scholarly journals Effects of Lower Temperature on Expression and Biochemical Characteristics of HCV NS3 Antigen Recombinant Protein

Catalysts ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1297
Author(s):  
Chen-Ji Huang ◽  
Hwei-Ling Peng ◽  
Anil Kumar Patel ◽  
Reeta Rani Singhania ◽  
Cheng-Di Dong ◽  
...  
Keyword(s):  
Elisa Test ◽  
Stability Study ◽  
Iptg Induction ◽  
Antigenic Activity ◽  
Long Term Storage ◽  
Different Temperatures ◽  
Commercial Kits ◽  
Lower Temperature ◽  
Term Storage

The nonstructural antigen protein 3 of the hepatitis C virus (HCV NS3), commonly-used for HCV ELISA diagnosis, possesses protease and helicase activities. To prevent auto-degradation, a truncated NS3 protein was designed by removing the protease domain. Firstly, it was overexpressed in E. coli by IPTG induction under two different temperatures (25 and 37 °C), and purified using affinity chromatography to attain homogeneity above 90%. The molecular mass of purified protein was determined to be approx. 55 kDa. While lowering the temperature from 37 to 25 °C, the yield of the soluble fraction of HCV NS3 was increased from 4.15 to 11.1 mgL−1 culture, which also improved the antigenic activity and specificity. The protein stability was investigated after long-term storage (for 6 months at −20 °C) revealed no loss of activity, specificity, or antigenic efficacy. A thermal stability study on both freshly produced and stored HCV NS3 fractions at both temperatures showed that the unfolding curve profile properly obey the three-state unfolding mechanism. In the first transition phase, the midpoints of the thermal denaturation of fresh NS3 produced at 37 °C and 25 °C, and that produced after long-term storage at 37 °C and 25 °C, were 59.7 °C, 59.1 °C, 55.5 °C, and 57.8 °C, respectively. Microplates coated with the fresh NS3 produced at 25 °C or at 37 °C that were used for the HCV ELISA test and the diagnosis outcome were compared with two commercial kits—Abbott HCV EIA 2.0 and Ortho HCV EIA 3.0. Results indicated that the specificity of the HCV NS3 produced fresh at 25 °C was higher than that of the fresh one at 37 °C, hence showing potential for application in HCV ELISA diagnosis.

scholarly journals Changes in stability and shelf-life of ultra-high temperature treated milk during long term storage at different temperatures

Heliyon ◽  
2019 ◽  
Vol 5 (9) ◽  
pp. e02431 ◽  
Author(s):  
Maria A. Karlsson ◽  
Maud Langton ◽  
Fredrik Innings ◽  
Bozena Malmgren ◽  
Annika Höjer ◽  
...  
Keyword(s):  
High Temperature ◽  
Shelf Life ◽  
Long Term Storage ◽  
Different Temperatures ◽  
Term Storage ◽  
Ultra High Temperature

scholarly journals Cryopreservation of Synchytrium solstitiale In Planta

Plant Disease ◽  
2006 ◽  
Vol 90 (4) ◽  
pp. 429-432 ◽  
Author(s):  
T. L. Widmer
Keyword(s):  
Propionic Acid ◽  
Skim Milk ◽  
Infected Tissue ◽  
In Planta ◽  
Long Term Storage ◽  
Zoospore Release ◽  
Biological Studies ◽  
Different Temperatures ◽  
Term Storage

The fungus Synchytrium solstitiale is a candidate for use as a biocontrol agent against Centaurea solstitialis. This obligate parasite can be propagated only in planta, which necessitates development of a method for preserving cultures for longer periods of time for routine biological studies and shipment to other laboratories. Normally, sporangia embedded within the plant tissue release zoospores when submersed in water at temperatures above freezing. To examine what chemicals might inhibit zoospore release, infected tissue was exposed to different suspensions of fungicides. Cycloheximide and benomyl completely inhibited zoospore release or immediately induced encystment from tissue stored in these two chemicals, respectively. A few zoospores were released in suspensions of iprodione and propionic acid but were not motile. However, when tissue stored in iprodione or propionic acid was transferred to fresh distilled water, abundant active zoospores were released. Freezing the infected tissue at different temperatures in different cryoprotectants also affected the release of motile zoospores. Infected C. solstitialis tissue was immersed in water, water plus iprodione, methanol, ethylene glycol, dimethyl sulfoxide, glycerol, skim milk, trehalose, or sucrose and subjected to different temperatures for various periods of time. Some treatments protected the viability of the fungus for a shorter period of time whereas other treatments completely inhibited release. The best results were obtained when infected tissue was stored at -2°C in 0.5 M sucrose where active zoospores were released after 12 weeks of storage. Overall, results obtained from this study demonstrate a technique for long-term storage of S. solstitiale.

Effect of low oxygen on quality attributes of ‘Barton’ pecan nuts after long-term storage at different temperatures

2020 ◽  
Vol 263 ◽  
pp. 109098
Author(s):  
Stephanie Reis Ribeiro ◽  
Quellen Machado Ribeiro ◽  
Bruna Klein ◽  
Ingrid Duarte dos Santos ◽  
Sarah Forgiarini ◽  
...  
Keyword(s):  
Quality Attributes ◽  
Low Oxygen ◽  
Long Term Storage ◽  
Different Temperatures ◽  
Term Storage

scholarly journals Highly Efficient Computationally Derived Novel Metagenome α-Amylase With Robust Stability Under Extreme Denaturing Conditions

2021 ◽  
Vol 12 ◽  
Author(s):  
Shohreh Ariaeenejad ◽  
Behrouz Zolfaghari ◽  
Seyedeh Fatemeh Sadeghian Motahar ◽  
Kaveh Kavousi ◽  
Morteza Maleki ◽  
...  
Keyword(s):  
Industrial Applications ◽  
Metagenomic Data ◽  
Harsh Environments ◽  
Long Term Storage ◽  
Stain Removal ◽  
Different Temperatures ◽  
Commercial Detergents ◽  
Ph Range ◽  
Term Storage

α-Amylases are among the very critical enzymes used for different industrial purposes. Most α-amylases cannot accomplish the requirement of industrial conditions and easily lose their activity in harsh environments. In this study, a novel α-amylase named PersiAmy1 has been identified through the multistage in silico screening pipeline from the rumen metagenomic data. The long-term storage of PersiAmy1 in low and high temperatures demonstrated 82.13 and 71.01% activities after 36 days of incubation at 4 and 50°C, respectively. The stable α-amylase retained 61.09% of its activity after 180 min of incubation at 90°C and was highly stable in a broad pH range, showing 60.48 and 86.05% activities at pH 4.0 and pH 9.0 after 180 min of incubation, respectively. Also, the enzyme could resist the high-salinity condition and demonstrated 88.81% activity in the presence of 5 M NaCl. PersiAmy1 showed more than 74% activity in the presence of various metal ions. The addition of the detergents, surfactants, and organic solvents did not affect the α-amylase activity considerably. Substrate spectrum analysis showed that PersiAmy1 could act on a wide array of substrates. PersiAmy1 showed high stability in inhibitors and superb activity in downstream conditions, thus useful in detergent and baking industries. Investigating the applicability in detergent formulation, PersiAmy1 showed more than 69% activity after incubation with commercial detergents at different temperatures (30–50°C) and retained more than 56% activity after incubation with commercial detergents for 3 h at 10°C. Furthermore, the results of the wash performance analysis exhibited a good stain removal at 10°C. The power of PersiAmy1 in the bread industry revealed soft, chewable crumbs with improved volume and porosity compared with control. This study highlights the intense power of robust novel PersiAmy1 as a functional bio-additive in many industrial applications.

scholarly journals Sequential Controlled Atmosphere Storage for `McIntosh' Apples

HortScience ◽  
2006 ◽  
Vol 41 (5) ◽  
pp. 1322-1324 ◽  
Author(s):  
P. Guy Lévesque ◽  
Jennifer R. DeEll ◽  
Dennis P. Murr
Keyword(s):  
Ambient Air ◽  
First Year ◽  
Controlled Atmosphere ◽  
Ripening Period ◽  
Long Term Storage ◽  
Lower Temperature ◽  
Atmosphere Storage ◽  
Term Storage

Sequential decreases or increases in the levels of O2 in controlled atmosphere (CA) were investigated as techniques to improve fruit quality of `McIntosh' apples (Malus ×sylvestris [L.] Mill. var. domestica [Borkh.] Mansf.), a cultivar that tends to soften rapidly in storage. Precooled fruit that were harvested at optimum maturity for long-term storage were placed immediately in different programmed CA regimes. In the first year, CA programs consisted of 1) `standard' CA (SCA; 2.5–3.0% O2 + 2.5% CO2 for the first 30 d, 4.5% CO2 thereafter) at 3 °C for 180 d; 2) low CO2 SCA (2.5–3.0% O2 + 2.5% CO2) at 3 °C for 60 d, transferred to low O2 (LO; 1.5% O2 + 1.5% CO2) at 0 or 3 °C for 60 d, and then to ultralow O2 (ULO; 0.7% O2 + 1.0% CO2) at 0 or 3 °C for 60 d; and 3) ULO at 3 °C for 60 d, transferred to LO at 0 or 3 °C for 60 d, and then to SCA or low CO2 SCA at 0 or 3 °C for 60 d. In the second year, the regimes sequentially decreasing in O2 were compared with continuous ULO and SCA. After removal from storage, apples were held in ambient air at 20 °C for a 1-week ripening period. Fruit firmness was evaluated after 1 and 7 d at 20 °C, whereas the incidence of physiological disorders was assessed only after 7 d. Lowering the temperature while decreasing O2 was the best CA program with significant increased firmness retention during storage and after the 1-week ripening period. Reduced incidence of low O2 injury in decreasing O2 programs and absence of core browning at the lower temperature were also observed.

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