The aim of this study was to further broaden the heterologous expression alginate lyase from Vibrio alginolyticus in a Bacillus subtilis vector. A B. WB600/pP43NMK-alg62 strain constructed. (NH4)2SO4 precipitation and Ni-affinity chromatography were performed purify enzyme. We then characterized Its molecular weight 57.64 kDa, it worked optimally at 30 °C with pH 8.0. Ca2+ markedly enhanced enz...

growing applications of phenylalanine dehydrogenase (phedh) enzyme in the medical and pharmaceutical industries encourages researchers to seek simple, fast and economical alternative purification methods. with goal of finding a new technique, the extraction and purification of recombinant bacillus badius phedh in polyethylene glycol 6000 (peg-6000) and ammonium sulfate aqueous two-phase systems...

Objective: To construct and identify recombinant expression of a therapeutic tumor vaccine with HBcAg as a vector. Methods: PCR primers were designed according to the gene sequences of restriction enzyme sites of recombinant pKK233.2-hepatitis B core antigen (HBcAg) and melanoma-associated antigen 3 (MAGE-A3: 112120aa). The target fragment of MAGE-A3 was synthesized and cloned to pKK233.2-HBcAg...

Recent crystal structures of xanthine dehydrogenase, xanthine oxidase and related enzymes have paved the way for a detailed structural and functional analysis of these enzymes. One problem encountered when working with these proteins, especially with recombinant protein, is that the preparations tend to be heterogeneous, with only a fraction of the enzyme molecules being active. This is due to ...

Human adenosine deaminase (EC 3.5.4.4), a key purine salvage enzyme essential for immune competence, has been overproduced in Spodoptera frugiperda cells and in Trichoplusia ni (cabbage looper) larvae infected with recombinant baculovirus. The coding sequence of human adenosine deaminase was recombined into a baculovirus immediately downstream from the strong polyhedrin gene promoter. Approxima...

conclusions the gp40/15 gene, which was cloned in the pet28a+, was successfully expressed and produced in e. coli. therefore, this protein can be used in future studies to develop recombinant vaccines and diagnostic kits. methods in this experimental study, the gp40/15 gene sequence was extracted from genbank (no. af155624) and cloned in the pet28a+ plasmid. colony polymerase chain reaction (pc...

Microbial enzymes are increasingly finding applications as therapeutics due to their targeted activity and minimal side effects. Serratiopeptidase, also known a miracle enzyme, has already proved its potential an anti-inflammatory, mucolytic, fibrinolytic, analgesic in many studies. A cost effective, bioreactor level production process been described here comprising of the fed-batch fermentatio...

In this study, the fermentation broth of recombinant Pichia pastoris strain ncy-2 was studied. After pretreatment, separation, and purification, lysozyme optimized using biofilm ion exchange separation. Finally, dry enzyme powder prepared by concentrating vacuum drying. The removal rate bacterial cells 99.99% when centrifuged at low temperature. optimum conditions were: transmembrane pressure 0...

dna amplification using taq dna polymerase is one of the most widely used techniques in molecular biology and biotechnology. the aim of this study was to amplify the gene of this enzyme from a thermophilic bacteria called thermus aqauticus and clone it into a vector for future use. using specific primers the cdna of taq dna polymerase was amplified and ligated into the cloning vector ptz57r usi...

The alpha1- and beta1-subunits of human soluble guanylate cyclase (sGC) were coexpressed in the Sf9 cells/baculovirus system. In addition to the native enzyme, constructs with hexahistidine tag at the amino and carboxyl termini of each subunit were coexpressed. This permitted the rapid and efficient purification of active recombinant enzyme on a nickel-affinity column. The enzyme has one heme p...