Over the years, culturable, cellulaseproducing bacteria have been isolated from a wide variety of sources such as composting heaps, decaying plant material from forestry or agricultural waste, the feces of ruminants such as cows, soil and organic matter, and extreme environments like hotsprings, to name a few 24. The residues were serially diluted using sterile distilled water and cultured on basal medium agar plates containing carboxymethylcellulose cmc as a substrate for cellulase production. The present investigation was undertaken to isolate and screen the cellulase. Screening, purification and characterization of cellulase. Screening of cellulase producing bacteria screening for bacterial cellulase activity in microbial isolates is typically performed on plates containing crystalline cellulose or microcrystalline cellulose such as avicel in the agar at a final concentration of 0. There has been increasing interest in cellulaseproducing bacteria because of their ability to grow fast. Deciphering cow dung for cellulase producing bacteria. Cellulolytic enzymes are synthesized by a number of microorganisms. Screening and identification of cellulase producing bacteria. Kaliwal department of studies in biotechnology and microbiology, karnatak university, dharwad 580 003, india corresponding author abstract introduction enzymes are delicate protein molecules necessary for life.
This study focuses on the isolation and screening of cellulaseproducing bacteria from sugar industry waste molasses and characterization by morphological and biochemical analysis. However, cellulase produced by bacteria is often more effective catalyst. Patagundi and basappa basawanneppa kaliwal, year2014. While the addition of cellulaseproducing bacteria increased. Cellulase is a multienzyme,system having 3 types of. Aerobic bacteria like thermobifida also produce all components of cellulolytic system including exo and endo glucanases 8. The present investigation deals with the isolation, screening and characterization of cellulose producing bacteria from termite gut and animal dung, as these are good source of the cellulose digesting environment. Cellulase production by bacillus subtilis isolated from. Microbial cellulasesproduction, applications and challenges article pdf available in journal of scientific and industrial research 6411. Bacterial cellulase usually lacks one of the three cellulase activities, that is fpase. The cellulolytic activity was measured both qualitatively and quantitatively. Microbial cellulases production, applications and challenges rajeev k sukumaran, reeta rani singhania and ashok pandey biotechnology division, regional research laboratory, industrial estate p o, trivandrum 695 019 microbial cellulases find applications in various industries and constitute a major group of the industrial enzymes.
Lekh ram1, kuldeep kaur2 and sandeep sharma3 department of biotechnology, arni university, kathgarh, indora, kangra, h. In the present study, the strain me271 produced the highest cmcase activity after 60 h of incubation. The name is also used for any naturally occurring mixture or complex of various such enzymes, that act serially or synergistically to decompose cellulosic material. Cellulase is any of several enzymes produced chiefly by fungi, bacteria, and protozoans that catalyze cellulolysis, the decomposition of cellulose and of some related polysaccharides. By contrast, most anaerobic microorganisms produce large 1. Bagasse powder is used as a substrate for cellulase production employing novel thermostable yeast. Cellulose is commonly degraded by an enzyme called cellulase. Further, purification of cellulase was carried by ammonium sulfate. In order to determine the effect of temperature on cellulase production, the selected bacterial isolate was grown in carboxymethyl cellulose and 65o c for 48 hours. The solid cellulosic waste can be treated efficiently. Characterization of some efficient cellulase producing.
Hemicellulaseproducing bacteria and engineering hemicellulases. This study focuses on the isolation of efficient cellulase producing bacteria found in organic fertilizers and paper mill sludges which can be considered for use in large scale biorefining. Isolation and characterization of cellulase producing bacteria from soil basavaraj i. Isolation and characterization of cellulaseproducing. Isolation and identification of cellulose degrading. For understanding the mechanism of cellulose degradation by cellulase, it is necessary to isolate, purify and characterize this enzyme. Characteristics, sources, production, and applications.
Microbial cellulases production, applications and challenges. Microorganisms that can produce cellulase enzymes cellulolytic microorganisms can degrade cellulose. Seven different bacterial strains were isolated and screened for cellulase production. Cellulosomes are protuberances on the cell wall of the bacteria, which harbor stable enzyme complexes. The aim of present work is to illustrate the screening and characterization of cellulytic bacteria from soil and waste molasses of sugar industry. The biochemical characterizations of the isolated strains c 1, c 2 and c 3 are presented in additional file 3. While cellulose is a basic structural material of most plants, it is also produced by bacteria, principally of the genera acetobacter, sarcina ventriculi and agrobacterium. Thermostable cellulase can increase the rate of reaction, decrease amount of enzyme needed, longer halflife and decrease. This study aimed to isolate and screen the cellulase producingbacteria from sugar industry wastes molasses and purify and characterize the cellulase produced by the isolated bacterial strains. Novel celluloseproducing bacteria including one capable of producing of a bacterial cellulose having a weightaverage degree of polymerization in terms of polystyrene of 1. Isolation and screening of cellulolytic bacteria from soil and optimization of cellulase production. In our investigation of cellulolytic bacteria, the isolates from oil palm meal were screened and identified based on the phenotypic characteristics.
The isolate were tentatively identified to be bacillus species based on cultural, morphological, biochemical analysis and labelled as cb3, cb4 and cb8. Cellulases are inducible enzymes that are synthesized by a large number of microorganisms during their growth on cellulosic materials. Isolation of cellulolytic bacteria cellulase producing bacteria were isolated from the sample by serial dilution followed by plating them on a cellulose congo red agar medium. Pdf identification of cellulaseproducing bacteria from soil in. This enzyme has various industrial applications and now considered as major. Cellulose is the most abundant component of plant biomass on the earth. They may also be less inhibited by the presence of material that has already been hydrolyzed feedback inhibition. Optimization of the fermentation medium for maximum cellulase production was carried out. Optimization of cellulase production from bacteria isolated from soil.
Isolation and molecular characterization of cellulase. Isolation and characterization of cellulase producing. Introduction cellulose are produced mainly in plants. Cellulase15 producing bacteria and fungi were isolated from variant sources such as agricultural wastes, soil, sediments. Pdf microbial cellulasesproduction, applications and.
Serial dilution was done by mixing 1gm of soil sample in 100ml of sterile distilled water and further proceeding up to 5 dilutions. Fungi are the main cellulase producing microorganisms, though a few bacteria and actinomycetes have also been reported to yield cellulase activity. Screening and identification of cellulase producing. Cellulase can also be isolated from cattle waste 15, woody biomass 16, cow manure 17 and compost 18. When compared to the anaerobic bacteria, aerobic bacteria can perform better cellulase production of around 90%95% of bacterial cellulose degradation.
Fungal cellulase is an abundant resources,is a potential raw material for the microbial production of food, fuel etc. It describes the current knowledge of cellulase production by submerged fermentation and solid state fermentation, properties of cellulase and cloning and expression of cellulase gene. This is produced by several microorganisms, commonly by bacteria and fungi 1,2. Soil and waste samples molasses from colony sugar mills phalia, punjab, pakistan were used for the.
Isolation, screening, and identification of the cellulaseproducing bacteria the sesame seed residues were collected in sterile plastic bags and numbered. Isolation and characterization of cellulase producing bacteria from. A microscopic examination revealed that the isolated strain c 1 and c 2 was rod shaped and found to be gram positive where as c 3 strain was short rod in shape and found to be. Cellulase enzymes are isolated from fungi present in higher termite soil. Effect of cellulaseproducing bacteria on the metabolic. In the light of this background, the present study focuses on the isolation and characterization of novel bacteria from this woodyard with potentials for producing cellulase. Ruminococcus and erwinia species strains are known as cellulaseproducing bacteria, 14 while aspergillus, penicillium, trichoderma and fusarium species are revealed to produce the enzyme in fungi. Pdf on mar 25, 2015, faiz rasul and others published isolation and characterization of cellulase producing bacteria. After incubation of a suitable period, a zone of clearing. Among bacteria, pseudomonas fluorescens is the best cellulase producer among the four followed by bacillus subtilis, e. When the addition of cellulaseproducing bacteria in fermenting grains was 106 cfug group 3, the average metabolic activity of bacterial communities was the highest. Screening, purification and characterization of cellulase from. Pdf the prospects of cellulaseproducing bacteria for.
Complexed cellulase systems cellulosomes on the other hand are native to anaerobic bacteria. Bacteria were further identified by morphological and biochemical tests and subjected to cellulase production in 250 ml erlenmayer flask using potato waste medium for 48h of fermentation period at 35oc with agitation speed of 140rpm. Results and discussion cellulaseproducing bacteria were isolated from soil. Further partial purification of cellulase was carried out by dialysis and ammonium phosphate precipitation and also to determine molecular weight by sdspage. Cellulaseproducing bacteria were isolated from soil and identified as pseudomonas fluorescens, bacillus subtilis, e. Bacterial isolated were grown on carboxy methyl cellulose cmc agar at various optimum conditions such as. Bacteria, which has high growth rate as compared to fungi has good potential to be used in cellulase production ariffin et al. Cellulase production by utilizing agricultural wastes. The wide variety of bacteria in the environment permits screening for more efficient cellulases to help overcome current challenges in biofuel production.
In the terrestrial environments, it is the primary product of photosynthesis and the most abundant renewable bioresource 100 billions dry tonsyear produced in the biosphere. Bacterial, or microbial, cellulose has different properties from plant cellulose and is characterized by high. Isolation and characterization of thermophilic cellulase. Phylogenetic diversity and characterization of novel and. Although the application of bacteria in producing cellulase is less investigated, cellulases produced by bacteria are often more e. Hemicellulose is the second most abundant renewable biomass, accounting for approximately 2535% of lignocellulosic biomass and therefore bacterial enzymes involved in its degradation have also been the focus of several hemicellulase engineering studies using either rational design or directed evolution. Research article optimization of cellulase production from. Cellulases provide a key opportunity for achieving tremendous benefits of biomass utilization wen et al. Inducible cellulase production from an organic solvent. The culture conditions like ph, temperature, carbon sources, and nitrogen sources were optimized. Pdf this study aimed to isolate and identify bacteria that can produce cellulase enzyme from nasinuan forest, kantarawichai district. Presence of some species of bacteria, yeast, and fungi in sugar industry wastes has been reported in a recent study 15.
The prospects of cellulaseproducing bacteria for the. Microorganisms of the genera trichoderma and aspergillus are thought to be cellulase producers, and crude enzymes produced by these microorganisms are commercially available for. Screening and identification of cellulase producing bacteria isolated from oil palm meal saowapar khianngam1, yupa pootaengon1, taweesak techakriengkrai2, somboon tanasupawat3 1faculty of animal sciences and agricultural technology, silpakorn university, phetchaburi it campus, phetchaburi 76120, thailand. Fungi and bacteria isolated from soil secrete several. Fungi are the main cellulaseproducing microorganisms, though a few bacteria and actinomycetes have also been recently reported to yield cellulase activity table 31.
The cellulase producing bacteria were isolated from industrial and agricultural areas in kerala. Based on the morphological and biochemical characteristics, the isolates were identi. The cellulase producing bacteria was isolated from cow dung and characterized by morphological and biochemical analysis. Cellulase production from species of fungi and bacteria. Bioethanol production with cellulase enzyme from bacillus. Production of cellulase was analyzed by dinitrosalicylic acid dns and carboxymethyl cellulase assay methods. The optimum conditions found for cellulase production were 40c at ph 10 with glucose as carbon source and ammonium sulphate as nitrogen source, and coconut cake stimulates the production of cellulase.
Isolation, screening and identification of cellulase producing bacteria. Potential isolates with cellulase production were identified by grams iodine dye staining method. Among bacteria, bacillus species produce a number of extracellular enzyme including amylases, proteinases, and polysaccharide hydrolases mawadza et al. Isolation and screening of cellulolytic bacteria from soil. Cellulase producing bacteria from the woodyards on kallai. Thus, present study was carried out to isolate cellulase producing bacteria from different environmental samples such as cow dung, compost soil and termite gut content. Screening isolation and characterization of cellulase producing microorganisms from soil. Optimization of cellulase production from bacteria. All the samples were tenfold diluted and pour plate and streak plate with carboxy methyl cellulose medium method were employed in order to isolate bacteria strains.972 335 1037 1483 672 628 1339 141 1379 481 1237 1106 1195 973 646 1340 1470 1007 1192 406 905 1159 1047 113 340 680 200 1181 377 1444 1423 1041 1229 752 1051 678 584 1462 865 870 236 1328 671 1138 695 42