scholarly journals The Effects of Temperature and Culture Medium Composition on Protein Recombinant JSU-pET Expression

2019 ◽  
Vol 17 (2) ◽  
pp. 31
Author(s):  
Indriawati Indriawati ◽  
Slamet Diah Volkandari ◽  
Endang Tri Margawati
Keyword(s):  
Escherichia Coli ◽  
Protein Expression ◽  
Culture Medium ◽  
Medium Composition ◽  
Protein Yield ◽  
Culture Temperature ◽  
Factors Influencing ◽  
Effects Of Temperature ◽  
The Right ◽  
Medium Culture

There are several factors influencing the protein expression. The study was aimed to increase the yield of protein recombinant JSU-pET by applying combination of temperature and medium culture composition. This research was composed in a 2x2 factorial design with two level temperatures of 37<sup>o</sup>C and 25<sup>o</sup>C and 2 medium compositions of LB and mLB (modified). <em>Escherichia coli</em> bearing JSU-pET plasmid were grown in two applied media and incubated in two applied temperatures. The cells were harvested by centrifugation the pellets were then lysed. The lysed cells were centrifuged, the supernatant then purified by Ni-NTA Resin Agarose. The purified JSU protein recombinant was characterized and quantified by spectrophotometer. The Result showed that the JSU protein recombinant was successfully expressed on the right size (37kDa) in all treatments even though the JSU-pET showed sharper band in m LB at 37<sup>o</sup>C. Statistically the protein yield was not significant different (P&gt;0.05), however culture temperature of 25<sup>o</sup>C showed higher yield (0.618±0.095 vs 0.704±0.094) than cultured at 37<sup>o</sup>C (0.598±0.137 vs 0.553±0.041), respectively either cultured in LB or mLB. This finding suggests that a lower culture temperature could increase protein yield both either in LB or modified LB.

Biosynthesis of Low Molecular Weight Bacterial Cellulose

2006 ◽  
Vol 309-311 ◽  
pp. 497-502 ◽  
Author(s):  
Yu Hong Feng ◽  
Xi Bin Wang ◽  
Qiang Lin ◽  
Zhou Xin Wu ◽  
Su Juan Pang ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Bacterial Cellulose ◽  
Culture Medium ◽  
Ph Value ◽  
High Yield ◽  
Low Molecular Weight ◽  
Culture Temperature ◽  
Culture Time ◽  
N Compounds ◽  
Medium Culture

The biosynthesis of low molecular weight bacterial cellulose (LMBC) by Static culture of Acetobacter xylinum HN001 has been investigated. The medium was prepared with coconut-water by adding sugar and N-compounds. Several conditions effected LMBC yields, such as start pH value of culture medium, culture temperature and culture time were investigated. In view of high yield of LMBC, the optimal culture time and culture temperature was about 70 hours and 32°C with yield of LMBC 1.2g/L. Molecular weight (MW) and molecular weight distribution (MWD) of LMBC were tested by GFC. Results showed that, though the start pH value of culture medium varied from 3.5 to 5.5, the MW of the product cellulose varied not notable and MWD of LMBC were all almost 1.3.

Expression of Foreign Proteins in Escherichia coli

2021 ◽  
Author(s):  
Ali Iftikhar
Keyword(s):  
Escherichia Coli ◽  
Protein Expression ◽  
Recombinant Proteins ◽  
Expression System ◽  
Maximum Yield ◽  
Protein Yield ◽  
Foreign Protein ◽  
Low Protein ◽  
Prokaryotic Expression System ◽  
Foreign Proteins

Abstract BackgroundOptimization of conditions for the recombinant production of proteins in a prokaryotic expression system is essential as the recombinant proteins impose a metabolic burden on cell's growth leading to low protein yield and low protein expression resulting from cell death.Main textThe concentration of media components is optimized to accommodate for depleted nutrients due to foreign protein expression. The temperature is optimized to reduce proteolytic degradation and accumulation of protein as inclusion bodies in Escherichia coli. The concentration of inducer and time of induction for high protein yield is also optimized. These optimization conditions depend on the promoter under which the gene of interest is present and the characteristics of the target protein.ConclusionIn the past few years, many optimization conditions for the production of recombinant proteins in Escherichia coli have been studied. These conditions depend mainly upon the promoter used to produce protein and the type of protein produced. Optimizing the expression parameters of protein produced in Escherichia coli ensures maximum yield of the desired protein.

scholarly journals Boosting Auto-induction of Recombinant Proteins in Escherichia coli with Glucose and Lactose Additives

2021 ◽  
Vol 28 ◽  
Author(s):  
Nariyasu Tahara ◽  
Itaru Tachibana ◽  
Kazuyo Takeo ◽  
Shinji Yamashita ◽  
Atsuhiro Shimada ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Proteins ◽  
Culture Medium ◽  
Thermus Thermophilus ◽  
Medium Composition ◽  
Final Concentration ◽  
Induction Medium ◽  
Lac Operon ◽  
Escherichia Coli Expression ◽  
Complex Culture

Background: Auto-induction is a convenient way to produce recombinant proteins without inducer addition using lac operon-controlled Escherichia coli expression systems. Auto-induction can occur unintentionally using a complex culture medium prepared by mixing culture substrates. The differences in culture substrates sometimes lead to variations in the induction level. Objectives: In this study, we investigated the feasibility of using glucose and lactose as boosters of auto-induction with a complex culture medium. Method: First, auto-induction levels were assessed by quantifying recombinant GFPuv expression under the control of the T7lac promoter. Effectiveness of the additive-containing medium was examined using ovine angiotensinogen (tac promoter-based expression) and Thermus thermophilus manganese-catalase (T7lac promoter-based expression). Results: Auto-induced GFPuv expression was observed with the enzymatic protein digest Polypepton, but not with another digest tryptone. Regardless of the type of protein digest, supplementing Terrific Broth medium with glucose (at a final concentration of 2.9 g/L) and lactose (at a final concentration of 7.6 g/L) was successful in obtaining an induction level similar to that achieved with a commercially available auto-induction medium. The two recombinant proteins were produced in milligram quantity of purified protein per liter of culture. Conclusion: The medium composition shown in this study would be practically useful for attaining reliable auto-induction for E. coli-based recombinant protein production.

Pertumbuhan Dan Perkembangan Anggrek Dendrobium anosmum Pada Media Kultur In Vitro Dengan Beberapa Konsentrasi Air Kelapa

Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Water Concentration ◽  
Medium Composition ◽  
Coconut Water ◽  
Orchid Species ◽  
Randomized Design ◽  
Wet Weight ◽  
Completely Randomized Design

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.

Pertumbuhan Cladocera jenis Chydoridae pada media kultur yang berbeda

2013 ◽  
Vol 1 (2) ◽  
Author(s):  
Denovis Sambode ◽  
Henneke Pangkey ◽  
Sartje Lantu
Keyword(s):  
Population Growth ◽  
Culture Medium ◽  
Quality Parameters ◽  
Organic Fertilizers ◽  
Cow Dung ◽  
Randomized Design ◽  
Period Data ◽  
Growth Development ◽  
Completely Randomized Design ◽  
Medium Culture

The aim of this study was to find out the effect of different organic fertilizers on the population growth of Chydorus sp. This research was conducted at Laboratorium of Nutrition and Food Technology, Faculty of Fishery and Marine Sciences, Sam Ratulangi University, from December, 2012 to January, 2013. The experiment was run in the Completely Randomized Design with 3 treatments and 3 replication. Chydorus sp. was cultured in 9 glass jars with a volume of 1 L each. Three treatments applied in this research included A: medium culture composed of 500ml of water and 50 grams of soil; B: medium culture composed of 500ml of water, 50 grams of soil and 10g of cow dung; C: medium culture composed of 500ml of water, 50 grams of soil and 10g of horse dung. The density of Cydorus sp. in each media was 10 individuals /500 ml water. Observation on the population growth, development of live preys, and water quality parameters was conducted for 20 days period. Data were statistically analized with Analysis of variance. The results showed the highest density of Chydorus sp. was reached by Chydorus cultured in medium with horse manure (2169 individual/500 ml), followed by medium culture with cow manure (1715 individual/500 ml), and the lowest in medium culture with soil which was 1065,33 individual/500 ml. However, Analysis of variances showed that Chydorus population growth were not significantly affected by different culture medium. Keywords: manure, liquid fertilizer, growth, Chydorus sp.

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