scholarly journals Production of Novel Polygalacturonase from Bacillus paralicheniformis CBS32 and Application to Depolymerization of Ramie Fiber

Polymers ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 1525 ◽  
Author(s):  
Md. Saifur Rahman ◽  
Yoon Seok Choi ◽  
Young Kyun Kim ◽  
Chulhwan Park ◽  
Jin Cheol Yoo
Keyword(s):  
Industrial Applications ◽  
Fermented Food ◽  
Ramie Fiber ◽  
16S Rrna Sequence ◽  
Application Study ◽  
Glycosidic Bonds ◽  
Bacillus Paralicheniformis ◽  
Sds Page ◽  
16S Rrna Sequence Analysis ◽  
Terminal Amino

Polygalacturonase (EC. 3.2.1.15) is an enzyme that hydrolyzes the alpha-1,4 glycosidic bonds between galacturonic acid. In this study, an alkaline polygalacturonase producer Bacillus paralicheniformis CBS32 was isolated from kimchi (conventional Korean fermented food). The 16S rRNA sequence analysis of the isolated strain revealed that it was 99.92% identical to B. paralicheniformis KJ 16LBMN01000156. The polygalacturonase from B. paralicheniformis CBS32 was named PN32, and the purified PN32 showed a 16.8% yield and a 33-fold purity compared to the crude broth. The molecular mass, 110 kDa, was determined by SDS-PAGE, and the active band was confirmed by zymography analysis. The N-terminal amino acid sequence residues of PN32 were determined to be Gly–Val–Lys–Glu–Val–X–Gln–Thr–Phe. In the sequence comparison, PN32 was suggested as a novel polygalacturonase, since the sequence was not matched with the previous reports. In an application study, enzymatic depolymerization of ramie was performed for fiber degumming, and the result showed that the PN32 had a 28% higher depolymerization compared to the commercial pectinase. Overall, based on the results, PN32 has high potential for industrial applications.

Purification, Properties and Mass Spectrometry Analysis of Two Novel Thermotolerant Endoglucanases from Bacillus akibai I-2

2011 ◽  
Vol 393-395 ◽  
pp. 911-915
Author(s):  
Sen Lin Liu ◽  
Miao Xing
Keyword(s):  
Mass Spectrometry ◽  
Soda Lakes ◽  
Maximum Activity ◽  
Mass Spectrometry Analysis ◽  
16S Rrna Sequence ◽  
Spectrometry Analysis ◽  
16S Rrna Sequence Analysis ◽  
Bacterium Strain ◽  
Terminal Amino ◽  
Alkaliphilic Bacterium

The alkaliphilic bacterium strain Ⅰ-2, which was isolated from soda lakes, was identified as Bacillus akibai by 16S rRNA sequence analysis and suggested to be a new subspecies of genus Bacillus. Two novel thermotolerant alkaline endoglucanases Ⅰ-2-A and Ⅰ-2-B were produced by this alkaliphilic strain. The purified Ⅰ-2-A and Ⅰ-2-B had molecular mass of approximately 60 and 90 kDa, respectively. The optimum pH of Ⅰ-2-A was about 9.0, while that of Ⅰ-2-B was about 8.0. Both enzymes exhibited maximum activity at around 50 °C and were stable up to 50 °C.The two enzymes were resistant to most metal ions and reagents examined. Mass spectrometry analysis indicated that Ⅰ-2-A was probably different from the endoglucanases reported. Ⅰ-2-B showed homology with those of family A5 endoglucanases but low similarity was found in C-terminal amino acid sequence.

scholarly journals EXPRESSION OF CELLULOSE-DEGRADING ENDOGLUCANASE FROM BACILLUS SUBTILIS USING PTOLT EXPRESSION SYSTEM IN ESCHERICHIA COLI

2021 ◽  
Vol 55 (5-6) ◽  
pp. 619-627
Author(s):  
HÜLYA KUDUĞ CEYLAN ◽  
YAKUP ULUSU ◽  
SEMA BILGIN ◽  
İSA GÖKÇE
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Expression System ◽  
Industrial Applications ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
E Coli ◽  
Glycosidic Bonds ◽  
Sds Page ◽  
Dna Fragment

Endoglucanases randomly hydrolyse the cellulose chains by acting upon internal β-1,4-D-glycosidic bonds and are used extensively in industrial applications. In this study, bacterial endoglucanase gene yhfE was obtained by PCR, using primers based on genomic sequences of Bacillus subtilis strains. 1041 bp DNA fragment of yhfE was cloned into Escherichia coli DH5α through the use of pTolT expression plasmid. PCR, restriction enzyme analysis and DNA sequencing were performed in order to confirm the cloning. E. coli BL21-AI cells expressed the yhfE after induction at 0.04% of arabinose concentration for 4 h. The expected 38.7 kDa size yhfE protein after digestion with thrombin of the His-tagged fusion protein (yhfE-TolAIII) was visualized by SDS-PAGE. The yhfE-TolAIII production yield was approximately 82 mg/L. The recombinant yhfE was characterized by MALDI-TOF mass spectrometry and CD analysis.

scholarly journals Identification of a MarineBacillusStrain C5 and Parathion-Methyl Degradation Characteristics of the Extracellular Esterase B1

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Jianhua Hao ◽  
Junzhong Liu ◽  
Mi Sun
Keyword(s):  
Ph Value ◽  
Degradation Products ◽  
16S Rrna Sequence ◽  
Native Page ◽  
Sds Page ◽  
16S Rrna Sequence Analysis ◽  
New Type ◽  
Effects Of Temperature ◽  
Marine Sludge ◽  
Degradation Effect

A bacterial strain C5 that can produce new type of marine esterase was isolated and screened from marine sludge. According to 16S rRNA sequence analysis and physiological and biochemical experiments, the strain was identified asBacillus subtilis. A single isozyme with a molecular weight of 86 kDa was observed by SDS-PAGE and native-PAGE. On this basis, the mechanism of esterase B1 secreted by strain C5 degrading parathion-methyl was explored, and the effects of temperature and pH on the degradation rate were investigated. From the results,p-nitrophenol was one of the degradation products of B1 degrading parathion-methyl, and the best degradation effect could be achieved at the temperature of 40°C and the neutral pH value.

scholarly journals Identification and Characterization of a Newly Isolated Chitinase-Producing Strain Bacillus licheniformis SSCL-10 for Chitin Degradation

Archaea ◽  
2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Abirami Sasi ◽  
Nagarajan Duraipandiyan ◽  
Kannan Marikani ◽  
Sugapriya Dhanasekaran ◽  
Noura Al-Dayan ◽  
...  
Keyword(s):  
Bacillus Licheniformis ◽  
Specific Activity ◽  
Wild Strain ◽  
Chitinase Activity ◽  
Chemical Mutagenesis ◽  
16S Rrna Sequence ◽  
Sds Page ◽  
Potential Applications ◽  
16S Rrna Sequence Analysis ◽  
Ethylmethane Sulfonate

Chitinases or chitinolytic enzymes have different applications in the field of medicine, agriculture, and industry. The present study is aimed at developing an effective hyperchitinase-producing mutant strain of novel Bacillus licheniformis. A simple and rapid methodology was used for screening potential chitinolytic microbiota by chemical mutagenesis with ethylmethane sulfonate and irradiation with UV. There were 16 mutant strains exhibiting chitinase activity. Out of the chitinase-producing strains, the strain with maximum chitinase activity was selected, the protein was partially purified by SDS-PAGE, and the strain was identified as Bacillus licheniformis (SSCL-10) with the highest specific activity of 3.4 U/mL. The induced mutation model has been successfully implemented in the mutant EMS-13 (20.2 U/mL) that produces 5-6-fold higher yield of chitinase, whereas the mutant UV-11 (13.3 U/mL) has 3-4-fold greater chitinase activity compared to the wild strain. The partially purified chitinase has a molecular weight of 66 kDa. The wild strain (SSCL-10) was identified as Bacillus licheniformis using 16S rRNA sequence analysis. This study explores the potential applications of hyperchitinase-producing bacteria in recycling and processing chitin wastes from crustaceans and shrimp, thereby adding value to the crustacean industry.

Characterization of Glycoprotein Fraction from Carp Pituitaries Isolated Using Concanavalin A as the Affinity Ligand

1998 ◽  
Vol 63 (3) ◽  
pp. 434-440 ◽  
Author(s):  
Irena Hulová ◽  
Jana Barthová ◽  
Helena Ryšlavá ◽  
Václav Kašička
Keyword(s):  
Concanavalin A ◽  
Reversed Phase ◽  
Amino Acid Sequences ◽  
Gel Chromatography ◽  
Affinity Ligand ◽  
Sds Page ◽  
Terminal Amino ◽  
Α And Β Subunits

Glycoproteins that have affinity to Concanavalin A were isolated from the acetone-dried pituitaries of common carp (Cyprinus carpio L.). Two fractions of glycoproteins were separated using gel chromatography on Superdex 75HR. The fraction with lower molecular weight (30 000) corresponding to the carp gonadotropin cGtH II was composed of two subunits as determined using SDS-PAGE. This protein fraction was further divided into four components using reversed-phase HPLC. Two fractions were pure α and β subunits of cGtH II as follows from immunodetection and from determination of N-terminal amino acid sequences. The other two were a mixture of α and β subunits as was also revealed by N-terminal analysis. Capillary electrophoresis was also used for characterization of isolated glycoproteins.

scholarly journals Pseudomonas eucalypticola sp. nov., a producer of antifungal agents isolated from Eucalyptus dunnii leaves

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yujing Liu ◽  
Zhang Song ◽  
Hualong Zeng ◽  
Meng Lu ◽  
Weiyao Zhu ◽  
...  
Keyword(s):  
Dna Hybridization ◽  
Antifungal Agents ◽  
Novel Species ◽  
Phytopathogenic Fungi ◽  
Strictly Aerobic ◽  
Rrna Sequence ◽  
16S Rrna Sequence ◽  
Eucalyptus Dunnii ◽  
16S Rrna Sequence Analysis

AbstractPseudomonas are ubiquitously occurring microorganisms and are known for their ability to produce antimicrobials. An endophytic bacterial strain NP-1 T, isolated from Eucalyptus dunnii leaves, exhibits antifungal properties against five tested phytopathogenic fungi. The strain is a Gram-negative rod-shaped bacterium containing a single polar flagellum. It is strictly aerobic, grows at 4–37 °C, 2–5% NaCl, and pH 3–7. The 16S rRNA sequence analysis showed that NP-1 T belongs to the Pseudomonas genus. Phylogenetic analysis based on four concatenated partial genes (16S rDNA, gyrB, rpoB and rpoD) and the phylogenomic tree indicated that NP-1 T belongs to Pseudomonas fluorescens lineage but is distinct from any known Pseudomonas species. The G + C mol % of NP-1 T genome is 63.96, and the differences between NP-1 T and related species are larger than 1. The digital DNA-DNA hybridization and tetranucleotide signatures are 23.8 and 0.97, which clearly separates strain NP-1 T from its closest neighbours, Pseudomonas coleopterorum and Pseudomonas rhizosphaerae. Its phenotypic and chemotaxonomic features confirmed its differentiation from related taxa. The results from this polyphasic approach support the classification of NP-1 T as a novel species of Pseudomonas, and the name of Pseudomonas eucalypticola is thus proposed for this strain, whose type is NP-1 T (= CCTCC M2018494T = JCM 33572 T).

scholarly journals Autoantibodies against triosephosphate isomerase. A possible clue to pathogenesis of hemolytic anemia in infectious mononucleosis.

1990 ◽  
Vol 171 (2) ◽  
pp. 565-570 ◽  
Author(s):  
K Ritter ◽  
H Brestrich ◽  
B Nellen ◽  
H Kratzin ◽  
H Eiffert ◽  
...  
Keyword(s):  
Infectious Mononucleosis ◽  
Triosephosphate Isomerase ◽  
Clinical Symptoms ◽  
Glycolytic Enzyme ◽  
Amino Acid Sequences ◽  
Cellular Proteins ◽  
51Cr Release ◽  
Terminal Amino Acid ◽  
Sds Page ◽  
Terminal Amino

In sera from patients with acute EBV, infection and the clinical symptoms of infectious mononucleosis antibodies of the Ig class M were found that are directed against two cellular proteins. The molecular mass of these proteins was determined to be 29 (p29) and 26 kD (p26), respectively, in SDS-PAGE. P29 was identified as part of the glycolytic enzyme triosephosphate isomerase (TPI) by comparison of the NH2-terminal amino acid sequences. A purified antibody against TPI induces a 51Cr release from human erythrocytes. Possibly, anti-TPI causes hemolysis, which is an infrequent but serious symptom of infectious mononucleosis.

Isolation and characterization of phosphate solubilizing bacteria from the rhizospheric soil of Ponthenpuzha forest, Pathanamthitta (District), Kerala

2021 ◽  
Vol 16 (8) ◽  
pp. 110-117
Author(s):  
Kannan Abhirami ◽  
K. Jayakumar
Keyword(s):  
Bacterial Isolate ◽  
Phosphate Solubilizing Bacteria ◽  
Cymbopogon Citratus ◽  
16S Rrna Sequence ◽  
Isolation And Characterization ◽  
16S Rrna Sequence Analysis ◽  
Phosphate Solubilizing ◽  
Level Identification

Phosphorous is considered as a major parameter for crop yield. Its availability to plant is independent of its abundance. For the plants to utilize phosphorous, it is to be converted to absorbable form. Here, the part rendered by phosphate solubilizing bacteria is significant for it plays a crucial role in the formation of plant usable phosphate from organic forms. In the present work, an effort had been made to isolate and identify phosphate solubilising bacterial isolate from the rhizhospheric soils of various plants in Ponthenpuzha forest. One of the isolate from Cymbopogon citrates responded positively to Pikovskaya’s medium by producing a halo zone during in vitro culture. Colony features and 16S rRNA sequence analysis identified the isolate as Burkholderia sps. We have reported the presence of genus Burkholderia in the rhizospheric zone of Cymbopogon citratus. Further studies are warranted for species level identification of the isolate.

scholarly journals Acquisition of an Agrobacterium Ri Plasmid and Pathogenicity by Other α-Proteobacteria in Cucumber and Tomato Crops Affected by Root Mat

2004 ◽  
Vol 70 (5) ◽  
pp. 2779-2785 ◽  
Author(s):  
S. A. Weller ◽  
D. E. Stead ◽  
J. P. W. Young
Keyword(s):  
Pathogenicity Test ◽  
Agrobacterium Radiobacter ◽  
Rrna Sequence ◽  
16S Rrna Sequence ◽  
Ri Plasmid ◽  
Fatty Acid Profiling ◽  
Transformation Rates ◽  
16S Rrna Sequence Analysis ◽  
Average Transformation

ABSTRACT Root mat of cucumbers and tomatoes has previously been shown to be caused by Agrobacterium radiobacter strains harboring a root-inducing Ri plasmid (pRi). Nine other pRi-harboring α-Proteobacteria have subsequently been isolated from root mat-infected crops. Fatty acid profiling and partial 16S rRNA sequence analysis identified three of these strains as being in the genus Ochrobactrum, five as being in the genus Rhizobium, and one as being in the genus Sinorhizobium. An in vitro pathogenicity test involving inoculation of cucumber cotyledons was developed. All pRi-harboring α-Proteobacteria induced typical root mat symptoms from the cotyledons. Average transformation rates for rhizogenic Ochrobactrum (46%) and Rhizobium (44%) strains were lower than those observed for rhizogenic A. radiobacter strains (64%). However, individual strains from these three genera all had transformation rates comparable to those observed from cotyledons inoculated with a rhizogenic Sinorhizobium strain (75%).

The effects of Pantoea sp. strain Y4-4 on alfalfa in the remediation of heavy-metal-contaminated soil, and auxiliary impacts of plant residues on the remediation of saline–alkali soils

2017 ◽  
Vol 63 (4) ◽  
pp. 278-286 ◽  
Author(s):  
Shuhuan Li ◽  
Jie Wang ◽  
Nanxiong Gao ◽  
Lizhu Liu ◽  
Yahua Chen
Keyword(s):  
Salt Tolerance ◽  
Contaminated Soil ◽  
Dry Mass ◽  
Plant Residue ◽  
Plant Residues ◽  
16S Rrna Sequence ◽  
Wheat Seedlings ◽  
Safe Strategy ◽  
Plant Growth Promoting ◽  
16S Rrna Sequence Analysis

The plant-growth-promoting rhizobacterium (PGPR) Y4-4 was isolated from plant rhizosphere soil and identified as Pantoea sp. by 16S rRNA sequence analysis. The effects of strain Y4-4 on alfalfa grown in heavy-metals-contaminated soil was investigated using a pot experiment. In a Cu-rich environment, the shoot dry mass and total dry mass of plants inoculated with strain Y4-4 increased by 22.6% and 21%, and Cu accumulation increased by 15%. In a Pb–Zn-rich environment, the shoot dry mass and total dry mass of plants inoculated with strain Y4-4 increased by 23.4% and 22%, and Zn accumulation increased by 30.3%. In addition, the salt tolerance and biomass of wheat seedlings could be improved by applying strain Y4-4 mixed with plant residue as a result of the Cu-rich plant residues providing copper nutrition to wheat. This study offers an efficient PGPR with strong salt tolerance and a safe strategy for the post-treatment of plant residue.

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