Large-Scale Production of a Soluble Human β-1,4-Galactosyltransferase Using a Saccharomyces cerevisiae Expression System

1995 ◽  
Vol 6 (1) ◽  
pp. 72-78 ◽  
Author(s):  
G.F. Herrmann ◽  
C. Krezdorn ◽  
M. Malissard ◽  
R. Kleene ◽  
H. Paschold ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Large Scale ◽  
Expression System ◽  
Scale Production ◽  
Large Scale Production

cDNA cloning and heterologous expression of a wheat proteinase inhibitor of subtilisin and chymotrypsin (WSCI) that interferes with digestive enzymes of insect pests

2005 ◽  
Vol 386 (4) ◽  
pp. 383-389 ◽  
Author(s):  
Simone Di Gennaro ◽  
Anna G. Ficca ◽  
Daniela Panichi ◽  
Elia Poerio
Keyword(s):  
Proteinase Inhibitor ◽  
Large Scale ◽  
Insect Pests ◽  
Expression System ◽  
Physiological Role ◽  
Scale Production ◽  
Insect Larvae ◽  
E Coli ◽  
Large Scale Production ◽  
Degenerate Oligonucleotide

Abstract A cDNA encoding the proteinase inhibitor WSCI (wheat subtilisin/chymotrypsin inhibitor) was isolated by RT-PCR. Degenerate oligonucleotide primers were designed based on the amino acid sequence of WSCI and on the nucleotide sequence of the two homologous inhibitors (CI-2A and CI-2B) isolated from barley. For large-scale production, wsci cDNA was cloned into the E. coli vector pGEX-2T. The fusion protein GST-WSCI was efficiently produced in the bacterial expression system and, as the native inhibitor, was capable of inhibiting bacterial subtilisin, mammalian chymotrypsins and chymotrypsin-like activities present in crude extracts of a number of insect larvae (Helicoverpa armigera, Plodia interpunctella and Tenebrio molitor). The recombinant protein produced was also able to interfere with chymotrypsin-like activity isolated from immature wheat caryopses. These findings support a physiological role for this inhibitor during grain maturation.

scholarly journals Secretory Expression of YY(3-36) Peptide in E. coli

2019 ◽  
Author(s):  
Sorush Niknamian
Keyword(s):  
Large Scale ◽  
Peptide Yy ◽  
Signal Sequence ◽  
Expression System ◽  
Complete Sequence ◽  
Scale Production ◽  
E Coli ◽  
Large Scale Production ◽  
Human Experiments ◽  
Clinical Experiments

Obesity is the prime suspect in a wide frequency of diabetes type II and cardiovascular diseases worldwide. Recombinant YY (tyrosine-tyrosine) peptide is a locally acting hormone, controlling secretion in the digestive tract. Interestingly, it was later shown that a truncated version of YY peptide, YY(3-36) peptide, has the potential as an important biopharmaceutical in a fight against obesity. This peptide has shown promising results in human clinical experiments in appetite reduction in human experiments. To develop an economical expression system for large-scale production of the peptide in gram-negative bacteria, we have developed a chimeric gene for extracellular expression of this peptide with the assistance of signal sequence of asparaginase II from Escherichia coli. This system has the advantage of producing the complete sequence of YY(3-36) without any extra tags that require further removal with the assistance of expensive specific proteases and reduce the downstream steps significantly. Our results pave the way for the recombinant production of YY(3-36) peptide and further proves the efficacy of asparaginase II signal sequence as a communicator of foreign peptides and proteins into extracellular space of E. coli.

Production of Recombinant Melittin by Auto-Induction in Escherichia coli

2013 ◽  
Vol 798-799 ◽  
pp. 1007-1012
Author(s):  
Wen He Zhu ◽  
Wei Zhang ◽  
Yan Li ◽  
Jun Jie Xu ◽  
Shi Jie Lv
Keyword(s):  
Fusion Protein ◽  
Large Scale ◽  
Expression Vector ◽  
Human Cancer ◽  
Expression System ◽  
Scale Production ◽  
Recognition Sequence ◽  
Large Scale Production ◽  
N Terminus ◽  
Gst Fusion Protein

Melittin is a novel peptide of biological activity isolated from bee venom. It has potential application value in medicine and agriculture. Here we encoded melittin gene with the EK recognition sequence in the N-terminus into expression vector pGEX-2T.The expressed fusion protein, which is about 29KDa, identified by Western Blot. To facilitate large-scale production of recombinant GST-fusion protein, we optimized different expression conditions to increase the overall production of the fusion protein. The production of the protein had increased about 10-fold when we used an auto-inducing medium. The GST fusion protein showed an equivalent activity with the natural melittin after digested by EK and can inhibited the proliferations of several human cancer lines. The expression system described in this study provides a feasible way for producing melittin in further studies.

scholarly journals Novel Expression System for Large-Scale Production and Purification of Recombinant Class IIa Bacteriocins and Its Application to Piscicolin 126

2004 ◽  
Vol 70 (6) ◽  
pp. 3292-3297 ◽  
Author(s):  
Gerard M. Gibbs ◽  
Barrie E. Davidson ◽  
Alan J. Hillier
Keyword(s):  
Fusion Protein ◽  
Large Scale ◽  
Expression System ◽  
Reversed Phase ◽  
Scale Production ◽  
Gene Encoding ◽  
Strong Activity ◽  
E Coli ◽  
Large Scale Production ◽  
Bacterial Cell Membrane

ABSTRACT Piscicolin 126 is a class IIa bacteriocin isolated from Carnobacterium piscicola JG126 that exhibits strong activity against Listeria monocytogenes. The gene encoding mature piscicolin 126 (m-pisA) was cloned into an Escherichia coli expression system and expressed as a thioredoxin-piscicolin 126 fusion protein that was purified by affinity chromatography. Purified recombinant piscicolin 126 was obtained after CNBr cleavage of the fusion protein followed by reversed-phase chromatography. Recombinant piscicolin 126 contained a single disulfide bond and had a mass identical to that of native piscicolin 126. This novel bacteriocin expression system generated approximately 26 mg of purified bacteriocin from 1 liter of E. coli culture. The purified recombinant piscicolin 126 acted by disruption of the bacterial cell membrane.

Large-scale production of VHH antibody fragments by Saccharomyces cerevisiae

2002 ◽  
Vol 30 (3) ◽  
pp. 273-278 ◽  
Author(s):  
Yvonne E. Thomassen ◽  
Wilmar Meijer ◽  
Laurens Sierkstra ◽  
C.Theo Verrips
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Large Scale ◽  
Antibody Fragments ◽  
Scale Production ◽  
Large Scale Production

scholarly journals Saccharomyces Cerevisiae Var. Boulardii: Valuable Probiotic Starter for Craft Beer Production

2019 ◽  
Vol 9 (16) ◽  
pp. 3250 ◽  
Author(s):  
Joaquín Mulero-Cerezo ◽  
Álvaro Briz-Redón ◽  
Ángel Serrano-Aroca
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Antioxidant Activity ◽  
Large Scale ◽  
Bioreactor Culture ◽  
Scale Production ◽  
Craft Beer ◽  
Yeast Viability ◽  
Large Scale Production ◽  
Lower Alcohol ◽  
Probiotic Yeast

The use of probiotic starters remaining viable in unpasteurized and unfiltered beers could significantly increase health benefits. Here, the probiotic Saccharomyces cerevisiae var. boulardii (Scb) and a commercial Saccharomyces cerevisiae (Sc) strain, which is commonly employed in the brewing industry, are compared as single starters. The healthy value of the produced beers and growth performance in a laboratory bioreactor are analysed by determining antioxidant activity, phenolic content and profile, alcohol, biomass growth modelling by the logistic and Gompertz equations, biovolume estimation from 2D microscopy images, and yeast viability after fermentation. Thus, in this study, the craft beer produced with the probiotic yeast possessed higher antioxidant activity, lower alcohol content, similar sensory attributes, much higher yeast viability and more acidification, which is very desirable to reduce contamination risks at large-scale production. Furthermore, Scb exhibited faster growth in the bioreactor culture and larger cell volumes than Sc, which increases the probiotic volume of the final craft beer.

Sign in / Sign up

Export Citation Format

Share Document