scholarly journals Characterization of the penicillin G acylase from Bacillus megaterium ATCC 14945

2005 ◽  
Vol 48 (spe) ◽  
pp. 105-111 ◽  
Author(s):  
Vanessa Ribeiro de Souza ◽  
Ana C. G. Silva ◽  
Laura Marina Pinotti ◽  
Heloísa Sobreiro Selistre Araújo ◽  
Raquel de Lima Camargo Giordano
Keyword(s):  
Bacillus Megaterium ◽  
Reaction Rate ◽  
Penicillin G Acylase ◽  
Hydrolysis Rate ◽  
Penicillin G ◽  
Average Molecular Mass ◽  
Influence Of Temperature ◽  
Sds Page ◽  
Page Electrophoresis

The purpose of this work was to characterize the enzyme penicillin G acylase (PGA) produced by Bacillus megaterium. Purification of the enzyme by ultra/diafiltration did not allow the detection of the PGA band by SDS-PAGE electrophoresis due to the high content of remaining proteins. However, using the DNA of the microorganism, it was possible to replicate the genes of the two B. megaterium PGA reported in literature, showing that the enzyme consisted of two sub-units, having 245 and 537 amino acids each and an average molecular mass of 26950 and 59070 Da, respectively. The parameters studied were: 1) the influence of temperature in the 25-60(0)C range, 2) pH in the 5-10 range and 3) substrate concentration, this was tested to obtain results on the Penicillin G hydrolysis reaction rate, using the initial velocities approach. The maximum hydrolysis rate was obtained at 37ºC and pH 8.0. The Michaelis-Menten model fitted well, resulting in estimated Km and Vmax parameters values of 1.83 mM and 0.165*10-3 mmol/min/UI, respectively.

Inoculum Studies in Production of Penicillin G Acylase by Bacillus megaterium ATCC 14945

2002 ◽  
pp. 679-686 ◽  
Author(s):  
Laura M. Pinotti ◽  
Rosineide G. Silva ◽  
Roberto C. Giordano ◽  
Raquel L. C. Giordano
Keyword(s):  
Bacillus Megaterium ◽  
Penicillin G Acylase ◽  
Penicillin G

Increasing synthetic performance of penicillin G acylase from Bacillus megaterium by site-directed mutagenesis

2007 ◽  
Vol 74 (5) ◽  
pp. 1023-1030 ◽  
Author(s):  
Jingang Wang ◽  
Qing Zhang ◽  
He Huang ◽  
Zhongyi Yuan ◽  
Dafu Ding ◽  
...  
Keyword(s):  
Bacillus Megaterium ◽  
Site Directed Mutagenesis ◽  
Penicillin G Acylase ◽  
Penicillin G ◽  
Directed Mutagenesis

Influence of temperature and concentration of inducers on the expression of penicillin G Acylase (PGA) by recombinant E. coli

2010 ◽  
Vol 150 ◽  
pp. 393-393
Author(s):  
A.M. Vélez ◽  
M.R.C. Iemma ◽  
E. Cueva ◽  
T.C. Zangirolami ◽  
R.L.C. Giordano
Keyword(s):  
Penicillin G Acylase ◽  
Penicillin G ◽  
Influence Of Temperature ◽  
E Coli

scholarly journals Identification of a novel mannose-capped lipoarabinomannan from Amycolatopsis sulphurea

2003 ◽  
Vol 372 (3) ◽  
pp. 821-829 ◽  
Author(s):  
Kevin J. C. GIBSON ◽  
Martine GILLERON ◽  
Patricia CONSTANT ◽  
Germain PUZO ◽  
Jérôme NIGOU ◽  
...  
Keyword(s):  
Cell Envelope ◽  
Original Structure ◽  
Average Molecular Mass ◽  
Ionization Time ◽  
Flight Mass Spectrometry ◽  
Sds Page ◽  
Intermediate Size ◽  
Genus Mycobacterium ◽  
Factor Α

The genus Amycolatopsis is a member of the phylogenetic group nocardioform actinomycetes, which also includes the genus Mycobacterium. Members of this group have a characteristic cell envelope structure, dominated by various complex lipids and polysaccharides. Amongst these, lipoglycans are of particular interest since mycobacterial lipoarabinomannans are important immunomodulatory molecules. In this study we report the isolation and structural characterization of Amycolatopsis sulphurea lipoarabinomannan, designated AsuLAM. SDS/PAGE analysis revealed that AsuLAM was of an intermediate size between Mycobacterium tuberculosis lipoarabinomannan and lipomannan, confirmed by matrix-assisted laser-desorption ionization–time-of-flight mass spectrometry that predicted an average molecular mass of 10 kDa. Using a range of chemical degradations, NMR experiments and capillary electrophoresis analysis, AsuLAM was revealed as an original structure. The mannosyl-phosphatidyl-myo-inositol anchor exhibits a single acyl-form, characterized by a diacylated glycerol moiety, and contains, as one of the main fatty acids, 14-methyl-pentadecanoate, a characteristic fatty acid of the Amycolatopsis genus. AsuLAM also contains a short mannan domain; and is dominated by a multi-branched arabinan domain, composed of an (α1→5)-Araf (arabinofuranose) chain substituted, predominately at the O-2 position, by a single β-Araf. The arabinan domain is further elaborated by manno-oligosaccharide caps, with around one per molecule. This is the first description of manno-oligosaccharide caps found in a non-mycobacterial LAM. AsuLAM was unable to induce the production of the pro-inflammatory cytokine tumour necrosis factor α when tested with human or murine macrophage cell lines, reinforcing the paradigm that mannose-capped LAM are poor inducers of pro-inflammatory cytokines.

scholarly journals Characterization of Gelatin Profile of Chicken Broiler (Gallus domestica) Bone Using SDS-PAGE Electrophoresis

ALCHEMY ◽  
2019 ◽  
Vol 7 (1) ◽  
pp. 7 ◽  
Author(s):  
Dewi Yuliani ◽  
Dhienda Risa Awalsasi ◽  
Akyunul Jannah
Keyword(s):  
Ammonium Sulfate ◽  
Protein Concentration ◽  
Peptide Fragments ◽  
Strong Base ◽  
Chicken Bone ◽  
Sds Page ◽  
Hydrolysis Of ◽  
Β Chain ◽  
Page Electrophoresis

<p>Gelatin, a proteinaceous additive, is obtained from hydrolysis of collagen in the bone, hide and skin of animals. As natural product, gelatin has been applied in many industries with various functions. This study attempt to characterize gelatin profile of broiler chicken (<em>Gallus domestica</em>) using SDS-PAGE electrophoresis. The chicken bone was pretreated using a strong base, sodium hydroxide, producing type B gelatin. The gelatin was purified through precipitation using the variation of ammonium sulfate concentrations (40-70%) and dialysis using cellophane membrane. The purified gelatin was characterized through SDS-PAGE electrophoresis. Based on electrophoresis visualization, reduction of band intensity by ammonium sulfate 40% showed removal of small peptide fragments. The remained gelatin showed two major bands, α-chains and a β-chain with the respective molecular weight of ~135 and ~245 kDa. The protein content of the unpurified gelatin (E1) was 71.65±0.60 mg/L.  The purified E1 gelatins by 40-70% of ammonium sulfate addition contained 61.42±3.90, 60.45±1.36, 59.89±0.24, and 55.32±1.05 mg/L of protein concentration, respectively.</p><p> </p><p>Keywords: chicken bone, gelatin profile, protein electrophoresis</p>

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