Journal: Microbial Cell Factories

Article Title: Microbial lipases and their industrial applications: a comprehensive review

doi: 10.1186/s12934-020-01428-8

Figure Lengend Snippet: Microbial source of Lipase and their industrial application

Article Snippet: The greatest extensively used in biodiesel production are free lipases between the commercially available lipases from P. fluorescens (Lipase AsK, Amano), B. cepacia (Lipase PS, Amano), and T. lanuginosus (Lipase LA201 and Lipopan 50BG, Novozymes), and immobilized lipases from T. lanuginosus (Lipozyme TL IM, Novozymes) and R. miehei (Lipozyme RM IM, Novozymes) [ , ].

Techniques: Synthesized, Produced, Functional Assay, Staining

Journal: Microbial Cell Factories

Article Title: Microbial lipases and their industrial applications: a comprehensive review

doi: 10.1186/s12934-020-01428-8

Figure Lengend Snippet: Electrochemical assays at lipase based Biosensor

Article Snippet: The greatest extensively used in biodiesel production are free lipases between the commercially available lipases from P. fluorescens (Lipase AsK, Amano), B. cepacia (Lipase PS, Amano), and T. lanuginosus (Lipase LA201 and Lipopan 50BG, Novozymes), and immobilized lipases from T. lanuginosus (Lipozyme TL IM, Novozymes) and R. miehei (Lipozyme RM IM, Novozymes) [ , ].

Techniques: Transformation Assay

Journal:

Article Title: Identification of Quorum-Sensing-Regulated Genes of Burkholderia cepacia

doi: 10.1128/JB.185.21.6456-6462.2003

Figure Lengend Snippet: (A) Schematic diagram (not to scale) of the cepR-based cloning plasmid used to identify QSC genes. Plasmid pSCR2 contains the B. cepacia ATCC 25146 cepR gene cloned in the broad-host-range plasmid pQF50 (12). The genetic components are as follows: cepR, the gene encoding the transcriptional activator CepR; lacZ, a promoterless β-galactosidase gene; T0, a transcriptional trpA terminator; bla, an ampicillin resistance marker. The indicated restriction sites in the polylinker (N, NcoI; H, HincII; B, BamHI; S, SmaI; Hi, HindIII) were used to clone DNA fragments obtained from genomic DNA of B. cepacia ATCC 25416 digested with compatible-end restriction enzymes. Positive clones on selective plates in the presence of C8-HSL were identified as blue colonies (see text for details). (B) E. coli clones were cross-streaked in close proximity to B. cepacia ATCC 25416 in LB plates containing 100 μg of ampicillin/ml and X-Gal and were incubated overnight at 30°C. The blue coloration developed only in proximity to the B. cepacia strain. pSCR2C and pSCon were the positive and negative controls, respectively. (C) E. coli clones were cross-streaked in close proximity to B. cepacia 25416-I on plates containing 100 μg of ampicillin/ml and X-Gal and were incubated overnight at 30°C. No blue coloration was observed either with clones or with the pSCR2C positive control, thus confirming the strict C8-HSL-dependent expression of the reporter β-galactosidase gene.

Article Snippet: Putative ORFs regulated by quorum sensing in Burkholderia cepacia DNA ligations between pSCR2 and the genomic restriction DNA fragments from B. cepacia (obtained using the restriction enzymes Nru I, Hae III, Hin cII, Alu I, and Sau 3AI independently) were transformed in E. coli DH5α and plated on selective medium containing 100 μg of ampicillin (Sigma-Aldrich)/ml, 20 μg of X-Gal (Sigma-Aldrich)/ml, and 100 nM C 8 -HSL.

Techniques: Clone Assay, Plasmid Preparation, Marker, Incubation, Positive Control, Expressing

Journal:

Article Title: Identification of Quorum-Sensing-Regulated Genes of Burkholderia cepacia

doi: 10.1128/JB.185.21.6456-6462.2003

Figure Lengend Snippet: Putative ORFs regulated by quorum sensing in Burkholderia cepacia

Article Snippet: Putative ORFs regulated by quorum sensing in Burkholderia cepacia DNA ligations between pSCR2 and the genomic restriction DNA fragments from B. cepacia (obtained using the restriction enzymes Nru I, Hae III, Hin cII, Alu I, and Sau 3AI independently) were transformed in E. coli DH5α and plated on selective medium containing 100 μg of ampicillin (Sigma-Aldrich)/ml, 20 μg of X-Gal (Sigma-Aldrich)/ml, and 100 nM C 8 -HSL.

Techniques: Activity Assay, Binding Assay, RNA Binding Assay, Clone Assay

Journal:

Article Title: Identification of Quorum-Sensing-Regulated Genes of Burkholderia cepacia

doi: 10.1128/JB.185.21.6456-6462.2003

Figure Lengend Snippet: Characterization of identified clones containing putative CepR-C8-HSL gene promoters. The DNA sequence of each identified clone was compared (using the BLASTN program [http://www.sanger.ac.uk/Projects/B_cepacia]) to sequences in the genome of B. cepacia J2315. For those clones that showed a high degree of identity at the DNA level (see text), a region of the genome surrounding 2 kbp of each sequence was subsequently annotated. In the majority of the cases, it was possible to identify a homologue of a gene coding for a protein of known function. In such cases, the names of the respective genes are indicated inside of the putative ORFs (represented as thick gray arrows). A black arrow represents the size, position, and orientation of the identified insert in each clone (in scale with each clone). Within some genetic regions shown, hypothetical proteins of unknown function were found (here designated ORF). Such was the case with P67 (ORF represents what was probably a lysR family transcriptional regulator from R. solanacearum), P80 (ORF represents a 4-hydroxybenzoate octaprenyl transferase from Xanthomonas axonopodis), P114 (ORF1 represents a hypothetical protein from B. fungorum; ORF2 represents a probable transcriptional regulator, PA1627, from P. aeruginosa), P111 (ORF2 represents a short chain dehydrogenase-reductase from Pseudomonas putida, KT2440; ORF1 represents a hypothetical protein, PA1829, from P. aeruginosa), P79 (ORF represents a protein of unknown function), P103 (ORF represents a protein of unknown function), P56 (ORF1 represents a probable efflux membrane protein from Mesorhizobium loti; ORF2 represents a probable nucleoside triphosphate pyrophosphohydrolase from Agrobacterium tumefaciens), P57 (ORF1 represents a probable porin transmembrane protein from R. solanacearum; ORF2 represents a putative lipoprotein from R. solanacearum), P105 (ORF represents a hypothetical protein from R. solanacearum), and P135 (ORF1 represents a human RNA binding protein; ORF2 represents a putative small heat shock protein from R . solanacearum). See text for details of all other ORFs mentioned.

Article Snippet: Putative ORFs regulated by quorum sensing in Burkholderia cepacia DNA ligations between pSCR2 and the genomic restriction DNA fragments from B. cepacia (obtained using the restriction enzymes Nru I, Hae III, Hin cII, Alu I, and Sau 3AI independently) were transformed in E. coli DH5α and plated on selective medium containing 100 μg of ampicillin (Sigma-Aldrich)/ml, 20 μg of X-Gal (Sigma-Aldrich)/ml, and 100 nM C 8 -HSL.

Techniques: Clone Assay, Sequencing, RNA Binding Assay

Journal:

Article Title: Identification of Quorum-Sensing-Regulated Genes of Burkholderia cepacia

doi: 10.1128/JB.185.21.6456-6462.2003

Figure Lengend Snippet: Primer extension analysis of three identified QSC promoters. Primer extension mapping of the mRNA 5′ end of clones P67 (A), P53 (B), and P110 (C) is shown. Total RNA was isolated from B. cepacia ATCC 25416 (WT) and the cepI mutant derivative (25416-I). Primer extensions were performed using 15 μg of RNA, and extension products were run adjacent to the sequencing ladder generated with the same primer used for reverse transcription. The approximate position of the mRNA 5′ end is depicted for each clone and indicated by an arrow.

Article Snippet: Putative ORFs regulated by quorum sensing in Burkholderia cepacia DNA ligations between pSCR2 and the genomic restriction DNA fragments from B. cepacia (obtained using the restriction enzymes Nru I, Hae III, Hin cII, Alu I, and Sau 3AI independently) were transformed in E. coli DH5α and plated on selective medium containing 100 μg of ampicillin (Sigma-Aldrich)/ml, 20 μg of X-Gal (Sigma-Aldrich)/ml, and 100 nM C 8 -HSL.

Techniques: Clone Assay, Isolation, Mutagenesis, Sequencing, Generated