Production, Physical Optimization, and Purification of Xylanase Enzyme from Bacillus sp. by Solid State Fermentation | Chapter 10 | Microbiology and Biotechnology Research: An Overview Vol. 1

Aim: Xylanases are extensively applied in paper and pulp industries as well as during the preparation of baked products to improve their quality. Additionally, it is also used in coffee, oil, and starch industries in order to increase their nutritional values.

Place and Duration of the Study: Azyme Biosciences Laboratory, Bangalore. An experiment was conducted between January 2021 and June 2021.

Methodology: In the present study, soil samples were collected near sawmills in various localities of urban Bangalore to isolate organisms for the production of xylanase using solid-state fermentation. Six organisms were isolated using selective media based on their morphological characters. Among them, one organism showed maximum production of xylanase enzyme identified as Bacillus sp based on their biochemical test and 16s RNA sequencing. Solid substrate fermentation was carried out using various agro wastes such as sugar cane bagasse; sawdust, paddy husk, wheat straw, and orange peel powder.

Results: In the results, production revealed that sugarcane bagasse showed maximum production of enzyme Bacillus sp compared to other substrates with different physical parameters such as pH 8, and temperature at 35 ^ºC after 72 hrs incubation. Trace element such as Mg⁺⁺ enhances the production of enzymes by more than 22% compared with other metal ions like Ca⁺⁺, Mn⁺⁺ and Fe⁺⁺. After production, the enzyme is purified by using three-step methods ammonium sulphate precipitation, dialysis, ion exchange, and gel filtration. Fold purification was increased up to 12 fold, yielding 36% and the molecular weight of the enzyme was 62 KDa determined using SDS PAGE.

Conclusion: Bacillus sp shown in this experiment is capable of producing xylanase enzyme when subjected to different physical parameters.

 

Author (s) Details

Shylesha, B.S.
Department of Chemistry, Sree Siddaganga College of Arts, Science and Commerce, Tumakuru-572102, Karnataka, India.

 

Mahesh, M.
Department of Biotechnology, Azyme Biosciences Private Limited, Bengaluru-560069, Karnataka, India.

 

Shubharani, R
Department of Biotechnology, Azyme Biosciences Private Limited, Bengaluru-560069, Karnataka, India.

 

V.N. Yoganandamurthy
Department of Biotechnology, Azyme Biosciences Private Limited, Bengaluru-560069, Karnataka, India.

 

Please see the book here:- https://doi.org/10.9734/bpi/mbrao/v1/4021

 

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

Extraction and Purification of C-Phycocyanin from Spirulina Powder and Evaluating its Anti-oxidant, Anti-coagulant and DNA Damage Prevention Properties | Chapter 2 | Advanced Concepts in Pharmaceutical Research Vol. 6

C-Phycocyanin (C-PC) is a phycobiliprotein found in blue-green algae, such as Spirulina platensis which is often used as a dietary nutritional supplement and exhibits a variety of pharmacological properties. Cyanobacteria and algae possess a wide range of colored compounds, including chlorophyll, carotenoids and phycobiliproteins. In this regard, extraction, partial purification, antioxidant, anticoagulation and prevention of DNA damage activity of C-PC were investigated. In the present study, a simple and efficient method to extract C-PC from Spirulina platensis dry powder is reported. Spirulina platensis dry powder was obtained from NB Laboratories Pvt. Ltd., Nagpur, India. The extractions were carried out using two different methods: cold maceration and sonication method. The extraction using the cold maceration method proved to be the most efficient method. The efficiency of extraction methods was determined by calculating the concentration and purity ratio of isolated C-PC. Obtained crud C-PC was purified by ammonium sulphate precipitation, dialysis and gel filtration and presented a final extraction yield of 3.27±0.09 mg/ml with a purity ratio of 2.317±0.08. When it was evaluated as an antioxidant in vitro, it was able for scavenging nitric oxide. SDS-PAGE results show that the 40% ammonium sulphate precipitated sample and dialyzed sample show two bands of C-PC i.e. α and β with 17 and 19kDa respectively along with other proteins. C-PC showed significant anticoagulation and prevention of DNA damage activity. C-PC is a significant contributor to DNA damage prevention by scavenging of –OH.

Author(s) Details:

Suresh P. Kamble,
Department of Biochemistry, Moolji Jaitha College (Autonomous), Jalgaon, Maharashtra, India.

Please see the link here: https://stm.bookpi.org/ACPR-V6/article/view/13412

Protein Pattern of Phoenix dactylifera Seeded Strains Grown in Basrah Region Using Cluster and Principal Component Analysis: Brief Overview | Chapter 10 | Advances and Trends in Agricultural Sciences Vol. 4

The present study was carried out on 17 seeded strains of date palm “Phoenix dactylifera L.” grown in Basrah region to investigate variations in protein pattern of leaflets and fruits among these strains. Cluster and principal component analysis were used to identify the degree of similarity and differences between date palm strains. The leaflet and fruit protein pattern on SDS-PAGE showed differences in band number, location, thickness and density leading to variations in protein quantity and quality among the studied strains. Cluster analysis data showed a high degree of similarity between leaflet and fruit samples of these strains recording an average similarity value of 0.76 and 0.67 for leaflet and fruit samples respectively. Cluster analysis also revealed that all strains occupied two main groups and then distributed into two subgroups under each main group. The principal component analysis showed that protein banding characters for leaflets and fruits could be used for identification of date palm seeded strains.

Author (s) Details

Ali H. M. Attaha
Department of Horticulture and Landscape, Agriculture College, Basrah University, Basrah, Iraq.

Taha Y. Mhoder
Department of Horticulture and Landscape, Agriculture College, Basrah University, Basrah, Iraq.

Abdul-Kareem M. Abd
Department of Horticulture and Landscape, Agriculture College, Basrah University, Basrah, Iraq

View Book :- http://bp.bookpi.org/index.php/bpi/catalog/book/168