Biotransformation of penicillin V to 6-aminopenicillanic acid using immobilized whole cells of E. coli expressing a highly active penicillin V acylase

2016 ◽  
Vol 47 (1) ◽  
pp. 52-57 ◽  
Author(s):  
Vellore Sunder Avinash ◽  
Palna Dinesh Chauhan ◽  
Shraddha Gaikwad ◽  
Archana Pundle
Keyword(s):  
Whole Cells ◽  
E Coli ◽  
Penicillin V ◽  
Highly Active ◽  
Penicillin V Acylase ◽  
Immobilized Whole Cells

scholarly journals Enzyme-catalyzed biodegradation of penicillin fermentation residues by β-lactamase OtLac from Ochrobactrum tritici

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Peng Wang ◽  
Chen Shen ◽  
Qinqin Cong ◽  
Kaili Xu ◽  
Jialin Lu
Keyword(s):  
Large Scale ◽  
In Vitro Study ◽  
Scale Removal ◽  
E Coli ◽  
Penicillin V ◽  
Highly Active ◽  
Steady State Kinetics ◽  
Biodegradation Process ◽  
Km Value

Abstract Background Biodegradation of antibiotics is a promising method for the large-scale removal of antibiotic residues in the environment. However, the enzyme that is involved in the biodegradation process is the key information to be revealed. Results In this study, the beta-lactamase from Ochrobactrumtritici that mediates the biodegradation of penicillin V was identified and characterized. When searching the proteins of Ochrobactrumtritici, the β-lactamase (OtLac) was identified. OtLac consists of 347 amino acids, and predicted isoelectric point is 7.0. It is a class C β-lactamase according to BLAST analysis. The coding gene of OtLac was amplified from the genomic DNA of Ochrobactrumtritici. The OtLac was overexpressed in E. coli BL21 (DE3) and purified with Ni2+ column affinity chromatography. The biodegradation ability of penicillin V by OtLac was identified in an in vitro study and analyzed by HPLC. The optimal temperature for OtLac is 32 ℃ and the optimal pH is 7.0. Steady-state kinetics showed that OtLac was highly active against penicillin V with a Km value of 17.86 μM and a kcat value of 25.28 s−1 respectively. Conclusions OtLac demonstrated biodegradation activity towards penicillin V potassium, indicating that OtLac is expected to degrade penicillin V in the future.

scholarly journals Enhanced Extracellular Synthesis of Gold Nanoparticles by Soluble Extracts from Escherichia coli Transformed with Rhizobium tropici Phytochelatin Synthase Gene

Metals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 472
Author(s):  
Qunying Yuan ◽  
Manjula Bomma ◽  
Zhigang Xiao
Keyword(s):  
Gold Nanoparticles ◽  
Gold Nanoparticle ◽  
Synthesis Method ◽  
Nanoparticle Synthesis ◽  
Morphological Properties ◽  
Phytochelatin Synthase ◽  
Rhizobium Tropici ◽  
Whole Cells ◽  
E Coli ◽  
Recombinant Cells

Phytochelatins, the enzymatic products of phytochelatin synthase, play a principal role in protecting the plants from heavy metal and metalloid toxicity due to their ability to scavenge metal ions. In the present study, we investigated the capacity of soluble intracellular extracts from E. coli cells expressing R. tropici phytochelatin synthase to synthesize gold nanoparticle. We discovered that the reaction mediated by soluble extracts from the recombinant E. coli cells had a higher yield of gold nanoparticles, compared to that from the control cells. The compositional and morphological properties of the gold nanoparticles synthesized by the intracellular extracts from recombinant cells and control cells were similar. In addition, this extracellular nanoparticle synthesis method produced purer gold nanoparticles, avoiding the isolation of nanoparticles from cellular debris when whole cells are used to synthesize nanoparticles. Our results suggested that phytochelatins can improve the efficiency of gold nanoparticle synthesis mediated by bacterial soluble intracellular extracts, and the potential of extracellular nanoparticle synthesis platform for the production of nanoparticles in large quantity and pure form is worth further investigation.

scholarly journals Identification of the Biosynthetic Gene Cluster for 3-Methylarginine, a Toxin Produced by Pseudomonas syringae pv. syringae 22d/93

2010 ◽  
Vol 76 (8) ◽  
pp. 2500-2508 ◽  
Author(s):  
S. D. Braun ◽  
J. Hofmann ◽  
A. Wensing ◽  
M. S. Ullrich ◽  
H. Weingart ◽  
...  
Keyword(s):  
Amino Acid ◽  
Gene Cluster ◽  
Pseudomonas Syringae ◽  
Biosynthetic Gene Cluster ◽  
Pentanoic Acid ◽  
Promising Candidate ◽  
Biosynthesis Gene Cluster ◽  
E Coli ◽  
Highly Active ◽  
Putative Aminotransferase

ABSTRACT The epiphyte Pseudomonas syringae pv. syringae 22d/93 (Pss22d) produces the rare amino acid 3-methylarginine (MeArg), which is highly active against the closely related soybean pathogen Pseudomonas syringae pv. glycinea. Since these pathogens compete for the same habitat, Pss22d is a promising candidate for biocontrol of P. syringae pv. glycinea. The MeArg biosynthesis gene cluster codes for the S-adenosylmethionine (SAM)-dependent methyltransferase MrsA, the putative aminotransferase MrsB, and the amino acid exporter MrsC. Transfer of the whole gene cluster into Escherichia coli resulted in heterologous production of MeArg. The methyltransferase MrsA was overexpressed in E. coli as a His-tagged protein and functionally characterized (Km , 7 mM; k cat, 85 min−1). The highly selective methyltransferase MrsA transfers the methyl group from SAM into 5-guanidino-2-oxo-pentanoic acid to yield 5-guanidino-3-methyl-2-oxo-pentanoic acid, which then only needs to be transaminated to result in the antibiotic MeArg.

Biochemical Energy Conversion by Immobilized Whole Cells.

1983 ◽  
Vol 413 (1 Biochemical E) ◽  
pp. 133-143 ◽  
Author(s):  
SHUICHI SUZUKI ◽  
ISAO KARUBE ◽  
HIDEAKI MATSUOKA ◽  
SATOSHI UEYAMA ◽  
HIROAKI KAWAKUBO ◽  
...  
Keyword(s):  
Energy Conversion ◽  
Whole Cells ◽  
Immobilized Whole Cells

scholarly journals ЕТІОЛОГІЧНА СТРУКТУРА ГОСТРИХ ШЛУНКОВО–КИШКОВИХ ЗАХВОРЮВАНЬ ТЕЛЯТ

2016 ◽  
Vol 18 (3(71)) ◽  
pp. 148-151
Author(s):  
A. Berezovskyi ◽  
T. Fotyna ◽  
L. Ulko ◽  
A. Nechyporenko ◽  
E. Tytov
Keyword(s):  
Biological Material ◽  
Gastrointestinal Disease ◽  
Gastrointestinal Diseases ◽  
Important Place ◽  
E Coli ◽  
Leading Role ◽  
Highly Active ◽  
Opportunistic Bacteria ◽  
Antibiotic Drugs ◽  
Low Efficiency

The results of tests of samples of biological material from the calves with acute gastrointestinal diseases presents in the article. It was found that the occurrence and development of acute gastrointestinal diseases the leading role played by opportunistic bacteria association: S. aureus, S. saprophiticus, S. agalactiae, S. faecalis, S. pyogenes, E. coli, E. cloacae, C. jejuni, P. vulgaris, P. mirabilis, P. aeruginosa and K. Pneumoniae. Of the 632 examinations we studied 2,786 samples of biological material. Analysis of the results on our diagnostic work shows that acute gastrointestinal disease in calves caused by microbial associations. At acute gastrointestinal disease isolated pathogen E. coli – 21.0%. Coccoid microorganisms group also had significant representation. Thus, a biomaterial of 15.5% was allocated S. faecalis, at 11.6% was allocated pathogen S. aureus. Although to a lesser extent, but large office in the species spectrum of microorganisms by acute gastrointestinal diseases of calves occupied S. agalactiae – 4.8%, S. saprophiticus – 4.6%, S. pyogenes – 3.7%. Among the important place occupied microorganisms culture P. vulgaris – 8.5% of the total allocation of the samples. The average discharge frequency was characteristic for Campylobacter jejuni – 6.5% of cases.In analyzing the sensitivity of microorganisms isolated from calves with acute gastro–intestinal diseases to different groups of antibiotic drugs found that E. coli has a low sensitivity to antibiotics almost all studied groups. The largest number of selected crops susceptible to cephalosporins – 74.8%. Most of the isolated cultures weaklysensitive or insensitive to antibiotics studied by us. These data are an indication of the spread of antibiotic–resistant strains of microorganisms among newborn calves in the farms of Sumy and Chernihiv regions, which explains the low efficiency of antibiotic therapy. Highly active towards crop S. aureus, S. saprophiticus, S. agalactiae, S. faecalis, S. pyogenes, E. coli, E. cloacae, C. jejuni, P. vulgaris, P. mirabilis, P. aeruginosa, and K. pneumoniae were drug «Ceftioklin» and combined antibacterial agents. 

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