Mannanase Production Research Articles

Evaluation of Aspergillus tamarii NRC 3 biomass as a biosorbent for removal and recovery of heavy metals from contaminated aqueous solutions

BackgroundBiomass produced as a byproduct from the β-mannanase production process by Aspergillus tamarii NRC 3was evaluated as a biosorbent for the removal and recovery of some heavy metal ions.ResultsUnder optimal conditions, the isolated strain recorded the highest β-mannanase activity (31.88 Uml−1). Thus, the biomass produced from mannanase production process as a byproduct was evaluated as a biosorbent for the removal and recovery of some heavy metal ions from aqueous solutions and an industrial wastewater. The fungal biomass was found to be efficient for the removal of Cu+2 and some heavy metal ions. The biosorption process of copper(II) by Aspergillus tamarii NRC 3 biomass was affected by changing of time, temperature, pH, metal ions concentration, the presence of some heavy metals, and biomass concentration. The rate of Cu+2 uptake from Cu+2solution proceeded rapidly, and it appeared to be virtually complete during the initial 5 min (92%); the maximum uptake of Cu+2 appeared at 30 °C, pH 5, and biomass concentration 5 g w/w. On the other hand, the fungal biomass was to remove considerable proportion of Pb2+, Co+2, Ni2+, Fe+3, and Cr3+ in addition to Cu2+. The uptake of Cu+2 by pretreated biomass was studied. Recovery of the sorbed metal ions by desorbing agents and the potential reuse of the regenerated biomass in metal ions uptake (reloading) were evaluated.ConclusionsAspergillus tamarii NRC 3 biomass seems to be quite feasible in the removal of heavy metal ions especially Cu+2 from aqueous solutions.

Screening and Optimization of Cultural Conditions for Production of Thermophilic Mannanase from Bacillus megaterium

Screening and Optimization of Cultural Conditions for Production of Thermophilic Mannanase from Bacillus megaterium

Mannanase Enzyme from Bacillus subtilis P2-5 with Waste Management

Nowadays, waste disposal has become a major of environment problem in Thailand because the volume of waste increase due to population growth and social behavior. It required optimal solutions in waste management. People should realize fundamental of 3R including reduce recycle and reuse in order to improve environment condition. This research interested in coffee grounds and tea waste because they were generated from household and industrial thus require to disposal or added value. The objective of this research was recycle coffee grounds and tea waste by utilize as a raw material for mannanase enzyme production. Mannanase enzyme can hydrolyze mannan structure, containing in plant such as konjac, ivory nut, locust bean gum, guar gum, palm kernel, coffee bean and copra meal. The methodology was a randomized complete block design to studied mannanase enzyme activity in different condition. There were two factors consist of inoculum percentage; Bacillus subtilis P2-5 had three levels (1, 3 and 5% (v/v) and three sources (locust bean gum, coffee grounds and tea waste) in producing medium (PM) for mannanase production. An analysis of variances (ANOVA) method was applied to determine the effect of inoculum percentage and regression analysis to predict mannanase enzyme activity. The test result found that inoculum percentage and three sources significantly affecting the mannanase enzyme activity. The prediction model had been formulated and prioritize source which produce the high mannanase enzyme activity as following tea waste, coffee grounds and locust bean gum, respectively. This research would be guidelines for waste management strategy to achieve sustainability approach.

Isolation and screening of thermophilic and thermotolerant fungi for production of hemicellulases from heated environments

ABSTRACTThermostable hemicellulases have potential to improve the quality of products of various industries including pulp and paper, food and feed, textile, etc. This study was aimed to isolate, screen, and identify potential xylanase and mannanase producers from heated environments. Sixty-eight thermophilic and thermotolerant fungi were isolated from various self-heated habitats based on their ability to grow at 45°C. With the aid of cultural and morphological observations, the fungi were identified and grouped into 19 fungal species. The enzyme production was evaluated by cultivating isolated fungi in submerged fermentation using wheat bran as carbon source. Screening of these isolates for hydrolysis of xylan and mannan was confirmed by Congo red plate assay method followed by assessment of xylanase and mannanase activity. Based on experimental analysis, we have found that all the isolates have exhibited xylanase activity, whereas only 22 isolates have found positive for mannanase activity. The highest xylanase and mannanase production was obtained by the cultivation of Malbranchea cinnamomea NFCCI 3724 (242 and 27 nkat/ml) followed by Melanocarpus albomyces (195 and 24 nkat/ml), Aspergillus terreus (165 and 21 nkat/ml), and Myceliophthora thermophila NFCCI 3725 (130 and 18 nkat/ml), respectively.

Production and Partial Purification of Beta-Mannanase from Aspergillus niger Associated with Ilaje Lake, Ondo State, Nigeria

The present study aimed at the isolation, screening and partial purification of beta-mannanase from fungi isolated from soil and water samples collected from Ilaje Lake, Ondo state, Nigeria. The associated fungi were isolated and counted by standard microbiological methods. Partial purification of crude mannanase was done by standard biochemical methods. Quantitatively, mannanase production was performed in mineral salt medium into which Locust Bean Gum (LBG) had been incorporated as the sole carbon source. Enzyme activity was determined by dinitrosalicylic acid (DNSA) method, while protein content was evaluated by Lowry’s method. The highest fungal counts were recorded for water sample collected from Ilaje Lake with 3.4 × 108 SFU/mL. The organisms encountered include Aspergillus flavus, Rhizopus stolonifer, A. fumigatus, A. niger, R. japonicus, Penicillum italicum, Fusarium solani and Candida albicans. All the fungal isolates encountered from these sources showed varied degrees of mannanase activities. The highest specific mannanase activity was recorded for isolate 4B1, while the lowest value was obtained with isolate 1B2. Purification of crude mannanase from A. niger was carried out by ammonium sulphate precipitation and gel filtration (Sephadex G-200). Fractionation of ammonium sulphate precipitated mannanase from A. niger on Sephadex G-200 produced two activity peaks. In this investigation, fungal isolates evaluated for mannanase production from this source gave appreciable mannanase activity and this could be applied in many industrial processes.

Screening, statistical optimized production, and application of β-mannanase from some newly isolated fungi.

Eighty-eight fungi isolated from soil and decaying organic matter were screened for mannanolytic activity. Twenty-eight fungi produced extracellular mannanase on locust bean gum as evidenced by zone of hydrolysis produced on mannan agar gel. Six prominent producers, including four Fusarium species namely Fusarium fusarioides NFCCI 3282, Fusarium solani NFCCI 3283, Fusarium equiseti NFCCI 3284, Fusarium moniliforme NFCCI 3287 with Cladosporium cladosporioides NFCCI 3285 and Acrophialophora levis NFCCI 3286 produced the β-mannanase in the range of 84-140 nkat/mL. All these grew well on particulate substrates in solid-state fermentation (SSF), producing relatively higher titers on mannan-rich palm kernel cake (PKC) and copra meal. Two high yielding strains, F. equiseti (1747 nkat/gds) and A. levis (897 nkat/gds) were selected for statistical optimization of mannanase on PKC. Interaction of two critical solid state fermentation parameters, pH and moisture on mannanase production by these two molds was studied by response surface method. Optimized production on PKC resulted in three- to fourfold enhancement in enzyme yield was observed in case of F. equiseti (5945 nkat/gds) and A. levis (4726 nkat/gds). HPLC analysis of mannan hydrolysate indicated that F. equiseti and A. levis mannanase performed efficient hydrolysis of konjac gum (up to 99%) with exclusive mannooligosaccahride (DP of 4) production. A seminative SDS-PAGE revealed that A. levis and F. solani produced three isoforms, F. moniliforme produced two isoforms while F. fusarioides, F. equiseti, and C. cladosporioides produced a single enzyme.

Experimental design of response surface methodology used for utilisation of palm kernel cake as solid substrate for optimised production of fungal mannanase

ABSTRACTThe results obtained from this work strongly indicate that the solid state fermentation (SSF) system using the palm kernel cake (PKC) as a substrate is an economical method for the production of β-mannanase at extremely low operational cost based on the fact that PKC is one of the cheap and abundant agro-waste by-products of the palm oil industry. Under initial conditions, i.e. 2 mm particle size of PKC, the moisture ratio of 1:1 of PKC:moistening agent and pH 7, Malbranchea cinnamomea NFCCI 3724 produced 109 U/gram distribution of the substrate (gds). The production of β-mannanase was optimised by the statistical approach response surface methodology (RSM) using independent variables, namely initial moisture (12.5), pH (9.0) and solka floc (100 mg). Noticeably, six fold enhancement of β-mannanase production (599 U/gds) was obtained under statistically optimised conditions. HPLC results revealed that β-mannanase is an endo-active enzyme that generated manno-oligosaccharides with a degree of polymerisation (DP) of 3 and 4. Semi-native PAGE analysis revealed that M. cinnamomea produced three isoforms of mannanase. Selective production of oligosaccharide makes M. cinnamomea β-mannanase an attractive enzyme for use in food and nutraceutical industries.

High level extracellular production of a truncated alkaline β-mannanase from alkaliphilic Bacillus sp. N16-5 in Escherichia coli by the optimization of induction condition and fed-batch fermentation.

To improve the extracellular production of alkaline β-mannanase from alkaliphilic Bacillus sp. N16-5 in Escherichia coli, two truncated recombinant mannanases (32a-ManAR2 and 22b-ManAR2) were obtained. Compared with the full-length mannanases (32a-ManAR1 and 22b-ManAR1), the truncated mannanases not only showed higher secretion rate, but also exhibited higher thermostability and alkalistability. The K m value (11mg/mL) of 32a-ManAR2 was higher than that (1.46mg/mL) of 32a-ManAR1. The specific activity of 22b-ManAR2 was 2.7 times higher than that of 22b-ManAR1. 22b-ManAR2 showed the highest k cat/K m value of 602.7ml/mgs. The parameters of induction for recombinant mannanase production of E. coli BL21 (pET32a-manAR2) and E. coli BL21 (pET22b-manAR2) were subsequently optimized. The yield of soluble mannanase was found to be enhanced with lower induction temperature (25°C), lower IPTG concentration (0.01-0.05mM), and Triton X-100 supplement (0.1%) in a shake flask. Moreover, a one-time feeding strategy and Triton X-100 supplement were applied in production of 22b-ManAR2 in a 10L fermentor. The productivity of the total soluble mannanase reached 9284.64U/mL with the extracellular rate of 74% at 46h of fermentation, which was the highest productive level of alkaline β-mannanase in recombinant E. coli to date.

Controlling filamentous fungi morphology with microparticles to enhanced β-mannanase production.

β-mannanase was produced mainly by Aspergillus species and can degrade the β-1,4-mannose linkages of galactomannans. This study was undertaken to enhance mannanase production using talcum and aluminum oxide as the microparticles, which control cell morphology of recombinant Aspergillus sojae in glucose and carob extract medium. Both microparticles improved fungal growth in glucose and carob pod extract medium. Aluminum oxide (1g/L) was the best agent for glucose medium which resulted in 514.0U/ml. However, the highest mannanase activity was found as 568.7U/ml with 5g/L of talcum in carob extract medium. Increase in microparticle concentration resulted in decreasing the pellet size diameter. Furthermore, more than 10g/L of talcum addition changed the filamentous fungi growth type from pellet to pellet/mycelium mixture. Results showed that right type and concentration of microparticle in fermentation media improved the mannanase activity and production rate by controlling the growth morphology.

Production and Properties of Mannanase and Xylanase by a Bacillus subtilis Isolate

Production and Properties of Mannanase and Xylanase by a Bacillus subtilis Isolate

English

The present study was carried out to investigate the optimal culture conditions for β-mannanase production by Trichosporonoides oedocephalis using cheap and easily available agro-wastes as the carbon source in solid state cultivation. The effect of process parameters like incubation time, carbon source, incubation temperature, pH, percentage moisture content and inoculum concentration were optimized for enhanced mannanase production. Maximum β-mannanase activity was observed after 96 h of incubation. Different agricultural wastes were screened as substrates for β-mannanase production in comparison with locust bean gum (LBG). Among tested carbon sources, orange peels proved to the best for β-mannanase production. Initial pH of the culture medium was also optimized and a pH of 8.0 was found to be the optimum pH value for β-mannanase production. The optimum temperature, percentage moisture content and inoculum concentration were 30°C, 60% and 1×107 (spores/ml), respectively. Key words: Beta-mannanase, process parameters, solid state fermentation, agricultural wastes, Trichosporonoides oedocephalis.

Identification of Suitable Condition for Mannanase Production by Bacillus sp. GA2(1)

Aims: The effects of 1% (w/v) supplementation of additional 5 agricultural wastes, corn cob, bagasses, coffee residues, soybean meal, and cop ra meal for mannanase production by Bacillus sp.GA2(1) were studied. Hence, partial characterization of mannanase was determined. Methodology: The 1%(v/v) overnight cultured of Bacillus sp. GA2(1) was transferred into the basal medium and shaken at 150 rpm for 18 h at 37 o C. The additional of 5 AWs, corn cob, bagasses, coffee residues, soybean meal, and copra meal for the mannanase production were investigated. The cell suspension was centrifuged, and the crude mannanases were collected and stored at – 20 o C f or enzyme assay. The mannanase activities were measured by the dinitrosalicylic acid method. The optimal pH of mannanase were studied by measuring enzyme activity at pH 3 - 10 using 50 mM of following buffers; citrate (pH 3.0 - 6.0), phosphate (pH 6.0 - 8.0), an d glycine - NaOH

Constitutive expression of alkaline β-mannanase in recombinant Pichia pastoris

Alkaline β-mannanase has important applications for specific industrial processes like pulp bleaching and the detergent industry. The low yield of alkaline β-mannanase produced from native microbes such as alkaliphilic Bacillus limits its applications. Pichia pastoris is the most efficient heterologous system to produce alkaline mannanase. However, the previous use of the AOX system required large amount of methanol and sophisticated operation strategy, which are undesirable in large scale production. In this study, we established a safe and simple constitutive expression process for mannanase production in P. pastoris. The mannanase gene was successfully expressed under the control of GAP promoter. Sequential optimization of the constructed strains was also performed including the copy number optimization and co-expression of chaperone genes. A two-stage feeding strategy was then applied for the finally optimized strain. After 96h fermentation, a production level of 2980U/mL was finally reached, illustrating the potential of the GAP constitutive expression system for industrial scale preparation of alkaline β-mannanase.

Optimization of flocculation conditions using chitosan in mannanase production

Optimization of flocculation conditions using chitosan in mannanase production

Production and Characterization of Mannanase from a Bacillus sp. YB-1401 Isolated from Fermented Soybean Paste

Production and Characterization of Mannanase from a Bacillus sp. YB-1401 Isolated from Fermented Soybean Paste

Improved mannanase production from Penicillium humicola and application for hydrolysis property

Objectives The aim of this research is to produce β-mannanases from a new microbial source using wastes in their nutrition medium. The enzyme produced can be used for the production of galactomanno-oligosaccharides, which are very useful in the health and medical fields. Materials and methods Seven fungal strains and five bacterial strains were tested for the production of β-mannanases. Enzyme activity, protein content, and biomass production were determined in all the cultures produced using standard methods. Optimization studies to maximize enzyme production from the most potent microorganisms, including culture conditions and medium compositions, were also carried out. Preliminary studies for the production of galactomanno-oligosaccharides from locust bean gum using partially purified enzymes were also carried out and followed by thin-layer chromatography techniques and Somogyi methods. Results and conclusion The highest mannanase activities were produced by Penicillium humicola (8.8 U/ml) and Penicillium spp. v (7.75 U/ml) in shaking cultures after 10 days using gum locust bean as a carbon source. Among 13 carbon sources examined, coffee residue and ceratonia seeds were the best carbon sources (10.3 and 8.9 U/ml, respectively) for P. humicola, whereas the best nitrogen source was a mixture of peptone, urea, and ammonium sulfate for the same microorganism. The optimum temperature and pH for enzyme reaction was 55 and 5.5°C, respectively. The enzyme was thermostable and retained 80% of its activity after 1 h at 50°C. The highest reducing sugar of 8900 μg/ml was obtained from locust bean gum hydrolytes after 28 h.

Production of Extremely Alkaliphilic, Halotolerent, Detergent, and Thermostable Mannanase by the Free and Immobilized Cells of Bacillus halodurans PPKS-2. Purification and Characterization

The alkaliphilic Bacillus halodurans strain PPKS-2 was shown to produce extracellular extreme alkaliphilic, halotolerent, detergent, and thermostable mannanase activity. The cultural conditions for the maximum enzyme production were optimized with respect to pH, temperature, NaCl, and inexpensive agro wastes as substrates. Mannanase production was enhanced more than 4-fold in the presence of 1% defatted copra meal and 0.5% peptone or feather hydrolysate at pH11 and 40°C. Mannanase was purified to 10.3-fold with 34.6% yield by ion exchange and gel filtration chromatography methods. Its molecular mass was estimated to be 22kDa by SDS-PAGE. The mannanase had maximal activity at pH11 and 70°C. This enzyme was active over a broad range of NaCl (0-16%) and thermostable retaining 100% of the original activity at 70°C for 3h. Immobilization of whole cells proved to be effective for continuous production of mannanase. Since the strain PPKS-2 grows on cheaper agro wastes such as defatted copra meal, corn husk, jowar bagasse, and wheat bran, these can be exploited for mannanase production on an industrial scale.

β‐Mannanase production by Aspergillus niger BCC4525 and its efficacy on broiler performance

Mannan is a hemicellulose constituent commonly found in plant-derived feed ingredients. The gum-like property of mannan can obstruct digestive enzymes and bile acids, resulting in impaired nutrient utilisation. In this study, β-mannanase production by Aspergillus niger strain BCC4525 was investigated using several agricultural residues under solid state condition. The biochemical properties of the target enzyme and the effects of enzyme supplementation on broiler performance and energy utilisation were assessed. Among five carbon sources tested, copra meal was found to be the best carbon source for β-mannanase production with the maximum yield of 1837.5 U g(-1) . The crude β-mannanase exhibited maximum activity at 80 °C within a broad range of pH from 2 to 6. In vitro digestibility assay, which simulates the gastrointestinal tract system of broilers, showed that β-mannanase could liberate reducing sugars from corn/soybean diet. Surprisingly, β-mannanase supplementation had no significant effect on broiler feed intake, feed conversion rate or energy utilisation. A high level of β-mannanase was produced by A. niger BCC4525 under solid state condition using copra meal as carbon source. Although the enzyme has the desired properties of an enzyme additive for improving broiler performance, it does not appear to be beneficial.

Isolation, Characterization and Production of Mannanase from Thermophilic Bacteria to Increase the Feed Quality

Isolation, Characterization and Production of Mannanase from Thermophilic Bacteria to Increase the Feed Quality

English

Mannanase production by a wild-type Aspergillus niger, isolated from spontaneously-degrading wood from South Western Nigeria was monitored at three day intervals for a period of 15 days in media containing pretreated wood shavings of Anogeissus leiocarpus, Gmelina arborea and Terminalia superba as carbon sources. Highest mannanase activity of 25.938 U/g of dry wood shavings was obtained on the 9th day in a medium containing G. arborea as sole carbon source. Maximum activity value for the medium containing A. leiocarpus was 14.110 U/g obtained on the 9th day. On the medium containing T. superba, a maximum activity value of 10.148 U/g was obtained on the 12th day of incubation. G. arborea substrate gave the highest total protein yield of 18.146 mg/ml and the specific mannanase activity of 1.201 U/mg on the crude enzyme extracts. A mannanase activity of 19.091 U/mg was subsequently achieved after purification on ion exchange chromatography with a purification fold of 15.894. Enzyme activities were at their peaks at pH 5.5 (0.4331 U/ml) and at temperature 30°C (1.8617 U/ml). A Km value of 0.754 mg/ml and Vmax value of 1.364 U/mg/min was achieved for the enzyme which was thermally stable up to 65°C. G. arborea wood shavings hold tremendous potential in mannanase production for several industrial uses by the fungus.   Key words: Aspergillus niger, enzyme characterisation, Gmelina arborea, mannanase, solid state fermentation, purification