Thermo-resistant enzyme-producing microorganisms isolated from composting

Organo-mineral fertilizers supplemented with biological additives are an alternative to chemical fertilizers. In this study, thermoresistant microorganisms from composting mass were isolated by two-step procedures. First, samples taken at different time points and temperatures (33 days at 52 ºC, 60 days at 63 ºC, and over 365 days at 26 ºC) were pre-incubated at 80 oC for 30 minutes. Second, the microbial selection by in vitro culture-based methods and heat shock at 60 oC and 100 oC for 2h and 4h. Forty-one isolates were able to grow at 60 °C for 4h; twenty-seven at 100 °C for 2h, and two at 100 °C for 4h. The molecular identification by partial sequencing of the 16S ribosomal gene using universal primers revealed that thirty-five isolates were from eight Bacillus species, one Brevibacillus borstelensis, three Streptomyces thermogriseus, and two fungi (Thermomyces lanuginosus and T. dupontii). Data from amylase, phytase, and cellulase activity assays and the enzymatic index (EI) showed that 38 of 41 thermo-resistant isolates produce at least one enzyme. For amylase activity, the highest EI value was observed in Bacillus licheniformis (isolate 21C2, EI= 4.11), followed by Brevibacillus borstelensis (isolate 6C2, EI= 3.66), Bacillus cereus (isolate 18C2, EI= 3.52), and Bacillus paralicheniformis (isolate 20C2, EI= 3.34). For phytase, the highest EI values were observed for Bacillus cereus (isolate 18C2, EI= 2.30) and Bacillus licheniformis (isolate 3C1, EI= 2.15). Concerning cellulose production, B. altitudinis (isolate 6C1) was the most efficient (EI= 6.40), followed by three Bacillus subtilis (isolates 9C1, 16C2, and 19C2) with EI values of 5.66, 5.84, and 5.88, respectively, and one B. pumilus (isolate 27C2, EI= 5.78). The selected microorganisms are potentially useful as a biological additive in organo-mineral fertilizers and other biotechnological processes.

Impact of Soil Chemical Properties on the Growth Promotion Ability of Trichoderma ghanense, T. tomentosum and Their Complex on Rye in Different Land-Use Systems

Microbial-based biostimulants that increase plant performance and ensure sustainable restoration of degraded soils are of great importance. The aim of the present study was to evaluate the growth promotion ability of indigenous Trichoderma ghanense, T. tomentosum and their complex on early rye seedlings in sustained grassland and arable soil. The impact of soil chemical properties on the ability of selected Trichoderma strains and their complex to promote plant growth was determined by the evaluation of the rye (Secale cereale L.) early seedling growth—measuring the length of shoots and roots as well as their dry weight. Trichoderma species were tested for their ability to produce extracellular degradative enzymes on solid media. Furthermore, the soil properties and CM-cellulase activity of soil were estimated. The indigenous Trichoderma strains possess the capacity to produce enzymes such as peroxidase, laccase, tyrosinase, and endoglucanase. The results indicated a significant (p < 0.05) increase in plant growth and the improvement of some soil chemical properties (total N, mobile humic and fulvic acids, exchangeable K2O, soil CM-cellulase activity) in inoculated soils when compared to the control. The growth of the roots of rye seedlings in sustained grassland was enhanced when T. tomentosum was applied (p = 0.005). There was an increase in total weight and shoot weight of rye seedlings when T. ghanense was used in the arable soil (p = 0.014 and p = 0.024). The expected beneficial effect of Trichoderma spp. complex on rye growth promotion was not observed in any tested soil. The results could find application in the development of new and efficient biostimulants, since not only do physiological characteristics of fungi play an important role but also the quality of the soil has an impact.

Development of a powerful synthetic hybrid promoter to improve the cellulase system of Trichoderma reesei for efficient saccharification of corncob residues

Background The filamentous fungus Trichoderma reesei is a widely used workhorse for cellulase production in industry due to its prominent secretion capacity of extracellular cellulolytic enzymes. However, some key components are not always sufficient in this cellulase cocktail, making the conversion of cellulose-based biomass costly on the industrial scale. Development of strong and efficient promoters would enable cellulase cocktail to be optimized for bioconversion of biomass. Results In this study, a synthetic hybrid promoter was constructed and applied to optimize the cellulolytic system of T. reesei for efficient saccharification towards corncob residues. Firstly, a series of 5’ truncated promoters in different lengths were established based on the strong constitutive promoter Pcdna1. The strongest promoter amongst them was Pcdna1-3 (− 640 to − 1 bp upstream of the translation initiation codon ATG), exhibiting a 1.4-fold higher activity than that of the native cdna1 promoter. Meanwhile, the activation region (− 821 to − 622 bp upstream of the translation initiation codon ATG and devoid of the Cre1-binding sites) of the strong inducible promoter Pcbh1 was cloned and identified to be an amplifier in initiating gene expression. Finally, this activation region was fused to the strongest promoter Pcdna1-3, generating the novel synthetic hybrid promoter Pcc. This engineered promoter Pcc drove strong gene expression by displaying 1.6- and 1.8-fold stronger fluorescence intensity than Pcbh1 and Pcdna1 under the inducible condition using egfp as the reporter gene, respectively. Furthermore, Pcc was applied to overexpress the Aspergillus niger β-glucosidase BGLA coding gene bglA and the native endoglucanase EG2 coding gene eg2, achieving 43.5-fold BGL activity and 1.2-fold EG activity increase, respectively. Ultimately, to overcome the defects of the native cellulase system in T. reesei, the bglA and eg2 were co-overexpressed under the control of Pcc promoter. The bglA-eg2 double expression strain QPEB70 exhibited a 178% increase in total cellulase activity, whose cellulase system displayed 2.3- and 2.4-fold higher saccharification efficiency towards acid-pretreated and delignified corncob residues than the parental strain, respectively. Conclusions The synthetic hybrid promoter Pcc was generated and employed to improve the cellulase system of T. reesei by expressing specific components. Therefore, construction of synthetic hybrid promoters would allow particular cellulase genes to be expressed at desired levels, which is a viable strategy to optimize the cellulolytic enzyme system for efficient biomass bioconversion.

Cellulase enzyme activity of the bacteria isolated from mangrove ecosystem in Aceh Besar and Banda Aceh

Cellulolytic bacteria is one of the beneficial bacteria that can found in mangrove ecosystem. The purposes of study were to analyse the cellulolytic index, and to analyse the cellulase activity of bacteria isolated from soil mangrove. Qualitatively, assessment of cellulase activity were carried out at the Microbiology laboratory of Fish Quarantine Station, Quality Control and Safety of Fishery Products (SKIPM) Aceh, while quantitatively was observed in microbiology laboratory, Biology Department, IPB. Assessment of qualitative cellulase activity is performed by growing the selected pure isolate on 1% CMC medium then spilled 1% congo red to test its cellulolytic potential. Cellulolytic potential was determined by clear zone performed around the colony after congo red flooded. The quantitative cellulase enzyme activity test carried out by DNS method tested on one selective isolate. There were 21 from 39 isolates showed a clear zone isolated from mangrove soil. The cellulolytic index (CI) obtained ranged from 0.07 to 0.80 classified as low cellulolytic index criteria. The cellulolytic index was higher in bacteria isolated from mangrove rehabilitated than mangrove unrehabilitated. The highest cellulase activity and specific cellulase activity of BTMD32 was at 48 hours with the value were 0.0012 U/ml, 0.077 U/mg, respectively. The result concluded that the bacteria cellulolytic isolated from mangrove soil had low cellulolytic index, low cellulase activity, and low specific cellulase activity.

Mangrove crab intestine and habitat sediment microbiomes cooperatively work on carbon and nitrogen cycling

Mangrove ecosystems, where litter and organic components are degraded and converted into detrital materials, support rich coastal fisheries resources. Sesarmid (Grapsidae) crabs, which feed on mangrove litter, play a crucial role in material flow in carbon-rich and nitrogen-limited mangrove ecosystems; however, the process of assimilation and conversion into detritus has not been well studied. In this study, we performed microbiome analyses of intestinal bacteria from three species of mangrove crab and five sediment positions in the mud lobster mounds, including the crab burrow wall, to study the interactive roles of crabs and sediment in metabolism. Metagenome analysis revealed species-dependent intestinal profiles, especially in Neosarmatium smithi, while the sediment microbiome was similar in all positions, albeit with some regional dependency. The microbiome profiles of crab intestines and sediments were significantly different in the MDS analysis based on OTU similarity; however, 579 OTUs (about 70% of reads in the crab intestinal microbiome) were identical between the intestinal and sediment bacteria. In the phenotype prediction, cellulose degradation was observed in the crab intestine. Cellulase activity was detected in both crab intestine and sediment. This could be mainly ascribed to Demequinaceae, which was predominantly found in the crab intestines and burrow walls. Nitrogen fixation was also enriched in both the crab intestines and sediments, and was supported by the nitrogenase assay. Similar to earlier reports, sulfur-related families were highly enriched in the sediment, presumably degrading organic compounds as terminal electron acceptors under anaerobic conditions. These results suggest that mangrove crabs and habitat sediment both contribute to carbon and nitrogen cycling in the mangrove ecosystem via these two key reactions.

Bioethanol Production From Hassawi Rice Straw Wastes Assisted by Aspergillus sp. NAS51 Cellulosic Enzyme Under SSF and Saccharification Process With in Silico Enzyme Structural Homology Modeling

With the distribution of exploitable non-renewable energy resources, the use of lignocellulosic wastes to make bioethanol and biogas has drawn great attention from researchers. In our effort to find a potent cellulase-producing fungal strain, the fungus NAS51 was isolated from a sponge collected from the Red Sea, Jeddah, among eight isolates and selected as it displayed potent cellulolytic activity. The fungus was identified morphologically and genetically by sequencing its 18SrRNA gene as Aspergillus sp. NAS51. The cellulase activity of Aspergillus sp. NAS51 was optimized and maximum enzyme production was obtained at initial pH7, temp 30oC, incubation period 11 days, moisture content 70%, urea as a nitrogen source, and K2HPO4 (2g/L). The cellulase gene has been sequenced and the protein 3D structure was generated via in silico homology modeling. Determination of binding sites and biological annotations of the constructed protein was carried out via COACH and COFACTOR based on the I-TASSER structure prediction. To reach the maximum enzyme hydrolysis, the rice straw collected from Al-Ahsa, Kingdom of Saudi Arabia was pretreated with NaOH 1.5% to remove lignin and to enhance the saccharification process by cellulase enzyme. The saccharified product was measured using HPLC, fermented by S. cerevisiae and the bioethanol yield produced from the fermentation was 0.454 ml ethanol/g fermentable sugars.

Cellulase activity of bacteria isolated from water of mangrove ecosystem in Aceh Province

Cellulolytic bacteria that produce cellulase enzymes play an essential role in degrading cellulose in their habitat. The presence of cellulolytic bacteria strongly supports the fertility and productivity in mangrove waters. The objectives of the study are to analyze the activity of cellulase enzyme qualitatively through the cellulolytic index and quantitatively through the activity and specific activity of the cellulase enzyme from bacteria isolated from the water of mangrove ecosystems in Aceh Province. The qualitative experiment of enzyme activity was carried out at the Microbiology laboratory SKIPM Aceh, and a quantitative experiment of enzyme activity was conducted at the Microbiology Laboratory, Biology Department, IPB. Isolation of cellulolytic bacteria isolated from mangrove water used Carboxy Methyl Cellulose (1% CMC) selective media and carried out by spread plate method. The ability of bacteria to produce cellulase was tested qualitatively using the spot technique, this test was carried out using 1% Congo Red. Furthermore, the quantitative testing of cellulase enzymes activity adopted the DNS spectrophotometric method. The specific activity of the cellulase enzyme can be determined by using the Lowry method. There were 21 isolates that had a clear zone and had the ability to produce cellulase enzymes from 49 isolates that were successfully purified. The highest cellulolytic index (CI) produced using BAM421 isolate with the value of 5.50 was included in the high category, followed by BAM326 and BAM132 isolates, with values of 1.55 and 1.05 were categorized into the medium category. The other isolates were in the low cellulolytic index category. The isolate with the highest CI value was further tested using the quantitative enzyme activity test. The highest cellulase enzyme activity of BAM421 occurred at 24hr (0.0029 U/ml). The highest specific cellulase activity of BAM421 was at 24hr with the value of 0.210 U/mg. The result concluded that the qualitative test showed CI values can be categorized into low, medium, and high. Moreover, the value of the quantitative assay described that the cellulase enzyme and the specific enzyme activities of the bacteria were low in the study area.Keywords:Cellulolytic indexQuantitative testMangrove watersCellulase enzymeMicroorganismTRANSLATE with x EnglishArabicHebrewPolishBulgarianHindiPortugueseCatalanHmong DawRomanianChinese SimplifiedHungarianRussianChinese TraditionalIndonesianSlovakCzechItalianSlovenianDanishJapaneseSpanishDutchKlingonSwedishEnglishKoreanThaiEstonianLatvianTurkishFinnishLithuanianUkrainianFrenchMalayUrduGermanMalteseVietnameseGreekNorwegianWelshHaitian CreolePersian // TRANSLATE with COPY THE URL BELOW Back EMBED THE SNIPPET BELOW IN YOUR SITE Enable collaborative features and customize widget: Bing Webmaster PortalBack//

PRODUCTION OF ENZYMES IN PREDOMINANT THERMOPHILIC FUNGI AVAILABLE FROM ORGANIC SUBSTRATES

Present investigation describes that the study site comes under Aurangabad Division Maharashtra and it falls in Deccan Plateau Zone of India. It was collected different types of organic substrates viz. vermiompost, poultary manure, baggase, farm yard manure (FYM), soil, Ash etc. Isolated thermophilic predominant fungi thermophilic fungi viz.Aspergillus niger, Mucor mucedo,Humicola insolens,Trichoderma harzianum,T. viride,Penicillium duponti,Fusarium oxysporun and Chaetomium thermophilum were carried out for the production of enzymes. Isolated predominant thermophilic fungi were evaluated on different types of enzymes. Among tested thermophilic fungi, the highest ativity was observed in C. thermophilium (20mm) followed by T. harzianum (19.50mm) In lipase, M. mucedo (15.40mm) was found maximum followed by F. oxysporun. Cellulase activity was found highest in A. nige (25mm) followed by others. In case of xylanase, catalase, peroxidase and esterase activities were found maximum, minimum and medium even negative in some fungi. Maximum pectinase activity was detected from H. insolens (52.26 @ 0 min) and (74.25 @ 10 min) and in case of M. mucedo, F. oxysporun and C. thermophilium was found most extreme while least in A. niger (30.12) and P. duponti (33.47) @ 0 minute. Key words: Organic Substrates, Thermophilic Fungi, Enzymes

Coniochaeta elegans sp. nov., Coniochaeta montana sp. nov. and Coniochaeta nivea sp. nov., three new species of endophytes with distinctive morphology and functional traits

A growing interest in fungi that occur within symptom-less plants and lichens (endophytes) has uncovered previously uncharacterized species in diverse biomes worldwide. In many temperate and boreal forests, endophytic Coniochaeta (Sacc.) Cooke (Coniochaetaceae, Coniochaetales, Sordariomycetes, Ascomycota) are commonly isolated on standard media, but rarely are characterized. We examined 26 isolates of Coniochaeta housed at the Gilbertson Mycological Herbarium. The isolates were collected from healthy photosynthetic tissues of conifers, angiosperms, mosses and lichens in Canada, Sweden and the United States. Their barcode sequences (nuclear ribosomal internal transcribed spacer and 5.8S; ITS rDNA) were ≤97% similar to any documented species available through GenBank. Phylogenetic analyses based on two loci (ITS rDNA and translation elongation factor 1-alpha) indicated that two isolates represented Coniochaeta cymbiformispora, broadening the ecological niche and geographic range of a species known previously from burned soil in Japan. The remaining 24 endophytes represented three previously undescribed species that we characterize here: Coniochaeta elegans sp. nov., Coniochaeta montana sp. nov. and Coniochaeta nivea sp. nov. Each has a wide host range, including lichens, bryophytes and vascular plants. C. elegans sp. nov. and C. nivea sp. nov. have wide geographic ranges. C. montana sp. nov. occurs in the Madrean biome of Arizona (USA), where it is sympatric with the other species described here. All three species display protease, chitinase and cellulase activity in vitro. Overall, this study provides insight into the ecological and evolutionary diversity of Coniochaeta and suggests that these strains may be amenable for studies of traits relevant to a horizontally transmitted, symbiotic lifestyle.

Exploring cellulolytic microorganisms from coffee industry by-products and their enzyme properties

Cellulolytic microorganism has immense potential due to their cellulase production, enzyme complexity and widespread habitat of life. This study was conducted to obtain microbial cellulase with vast industrial applicability from the coffee industry by-product in East Java, Indonesia. Fifty-four isolates with significant clear zone formation were obtained by Congo red staining in CMC agar plates. Eighteen bacteria, two yeasts and two moulds with high cellulolytic index were subjected to protein content determination as well as reducing sugar analysis in various conditions such as pH, temperature, addition of metal ions, surfactant and inhibitor agent. The specific activity measurements of all the crude enzymes result in the highest value of cellulase activity produced by isolate C12 which was 0.401 ± 0.018 U/mg. This enzyme activity was known to be optimum at 50°C and pH 9. It was also stimulated by K+, Na+, Mg2+, Fe3+, and SDS. However, the enzyme activity was inhibited by EDTA at 10 mM concentration. The use of coffee industry by-products as the source of cellulolytic microorganisms offers a promising approach for its various types of indigenous microorganisms and their unique property of cellulase produced that is useful for industrial application.