Cellulase Enzyme Production Research Articles

Optimization of Culture Conditions for the Production of Alkaline Cellulase Enzyme Produced from Fusarium oxysporum VSTPDK

Alkaline cellulase producing Fusarium oxysporum VSTPDK was isolated and screened from the soil of Kapurthala district Punjab, India. This organism produced endoglucanase (CMCase) and exoglucanase (FPase) when grown at a different range of parameters pH (6, 7, 8, 9, 10 and 11), temperature (30, 40, 50 and 60OC), incubation time (4th 6th 8th 10th and 12th day) and nitrogen source (NH4SO4, NH4Cl, NaNO3 and NH4HCO3). Carboxymethylcellulose (CMC) and cellulose powder were used as the sole carbon source. In this research, statistical tools called Response Surface Methodology (RSM) was used for the optimization of cellulase enzyme by selecting three important parameters after one factor at a time (OFAT) approach. Using OFAT, optimum production of both CMCsase (3.52U/mL) and FPase (4.07U/mL) were achieved after 8 days incubation at pH 8, temperature 30OC and 1.0g/L ammonium sulphate while RSM produced CMCase 3.91U/mL and FPase 4.26U/mL respectively when incubated for 8 days at pH 8.5, temperature 45 and 3% ammonium sulphate concentration. Optimization of the culture conditions using RSM leads to an increase of 0.39U/mL (CMCase) and 0.19U/mL (FPase). The use of RSM has gained considerable attention in the past decade in the optimization of various physicochemical parameters and nutritional factors. Its application in different industries may find ways of selecting different factors influencing cellulase activity. The fungus can produce a considerable amount of cellulase enzyme at a pH of up to 10 and 50OC. To our knowledge, this is the first report of alkalothermophilic oxysporum VSTPDK from Punjab, India.

Isolation, Screening and Production of Bacterial Cellulase from Cow Dung

Cellulases are the most important industrial enzyme due to their potential application in various industries including pulp and paper, textile, laundry, biofuel production, food and feed industry, brewing and agriculture. The present study was carried out to isolate and characterize cellulase producing microorganisms from different cow dung samples. Four different cow dung samples were collected from variety of cows namely Gir, Holstein, Jersey and Desi and isolation and screening was done to check cellulase producing microorganisms. The obtained 11 isolates were screened for their cellulase activity by using CMC (carboxymethyl cellulose) agar medium. In the current study, 10 cellulase producing isolates were obtained and were characterized morphologically from which 8 isolates were found Gram negative and 2 were found Gram positive. All the 10 cellulase producers were further confirmed for their cellulase producing ability by performing turbidity test. Out of these 10 isolates, GN4 and HN2 with optical density 0.35 and 0.28, respectively were found to be best cellulase producer and were selected for cellulase enzyme production and other further studies. Botssh the isolates were tested for their enzymatic activities by performing DNSA method and protein estimation by Lowry’s method. Cellulase activities were found to be 7.33 µg/ml/min and 3.66 µg/ml/min and the total protein concentrations were found to be 40 µg/ml and 32 µg/ml for GN4 and HN2, respectively. Thus, dungs cow dung can be considered as the excellent source for cellulase producing bacteria.

Nickel ferrite nanoparticles induced improved fungal cellulase production using residual algal biomass and subsequent hydrogen production following dark fermentation

The present study reports nickel ferrite nanoparticles (NiFe2O4 NPs) induced enhanced production of crude cellulase enzyme using residual algal biomass of cyanobacteria Lyngbya limnetica as substrate. It is noticed that the residual algal substrate and NiFe2O4 NPs mediated crude cellulase exhibits nearly 2.5 fold enhanced filter paper activity after 72 h along with better efficiency in terms of pH and thermal stability as compared to the control system. Further, NiFe2O4 NPs mediated crude cellulase enzyme was employed for the enzymatic hydrolysis of rice straw to produce sugar hydrolyzate. Subsequently, using bacterial strains Bacillus subtilisPF_1 the cumulative hydrogen ~ 1820 mL/L has been produced under the dark fermentation.

Screening and identification of cellulolytic fungi at rhizosphere of safira taro plant

Agriculture waste and pathogen cell wall of plant contain a lot of cellulose which are very difficult to degrade. One of the ways to hydrolyse them is through utilizing cellulase, an enzyme produced from secondary metabolite of cellulolytic fungi. This research was aimed at isolating fungi from rhizosphere of safira taro plant and testing the isolate capability to produce cellulase enzyme to be able to act as decomposer and biocontrol of the plant. The research methods are isolating rhizosphere fungi, testing isolate capability in producing cellulase enzyme and identifying potential isolate producing cellulolytic fungi. It obtained 10 fungi isolates, 4 were negative and 6 positive in producing cellulase enzyme with cellulolytic index 1.16 – 1.66. The highest index was RTB14 isolate which belongs to Aspergillus flavus.

Effect of ultrasonic technology on cellulase enzyme activity produced by local bacterial isolate

The research aimed to obtain new bacterial isolate producing cellulase enzyme, and ultrasonic effect on enzyme activity. Three bacterial isolates were isolated from different agricultural wastes samples. Their effectiveness in cellulolytic was detected by measurement of the clear zone diameter around bacterial growth. The enzyme was extracted after the growtht of local bacterial isolate which has highest enzyme activity. The enzyme was exposed to ultrasound intensity of 40 KH, low, medium and high power, and exposure time (10, 20, 30, 40, 50, and 60) min, enzymatic activity was measured after each treatment compared with control. It was found at low level that an increase in the cellulase activity with increased ultrasonic exposure time, the higher activity at 60(min). The effect of ultrasonic treatment at mid-level, showed higher activity at 20 (min), while at high level the activity decrease with exposure time when compared with control. Protein concentration was estimated, there was no significant difference on protein content after ultrasonic treatment in samples at (10, 20, 30, 40) min while decrease in protein content in samples at (50, 60) min, compared with with control.

Isolation and Screening of Multifunctional Plant Growth Promoting Rhizobacteria from Rhizopshere of Different Crop Fields

Rhizospheric bacteria are naturally occurring soil microbes that are aggressively found in the plant rhizosphere, at root surface and in association with roots. They give satisfactory benefit plants by several mechanisms such as nitrogen fixation, phosphate solubilization, potassium decomposition, IAA production, antagonism against phytopathogenic microorganisms by production of siderophore, antibiotics and cell wall degrading enzymes. The total number of beneficial bacteria were isolated from different rhizospheric soil in agricultural lands. The isolated bacterial strains were screened for their plant growth promoting factors such as production of ammonia, siderophore, cellulase, chitinase, and pectinase enzyme. All of the isolates produced ammonia and 79% of the isolates produced siderophore on chrome azurole S agar plates. Furthermore, the bacterial isolates produced cell wall degrading enzyme; pectinase (69%), cellulase (94%), chitinase (51%), amylase (61%) and glucanase enzyme (59%) on agar plate method. The isolates also produced auxin type plant hormone (IAA), all the isolates produced IAA and the highest IAA producing strain is W1 and the produce amount was 21.91mg/L. Among the isolated bacteria, only two strains could produce HCN with the use of Feigl-Anger paper method. The recent study suggests that the use of these PGPR isolates as inoculants might be a promising source for sustainable agricultural use.

The Effect of Addition Mn2+ Metal Ions and Incubation Time to Bacillus cereus Cellulase Enzyme Activity from Endophytic Bacteria of Curcuma Rhizome (Curcuma zanthorrizha Roxb.)

Cellulase is one of the most widely used enzymes in the industrial world and wastes decomposition process. Bacillus cereus is one of the bacteria that can produce cellulase enzymes that can hydrolyze cellulose to glucose. The addition of cofactors and incubation time can help determine the optimum conditions needed by cellulase enzymes to work optimally. This study aims to determine the effect of adding Mn2+ metal ions and incubation time to the activity of cellulase enzymes from Bacillus cereus endophytic bacteria. This research is experimentally used a Completely Randomized Design (CRD) factorial design with two factor treatments and 3 times repetitions. The first factor is variation of addition Mn2+ metal ions which are 5 mM, 10 mM, and 15 mM, the second is variation of incubation time which are 1 hour, 2 hours, 3 hours, 4 hours, and 5 hours. The data were analyzed using Analysis Of Variance (ANOVA) and if the data significantly affected the parameter, then it would be continued by Duncan Multiple Range Test (DMRT) with the fault level of 5%. The result showed that the interaction of addition Mn2+ metal ions and incubation time affected the cellulase enzyme activity of Bacillus cereus from endophytic bacteria. The highest cellulase enzyme activity obtained from interaction addition Mn2+ metal 10 mM and incubation time 3 hours with an activity 0,335 U/mL, while the lowest cellulase enzyme activity obtained from interaction addition Mn2+ metal 15 mM and incubation time 5 hours with an activity of 0,073 U/mL

PRODUCTION, OPTIMIZATION AND DEINKING CAPACITY OF ALKALINE CELLULASE PRODUCED FROM MUCOR CIRCINELLOIDES WSSDBS2F1

An alkaliphilic Mucor circinelloides WSSDBS2F1 strain, with the ability to produce endoglucanase and exoglucanase enzymes, was isolated from decomposing bagasse, originating from a sugar processing mill in Punjab, India, and then screened. A pH range of 7-11, temperature of 30-60 °C, incubation period of 4-12 days, different lignocellulosic waste and nitrogen sources were investigated for optimum production of cellulase enzyme. Response Surface Methodology (RSM) was the statistical tool used for optimization of endoglucanase and exoglucanase production, following the one-factor-at-a-time approach. The optimized endoglucanase was further evaluated for its ability to deink waste paper, compared to chemical deinking. It was found that the enzymatically deinked pulp had high ISO brightness, tensile, burst and tearing strength, as compared to chemically deinked and control pulps. Scanning electron microscopy and Fourier transform infrared spectroscopy (FTIR) were also used to confirm the action of the endoglucanase enzyme on waste paper.

Integrated production of optically pure l-lactic acid from paper mill sludge by simultaneous saccharification and co-fermentation (SSCF).

Paper mill sludge (PMS) raises critical environmental issues due to its disposal problem, but its high sugar content and well-dispersed structure make it a great feedstock for biochemical production. The technical feasibility of integrating cellulase enzyme production into lactic acid (LA) fermentation from PMS was investigated in this study. The low ash content of PMS suggests a great potential for cellulase production. The enzyme produced using PMS without any treatment gave an activity of 7.8 FPU/ml, a performance comparable to the commercial enzyme, Cellic CTec 2. The LA yield from PMS with in-house enzyme was 64.7% and 73.7% at the enzyme loading of 10 and 15 FPU/g-glucan, respectively. The LA obtained was optically pure L- isomer with over 99% purity. The optimal condition of LA production by Bacillus coagulans was found to be 50 °C and pH 5.3 (with 50g/L CaCO3). The nutrient effect of yeast extract (YE) and corn steep liquor (CSL) was substrate dependent, and CSL could substitute YE as an inexpensive nutrient when using PMS as a substrate.

Screening of Cellulolytic Bacteria from Biological Education and Research Forest Floor Andalas University, Indonesia.

<b>Background and Objective:</b> Organic waste dump is a problem that needs to be solved, one of which is by using microbe. Cellulolytic bacteria's ability to produce cellulase enzymes that can hydrolyze cellulose. Cellulose is the major component of the plant cell walls that are difficult to endure degradation naturally. This study aimed to find cellulolytic bacterial isolates on Biological Education and Research Forest floor Andalas University and characterize the cellulolytic bacteria found. Material and Methods: CMC (Carboxymethyl Cellulose) medium is used as a screening for bacteria isolate and this study used the survey method and also conducted catalase, glucose and lactose test for characterizations. <b>Results:</b> We found 16 bacterial isolates on Biological Education and Research Forest floor where 9 isolates were in a shaded area and 7 isolates were in an unshaded area. There were 12 isolates from 16 isolates that have the positive cellulolytic ability with a variety of clear zone sizes, where there were 8 isolates with the large clear zone and 4 isolates producing very small clear zones. Characteristics of bacterial isolates with the large clear zone obtained were 2 gram-positive coccus isolates with positive catalase test, 3 gram-negative coccus isolates with positive glucose test and 3 gram-negative bacilli isolates with negative fructose test. <b>Conclusion:</b> We identified 2 potential isolates with a cellulolytic index value greater than 2, isolate UCB 4 with a cellulolytic index value of 3.5 and UCB 6 with a cellulolytic index value of 2.2.

Technological advances for improving fungal cellulase production from fruit wastes for bioenergy application: A review

Fruit wastes can be imperative to elevate economical biomass to biofuels production process at pilot scale. Because of the renewable features, huge availability, having low lignin content organic nature and low cost; these wastes can be of much interest for cellulase enzyme production. This review provides recent advances on the fungal cellulase production using fruit wastes as a potential substrate. Also, the availability of fruit wastes, generation and processing data and their potential applications for cellulase enzyme production have been discussed. Several aspects, including cellulase and its function, solid-state fermentation, process parameters, microbial source, and the application of enzyme in biofuels industries have also been discussed. Further, emphasis has been made on various bottlenecks and feasible approaches such as use of nanomaterials, co-culture, molecular techniques, genetic engineering, and cost economy analysis to develop a low-cost based comprehensive technology for viable production of cellulase and its application in biofuels production technology.

12. Isolate Of Bacteriua Cellulase Enzyme Production In The Rumen Of Aceh Cattle Base On Analysis Homology 16S rRNA

The rumen of cattle is well known one of a bacterium cellulolitic production that have ability to degrade cellulose for produce cellulase enzymes. The aims of the present study is to determine the type of bacteria produced cellulase enzymes in the rumen of cattle aceh. The diversity of cellulolitic bacteria is determined by analysis of 16S rRNA genes. Isolation of cellulolytic bacteria, total DNA isolation, agarose gel electrophoresis, amplification of the 16S rRNA gene, determining the DNA sequence, DNA sequence homology and alignment of DNA sequences. Isolation of bacterium cellulolitic, isolating total DNA, gel electroforesis agarosa, amplified of gen 16S rRNA, determination the DNA barcode, homology DNA barcode, and alignment of DNA sequences. The sample of rumen liquid was taked from RPH , then maked isolation of total DNA to get pure DNA from bacterium whos result and amplification DNA of bacterium cellulolitic separated by electroforesis gel agarosa. Gene of bacterium cellulolitic was isolated based Polymerase Chain Reaction methode. Alignment is done by using BlastN program, then use CLustalW program. It can be concluded that the species of bacteria (FKH_USK_6) in rumen has a closeness with Coronabacter sakazakii bacteria strain PNP8 of 97% according to the sizeof the DNA sequence and the same position on an existing gene in GenBank.

Application of yeasts isolated from fermented cocoa beans for biocontrol of pathogenic mold in chocolate fruit

Contamination by pathogenic mold in postharvest cocoa beans becomes a significant concern by most Indonesian farmers. Pathogenic mold can cause damage to cocoa beans by such as rotting diseases in fruit. One alternative that can be used to control pathogenic mold is using biological agents such as yeasts. Some group of yeasts can produce cellulase enzyme that can degrade cellulose, and it can possibly break the cell wall with of mold which composed of semi-crystalline chitin, p-need, and cellulose. This study aims to determine the yeast originated from fermented cocoa beans which can produce cellulase enzymes and their potential ability as a biocontrol for pathogenic molds in chocolate fruit. This study includes yeast isolation from fermented beans, screening of yeast isolates that produce cellulase enzymes, and in-vitro antagonistic testing against pathogenic molds on chocolate fruit. The results showed that there were 21 yeast isolates from fermented cocoa beans, and among all, there were five isolates which can produce cellulase enzymes, namely isolate C4.-3.3, C4.-3.13, C4.-4.9, C4.-4.10, and C4.-5.9. Yeast isolate C4.-4.10 can produce cellulase enzymes with an index of 0.32 U/mL. This research showed that the 5 yeast isolates have the low category of cellulase enzyme, and further study is needed to be done to confirm their ability to act as a biocontrol agent.

Isolation and evaluation of the ability to decompose sugarcane leaves of bacterial strains from acid sulfate soils in the Mekong River Delta

The objectives of this study were to isolate and determine the ability of bacterial strains to decompose sugarcane leaves from acid sulfate soils in the Mekong River Delta (MRD). Soil samples were collected from three acid sulfate soils in Ben Luc district, Long An province, Phung Hiep district, Hau Giang province, and Hon Dat district, Kien Giang province where large sugarcane areas cultivated. Six soil samples were collected and coded: LA1, LA2, HG1, HG2, KG1, and KG2. The results of the study have isolated 18 strains of bacteria are capable of producing cellulase enzyme that breaks down cellulose including LA2-4b, LA2-1, LA2-4a, LA2-2, KG2-1, KG2-2a, KG2-2b, KG2-3, KG2-20, KG2-21, KG2-22, KG2-24, KG2-26, KG2-27, LA1-1, LA1-2, LA1-3, LA1-7. All isolated bacterial strains have the ability to effectively decompose sugarcane leaves, have the potential to apply in practical production, and contribute to improving the quality of acid sulfate soils in the MRD. In which, five selected bacterial strains (LA1-1, LA2-4a, LA2-4b, KG2-2b, and KG2-24) were significantly higher (p<0.05) in the decomposition of sugarcane leaves than the treatment without bacteria.

Isolation and Characterization of Yeasts from Rumen Fluids for Potential Use as Additives in Ruminant Feeding.

Saccharomyces cerevisiae is a yeast strain often used to improve the feed quality of ruminants. However, S. cerevisiae has limited capacity to provide biomass when inoculated with carbon sources and a low ability to produce cellulase enzymes. Here, we hypothesized that yeast in the rumen produces a large amount of biomass and could release cellulase enzymes to break down fiber content. Therefore, the aim of this study was to screen, isolate and identify yeast from the rumen fluids of Holstein Friesian steers and measure the efficiency of biomass production and cellulase activity. A fermentation medium containing sugarcane molasses as a carbon source and urea as a nitrogen source was optimized. Two fistulated–crossbred Holstein Friesian steers averaging 350 ± 20 kg body weight were used to screen and isolate the ruminal yeast. Two experiments were designed: First, a 12 × 3 × 3 factorial was used in a completely randomized design to determine biomass and carboxymethyl cellulase activity. Factor A was the isolated yeast and S. cerevisiae. Factor B was sugarcane molasses (M) concentration. Factor C was urea (U) concentration. In the second experiment, potential yeasts were selected, identified, and analyzed for 7 × 4 factorial use in a completely randomized design. Factor A was the incubation times. Factor B was the isolated yeast strains, including codes H-Khon Kaen University (KKU) 20 (as P. kudriavzevii-KKU20), I-KKU20 (C. tropicalis-KKU20), and C-KKU20 (as Galactomyces sp.-KKU20). Isolation was imposed under aerobic conditions, resulting in a total of 11 different colonies. Two appearances of colonies including asymmetric colonies of isolated yeast (indicated as A, B, C, E, and J) and ovoid colonies (coded as D, F, G, H, I, and K) were noted. Isolated yeast from the rumen capable of providing a high amount of biomass when inoculant consisted of the molasses 15% + urea 3% (M15 + U3), molasses 25% + urea 1% (M25 + U1), molasses 25% + urea 3% (M25 + U3), and molasses 25% + urea 5% (M25 + U5) when compared to the other media solution (p < 0.01). In addition, 11 isolated biomass-producing yeasts were found in the media solution of M25 + U1. There were 4 isolates cellulase producing yeasts discovered in the media solution of M25 + U1 and M25 + U5 whereas molasses 5% + urea 1% (M5 + U1), molasses 5% + urea 3% (M5 + U3), molasses 5% + urea 5% (M5 + U5), molasses 15% + urea 1% (M15 + U1), molasses 15% + urea 3% (M5 + U3), and M25 + U3 were found with 2, 3, 1, 2, 1, and 2 isolates, respectively. Ruminal yeast strains H-KKU20, I-KKU20, and C-KKU20 were selected for their ability to produce biomass. Identification of isolates H-KKU20 and I-KKU20 revealed that those isolates belonged to Pichia kudriavzevii-KKU20 and Candida tropicalis-KKU20 while C-KKU20 was identified as Galactomyces sp.-KKU20. Two strains provided maximum cell growth: P. kudriavzevii-KKU20 (9.78 and 10.02 Log cell/mL) and C. tropicalis-KKU20 (9.53 and 9.6 Log cells/mL) at 60 and 72 h of incubation time, respectively. The highest ethanol production was observed in S. cerevisiae at 76.4, 77.8, 78.5, and 78.6 g/L at 36, 48, 60, and 72 h of incubation time, respectively (p < 0.01). The P. kudriavzevii-KKU20 yielded the least reducing sugar at about 30.6 and 29.8 g/L at 60 and 72 h of incubation time, respectively. The screening and isolation of yeasts from rumen fluids resulted in 11 different yeasts being obtained. The potential yeasts discovered in the rumen fluid of cattle were Pichia kudriavzevii-KKU20, Candida tropicalis-KKU20, and Galactomyces sp.-KKU20. P. kudriavzevii-KKU20 had higher results than the other yeasts in terms of biomass production, cellulase enzyme activity, and cell number.

Isolation, Identification and In Silico Study of Native Cellulase Producing Bacteria

Aims: The purpose of this study was to screen the bacteria producing cellulase enzymes and their bioinformatics studies. Background: Cellulose is a long-chain polymer of glucose that hydrolyzes by cellulases to glucose molecules. In order to design the new biotechnological applications, some strategies have been used as increasing the efficiency of enzyme production, generating cost-effective enzymes, producing stable enzymes and identification of new strains. Objective: On the other hand, some bacteria special features have made them suitable candidates for the identification of the new source of enzymes. In this regard, some native strains of bacteria were screened. Methods: These bacteria were grown on a culture containing the liquid M9 media containing CMC to ensure the synthesis of cellulase. The formation of a clear area in the culture medium indicated decomposition of cellulose. In the following, the DNA of these bacteria were extracted and their 16S rDNA genes were amplified. Result: The results show that nine samples were able to synthesize cellulase. In following, these strains were identified using 16S rDNA. The results show that these screened bacteria belonged to the Bacillus sp., Alcaligenes sp., Alcaligenes sp., and Enterobacter sp. Conclusion: The enzyme activity analysis shows that the Bacillus toyonensis, Bacillus sp. strain XA15-411 Bacillus cereus have produced the maximum yield of cellulases. However, these amounts of enzyme production in these samples are not proportional to their growth rate. As the bacterial growth chart within 4 consecutive days shows that the Alcaligenes sp. Bacillus cereus, Bacillus toyonensis, Bacillus sp. strain XA15-411 have a maximum growth rate. The study of the phylogenetic tree also shows that Bacillus species are more abundant in the production of cellulase enzyme. These bioinformatics analyses show that the Bacillus species have different evolutionary relationships and evolved in different evolutionary time. However, for maximum cellulase production by this bacteria, some information as optimum temperature, optimum pH, carbon and nitrogen sources are needed for the ideal formulation of media composition. The cellulase production is closely controlled in microorganisms and the cellulase yields appear to depend on a variety of factors. However, the further studies are needed for cloning, purification and application of these new microbial cellulases in the different commercial fields as in food, detergent, and pharmaceutical, paper, textile industries and also various chemical industries. However, these novel enzymes can be further engineered through rational design or using random mutagenesis techniques.

Literature study of production dry cellulase from Trichoderma reesei, Aspergillus niger, and Bacillus subtilis

Cellulase is an enzyme that can degrade cellulose through a catalytic process. The enzyme that works synergistically to release sugar (glucose). This review aims to determine cellulase enzymes’ activation from three different microorganisms, Aspergillus niger, Bacillus subtilis, and Trichoderma reesei. Enzymes’ activation using various kinds of lignocellulosic substrates and the effect of adding carrier agents on the freeze-drying process. The cellulase enzyme from Trichoderma reesei has an activity of 3.4 IU / mL. This enzyme is produced by pre-treatment 1% H2O2 in lignocellulosic media. Aspergillus niger has a cellulase enzyme activity of 0.229 IU / mL using a sugarcane bagasse substrate that has been pre-treated by shiitake mushrooms. The cellulase enzyme from Bacillus subtilis has an activity of 0.907 IU / mL, with the addition of CMC levels of 5% as an inducer. Cellulase enzymes in liquid form are susceptible to denaturation during storage. The production of cellulase enzymes in solid form is expected to maintain the stability of the resulting cellulase enzyme activity longer. The addition of non-reducing sugar as a carrier agent in the freeze-drying process is reported to preserve the biological activity. The addition of sucrose and trehalose, each with a concentration of 300 mM, maintained the amylase enzyme activity up to 90%. The addition of sucrose with an optimum concentration of 60 mM protects both total protein stability and good lysozyme activity. It is suspected that a carrier agent with an optimal concentration can maintain protein stability in cellulase enzyme activity in this review.

Isolation and optimization of the fermentation condition of cellulolytic microbial isolates from cassava waste water

The study was conducted to isolate and identify cellulose producing microorganisms from cassava waste water. Cassava waste water sample was obtained from a cassava processing factory at Lokogoma town in Wushishi Local government area of Niger State. The microorganisms were isolated, identified and counted by standard microbiological methods. The mean bacteria count ranges from 6.8 x 103 cfu/mL-1 to 2.1 x 103 cfu/mL-1 while the fungi count ranges from 3.2 x 103 cfu/mL-1 to 1.2 x 103 cfu/mL-1. A total of eight (8) bacterial; Staphylococcus aureus, Bacillus anthrax, Bacillus subtilis, Escherichia coli, Klebsilla sp, Bacillus megaterus, Staph. Epidermidis and Pseudomonas aeruginosa, and six (6) fungi; Saccharomyce serivicea, Aspergillus niger, Penecillium sp., Muccor sp., Aspergilus flavus and Aspergilus fumigetus isolates were identified in the waste water. Among these organism, the best cellulase activity was recorded for Bacillus subtilis (10.39 x 10-4 mg/ml/sec) and Aspergillus niger (11.21 x 10-4 mg/ml/sec). However, maximum activity was obtained at pH ranges from 3 ~ 9, temperature ranges from 30 oC ~ 80oC and substrate concentrations ranges from 1.5% ~3.0%. In conclusion, cassava processing water regarded as waste water could be an alternative source of microorganisms capable of producing cellulase enzyme for industrial purposes.

Current perspective on improved fermentative production and purification of fungal cellulases for successful biorefinery applications: a brief review

Cellulolytic enzymes play the most indispensable and centralized role in the hydrolysis of cellulosic material. Effective enzymatic hydrolysis is one of the major prerequisites in the development of a successful lignocellulosic biorefinery. However, the high cost of cellulase production yet remains to be the major bottleneck for large-scale commercial realization of lignocellusic bio-refinery. In addition, the lack of specificity and inconsistent bioconversion efficiencies of enzymes have restricted their applications towards bioconversion of narrow range of substrates. In view of addressing such bottlenecks associated with cellulase applications and further to improve the applicability of cellulase enzyme in various industrial processes, the current review essentially captures up-to-date information of various strategies adopted for the prospective production and purification of cellulase enzymes. Some holistic strategies deliberated in this review, which evolved over time and have brought about notable improvements in the final titer, yield, specificity, and productivity of the cellulolytic enzymes. It includes utilization of low-cost industrial or agricultural wastes for onsite production, application of enzyme, strain improvement through mutagenic, metabolic engineering and genetic engineering approaches, application of suitable bioprocess optimization techniques, development of ideal bioreactor configuration based on mode of fermentation process, and adaptation of the most intensive purification schemes. Additionally, to address the present existing challenges, more emphasizes have been concentrated towards economically sustainable and comprehensive production strategies of the enzyme.

α-Amylase and cellulase production by novel halotolerant Bacillus sp.PM06 isolated from sugarcane pressmud.

A novel Bacillus sp.PM06 isolated from sugarcane waste pressmud was tested for extracellular α-amylase and cellulase enzyme production. The effect of different substrates, nitrogen sources, pH, and temperature on growth and extracellular enzyme production was examined. Bacillus sp.PM06 was able to grow with starch and carboxymethyl cellulose (CMC) as a sole source of carbon and ammonium chloride was found to be the best nitrogen source. Maximum enzyme production was obtained at 48H for both α-amylase and cellulase. The optimal condition for measuring enzyme activity was found to be pH 5.5 at 50°C for α-amylase and pH 6.4 at 60°C for cellulase respectively. It was found that Bacillus sp.PM06 exhibited halotolerance up to 2M Sodium chloride (NaCl) and Potassium chloride (KCl). The isolate could produce α-amylase in the presence of 2M NaCl and 1M KCl. However, the strain produced cellulase even in the presence of 2M NaCl and KCl. Concomitant production of both enzymes was observed when the medium was supplemented with starch and CMC. A maximum of 31±1.15U/mL of amylase and 15±1.5U/mL of cellulase was produced in 48H. The enzyme was partially purified by Ammonium sulphate (NH4 )2 SO4 precipitation with 2.2 and 2.3-fold purification.