scholarly journals Screening and identification of thermophilic cellulolytic bacteria isolated from sawdust compost

2021 ◽  
pp. 348-357
Author(s):  
D. T. H. Phuong ◽  
D. T. Tuyen ◽  
L. V. Thang
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Megaterium ◽  
Metabolic Pathways ◽  
Rrna Gene ◽  
Cellulolytic Bacteria ◽  
16S Rrna Gene Analysis ◽  
Wide Range ◽  
Commercial Enzymes ◽  
Screening And Identification ◽  
Physiological And Biochemical

Composting process mainly depends on the metabolic pathways of the microorganism and involves the activity of different enzymes. Thermophilic cellulase-producing bacteria isolated from sawdust compost were tested for formation of a visible zone around the colonies on the agar plates medium containing carboxymethyl cellulose at 50ºC. Screening of carboxymethyl cellulase producing isolates was further realized on the basis in liquid medium by DNS method. Among 29 isolates investigated, V1 and V11 strains exhibited maximum enzyme activity of 1.9 and 2.3 U/mL, respectively. These isolates were selected for morphological, physiological and biochemical studies and 16S rRNA gene analysis. They were found a Gram-positive, rod-shaped spore forming cells, which were identified as Bacillus megaterium (V1) and Bacillus subtilis (V11) based on cell morphology, nucleotide homology and phylogenetic analysis. The optimal temperature for activity of endoglucanases (CMCase) ranged from 35–45°C (strain V1) and 40– 50ºC (strain V11). Our findings showed that Bacillus megaterium (V1) and Bacillus subtilis (V11) cellulase demonstrate thermophilic characteristics within wide range of temperature and meets the requirements for commercial enzymes. 

scholarly journals Isolation of Cellulolytic Bacillus subtilis Strains from Agricultural Environments

2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Yu-Kyoung Kim ◽  
Shin-Chan Lee ◽  
Young-Yun Cho ◽  
Hyun-Jeong Oh ◽  
Young Hwan Ko
Keyword(s):  
Bacillus Subtilis ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Biochemical Characterization ◽  
Soluble Sugars ◽  
Cellulolytic Enzyme ◽  
Rrna Gene ◽  
Human Society ◽  
Cellulolytic Bacteria ◽  
16S Rrna Gene Analysis

The bioconversion of cellulose and hemicellulose to soluble sugars is important for global stabilization and a sustainable human society. Here, hundreds of cellulolytic bacteria were screened and isolated from soil, compost, and animal waste slurry in Jeju Island, South Korea. Among the isolates, three strains, SL9-9, C5-16, and S52-2, showing higher potential for practical uses were purified on carboxymethyl cellulose (CMC) agar plates and identified as Bacillus subtilis strains by morphological, physiological, and biochemical characterization and 16S rRNA gene analysis. The production patterns of cellulose or hemicellulose-degrading enzymes were investigated during cell culture. All three isolated strains produced CMCase, Avicelase, β-glucosidase, and xylanase enzymes, which suggested synergic cellulolytic systems in Bacillus subtilis. The enzymes showing CMCase, Avicelase, and xylanase activities existed in cell-free culture supernatant, meanwhile β-glucosidase activity was detected in cell debris suggesting that three of the enzymes, including CMCase, Avicelase, and xylanase, were extracellular, and β-glucosidase was cell membrane bound. The three isolates, SL9-9, C5-16, and S52-2, were not the same strains, presenting slight differences in biochemical characteristics, 16S rRNA gene sequences, and cellulolytic enzyme activities.

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

2014 ◽  
Vol 2014 ◽  
pp. 1-13 ◽  
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.

scholarly journals Polaromonas hydrogenivorans sp. nov., a psychrotolerant hydrogen-oxidizing bacterium from Alaskan soil

2007 ◽  
Vol 57 (3) ◽  
pp. 616-619 ◽  
Author(s):  
Maria Sizova ◽  
Nicolai Panikov
Keyword(s):  
Gene Sequence ◽  
Sequence Similarity ◽  
Biochemical Properties ◽  
Rrna Gene ◽  
Organic Substrates ◽  
The Novel ◽  
Membrane Phospholipids ◽  
Wide Range ◽  
Physiological And Biochemical ◽  
Physiological And Biochemical Properties

Psychrotolerant (0–25 °C), chemolithotrophic Gram-negative cocci were isolated from Alaskan forest soil. The novel isolate was found to grow autotrophically on H2 : CO2 mixtures and to switch to heterotrophic growth on media containing organic substrates. The novel strain utilized a wide range of organic acids, some simple sugars and alcohols. Naphthalene vapour did not support growth. On the basis of 16S rRNA gene sequence similarity, the novel strain is affiliated to the genus Polaromonas, of the class Betaproteobacteria, and is related to Polaromonas naphthalenivorans (99.6 % gene sequence similarity), Polaromonas aquatica (97.4 %) and Polaromonas vacuolata (96.1 %). The membrane phospholipids contained 16 : 1ω7c/16 : 1ω6c, 16 : 0 and 18 : 1ω7c, similar to the fatty acids found for P. naphthalenivorans, P. aquatica and P. vacuolata. On the basis of DNA–DNA hybridization, physiological and biochemical properties, the hydrogen-oxidizing mixotrophic isolate represents a novel species, for which the name Polaromonas hydrogenivorans sp. nov. is proposed. The type strain is DSM 17735T (=NRRL B-41369T).

scholarly journals Chitinolytic Bacillus subtilis Ege-B-1.19 as a biocontrol agent against mycotoxigenic and phytopathogenic fungi

2018 ◽  
Vol 44 (3) ◽  
pp. 323-331
Author(s):  
Ozlem Oztopuz ◽  
Nermin Sarigul ◽  
Fakhra Liaqat ◽  
Ro-Dong Park ◽  
Rengin Eltem
Keyword(s):  
Molecular Weight ◽  
Bacillus Subtilis ◽  
Biocontrol Agent ◽  
Pathogenic Fungi ◽  
Phytopathogenic Fungi ◽  
Aspergillus Ochraceus ◽  
Lytic Enzyme ◽  
Rrna Gene ◽  
Lytic Enzymes ◽  
16S Rrna Gene Analysis

Abstract Background Biological control of pathogenic fungi is a possible alternate to the chemical control, which is harmful to humans and environment. Soil-borne Bacillus strains can be potential biocontrol agents and a source of lytic enzymes. Aim This study aimed to examine biocontrol potential and lytic enzyme activities of a soil isolate Bacillus subtilis Ege-B-1.19. Materials and methods Strain was identified by biochemical and 16S rRNA gene analysis and its biocontrol activity was investigated against Aspergillus niger EGE-K-213, Aspergillus foetidus EGE-K-211, Aspergillus ochraceus EGE-K-217, Fusarium solani KCTC6328, Rhizoctonia solani KACC40111 and Colletotrichum gloeosporioides KACC40689. Chitinase, chitosanase, N-acetyl-β-hexosaminidase and protease activities of B. subtilis Ege-B-1.19 were also determined. Chitosanase was purified using Sephadex G-150 column and its molecular weight was determined by SDS-PAGE. Chitooligosaccharides production using chitosanase was carried out and analysed by TLC and HPLC. Results Results depicted that B. subtilis Ege-B-1.19 has shown inhibitory effects against all the test fungi. Chitinase, chitosanase, N-acetyl-β-hexosaminidase and protease activities were determined as 2.7 U mL−1, 7.2 U mL−1, 6.2 U mL−1 and 12.2 U mL−1, respectively. Molecular weight of purified chitosanase was 44 kDa. Chitosanase hydrolysed chitosan to glucosamine (GlcN), dimers (GlcN)2 and trimers (GlcN)3. Conclusion Bacillus subtilis Ege-B-1.19 can be anticipated as useful biocontrol agent and its chitosanase can be utilized for enzymatic synthesis of chitooligosaccharides.

scholarly journals The Use of Ascophyllum nodosum and Bacillus subtilis C-3102 in the Management of Canine Chronic Inflammatory Enteropathy: A Pilot Study

Animals ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3417
Author(s):  
Marco Isidori ◽  
Fabrizio Rueca ◽  
Francesca Romana Massacci ◽  
Manuela Diaferia ◽  
Andrea Giontella ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Activity Index ◽  
Disease Activity Index ◽  
Ascophyllum Nodosum ◽  
Rrna Gene ◽  
16S Rrna Gene Analysis ◽  
Positive Effects ◽  
Inflammatory Bowel ◽  
Bacillus Genus ◽  
Compositional Variations

The aim was to assess the effects of Ascophyllum nodosum (AN) with/without Bacillus subtilis C-3102 as alternative treatments for Chronic Inflammatory Enteropathy (CIE) of dogs. Fourteen CIE patients, which had received the same control (CTR) diet, were enrolled to serially receive three diets: (1) hydrolysed protein (HP) diet; (2) 4.0% AN supplemented HP (HPA) food, (3) HPA diet fortified with 125 billion B. subtilis C-3102 spores/10 kg body weight (HPAB diet). Clinical outcome was assessed by Canine Inflammatory Bowel Disease Activity Index (CIBDAI), whereas gut microbiota compositional variations were investigated via 16S rRNA gene analysis, and faecal fermentation end-products by liquid chromatography. Higher abundances of the Ruminococcaceae and Rikenellaceae families were shown in HPA relative to CTR treatment, with Bacillus genus being differentially abundant on HPAB diet. Concentrations of acetate were higher (p < 0.05) in dogs fed HPA compared to CTR diet, and amounts of isovalerate and isobutyrate were greater (p < 0.05) in HPA compared to HP food. A tendency for higher amounts of faecal butyrate was found for the HPAB treatment (p = 0.06). Comprehensively, while displaying potentially positive effects on faecal fermentations, the tested substances failed to improve CIBDAI scores and microbial richness in CIE dogs.

scholarly journals ISOLATION OF CELLULOLYTIC BACTERIA FROM SOIL AND VALORIZATION OF DIFFERENT LIGNOCELLULOSIC WASTES FOR CELLULASE PRODUCTION BY SUBMERGED FERMENTATION

2021 ◽  
Vol 55 (7-8) ◽  
pp. 821-828
Author(s):  
MISBAH GHAZANFAR ◽  
MUHAMMAD IRFAN ◽  
MUHAMMAD NADEEM ◽  
HAFIZ ABDULLAH SHAKIR ◽  
MUHAMMAD KHAN ◽  
...  
Keyword(s):  
Bacillus Megaterium ◽  
Submerged Fermentation ◽  
Pseudomonas Stutzeri ◽  
Cellulase Production ◽  
Rrna Gene ◽  
Cellulolytic Bacteria ◽  
Bacterial Strains ◽  
Eucalyptus Leaves ◽  
Lignocellulosic Wastes ◽  
Bacillus Flexus

Cellulases are known to convert cellulose into monomeric or dimeric structures, hence playing an important role in bioethanol production, along with their applications in textile and paper industries. This study was directed towards the isolation and screening of cellulase producing bacteria from different soil samples on CMC (carboxymethyl cellulose) agar plates, followed by Gram’s iodine staining. Six strains showed clear zones of hydrolysis on CMC agar plates. Isolates were identified as Bacillus megaterium, Pseudomonas stutzeri, Bacillus aerius, Bacillus paralichniformis, Bacillus flexus, and Bacillus wiedmanni by 16S rRNA gene sequencing. These strains were cultivated by submerged fermentation for cellulase production using various lignocellulosic wastes, such as corn cob, rice husk, wheat straw, seed pods of Bombax ceiba and eucalyptus leaves. Results showed that Pseudomonas stutzeri is the best cellulase producer among these strains. It offered the highest cellulase activity of 170.9±4.1 (IU/mL/min) in media containing eucalyptus leaves after 24 h of incubation at 37 °C, followed by Bacillus paralichniformis, Bacillus wiedmanni, Bacillus flexus, Bacillus aerius and Bacillus megaterium. These bacterial strains and lignocellulosic wastes could be potentially used for industrial exploitation, particularly in biofuels and textiles.

scholarly journals Identification of SGS 1609 Cellulolytic Bacteria Isolated from Sargassum spec. and Characterization of The Cellulase Produced

2013 ◽  
Vol 8 (2) ◽  
pp. 57
Author(s):  
Yusro Nuri Fawzya ◽  
Stenny Putri ◽  
Nita Noriko ◽  
Gintung Patantis
Keyword(s):  
Cellulase Production ◽  
Rrna Gene ◽  
Cellulolytic Bacteria ◽  
16S Rrna Gene Analysis ◽  
Optimum Ph ◽  
Temperature Heat ◽  
Ultra Filtration ◽  
Na Ions ◽  
Optimum Production

Bacterial isolate from seaweed designated as SGS 1609 was previously found to be able to produce cellulase represented by formation of clear zone on solid medium containing carboxymethylcellulose (CMC). This research was conducted to identify the isolate and determine optimum production time as well as characterize the cellulase produced. The isolate was identified using  16s-rRNA gene analysis. Cellulase production was conducted by cultivating the isolate in the liquid medium containing CMC followed by centrifuging to get supernatant as the crude enzyme. The enzyme was then concentrated using ammonium sulfate precipitation and ultra filtration. The concentrated enzyme having higher activity produced from the concentration process was then characterized  to determine its optimum pH and temperature, heat stabilization, metal ions effect and substrate specificity. The result showed that the SGS 1609 isolate was identified as Serratia marcescens with 99%  similarity. The isolate produced cellulase optimally at 4 days incubation. Ultra filtration produced higher enzyme activity compared to NH4-sulfate precipitation. The enzyme concentrated by ultra filtration worked optimally at the  pH of 7, temperature of 50 oC, stable at the temperature of 60 oC for 240 minutes and was increased its activity by Ca2+ and Mg2+ ions. On the other hand, the enzyme was inhibited by Fe3+, Zn2+ and Na+ ions, but was not relatively affected by K+ and EDTA. The use of conventional agar producer waste  treated with 6% NaOH gave highest activity compared to other substrates.

scholarly journals Biodegradation of Di (2-Ethylhexyl) Phthalate by a novel Enterobacter spp. Strain YC-IL1 Isolated from Polluted Soil, Mila, Algeria

2020 ◽  
Vol 17 (20) ◽  
pp. 7501
Author(s):  
Imane Lamraoui ◽  
Adel Eltoukhy ◽  
Junhuan Wang ◽  
Messaouda Lamraoui ◽  
Amer Ahmed ◽  
...  
Keyword(s):  
Contaminated Soil ◽  
Acid Ester ◽  
Phthalic Acid ◽  
Degradation Pathway ◽  
Rrna Gene ◽  
16S Rrna Gene Analysis ◽  
Phthalic Acid Ester ◽  
Wide Range ◽  
Ring Structures ◽  
Ethylhexyl Phthalate

Di-(2-ethylhexyl) phthalate (DEHP) is one of the phthalic acid ester representatives and is mainly used as a plasticizer to endow polyvinyl chloride plastics with desirable physical properties. It is synthesized in massive amounts worldwide. Many studies have proved the adverse effects of DEHP on human health and wildlife. DEHP is labeled as an endocrine disruptor which causes human reproductive problems. Enterobacter spp. YC-IL1, a novel isolated strain from contaminated soil, was identified by 16S rRNA gene analysis and electronic microscope. It is capable of efficiently degrading DEHP (100%) and a wide range of phthalic acid ester PAEs, particularly those containing side chains with branches, or ring structures such as dutylbenzyl phthalate and dicyclohexyl phthalate, which are hard to degrade, with, respectively, 81.15% and 50.69% degradation after 7 days incubation. YC-IL1 is an acido-tolerant strain which remained in pH values lower than pH 5.0 with the optimum pH 7.0 and temperature 30 °C. The DEHP metabolites were detected using HPLC-QQQ and then the degradation pathway was tentatively proposed. Strain YC-IL1 showed high DEHP degradation rate in artificially contaminated soil with 86% removed in 6 days. These results indicate the application potential of YC-IL1 in bioremediation of PAE-polluted sites, even the acidic ones.

Isolation, development and identification of salt-tolerant bacterial consortium from crude-oil-contaminated soil for degradation of di-azo dye Reactive Blue 220

2015 ◽  
Vol 72 (2) ◽  
pp. 311-321 ◽  
Author(s):  
Vipul R. Patel ◽  
Nikhil Bhatt
Keyword(s):  
Oxygen Demand ◽  
Bacterial Consortium ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
16S Rrna Gene Analysis ◽  
Cod Reduction ◽  
Spectrometry Analysis ◽  
Wide Range

The objective of this study was development and characterization of a halophilic bacterial consortium for rapid decolorization and degradation of a wide range of dyes and their mixtures. The 16S rRNA gene analysis of developed halophilic consortium VN.1 showed that the bacterial consortium contained six bacterial strains, which were identified as Pseudomonas fluorescens HM480360, Enterobacter aerogenes HM480361, Shewanella sp. HM589853, Arthrobacter nicotianae HM480363, Bacillus beijingensis HM480362 and Pseudomonas aeruginosa JQ659549. Halophilic consortium VN.1 was able to decolorize up to 2,500 mg/L RB220 with &gt;85% chemical oxygen demand (COD) reduction under static condition at 30 °C and pH 8.0 in the presence of 7% NaCl. VN.1 also exhibited more than 85% COD reduction with &gt;25 mg/(L h) rate of decolorization in the case of different reactive dye mixtures. We propose the symmetric cleavage of RB220 using Fourier transform infrared, high-performance liquid chromatography (HPLC), nuclear magnetic resonance and gas chromatography-mass spectrometry analysis, and confirmed the formation of sodium-4-aminobenzenesulfonate, sodium-6-aminonepthalenesulfonate, and sodiumbenzene/nepthalenesulfonate. Toxicity studies confirm that the biodegraded products of RB220 effluent stimulate the growth of plants as well as the bacterial community responsible for soil fertility.

scholarly journals Assembly of Minicellulosomes on the Surface of Bacillus subtilis

2011 ◽  
Vol 77 (14) ◽  
pp. 4849-4858 ◽  
Author(s):  
Timothy D. Anderson ◽  
Scott A. Robson ◽  
Xiao Wen Jiang ◽  
G. Reza Malmirchegini ◽  
Henri-Pierre Fierobe ◽  
...  
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Cell Surface ◽  
Noncovalent Interactions ◽  
Genetic System ◽  
Cellulolytic Bacteria ◽  
Content Type ◽  
Multienzyme Complexes ◽  
Wide Range ◽  
A Cell

ABSTRACTTo cost-efficiently produce biofuels, new methods are needed to convert lignocellulosic biomass into fermentable sugars. One promising approach is to degrade biomass using cellulosomes, which are surface-displayed multicellulase-containing complexes present in cellulolyticClostridiumandRuminococcusspecies. In this study we created cellulolytic strains ofBacillus subtilisthat display one or more cellulase enzymes. Proteins containing the appropriate cell wall sorting signal are covalently anchored to the peptidoglycan by coexpressing them with theBacillus anthracissortase A (SrtA) transpeptidase. This approach was used to covalently attach the Cel8A endoglucanase fromClostridium thermocellumto the cell wall. In addition, a Cel8A-dockerin fusion protein was anchored on the surface ofB. subtilisvia noncovalent interactions with a cell wall-attached cohesin module. We also demonstrate that it is possible to assemble multienzyme complexes on the cell surface. A three-enzyme-containing minicellulosome was displayed on the cell surface; it consisted of a cell wall-attached scaffoldin protein noncovalently bound to three cellulase-dockerin fusion proteins that were produced inEscherichia coli.B. subtilishas a robust genetic system and is currently used in a wide range of industrial processes. Thus, grafting larger, more elaborate minicellulosomes onto the surface ofB. subtilismay yield cellulolytic bacteria with increased potency that can be used to degrade biomass.

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