Isolation, Screening and Identification of Cellulolytic Bacteria from Soil

Background: Cellulose is the most abundant carbohydrate on earth and is considered as a good candidate for production of second generation biofuel (ethanol) and many other products of routine use. For degradation, cellulases are used which are mostly secreted by microbes such as fungi. Cellulases also play an important role in senescence of plants and in host-parasite relationship for invading the plant cell wall. However, comparatively lesser studies have been carried out on cellulase producing bacteria. Therefore, present study was aimed to isolate cellulase (Endo-β-1,4-D-glucanase; EC. 3.2.1.4.) from bacterial sources. Methodology: To isolate thermophilic/ mesophilic cellulase producing bacteria, soil samples were collected from wood furnishing area and agricultural farm around Indore. Besides, soil sample was also collected from the vicinity of Amlai Paper Mill in Budhar district, Madhya Pradesh. These soil samples after suitable dilutions were streaked on different nutrients agar petri-dishes having carboxymethyl cellulose (CMC) as an inducer. After screening, four colonies were isolated capable of producing good amount of cellulase. Screening was done using Congo red staining and confirmation was done after growth of the bacteria in liquid nutrient medium having CMC. These colonies individually were grown in suitable nutrient media having CMC as an inducer and enzyme activity was determined in the nutrient media after harvesting bacterial cells by centrifugation. Results: The highest enzyme producing bacteria were identified as Bacillus lichenoformis and Ochrobactrum anthropi after biochemical analyses, 16S rRNA sequencing and subsequently phylogenetic tree analysis.

Download Full-text

Screening and Identification of Laccase Producing Fungi from Environmental Samples

UMYU Journal of Microbiology Research (UJMR) ◽

10.47430/ujmr.2161.012 ◽

2021 ◽

Vol 6 (1) ◽

pp. 91-98

Author(s):

A. Bello ◽

◽

J. B. Ameh ◽

D. A. Machido ◽

A. I. Mohammed-Dabo

Keyword(s):

Tannic Acid ◽

Fungal Species ◽

Soil Samples ◽

Potato Dextrose Agar ◽

Laccase Production ◽

Curvularia Lunata ◽

Main Campus ◽

Fungal Isolates ◽

Screening And Identification ◽

Decaying Wood

Laccases are oxidases with broad substrate specificity and ability to oxidize various phenolic and non-phenolic compounds. This study was carried out to isolate and characterizes laccase producing fungi from environment samples. Soil and decaying wood samples were collected from different locations within Ahmadu Bello University, Zaria Main campus. Suspensions of the samples (1 g in 10 mL sterile distilled water) were serially diluted, inoculated onto Potato Dextrose Agar (PDA) containing 0.01% Chloramphenicol and incubated for 7 days at 30oC.The fungal isolates were characterized macroscopically and microscopically with the aid of an atlas. The identified fungal isolates were screened for laccase production by inoculating onto PDA containing 0.02% Guaiacol, 1mM ABTS (2 2’-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) and 0.5% Tannic acid as indicator compounds and incubated at 250C for 7 days. The laccase producing isolates were confirmed molecularly by ITS rDNA sequence analysis using the FASTA algorithm with the Fungus database from the European Bioinformatics Institute (EBI).A total of 25 fungal species (11 from soil and 14 from decaying wood samples) were isolated. Two isolates from the soil origin identified as Curvularia lunata SSI7 (Accession No. QIE06317.1) and Fusarium clade VII SSI3 (Accession No. GQ505677) were found to produce laccase where Curvularia lunata SSI7 was able to oxidize all the indicator compounds used for the screening. Fusarium clade VII SSI3 was able to oxidize only 0.5% Tannic acid. Laccase producing Curvularia lunata and Fusarium clade VII were isolated from soil samples collected from ABU Zaria Main Campus. Keywords: laccase, fungi, soil, decaying wood

Download Full-text

Evaluation of commercially available nutrient media for isolation of agents of endemic and ivasive mycoses from soil samples

Immunopathology Allergology Infectology ◽

10.14427/jipai.2019.3.83 ◽

2019 ◽

pp. 83-86

Author(s):

N Polovets ◽

A Lipnitsky ◽

R Surkova ◽

O Shergina ◽

D Victorov ◽

...

Keyword(s):

Soil Samples ◽

Nutrient Media

Download Full-text

Interactions between Treponema bryantii and cellulolytic bacteria in the in vitro degradation of straw cellulose

Canadian Journal of Microbiology ◽

10.1139/m87-041 ◽

1987 ◽

Vol 33 (3) ◽

pp. 244-248 ◽

Cited By ~ 98

Author(s):

H. Kudo ◽

K.-J. Cheng ◽

J. W. Costerton

Keyword(s):

Plant Cell Wall ◽

Bacterial Species ◽

Barley Straw ◽

Cellulolytic Bacteria ◽

Morphological Examination ◽

Cellulose Digestion ◽

Acid Generation ◽

Pure Cultures ◽

Treponema Bryantii

To assess the contribution of individual bacterial species to the overall process of cellulose digestion in the rumen, cellulolytic bacteria (Bacteroides succinogenes and Ruminococcus albus) were tested as pure cultures and as cocultures with noncellulolytic Treponema bryantii. In studies of in vitro barley straw digestion, Treponema cocultures surpassed pure cultures of the cellulolytic organisms in dry matter disappearance, volatile fatty acid generation, and in the production of succinic acid, lactic acid, and ethanol. Morphological examination, by electron microscopy, showed that cells of T. bryantii associate with the plant cell wall materials in straw, but that cellulose digestion occurs only when these organisms are present with cellulolytic species such as B. succinogenes. These results show that cellulolytic bacteria interact with noncellulolytic Treponema to promote the digestion of cellulosic materials.

Download Full-text

Isolation, Screening, and Identification of Cellulolytic Bacteria from Natural Reserves in the Subtropical Region of China and Optimization of Cellulase Production byPaenibacillus terraeME27-1

BioMed Research International ◽

10.1155/2014/512497 ◽

2014 ◽

Vol 2014 ◽

pp. 1-13 ◽

Cited By ~ 44

Author(s):

Yan-Ling Liang ◽

Zheng Zhang ◽

Min Wu ◽

Yuan Wu ◽

Jia-Xun Feng

Keyword(s):

Cellulase Production ◽

Biochemical Properties ◽

Rrna Gene ◽

Cellulolytic Bacteria ◽

Bacterial Strains ◽

Subtropical Region ◽

Cmcase Activity ◽

Degrading Bacteria ◽

Screening And Identification ◽

Natural Reserves

From different natural reserves in the subtropical region of China, a total of 245 aerobic bacterial strains were isolated on agar plates containing sugarcane bagasse pulp as the sole carbon source. Of the 245 strains, 22 showed hydrolyzing zones on agar plates containing carboxymethyl cellulose after Congo-red staining. Molecular identification showed that the 22 strains belonged to 10 different genera, with theBurkholderiagenus exhibiting the highest strain diversity and accounting for 36.36% of all the 22 strains. Three isolates among the 22 strains showed higher carboxymethyl cellulase (CMCase) activity, and isolate ME27-1 exhibited the highest CMCase activity in liquid culture. The strain ME27-1 was identified asPaenibacillus terraeon the basis of 16S rRNA gene sequence analysis as well as physiological and biochemical properties. The optimum pH and temperature for CMCase activity produced by the strain ME27-1 were 5.5 and 50°C, respectively, and the enzyme was stable at a wide pH range of 5.0–9.5. A 12-fold improvement in the CMCase activity (2.08 U/mL) of ME27-1 was obtained under optimal conditions for CMCase production. Thus, this study provided further information about the diversity of cellulose-degrading bacteria in the subtropical region of China and foundP. terraeME27-1 to be highly cellulolytic.

Download Full-text

Screening and Identification of Ligninolytic Bacteria for the Treatment of Pulp and Paper Mill Effluent

Water Air & Soil Pollution ◽

10.1007/s11270-015-2535-y ◽

2015 ◽

Vol 226 (9) ◽

Cited By ~ 16

Author(s):

Rajni Hooda ◽

Nishi K. Bhardwaj ◽

Pamela Singh

Keyword(s):

Paper Mill ◽

Pulp And Paper ◽

Pulp And Paper Mill ◽

Paper Mill Effluent ◽

Screening And Identification

Download Full-text

Studies of the Extracellular Glycocalyx of the Anaerobic Cellulolytic Bacterium Ruminococcus albus 7

Applied and Environmental Microbiology ◽

10.1128/aem.01632-06 ◽

2006 ◽

Vol 72 (12) ◽

pp. 7559-7566 ◽

Cited By ~ 31

Author(s):

Paul J. Weimer ◽

Neil P. J. Price ◽

Otini Kroukamp ◽

Lydia-Marie Joubert ◽

Gideon M. Wolfaardt ◽

...

Keyword(s):

Energy Demand ◽

Extracellular Polymeric Substances ◽

Cellulose Microfibrils ◽

Variable Pressure ◽

Cellulolytic Bacterium ◽

Detergent Solution ◽

Bacterial Cells ◽

Cellulolytic Bacteria ◽

Ruminococcus Albus ◽

Almost All

ABSTRACT Anaerobic cellulolytic bacteria are thought to adhere to cellulose via several mechanisms, including production of a glycocalyx containing extracellular polymeric substances (EPS). As the compositions and structures of these glycocalyces have not been elucidated, variable-pressure scanning electron microscopy (VP-SEM) and chemical analysis were used to characterize the glycocalyx of the ruminal bacterium Ruminococcus albus strain 7. VP-SEM revealed that growth of this strain was accompanied by the formation of thin cellular extensions that allowed the bacterium to adhere to cellulose, followed by formation of a ramifying network that interconnected individual cells to one another and to the unraveling cellulose microfibrils. Extraction of 48-h-old whole-culture pellets (bacterial cells plus glycocalyx [G] plus residual cellulose [C]) with 0.1 N NaOH released carbohydrate and protein in a ratio of 1:5. Boiling of the cellulose fermentation residue in a neutral detergent solution removed almost all of the adherent cells and protein while retaining a residual network of adhering noncellular material. Trifluoroacetic acid hydrolysis of this residue (G plus C) released primarily glucose, along with substantial amounts of xylose and mannose, but only traces of galactose, the most abundant sugar in most characterized bacterial exopolysaccharides. Linkage analysis and characterization by nuclear magnetic resonance suggested that most of the glucosyl units were not present as partially degraded cellulose. Calculations suggested that the energy demand for synthesis of the nonprotein fraction of EPS by this organism represents only a small fraction (<4%) of the anabolic ATP expenditure of the bacterium.

Download Full-text

Screening and identification of microbial strains for removal of colour and adsorbable organic halogens in pulp and paper mill effluent

Process Biochemistry ◽

10.1016/s0032-9592(03)00303-0 ◽

2004 ◽

Vol 39 (11) ◽

pp. 1693-1699 ◽

Cited By ~ 52

Author(s):

Indu Shekhar Thakur

Keyword(s):

Paper Mill ◽

Pulp And Paper ◽

Pulp And Paper Mill ◽

Paper Mill Effluent ◽

Screening And Identification ◽

Microbial Strains ◽

Organic Halogens ◽

Adsorbable Organic Halogens

Download Full-text

Screening and identification of indigenous cellulolytic bacteria from indonesian coffee pulp and investigation of its caffeine tolerance ability

Malaysian Journal of Microbiology ◽

10.21161/mjm.86416 ◽

2017 ◽

Author(s):

Arimurti, S. ◽

Nurani, Y. ◽

Ardyati, T. ◽

Suharjono, S.

Keyword(s):

Cellulolytic Bacteria ◽

Coffee Pulp ◽

Screening And Identification

Download Full-text

Features of isolation of the anthrax pathogen depending on the type of nutrient medium

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v11i3.2647 ◽

2020 ◽

Vol 11 (3) ◽

pp. 4318-4322

Author(s):

Nariman S. Sadykov ◽

Ramzi N. Nizamov ◽

Elmira N. Mustafina ◽

Marina Yu. Gallyamova ◽

Timur R. Mustafin ◽

...

Keyword(s):

Nutrient Medium ◽

External Environment ◽

Soil Samples ◽

Complete Dissolution ◽

Nutrient Media ◽

Bacillus Genus ◽

Positive Evaluation

The purpose of the work is to develop a nutrient medium for differentiation of bacillus from soil aerobic bacilli. In order to achieve the set goal, we used the method of introduction into the environment of cultivation of microorganisms separated from animals and objects of external environment (water, soil, feed, air, scrapes from different surfaces suspected of contamination by their bacillus ) of nutrient substrate sucrose, used by bacteria of Bacillus genus for synthesis of product of their metabolism, a sign absent in bacillus . This feature is essential for identification and differentiation of bacillus from closely related saprophytes. To identify and differentiate the bacillus , the microbes isolated from the external environment were cultivated in a nutrient medium consisting of agar (MPA) and () synthesis of sucrose in the amount of 10% to 100 ml of melted agar. The proposed nutrient media was prepared as follows. agar (500 ml) was melted at 1000C, 10 g of sucrose was added per 100 ml of medium and after the complete dissolution of sucrose, the nutrient medium was poured into dishes and used for sowing the studied material for identification and differentiation of grown crops. The efficiency of the method has been tested in production experiments with positive evaluation. For this purpose, soil samples taken from the territory of old cattle cemeteries were fractionally sown on MPA and for 16-18 hours at 370C and examined crops for the presence of matte and rough (R-form) colonies.

Download Full-text