples were collected from woodlands and pastures in the vicinities of Baton Rouge, New Orleans, and Lake Charles, LA, from which six Metarhizium anisopliae (Metsch.) Sorokin and nine Beauveria bassiana (Balsamo) Vuillemin isolates were obtained. Numbers of fungal isolates from the three sampling locations did not differ, but more isolates were found in woodlands than in pastures. Median lethal doses (LD50s) of these fungal species to C. formosanus were interspersed, indicating that fungal isolates rather than species had the greatest effect on virulence. Among nine Louisiana and two USDA isolates of B. bassiana, LD50s ranged from 4.95 × 103 to 4.96 × 105 conidia/termite, a difference of 100×. LD50s of six Louisiana and four USDA isolates of M. anisopliae ranged from 7.89 × 103 to 1.22 × 105 conidia/termite. Survival time also was used to compare virulence; M. anisopliae infections caused significantly shorter host survival times than B. bassiana. In vitro growth characteristics were significantly correlated with virulence against termites, suggesting that the characteristics of a fungus growing on agar might contribute to estimating the fungal virulence in vivo.
Identification of novel enzymes from lignin degrading microorganisms will help to develop biotech... more Identification of novel enzymes from lignin degrading microorganisms will help to develop biotechnologies for biomass valorization and aromatic hydrocarbons degradation. Bacillus ligniniphilus L1 grows with alkaline lignin as the single carbon source and is a great candidate for ligninolytic enzyme identification. The first dioxygenase from strain L1 was heterologously expressed, purified, and characterized with an optimal temperature and pH of 32.5 °C and 7.4, respectively. It showed the highest activity with 3-ethylcatechol and significant activities with other substrates in the decreasing order of 3-ethylcatechol > 3-methylcatechol > 3-isopropyl catechol > 2, 3-dihydroxybiphenyl > 4-methylcatechol > catechol. It did not show activities against other tested substrates with similar structures. Most reported catechol 2,3-dioxygenases (C23Os) are Fe2+-dependent whereas Bacillus ligniniphilus catechol 2,3-dioxygenase (BLC23O) is more Mn2+- dependent. At 1 mM, Mn2+ led t...
Background Cotton bollworm, Helicoverpa armigera is a widely distributed, devastating pest of ove... more Background Cotton bollworm, Helicoverpa armigera is a widely distributed, devastating pest of over 200 crop plants that mainly consist of some cellulosic materials. Despite its economic importance as a pest, little is known about the diversity and community structure of gut symbiotic bacteria potentially functioned in cellulose digestion in different gut-sections of H. armigera. In view of this lacuna, we attempted to evaluate and characterize cellulose-degrading bacteria (CDB) from foregut, midgut, and hindgut -regions of H. armigera by using a culture-dependent approach. Methodology The symbiotic bacteria were isolated from different gut-systems of H. armigera by enrichment techniques using Carboxymethyl cellulose sodium salt (CMC) as carbon source. The isolated bacteria were purified and subsequently screened for cellulose-degradation by plate-based method to display the zones of CMC clearance around the colonies. The identification and phylogeny of the gut-bacteria were reconstr...
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