Bacillus velezensis Identification and Recombinant Expression, Purification and Characterization of Its Alpha-Amylase

Fermentation ◽  
2021 ◽  
Vol 7 (4) ◽  
pp. 227
Author(s):  
Xiaodong Zhang ◽  
Caixia Li ◽  
Xuantong Chen ◽  
Chonlong Chio ◽  
Sarita Shrestha ◽  
...  

Amylases account for about 30% of the global market of industrial enzymes, and the current amylases cannot fully meet industrial needs. This study aimed to identify a high α-amylase producing bacterium WangLB, to clone its α-amylase coding gene, and to characterize the α-amylase. Results showed that WangLB belonged to Bacillus velezensis whose α-amylase gene was 1980 bp coding 659 amino acids designated as BvAmylase. BvAmylase was a hydrophilic stable protein with a signal peptide and a theoretical pI of 5.49. The relative molecular weight of BvAmylase was 72.35 kDa, and was verified by SDS-PAGE. Its modeled structure displayed that it was a monomer composed of three domains. Its optimum temperature and pH were 70 °C and pH 6.0, respectively. It also showed high activity in a wide range of temperatures (40–75 °C) and a relatively narrow pH (5.0–7.0). It was a Ca2+-independent enzyme, whose α-amylase activity was increased by Co2+, Tween 20, and Triton X-100, and severely decreased by SDS. The Km and the Vmax of BvAmylase were 3.43 ± 0.53 and 434.19 ± 28.57 U/mg. In conclusion, the α-amylase producing bacterium WangLB was identified, and one of its α-amylases was characterized, which will be a candidate enzyme for industrial applications.

2013 ◽  
Vol 794 ◽  
pp. 380-390
Author(s):  
Krishnan Sivaraman ◽  
Dileep Kulkarni

Stainless steels are engineering materials capable of meeting a wide range of design criteria. They exhibit excellent corrosion resistance, strength at elevated temperature, toughness at cryogenic temperature and fabrication characteristics, and they are selected for a broad range of consumer, commercial, and industrial applications. In the fabrication of stainless steel products, components, or equipment, manufacturers employ welding as the principal joining method. Stainless steels possess good weldability and a welded joint can provide optimum corrosion resistance, strength, and fabrication economy provided reasonable care is exercised during welding. L&T's Heavy Engineering (HE) has established a reputation for quality products in the global market with its strong engineering capabilities and state-of-the-art manufacturing facilities. It manufactures and supplies various critical equipments like reactors, vessels, heat exchangers and inter-connecting piping to Fertilizer, Refinery, Petrochemical, Chemical, Oil & Gas, Power, Nuclear and allied Strategic sectors. The wide spectrum of equipments mentioned involves fabrication of various grades of Stainless Steel (SS)like Austenitic, Ferritic, Martensitic, Duplex, Super Duplex etc. This paper discusses some of the high productivity welding processes and the techniques being used in manufacturing Stainless Steel vessels at Larsen & Toubro’s Heavy Engineering such as: Narrow groove welding of high thickness SS joints by Submerged Arc Welding(SAW), High deposition SS weld surfacing using Electro Slag Strip Cladding (ESSC), Hotwire GTAW for joining & surfacing of SS, SS Liner welding by GTAW for critical Urea Service applications, Automatic Tube to Tube sheet Welding etc.


2021 ◽  
Vol 28 ◽  
Author(s):  
Barış Enez

Background: Amylases are used in several industrial and biotechnological sectors, including those producing textiles, detergents, paper and bakery products. Objective: This study aimed to purify an industrially important α-amylase from Bacillus sp. For this purpose, a single and rapid α-amylase purification was performed using the starch affinity method. Methods: Characterization of the purified enzyme was determined by investigating temperature, pH stability, detergents, and metal ions. Results: The purification coefficient of 29.8-fold with a yield of 9.2% was found. The molecular weight of the purified α-amylase was determined to be 53 kDa by SDS-PAGE, and thermostability was confirmed with 100% activity at 30ºC and 40ºC after 1 h. The purified enzyme was stable over a wide range of pH values, with optimum activity at pH 6.0, 7.0 and 8.0 after 2 h. The study also investigated the effects of the metal ions and detergents on the purified amylase and found that Mg2+ and Ca2+ ions were the activators of the enzyme, while Zn2+, Co2+ and Na+ ions decreased the activity. Furthermore, Hg2+ indicated complete inhibition of amylase activity. The detergents Triton X-100 and Tween 20 increased the α-amylase activity, while sodium dodecyl sulfate inhibited the activity. Conclusion: The purified α-amylase obtained from Bacillus sp. is considered to be environmentally friendly, can be processed in a short time, and has a low cost.


2017 ◽  
Vol 2017 ◽  
pp. 1-7 ◽  
Author(s):  
Oliyad Jeilu Oumer ◽  
Dawit Abate

The demand for enzymes in the global market is projected to rise at a fast pace in recent years. There has been a great increase in industrial applications of pectinase owing to their significant biotechnological uses. For applying enzymes at industrial scale primary it is important to know the features of the enzyme. Thus, this study was undertaken with aims of characterizing the pectinase enzyme from Bacillus subtilis strain Btk27 and proving its potential application in demucilisation of coffee. In this study, the maximum pectinase activity was achieved at pH 7.5 and 50°C. Also, the enzyme activity was found stimulated with Mg2+ and Ca2+ metal ions. Moreover, it was stable on EDTA, Trixton-100, Tween 80, and Tween 20. Since Bacillus subtilis strain Btk27 was stable in most surfactants and inhibitors it could be applicable in various industries whenever pectin degradation is needed. The enzyme Km and Vmax values were identified as 1.879 mg/ml and 149.6 U, respectively. The potential application of the enzyme for coffee processing was studied, and it is found that complete removal of mucilage from coffee beans within 24 hours of treatment indicates the potential application in coffee processing.


2019 ◽  
Author(s):  
James Ewen ◽  
Carlos Ayestaran Latorre ◽  
Arash Khajeh ◽  
Joshua Moore ◽  
Joseph Remias ◽  
...  

<p>Phosphate esters have a wide range of industrial applications, for example in tribology where they are used as vapour phase lubricants and antiwear additives. To rationally design phosphate esters with improved tribological performance, an atomic-level understanding of their film formation mechanisms is required. One important aspect is the thermal decomposition of phosphate esters on steel surfaces, since this initiates film formation. In this study, ReaxFF molecular dynamics simulations are used to study the thermal decomposition of phosphate esters with different substituents on several ferrous surfaces. On Fe<sub>3</sub>O<sub>4</sub>(001) and α-Fe(110), chemisorption interactions between the phosphate esters and the surfaces occur even at room temperature, and the number of molecule-surface bonds increases as the temperature is increased from 300 to 1000 K. Conversely, on hydroxylated, amorphous Fe<sub>3</sub>O<sub>4</sub>, most of the molecules are physisorbed, even at high temperature. Thermal decomposition rates were much higher on Fe<sub>3</sub>O<sub>4</sub>(001) and particularly α-Fe(110) compared to hydroxylated, amorphous Fe<sub>3</sub>O<sub>4</sub>. This suggests that water passivates ferrous surfaces and inhibits phosphate ester chemisorption, decomposition, and ultimately film formation. On Fe<sub>3</sub>O<sub>4</sub>(001), thermal decomposition proceeds mainly through C-O cleavage (to form surface alkyl and aryl groups) and C-H cleavage (to form surface hydroxyls). The onset temperature for C-O cleavage on Fe<sub>3</sub>O<sub>4</sub>(001) increases in the order: tertiary alkyl < secondary alkyl < primary linear alkyl ≈ primary branched alkyl < aryl. This order is in agreement with experimental observations for the thermal stability of antiwear additives with similar substituents. The results highlight surface and substituent effects on the thermal decomposition of phosphate esters which should be helpful for the design of new molecules with improved performance.</p>


Alloy Digest ◽  
1970 ◽  
Vol 19 (11) ◽  

Abstract PLATINUM is a soft, ductile, white metal which can be readily worked either hot or cold. It has a wide range of industrial applications because of its excellent corrosion and oxidation resistance and its high melting point. This datasheet provides information on composition, physical properties, hardness, elasticity, and tensile properties. It also includes information on high temperature performance and corrosion resistance as well as forming, heat treating, machining, and joining. Filing Code: Pt-1. Producer or source: Matthey Bishop Inc..


2020 ◽  
Vol 12 ◽  
Author(s):  
Nihar Ranjan Biswal

Background: Surfactant adsorption at the interfaces (solid–liquid, liquid–air, or liquid–liquid) is receiving considerable attention from a long time due to its wide range of practical applications. Objective: Specifically wettability of solid surface by liquids is mainly measured by contact angle and has many practical importances where solid–liquid systems are used. Adsorption of surfactants plays an important role in the wetting process. The wetting behaviours of three plant-based natural surfactants (Reetha, Shikakai, and Acacia) on the glass surface are compared with one widely used nonionic synthetic surfactant (Triton X-100) and reported in this study. Methods: The dynamic contact angle study of three different types of plant surfactants (Reetha, Shikakai and Acacia) and one synthetic surfactant (Triton X 100) on the glass surface has been carried out. The effect of two different types of alcohols such as Methanol and amyl alcohol on wettability of shikakai, as it shows little higher value of contact angle on glass surface has been measured. Results: The contact angle measurements show that there is an increase in contact angle from 47° (pure water) to 67.72°, 65.57°, 68.84°, and 68.79° for Reetha, Acacia, Shikakai, and Triton X-100 respectively with the increase in surfactant concentration and remain constant at CMC. The change in contact angle of Shikakai-Amyl alcohol mixtures are slightly different than that of methanol-Shikakai mixture, mostly there is a gradual increase in contact angle with the increasing in alcohol concentration. Conclusion: There is no linear relationship between cos θ and inverse of surface tension. There was a linear increase in surface free energy results with increase in concentration as more surfactant molecules were adsorbing at the interface enhancing an increase in contact angle.


2020 ◽  
Vol 15 (1) ◽  
pp. 787-796 ◽  
Author(s):  
Marek Kieliszek ◽  
Kamil Piwowarek ◽  
Anna M. Kot ◽  
Katarzyna Pobiega

AbstractCellular biomass of microorganisms can be effectively used in the treatment of waste from various branches of the agro-food industry. Urbanization processes and economic development, which have been intensifying in recent decades, lead to the degradation of the natural environment. In the first half of the 20th century, problems related to waste management were not as serious and challenging as they are today. The present situation forces the use of modern technologies and the creation of innovative solutions for environmental protection. Waste of industrial origin are difficult to recycle and require a high financial outlay, while the organic waste of animal and plant origins, such as potato wastewater, whey, lignin, and cellulose, is dominant. In this article, we describe the possibilities of using microorganisms for the utilization of various waste products. A solution to reduce the costs of waste disposal is the use of yeast biomass. Management of waste products using yeast biomass has made it possible to generate new metabolites, such as β-glucans, vitamins, carotenoids, and enzymes, which have a wide range of industrial applications. Exploration and discovery of new areas of applications of yeast, fungal, and bacteria cells can lead to an increase in their effective use in many fields of biotechnology.


Polymers ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1711
Author(s):  
Heba A. Gad ◽  
Autumn Roberts ◽  
Samirah H. Hamzi ◽  
Haidy A. Gad ◽  
Ilham Touiss ◽  
...  

Jojoba is a widely used medicinal plant that is cultivated worldwide. Its seeds and oil have a long history of use in folklore to treat various ailments, such as skin and scalp disorders, superficial wounds, sore throat, obesity, and cancer; for improvement of liver functions, enhancement of immunity, and promotion of hair growth. Extensive studies on Jojoba oil showed a wide range of pharmacological applications, including antioxidant, anti-acne and antipsoriasis, anti-inflammatory, antifungal, antipyretic, analgesic, antimicrobial, and anti-hyperglycemia activities. In addition, Jojoba oil is widely used in the pharmaceutical industry, especially in cosmetics for topical, transdermal, and parenteral preparations. Jojoba oil also holds value in the industry as an anti-rodent, insecticides, lubricant, surfactant, and a source for the production of bioenergy. Jojoba oil is considered among the top-ranked oils due to its wax, which constitutes about 98% (mainly wax esters, few free fatty acids, alcohols, and hydrocarbons). In addition, sterols and vitamins with few triglyceride esters, flavonoids, phenolic and cyanogenic compounds are also present. The present review represents an updated literature survey about the chemical composition of jojoba oil, its physical properties, pharmacological activities, pharmaceutical and industrial applications, and toxicity.


Polymers ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1058
Author(s):  
Hikaru Okubo ◽  
Haruka Kaneyasu ◽  
Tetsuya Kimura ◽  
Patchiya Phanthong ◽  
Shigeru Yao

Each year, increasing amounts of plastic waste are generated, causing environmental pollution and resource loss. Recycling is a solution, but recycled plastics often have inferior mechanical properties to virgin plastics. However, studies have shown that holding polymers in the melt state before extrusion can restore the mechanical properties; thus, we propose a twin-screw extruder with a molten resin reservoir (MSR), a cavity between the screw zone and twin-screw extruder discharge, which retains molten polymer after mixing in the twin-screw zone, thus influencing the polymer properties. Re-extruded recycled polyethylene (RPE) pellets were produced, and the tensile properties and microstructure of virgin polyethylene (PE), unextruded RPE, and re-extruded RPE moldings prepared with and without the MSR were evaluated. Crucially, the elongation at break of the MSR-extruded RPE molding was seven times higher than that of the original RPE molding, and the Young’s modulus of the MSR-extruded RPE molding was comparable to that of the virgin PE molding. Both the MSR-extruded RPE and virgin PE moldings contained similar striped lamellae. Thus, MSR re-extrusion improved the mechanical performance of recycled polymers by optimizing the microstructure. The use of MSRs will facilitate the reuse of waste plastics as value-added materials having a wide range of industrial applications.


2021 ◽  
Vol 9 (2) ◽  
pp. 336
Author(s):  
Laura Matarredona ◽  
Mónica Camacho ◽  
Basilio Zafrilla ◽  
Gloria Bravo-Barrales ◽  
Julia Esclapez ◽  
...  

Haloarchaea can survive and thrive under exposure to a wide range of extreme environmental factors, which represents a potential interest to biotechnology. Growth responses to different stressful conditions were examined in the haloarchaeon Haloferax mediterranei R4. It has been demonstrated that this halophilic archaeon is able to grow between 10 and 32.5% (w/v) of sea water, at 32–52 °C, although it is expected to grow in temperatures lower than 32 °C, and between 5.75 and 8.75 of pH. Moreover, it can also grow under high metal concentrations (nickel, lithium, cobalt, arsenic), which are toxic to most living beings, making it a promising candidate for future biotechnological purposes and industrial applications. Inductively Coupled Plasma Mass Spectrometry (ICP-MS) analysis quantified the intracellular ion concentrations of these four metals in Hfx. mediterranei, concluding that this haloarchaeon can accumulate Li+, Co2+, As5+, and Ni2+ within the cell. This paper is the first report on Hfx. mediterranei in which multiple stress conditions have been studied to explore the mechanism of stress resistance. It constitutes the most detailed study in Haloarchaea, and, as a consequence, new biotechnological and industrial applications have emerged.


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