Skip to main content

Purification and Characterization of Pectin Methylesterase Produced in Solid State Fermentation by Aspergillus tubingensis | Chapter 03 | Advances and Trends in Biotechnology and Genetics Vol. 2

Aim: Purification and characterization of pectin methylesterase produced by Aspergillus tubingensis in solid state fermentation.

Study Design:  Pectin methylesterase enzyme produced by A. tubingensis was extracted from the fermented solid medium and purified using chromatographic techniques. The purified enzyme was characterized for physico-chemical and kinetic properties.

Place and Duration of Study: Experiments were performed at the School of Biotechnology, Devi Ahilya University, Indore, INDIA and Maharaja Ranjit Singh College of Professional Sciences, Indore, INDIA, between October, 2014 and August, 2015.

Methodology: The enzyme was extracted and purified using ammonium sulphate fractionation, ion exchange chromatography (IEC) using CM- cellulose and gel filtration chromatography (GFC) using Sephadex G-100. The molecular weight of the purified enzyme was determined using native polyacrylamide gel electrophoresis (Native PAGE) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE). The purified enzyme was characterized to determine the pH and temperature optima. Thermostability, pH stability and substrate kinetics were studied for purified pectin methylesterase.

Results: The acidic pectin methylesterase of Aspergillus tubingensis was purified to 20.3 fold with a 47.7% recovery through IEC on CM- cellulose and GFC using Sephadex G-100. The purified enzyme had a specific activity, 112.6 U/mg. The SDS-PAGE revealed that the enzyme was monomeric with a molecular weight of 45.7 kDa. The optimum pH and temperature were 4.6 and 50°C, respectively. This enzyme was stable over a wide pH range (3.0–8.0) and at relatively high temperature at 50°C for 1 h. The Km and Vmax values of pectin methylesterase towards citrus pectin were 33.3 mg/l and 251.2 µmol/ml/min, respectively. In addition, the enzyme activity increased by about 16% in the presence of 5 mM Mg2+.

Conclusion: The pectin methylesterase enzyme of A. tubingensis has been purified up to homogeneity and found to be monomeric on SDS-PAGE. Enzyme characterization revealed that purified enzyme worked optimally in acidic conditions and was stable at wider pH range. 

Author(s) Details

Dr. Mukesh Kumar Patidar
Department of Biosciences, Maharaja Ranjit Singh College of Professional Sciences, Indore, India.

Dr. Anand Nighojkar
Department of Biosciences, Maharaja Ranjit Singh College of Professional Sciences, Indore 452001, India.

Dr. Sadhana Nighojkar
Department of Biotechnology, Mata Gujri College of Professional Studies, Indore 452001 India.

Dr. Anil Kumar
School of Biotechnology, Devi Ahilya University, Khandwa Road, Indore 452001, India.

Read full article: http://bp.bookpi.org/index.php/bpi/catalog/view/82/1164/807-1
View Volume: https://doi.org/10.9734/bpi/atbg/v2
Labels:

Comments

Post a Comment

Popular posts from this blog

Greening Regional Airports: A Vision for Carbon Neutral Infrastructure | Chapter 12 | Contemporary Perspective on Science, Technology and Research Vol. 3

 This study provides an overview of the energy demand of a regional airport, divided into individual time horizons. The electrification of aircraft systems raises the question of whether airports will be among the largest electricity consumers in our infrastructure in the future. Sustainability and especially emission reductions are significant challenges for airports that are currently being addressed. The Clean Sky 2 project GENESIS addresses the environmental sustainability of hybrid-electric 50-passenger aircraft systems in a life cycle perspective to support the development of a technology roadmap for the transition to sustainable and competitive electric aircraft systems. This article originates from the GENESIS research and describes various options for ground power supply at a regional airport. Potential solutions for airport infrastructure with a short (2030), medium (2040), and long (2050) time horizon are proposed. In addition to the environmental and conservation benefi...
Post a Comment
Read more

Risk Factors for Postpartum Psychiatric Disorders. A Review of the Literature | Chapter 8 | New Visions in Medicine and Medical Science Vol. 4

  Objective: The aim of this study was to explore the risk factors for the development of postpartum psychiatric disorders through international literature. Materials and Methods: Throughout many articles in PubMed, Google scholar and PsycInfo, a great amount of recent data was gathered to identify the disorders that are most common as well as their risk factors. Results: After childbirth, most commonly women experience postpartum depression, anxiety disorders, post-traumatic stress disorder and postpartum psychosis. All the disorders have many similar risk-factors with the main one being preexisting psychiatric disorder and many similar symptoms too. Conclusions: Women after childbirth are at risk of experience many psychiatric disorders, such as postpartum distress, postpartum post traumatic stress disorder and even more rarely postpartum psychosis. It is important to provide comprehensive support to ensure the well-being of both the mother and the infant and this will b...
Post a Comment
Read more

Alkali Element Modification of Glucose Molecules as a Method to Dissolve Cancer Cells | Chapter 12 | New Visions in Medicine and Medical Science Vol. 4

  The present study highlights about alkali element modification of glucose molecules as a method to dissolve cancer cells. The central regulation of the mechanisms governing cell proliferation has little effect on cancer cells. Cancer cells are entirely independent of the central command and divide and proliferate on their own, making it challenging to activate their response mechanism. Precisely, this is the reason why they are at risk to the health of humans and/or any biological entities. Instead of trying to reconnect the central command of the growth control mechanism to cancer cells that are already out of the range, we present a method of using the cancer cell’s own irresponsive and uncontrolled growth mechanism to their disadvantage and destroy the cancer cells. We found that this is achievable in an atomic/molecular level study of the glucose molecule, which is the primary food source used for growth and energy generation by all cells in the body, including the cancer cel...
Post a Comment
Read more