Exploring the Potential of α-Amylase: Genetic Identification, Purification Techniques and Its Applications | Chapter 7 | Contemporary Research and Perspectives in Biological Science Vol. 5

α-Amylase is a key enzyme widely employed in various industrial applications, including foodstuffs, fabric goods, and biofuel production. In our review study, we focused on genetically characterized microbial isolates that produce α-amylase from diverse sources. DNA isolation was performed using an alkaline lysis protocol followed by PCR amplification of the 16S rRNA gene. After isolating DNA samples, the 16S rRNA gene was PCR-amplified, and the phylogenetic relationships were determined through sequencing and subsequent analysis. To optimize enzyme production, various parameters such as pH, temperature, and incubation time were investigated. To purify the α-amylase enzyme, several purification techniques were employed. The crude amylase enzyme preparation was precipitated using ammonium sulfate. After dialysis and size exclusion chromatography, the purified enzyme preparation was analyzed further. To assess purity, the dialyzed samples were analyzed using a suitable chromatographic method to separate and evaluate the sample components. α-amylase characterization involved molecular weight determination by using SDS-PAGE, size determination by Size exclusion chromatography, and comprehensive examination was done by using mass spectrometry. In conclusion, this study successfully compared the genetically identified and characterized α-amylase producing microbial isolates. The optimized incubation conditions and purification techniques provided valuable insights into the production and purification of this industrially important enzyme. α-Amylase enzymes play an essential role in several industrial processes, improving and facilitating eco-friendly methods in areas such as textile, food, pharmaceutical, and alcohol manufacturing. Their adaptability drives cross-sector, sustainable innovations.

Further studies can focus on enhancing the enzyme's characteristics and exploring its potential applications in various industries.

 

Author (s) Details

 

Arvind Sharma
Department of Zoology, B.K.S.N. Govt. College Shajapur (M.P.), India.

 

R.K. Sharma
Department of Botany, B.K.S.N. Govt. College Shajapur (M.P.), India.

 

Please see the book here:- https://doi.org/10.9734/bpi/crpbs/v5/2929

Seasonal Variation in Physico-chemical Parameters of Groundwater in Yerraguntla Area of YSR Kadapa DT, Andhra Pradesh, India | Chapter 4 | Chemistry and Biochemistry: Research Progress Vol. 1

People around the globe are in tremendous trouble because of unexpected changes in the physical, chemical, and natural states of the air, water, and soil. Due to the increased mortal population, industrialization, use of diseases, and man-made exertion, water is heavily contaminated with different dangerous pollutants. Disposing of contaminated jewels, filtering soils, and mining processing lead to contamination of natural water. The quality of drinking water must be checked regularly. Because of this contaminated drinking water, the mortal population suffers from varied forms of waterborne diseases. The vacuity of good quality water is a necessary point for precluding the conditions and perfecting the quality of life. It is necessary to know the details about different Physicochemical parameters such as Hydrogen ion concentration, Electrical Conductivity, Total Dissolved Solids, Total Hardness, Calcium, Magnesium, Sodium, Potassium, Bicarbonate, Chloride, Sulphate, Phosphate, Dissolved Oxygen, Biochemical Oxygen Demand, Chemical Oxygen Demand, etc. for testing water quality. Yerraguntla area of YSR Kadapa district was selected to determine the groundwater quality in both rainy and summer seasons. In the present study, the results showed that most of the Physico-chemical parameters exceeded the admissible limits of WHO. The water Quality Index showed that groundwater is not suitable for drinking purposes in all locales of the study area. The correlation measure showed the inter-relevance between various parameters that impact groundwater quality. Piper tri direct illustration showed that the groundwater of the area contains Ca-HCO₃ type of mariners. So, it is necessary to adopt sound water sanitation methods in the study area.

 

Author (s) Details

 

Suresh Pulapaka

Department of Chemistry, SRR & CVR Govt. Degree College(A), Vijayawada, A.P., India.

 

Please see the book here:- https://doi.org/10.9734/bpi/cbrp/v1/3918

An Overview of the Research and Application of Various Activated Carbons and Ash Used in Water Purification Technology | Chapter 7 | Emerging Issues in Environment, Geography and Earth Science Vol. 1

 Water is an essential aptitude that nature gives to all living individual. In order to keep all history safe on soil, it is necessary to keep the amount of water in the corpse adequate and enough at all times. Water pollution is growing as a result of differing factors such as study of human population, large-scale urbanization, clear-cutting, and unfair activities in waterways and other water beginnings. Various experts have worked on the result of activated element from renewable energy. This comprises economical technology and merchandise in an eco-friendly way for a assortment of uses. This review article discusses the differing methods secondhand by experts to develop triggered carbon so that minimize water pollution. Peanut coverings, Moringa oleifera seeds, Coconut structures, Rice husk, Lemon peel, Pomegranate peel and Banana peel are all excellent natural resources for treating poisoned water. It was demonstrated that pollutants to a degree pH, colour, DO, conductivity, turbidity, chloride, fluoride, TSS, TDS, BOD, COD, nitrate, phosphate, burdensome metals, and total coliforms can be isolated from water utilizing physical, machinelike, and biological techniques.

Author(s) Details:

Ajinkya Ravindra Telgote,
Department of Environmental Sciences, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad, India.

Satish S. Patil,
Department of Environmental Sciences, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad, India.

Please see the link here: https://stm.bookpi.org/EIEGES-V1/article/view/11827

Purification and Characterization of Organic Solvent Tolerant Lipase from Newly Isolated Pseudomonas aeruginosa FW_SH-1 | Chapter 3 | Novel Aspects on Chemistry and Biochemistry Vol. 3

 In the work, a novel solvent easygoing extracellular lipase from newly unique Pseudomonas aeruginosa FW_SH-1 was purified by acetone sleet and ion exchange chromatography. The purified lipase was characterized and distinguished with lipases got from other bacterial strains.  A recently isolated strain that can produce basic solvent open-minded lipase was the aim of the present study, the strain was named as FW_SH-1, that was identified as a Pseudomonas sp. The extracellular lipase from Pseudomonas aeruginosa FW_SH-1 was purified through acetone storm followed by Q-sepharose anion exchange chromatography. A ending specific exercise of 572 U/mg was achieved accompanying 26.12 fold purification and an overall improvement of 10.21%. The molecular pressure of the purified lipase was raise to be 66 kDa using SDS polyacrylamide coagulate electrophoresis. The optimum pH and hotness for the activity of the freed lipase was found expected 8.0 and 45 oC. In the presence of hydrophilic solvent like methanol, flammable liquid, and propanol, the purified lipase remained resistant. The presence of Ca2+, Mg2+, Na+, K+ and Tween 20 aroused the activity of the freed lipase.  The metal ions to a degree Ca2+, Mg2+, Na+, K+ and Tween (20 and 80) as surfactants were found to reinforce the relative activity. The basic solvents in the way that methanol, ethanol and propanol had a very little effect in deactivating the lipase.

Author(s) Details:

Chaudhry Haider Ali,
Department of Chemical Polymer and Composite Materials Engineering, University of Engineering & Technology, New Campus, KSK Lahore-54890, Pakistan and State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai, China.

Abdul Sattar Qureshi,
Institute of Biotechnology and Genetic Engineering, University of Sindh, Jamshoro, Sindh, Pakistan.

Serge Maurice Mbadinga,
State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai, China.

Jin-Feng Liu,
State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai, China.

Shi-Zhong Yang,
State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai, China.

Bo-Zhong Mu,
State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai, China.

Please see the link here: https://stm.bookpi.org/NACB-V3/article/view/11022

Aspergillus fumigatus Catalase | Chapter 3 | Research Advances in Microbiology and Biotechnology Vol. 6

 Catalases are antioxidant enzymes that movement the cleavage of hydrogen whiten into water and molecular oxygen and are widely distributed in type. There is little information in the composition on the kinetic and biochemical properties of catalases created by fungi. Therefore, this study aimed to disengage the catalase enzyme from Aspergillus fumigatus, that is known to be a good catalase builder, and to determine allure properties. Optimal catalase production was worked out on the seventh day of culturing containers grown at 37o C and 155 rpm in excellent-liter YpSs medium containing 1% (w/v) glucose and 0.5 mM H2O2. The catalyst was successfully freed 24-fold with a 55% recovery. The microscopic weight was driven to be ~70 kDa at SDS-PAGE. The optimum reaction temperature of the something which incites activity was set at 60°C and pH at 7.0. Km and Vmax principles were calculated as 7.4 mM and 1250 μ M min-1, individually. Stability tests showed that the something which incites activity could wait active in a wide pH range (4.0-9.0). The warm stability of catalase was 'tween 30o C and 50o C. The enzyme was likewise stable to various solvent such as intoxicating, methanol, acetone, and dimethyl sulfoxide depending on the aggregation and incubation opportunity. The biochemical properties of the something which incites activity (low Km, stability towards various pH and organic solvent, etc.) indicate that A. fumigatus catalase can serve as a good biocatalyst for differing industrial uses.

Author(s) Details:

Ceren Aggez,
Department of Biology, Institute of Natural and Applied Sciences, Kocaeli University, Kocaeli-41001, Turkey.

Yonca Yuzugullu Karakus,
Department of Biology, Faculty of Arts and Sciences, Kocaeli University, Kocaeli-41001, Turkey.

Please see the link here: https://stm.bookpi.org/RAMB-V6/article/view/10920

Increase in Laccases Production by an Improved Strain of White Rot Fungi Pycnoporus cinnabarinus with an Additional Improvident in the Production Medium | Chapter 8 | Research Advances in Microbiology and Biotechnology Vol. 2

 Several microorganisms containing fungi and bacteria produce ultimate important industrially applicable catalyst “laccases”. The present work was aimed to request mutagenesis on the test fungus Pycnoporus cinnabarinus for enhancement of the catalyst production and further increase in the result by supplying an improvident in the television. The efficiency of Laccases result by the wild fungal strains Pycnoporus cinnabarinus was examined by the situation with physical cancer-causing agent [ultraviolet radiation (UV) and X-beams] and chemical mutagens [Ethidium bromide, Colchicine and Hydrogen whiten]. The effect of X-rays presented an increase in production with growing exposure (Max. at 8 brief time period.). Beyond 8 seconds there was a decrease in result. UV irradiation influenced lowering the enzyme result and the maximum dosage is lethal to the tumor. Among the three chemical mutagens, hydrogen whiten was found to have lethal belongings to the fungus and reduced enzyme production even in minimum concentrations. Colchicine and Ethidium platitude showed an increase in substance causing chemicals to split into simpler substances production with growing concentrations (Max. at 4 and 7 g respectively). The upgraded strain of Pycnoporus cinnabarinus showed a 15% of increase in the yield. The increase in production of laccases in a low production medium planned by using agricultural and technical wastes may be advantageous industrially when compared to the different costly conventional radio. Further, mutants obtained by original fusion resulted in even more result.Since ancient times, land and industrial wastes have been secondhand for production of laccases by Basidiomycetes in larger quantity, but the usage of aforementioned wastes is still not up to the mark. In this study, Ghee slag (Dairy waste) and wheat bran (land waste) in the laccases production medium (Potato organic compound composed of carbon broth) which is inferior and efficient when distinguished to the costly chemical located conventional television with an upgraded strain of Pycnoporus cinnabarinus (KX8, Accession number 56259) obtained by physical or synthetic mutagenesis followed by intraspecific original fusion found expected beneficial. In vitro bioassay of the paternal P.cinnabarinus (154.65U/ml/min), its best mutation (165.219U/ml/min) and fusant (176.14U/ml/brief time period) indicated a gradual increase in result of laccases by 17.59%. It was purified utilizing ammonium sulphate precipitation, dialysis, and anion exchange chromatography, flexible 23.86%. Thus, intraspecific protoplast mixture techniques can be working to develop superior composite strains of laccases producing fungi as effective forms for increasing modern productivity.

Author(s) Details:

Rasheeda Khanam,
Department of Microbiology, GITAM Institute of science, GITAM University, Visakhapatnam, India.

Please see the link here: https://stm.bookpi.org/RAMB-V2/article/view/9422

Study on Purification and Some Properties of Endo-1,4-β-Glucanases of Trichoderma harzianum UzCF-28| Chapter 2 | Current Overview on Science and Technology Research Vol. 9

 The purpose concerning this study was to isolate, free, and study the biochemical properties of cellulolytic enzymes synthesised apiece Trichoderma harzianum UzCF-28 strain. During submerged cultivation in liquid civilization on modified Mandels fiber medium with wheat hay as a carbon beginning, strain UzCF-28 demonstrated high cellulolytic exercise. When enzyme exercise was studied dynamically for 10 days, maximum activity endo-1,4-β-glucanase (5.2 wholes/ml) and xylanase (7.65 units/ml) of the gelled waste detected after 3 days of culturing. High action of exo-1,4-β-glucanase was observed eventually 4 and was found to be 2.96 wholes/ml. As a result of purification by precipitation accompanying ammonium sulfate and further ion exchange chromatography, two isoforms of endo-1,4-β-glucanase-EG II and EG III with microscopic weight of 135 and 75 kDa respectively were told. The pH optimum for EG I and EG III was 4.5, while EG II was 4.7, however the substrate used—CMC or "Whatman filter" paper.  Heating up to 40oC of EG III acted not result in allure inactivation, but rather increased allure activity by in addition three times when compared to the beginning activity of the substance causing chemicals to split into simpler substances, indicating that the thermostability of EG III among proven enzymes varied considerably.

Author(s) Details:

N. Sh. Azimova,
Institute of Microbiology of Uzbekistan Academy of Sciences, Tashkent, Republic of Uzbekistan.

Kh. M. Khamidova,
Institute of Microbiology of Uzbekistan Academy of Sciences, Tashkent, Republic of Uzbekistan.

M. B. Djumagulov,
Institute of Microbiology of Uzbekistan Academy of Sciences, Tashkent, Republic of Uzbekistan.

Z. S. Shakirov,
Institute of Microbiology of Uzbekistan Academy of Sciences, Tashkent, Republic of Uzbekistan.

Please see the link here: https://stm.bookpi.org/COSTR-V9/article/view/8759

Study about Isolation, Purification and Partial Characterization of Thermostable Serine AlkalineProtease from a Newly Isolated Bacillus thuringinsis | Chapter 8 | Innovations in Microbiology and Biotechnology Vol. 5

The isolation, purification, and partial characterisation of thermostable serine alkaline protease generated by Bacillus thuringiensis were described in this study. Using ribosomal sequence analysis, the culture was isolated from slaughterhouse waste soil and identified. Using ammonium sulphate precipitation, dialysis, and Sephadex G-200 gel permeation chromatography, the crude enzyme was purified up to 17.04 fold with an 8.47 percent recovery. The relative molecular weight of the isolated enzyme was determined using SDS-PAGE (67 kDa). After 48 hours of incubation at 45°C, the maximal enzyme and cell biomass output was reported. At pH 10 to 11, enzyme activity was high, and alkaline protease was confirmed after incubation for up to 2 and 20 hours at the same pH range. The best temperature for protease activity was 45°C, with thermal stability of 100 percent evaluated after 350 minutes of incubation. Among the natural substrates studied, casein was shown to be the best. Metal ions such as Ca+2, Mg+2, and Mn+2 significantly increased enzyme activity, but phenylmethylsulphonyl fluoride (PMSF) reduced enzyme activity completely, and diisopropyl fluorophosphates (DFP) decreased enzyme activity up to 92 percent, confirming serine protease. The detergent compatibility of the enzyme was tested at 45°C with 10 mM CaCl2 and 1 M glycine. This indicates 80 to 100 percent stability after 0.5 to 2.5 hours of incubation. The washing performance and removal of blood spots from the cotton material improved after using the detergent Surf excel with enzyme. Overall, the isolated protease's properties justify commercial use in the detergent and leather industries. in a bioreactor for the removal of POPs from wastewater

Author (s) Details

Sunil L. Harer
Department of Pharmaceutical Chemistry, Dattakala College of Pharmacy, Swami Chincholi, Daund, Pune 413130 (MS), India.

Priya S. Harer
Department of Pharmaceutics, PES’s, Modern College of Pharmacy, Yamunanagar, Nigdi, Pune 411044 (MS), India.

View Book :- https://stm.bookpi.org/IMB-V5/article/view/6569

Plant Genomic DNA Isolation: An Important Technology for Marker Assisted Selection | Chapter 7 | Recent Progress in Plant and Soil Research Vol. 4

 Plant biotechnology is required to increase the chances of crop enhancement projects succeeding. DNA marker technology, which is derived from molecular genetics and genomics research, holds a lot of potential for plant breeding. In plant breeding, marker-assisted selection improves efficiency and precision. Mapping and marker-assisted selection of a certain characteristic necessitates high-quality DNA from a large number of plants in a short amount of time. Many techniques for extracting plant DNA have been developed and described, with the primary goal of developing a relatively rapid, inexpensive, and repeatable way to extract good quality DNA. All of these DNA extraction protocols follow the same general principle of disrupting the cell wall, cell membrane, and nuclear membrane to release highly intact DNA into solution while ensuring that contaminating biomolecules such as proteins, polysaccharides, lipids, phenols, and other secondary metabolites are removed using enzymatic or chemical methods. For marker assisted selection applications in crop improvement as well as sequencing-based applications, high quality DNA is a must. Plant Genomic DNA isolation and quantification procedures for several crops were included in this article.

Author(S) Details

Sushma Tiwari
Department of Plant Molecular Biology and Biotechnology, College of Agriculture, Rajmata Vijayraje Scindia Agricultural University, Gwalior, Madhya Pradesh, India.

M. K. Tripathi
Department of Plant Molecular Biology and Biotechnology, College of Agriculture, Rajmata Vijayraje Scindia Agricultural University, Gwalior, Madhya Pradesh, India.

R. S. Tomar
Rani Laxmibai Central Agricultural University, Jhansi, India.

Ashok Ahuja
Department of Plant Molecular Biology and Biotechnology, College of Agriculture, Rajmata Vijayraje Scindia Agricultural University, Gwalior, Madhya Pradesh, India.

View Book:- https://stm.bookpi.org/RPPSR-V4/article/view/5198

The Influence of Concentration and Acid Type on the Acidification Process for Increasing Glycerol Concentration and Its Use as a Bio Additive | Chapter 2 | Advanced Aspects of Engineering Research Vol. 15

 As the demand for biodiesel grew, so did the amount of glycerol produced as a by-product. Purifying crude glycerol and converting it into a more valuable product takes time and effort. Crude glycerol as a by-product could not be isolated from the rise in biodiesel production. One of these is by the acetylation process, which converts glycerol into triacetin, which can be utilised as a bio additive in gasoline. The goal of this research is to see how the concentration and kind of acid used in glycerol purification affect the results. Acidification, adsorption, filtration, distillation, and evaporation were used to purify the water. H3PO4, H2SO4, and HCl were used in the acidification processes in percentages of 0.5 percent, 1 percent, 2.5 percent, 5 percent, 7.5 percent, and 10%. The purified glycerol was analysed for glycerol content, water content, density, and viscosity before being converted using Ni/Zeolite as a catalyst in an acetylation reaction. The reaction products were analysed using a NaOH titration to determine the amount of unreacted acetic acid that was converted to reaction conversion. It can be concluded that the addition of 5% H3PO4 resulted in the highest glycerol concentration, with a glycerol content of 79.59 percent and a glycerol conversion in the acetylation reaction of 82.39 percent. It can also boost the Octane Number of commercial gasoline by 6.5 percent when used as a bio additive.

Author (S) Details

H. Dewajani
Chemical Engineering Program, Politeknik Negeri Malang, Jalan Sukarno Hatta 9, Malang, 65141, Indonesia.

T. Susanti
Chemical Engineering Program, Politeknik Negeri Malang, Jalan Sukarno Hatta 9, Malang, 65141, Indonesia.

D. Pratiwi
Chemical Engineering Program, Politeknik Negeri Malang, Jalan Sukarno Hatta 9, Malang, 65141, Indonesia.

View Book :- https://stm.bookpi.org/AAER-V15/article/view/1738