scholarly journals Characterization of the bacteriocin-producing strain Lactobacillus paracasei subsp. paracasei BGUB9

2010 ◽  
Vol 62 (4) ◽  
pp. 889-899 ◽  
Author(s):  
Maja Tolinacki ◽  
M. Kojic ◽  
Jelena Lozo ◽  
Amarela Terzic-Vidojevic ◽  
L. Topisirovic ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Biochemical Characterization ◽  
Proteolytic Enzymes ◽  
Lactobacillus Paracasei ◽  
Plasmid Curing ◽  
Closely Related Species ◽  
Antimicrobial Spectrum ◽  
Activity Test ◽  
Ph Range

The strain Lactobacillus paracasei subsp. paracasei BGUB9 that was isolated from traditionally homemade hard cheese produces bacteriocin designated as BacUB9, with an approximate molecular mass of 4 kDa. Biochemical characterization and the antimicrobial activity test of BacUB9 were performed. The onset of BacUB9 biosynthesis was observed at the end of an exponential phase of growth. Bacteriocin UB9 retained the antimicrobial activity within the pH range from 1 to 10 and after treatment at 100oC for 30 min. The bacteriocin is susceptible to the activity of proteolytic enzymes. Bacteriocin BacUB9 has a very narrow antimicrobial spectrum, limited to several strains that belong to closely related species. The effect of BGUB9 on the growth of the strain Lactobacillus paracasei subsp. paracasei BGHN14 in a mixed culture was monitored. The mode of action of BacUB9 on the strain BGHN14 was identified as bacteriostatic. Plasmid curing results indicated that a plasmid, designated as pUB9, seemed to be responsible for both bacteriocin BacUB9 production and host immunity.

Identification and Characterization of Two Bacteriocin-Producing Bacteria Isolated from Garlic and Ginger Root†

1999 ◽  
Vol 62 (8) ◽  
pp. 899-904 ◽  
Author(s):  
M. E. JANES ◽  
R. NANNAPANENI ◽  
M. G. JOHNSON
Keyword(s):  
Leuconostoc Mesenteroides ◽  
Proteolytic Enzymes ◽  
Bacterial Strains ◽  
16S Rrna Sequence ◽  
Antimicrobial Spectrum ◽  
Physiochemical Properties ◽  
Overlay Method ◽  
Ph Range ◽  
Identification And Characterization

Two bacteriocin-producing bacterial strains were isolated from garlic and ginger root by the agar overlay method. The bacteria were identified by 16S rRNA sequence analyses and fermentation patterns as Leuconostoc mesenteroides (garlic isolate) and Lactococcus lactis (ginger isolate). The bacteriocins were assigned the names leucocin BC2 and lactocin GI3, respectively. Physiochemical properties and antimicrobial spectra of the bacteriocins were determined by the spot-on-lawn method. Both bacteriocins were inhibited by proteolytic enzymes. Leucocin BC2 exhibited a narrow antimicrobial spectrum, inhibiting only Bacillus, Enterococcus, and Listeria species. Lactocin GI3 had a broader spectrum, inhibiting Bacillus, Clostridium, Listeria, Enterococcus, Leuconostoc, Pediococcus, and Staphylococcus species. Both bacteriocins remained active when heated at 90°C for 15 min or 120°C for 20 min. Leucocin BC2 assayed at 37°C showed an inhibitory activity of 1,600 AU/ml, whereas at 8°C the activity was 12,800 AU/ml. Conversely, lactocin GI3 activity was the same at both assay temperatures. Both bacteriocins remained active over a pH range of 2.0 to 9.0 and in various organic solvents. The activity of leucocin BC2 was increased when treated with 0.5% acetic acid and 0.5% lactic acid, whereas lactocin GI3 activity was decreased with either acid. The molecular mass values were 3.7 kDa for leucocin BC2 and 3.9 kDa for lactocin GI3. These results show that the inhibitory substances produced by the bacteria isolated from garlic and ginger are bacteriocins that appear to be different in some characteristics from previously reported bacteriocins.

Characterization and Antimicrobial Activity of Bacteriocin 217 Produced by Natural Isolate Lactobacillus paracasei subsp. paracasei BGBUK2-16

2004 ◽  
Vol 67 (12) ◽  
pp. 2727-2734 ◽  
Author(s):  
JELENA LOZO ◽  
MAJA VUKASINOVIC ◽  
IVANA STRAHINIC ◽  
LJUBISA TOPISIROVIC
Keyword(s):  
Antimicrobial Activity ◽  
Pathogenic Bacteria ◽  
Biochemical Characterization ◽  
Proteolytic Enzymes ◽  
Inhibitory Effect ◽  
Lactobacillus Paracasei ◽  
Logarithmic Phase ◽  
Short Period ◽  
Fermented Dairy Products ◽  
Fermented Dairy

The strain Lactobacillus paracasei subsp. paracasei BGBUK2-16, which was isolated from traditionally homemade white-pickled cheese, produces bacteriocin 217 (Bac217; ∼7 kDa). The onset of Bac217 biosynthesis was observed in the logarithmic phase of growth, and the production plateau was reached after 9 or 12 h of incubation at 37 and 30°C, respectively, when culture entered the early stationary phase. Biochemical characterization showed that Bac217 retained antimicrobial activity within the range of pH 3 to 12 or after treatment at 100°C for 15 min. Bac217 antimicrobial activity also remained unchanged after storage at 4°C for 6 months or −20°C for up to 12 months. However, Bac217 activity was completely lost after treatment with different proteolytic enzymes. BGBUK2-16 contains only one plasmid about 80 kb in size. Plasmid curing indicated that genes coding for Bac217 synthesis and immunity seem to be located on this plasmid. Bac217 exhibited antimicrobial activity against some pathogenic bacteria, such as Staphylococcus aureus and Bacillus cereus. Interestingly, Bac217 showed activity against Salmonella sp. and Pseudomonas aeruginosa ATCC27853. The inhibitory effect of BGBUK2-16 on the growth of S. aureus in mixed culture was observed. S. aureus treatment with Bac217 led to a considerable decrease (CFU/ml) within a short period of time. The mode of Bac217 action on S. aureus was identified as bactericidal. It should be noted that the strain BGBUK2-16 was shown to be resistant to bacteriocin nisin, which is otherwise widely used as a food additive for fermented dairy products.

Biochemical characterization of a glucoamylase from Saccharomycopsis fibuligera R64

Biologia ◽  
2011 ◽  
Vol 66 (1) ◽  
Author(s):  
Dessy Natalia ◽  
Keni Vidilaseris ◽  
Pasjan Satrimafitrah ◽  
Wangsa Ismaya ◽  
Purkan ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Amino Acid ◽  
Amino Acid Sequence Identity ◽  
Biochemical Characterization ◽  
Saccharomycopsis Fibuligera ◽  
Sequence Identity ◽  
Ic50 Value ◽  
High Amino Acid ◽  
Ph Range

AbstractGlucoamylase from the yeast Saccharomycopsis fibuligera R64 (GLL1) has successfully been purified and characterized. The molecular mass of the enzyme was 56,583 Da as determined by mass spectrometry. The purified enzyme demonstrated optimum activity in the pH range of 5.6–6.4 and at 50°C. The activity of the enzyme was inhibited by acarbose with the IC50 value of 5 μM. GLL1 shares high amino acid sequence identity with GLU1 and GLA1, which are Saccharomycopsis fibuligera glucoamylases from the strains HUT7212 and KZ, respectively. The properties of GLL1, however, resemble that of GLU1. The elucidation of the primary structure of GLL1 contributes to the explanation of this finding.

scholarly journals Biochemical Characterization of a New β-Agarase from Cellulophaga Algicola

2019 ◽  
Vol 20 (9) ◽  
pp. 2143 ◽  
Author(s):  
Han ◽  
Zhang ◽  
Yang
Keyword(s):  
Structural Model ◽  
Biochemical Characterization ◽  
Maximum Activity ◽  
Optimal Temperature ◽  
Salt Tolerant ◽  
Wide Ph Range ◽  
Specific Activities ◽  
Ph Range ◽  
Layer Chromatography

Cellulophaga algicola DSM 14237, isolated from the Eastern Antarctic coastal zone, was found to be able to hydrolyze several types of polysaccharide materials. In this study, a predicted β-agarase (CaAga1) from C. algicola was heterologously expressed in Escherichia coli. The purified recombinant CaAga1 showed specific activities of 29.39, 20.20, 14.12, and 8.99 U/mg toward agarose, pure agar, and crude agars from Gracilaria lemaneiformis and Porphyra haitanensis, respectively. CaAga1 exhibited an optimal temperature and pH of 40 oC and 7, respectively. CaAga1 was stable over a wide pH range from 4 to 11. The recombinant enzyme showed an unusual thermostability, that is, it was stable at temperature below or equal to 40oC and around 70 oC, but was thermolabile at about 50 oC. With the agarose as the substrate, the Km and Vmax values for CaAga1 were 1.19 mg/mL and 36.21 U/mg, respectively. The reducing reagent (dithiothreitol) enhanced the activity of CaAga1 by more than one fold. In addition, CaAga1 was salt-tolerant given that it retained approximately 70% of the maximum activity in the presence of 2 M NaCl. The thin layer chromatography results indicated that CaAga1 is an endo-type β-agarase and efficiently hydrolyzed agarose into neoagarotetraose (NA4) and neoagarohexaose (NA6). A structural model of CaAga1 in complex with neoagarooctaose (NA8) was built by homology modeling and explained the hydrolysis pattern of CaAga1.

scholarly journals An antimicrobial Staphylococcus sciuri with broad temperature and salt spectrum isolated from the surface of the African social spider, Stegodyphus dumicola

2021 ◽  
Author(s):  
Seven Nazipi ◽  
Sofie Gerdes Vangkilde-Pedersen ◽  
Mette Marie Busck ◽  
Dorthe Kirstine Lund ◽  
Ian P.G. Marshall ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Related Species ◽  
Phenotypic Characterization ◽  
Phylogenomic Analysis ◽  
Closely Related Species ◽  
Social Spider ◽  
Social Spiders ◽  
Mutualistic Symbiosis ◽  
The Social

Abstract Some social arthropods engage in mutualistic symbiosis with antimicrobial compound-producing microorganisms that provide protection against pathogens. Social spiders live in communal nests and contain specific endosymbionts with unknown function. Bacteria are also found on the spiders' surface, including prevalent staphylococci, which may have protective potential. Here we present the genomic and phenotypic characterization of strain i1, isolated from the surface of the social spider Stegodyphus dumicola. Phylogenomic analysis identified i1 as novel strain of Staphylococcus sciuri within subgroup 2 of three newly defined genomic subgroups. Further phenotypic investigations showed that S. sciuri i1 is an extremophile that can grow at a broad range of temperatures (4°C-45°C), high salt concentrations (up to 27%), and has antimicrobial activity against closely related species. We identified a lactococcin 972-like bacteriocin gene cluster, likely responsible for the antimicrobial activity, and found it conserved in two of the three subgroups of S. sciuri. These features indicate that S. sciuri i1, though not a specific symbiont, is well-adapted to survive on the surface of social spiders and may gain a competitive advantage by inhibiting closely related species.

scholarly journals Purification and Characterization of a Novel Class IIa Bacteriocin, Piscicocin CS526, from Surimi-Associated Carnobacterium piscicola CS526

2005 ◽  
Vol 71 (1) ◽  
pp. 554-557 ◽  
Author(s):  
Koji Yamazaki ◽  
Minako Suzuki ◽  
Yuji Kawai ◽  
Norio Inoue ◽  
Thomas J. Montville
Keyword(s):  
Molecular Mass ◽  
Mode Of Action ◽  
Proteolytic Enzymes ◽  
Terminal Sequence ◽  
Purification And Characterization ◽  
Consensus Motif ◽  
Carnobacterium Piscicola ◽  
Ph Range

ABSTRACT The bacteriocin piscicocin CS526 was inactivated by proteolytic enzymes, was stable at 100�C for 30 min, had a pH range of 2 to 8, and was active against Enterococcus, Listeria, Pediococcus, and Leuconostoc. The N-terminal sequence was YGNG L , not the YGNG V consensus motif common in class IIa bacteriocins (alternate residues underlined). The molecular mass of piscicocin CS526, which had a bactericidal mode of action, was ∼4,430 Da.

scholarly journals Purification and biochemical characterization of a poly(ADP-ribose) polymerase-like enzyme from the thermophilic archaeon Sulfolobus solfataricus

1998 ◽  
Vol 335 (2) ◽  
pp. 441-447 ◽  
Author(s):  
Maria Rosaria FARAONE-MENNELLA ◽  
Agata GAMBACORTA ◽  
Barbara NICOLAUS ◽  
Benedetta FARINA
Keyword(s):  
Biochemical Characterization ◽  
Sulfolobus Solfataricus ◽  
Enzymic Activity ◽  
Sds Page ◽  
Thermophilic Archaeon ◽  
Displacement Assay ◽  
Elongation Step ◽  
Ph Range ◽  
Good Agreement

A poly(ADP-ribose) polymerase-like enzyme, detected in a crude homogenate from Sulfolobus solfataricus by means of activity and immunoblot analyses, was purified to electrophoretic homogeneity by a rapid procedure including two sequential affinity chromatographies, on NAD+-agarose and DNA-Sepharose. The latter column selected specifically the poly(ADP-ribosyl)ating enzyme with a 17% recovery of enzymic activity and a purification of more than 15000-fold. The molecular mass (54–55 kDa) assessed by SDS/PAGE and immunoblot was definitely lower than that determined for the corresponding eukaryotic protein. The enzyme was proved to be thermophilic, with a temperature optimum of approx. 80 °C, and thermostable, with a half-life of 204 min at 80 °C, in good agreement with the requirements of a thermozyme. It displayed a Km towards NAD+ of 154±50 µM; in the pH range 6.5–10.0 the activity values were similar, not showing a real optimum pH. The enzyme was able to bind homologous DNA, as evidenced by the ethidium bromide displacement assay. The product of the ADP-ribosylating reaction co-migrated with the short oligomers of ADP-ribose (less than 6 residues) from a eukaryotic source. Reverse-phase HPLC analysis of the products, after digestion with phosphodiesterase I, gave an elution profile reproducing that obtained by the enzymic digestion of the rat testis poly(ADP-ribose). These results strongly suggest that the activities of the purified enzyme include the elongation step.

scholarly journals Arthrobacter equi sp. nov., isolated from veterinary clinical material

2011 ◽  
Vol 61 (9) ◽  
pp. 2089-2094 ◽  
Author(s):  
A. F. Yassin ◽  
C. Spröer ◽  
C. Siering ◽  
H. Hupfer ◽  
P. Schumann
Keyword(s):  
Biochemical Characterization ◽  
Rrna Gene ◽  
Positive Staining ◽  
Closely Related Species ◽  
Hybridization Data ◽  
Gene Sequence Analysis ◽  
Bacterium Strain ◽  
Type Strains ◽  
Sequence Similarities

A Gram-positive-staining, catalase-positive, non-spore-forming, rod-shaped bacterium, strain IMMIB L-1606T, isolated from genital swabs of a horse, was characterized using a polyphasic approach. Comparative 16S rRNA gene sequence analysis showed that the organism was related to members of the genus Arthrobacter, displaying sequence similarities of 93.5–99.1 % with the type strains of recognized species of the genus. Cell-wall analysis revealed peptidoglycan type A3α l-Lys–l-Ser–l-Thr–l-Ala. DNA–DNA hybridization data and biochemical characterization of strain IMMIB L-1606T enabled the isolate to be differentiated genotypically and phenotypically from phylogenetically closely related species of the genus Arthrobacter. Therefore, it is concluded that strain IMMIB L-1606T represents a novel species of the genus Arthrobacter, for which the name Arthrobacter equi sp. nov. is proposed. The type strain of Arthrobacter equi sp. nov. is IMMIB L-1606T ( = DSM 23395T = CCUG 59597T).

scholarly journals A Haloalkane Dehalogenase from a Marine Microbial Consortium Possessing Exceptionally Broad Substrate Specificity

2017 ◽  
Vol 84 (2) ◽  
Author(s):  
Tomas Buryska ◽  
Petra Babkova ◽  
Ondrej Vavra ◽  
Jiri Damborsky ◽  
Zbynek Prokop
Keyword(s):  
Substrate Specificity ◽  
Active Site ◽  
Biochemical Characterization ◽  
Environmental Pollutants ◽  
Haloalkane Dehalogenase ◽  
Broad Substrate Specificity ◽  
Organic Cosolvents ◽  
Marine Metagenome ◽  
Ph Range

ABSTRACTThe haloalkane dehalogenase enzyme DmmA was identified by marine metagenomic screening. Determination of its crystal structure revealed an unusually large active site compared to those of previously characterized haloalkane dehalogenases. Here we present a biochemical characterization of this interesting enzyme with emphasis on its structure-function relationships. DmmA exhibited an exceptionally broad substrate specificity and degraded several halogenated environmental pollutants that are resistant to other members of this enzyme family. In addition to having this unique substrate specificity, the enzyme was highly tolerant to organic cosolvents such as dimethyl sulfoxide, methanol, and acetone. Its broad substrate specificity, high overexpression yield (200 mg of protein per liter of cultivation medium; 50% of total protein), good tolerance to organic cosolvents, and a broad pH range make DmmA an attractive biocatalyst for various biotechnological applications.IMPORTANCEWe present a thorough biochemical characterization of the haloalkane dehalogenase DmmA from a marine metagenome. This enzyme with an unusually large active site shows remarkably broad substrate specificity, high overexpression, significant tolerance to organic cosolvents, and activity under a broad range of pH conditions. DmmA is an attractive catalyst for sustainable biotechnology applications, e.g., biocatalysis, biosensing, and biodegradation of halogenated pollutants. We also report its ability to convert multiple halogenated compounds to corresponding polyalcohols.

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