Multifaceted Protocols in Biotechnology, Volume 2

Contents
Abbreviations
1 Graviola Fruit Extraction by Ionic Liquid Microwave-Assisted Extraction (IL-MAE)
	1.1 Introduction
	1.2 Ionic Liquid Microwave-Assisted Extraction (MAE)
		1.2.1 Principle
		1.2.2 Objective of Experiment
	1.3 Materials
	1.4 Methodology
		1.4.1 Statistical Optimization Set-Up
		1.4.2 One-Factor-at-a-Time (OFAT) Experimental Design
		1.4.3 Optimization of Extraction Conditions
		1.4.4 Validation of the Experimental Model
	1.5 Results and Discussion
		1.5.1 Extraction of Graviola Fruit Extract
		1.5.2 The Best Ionic Liquid Solution by OFAT
		1.5.3 Optimization of the IL-MAE Parameters by RSM
	1.6 Discussion
		1.6.1 Analysis of Variance
		1.6.2 Analysis of Variance of the Extraction Yield
		1.6.3 Test for Significance of the Regression
		1.6.4 The Interaction Response Effects
		1.6.5 Validation of the Model
		1.6.6 Analysis of Variance of the IC50
		1.6.7 Test for Significance of the Regression
		1.6.8 The Interaction Response Effects
		1.6.9 Validation of the Model
	1.7 Conclusion
	References
2 Role of Ionic Liquids in the Enzyme Stabilization: A Case Study with Trichoderma Ressie Cellulase
	2.1 Introduction
	2.2 The Role of ILs in Enzyme-Catalyzed Hydrolysis
		2.2.1 Principle
		2.2.2 Objective of Experiment
	2.3 Materials
	2.4 Methodology
		2.4.1 Synthesis of Ionic Liquids
		2.4.2 Cellulase Production
		2.4.3 Stability of Cellulase ILs
		2.4.4 Cellulase Assay
	2.5 Results and Discussion
		2.5.1 Cellulase Activity
		2.5.2 Stability of Cellulase in ILs
		2.5.3 Discussion
	2.6 Conclusion
	References
3 Role of Ionic Liquids in the Processing of Lignocellulosic Biomass
	3.1 Introduction
	3.2 A Brief Perspective of Ionic Liquid
		3.2.1 Pretreatment Process for Lignocellulosic Biomass
		3.2.2 Ionic Liquid–Cellulase Compatible Systems
	3.3 Objective of Experiment
	3.4 Materials
	3.5 Methodology
		3.5.1 Production of PKC-Cel
		3.5.2 Enzymatic Hydrolysis of EFB
		3.5.3 Characterization of the Native and Treated EFB for IL-E Evaluation
		3.5.4 Structural Carbohydrates and Sugar Determination
	3.6 Results and Discussion
		3.6.1 Characterization of the Native (Untreated) and Treated Substrate to Evaluate the Effect of IL-E
		3.6.2 Discussion
	3.7 Conclusion
	References
4 Proliferation of Rat Amniotic Stem Cell (AFSC) on Modified Surface Microcarrier
	4.1 Introduction
		4.1.1 Principle
	4.2 Objective of Experiment
	4.3 Methodology
		4.3.1 Cell Line
		4.3.2 Media Preparation
		4.3.3 Embryonic Stem Media (ESM)
		4.3.4 Embryoid Bodies Media (EBM)
		4.3.5 Cell Propagation in a 2D Cultured Flask
		4.3.6 Microcarrier Spinner Vessel Culture
	4.4 Result and Discussion
		4.4.1 Spontaneous Differentiation of AFSC
	4.5 Conclusion
	References
5 Application of Spectroscopic and Chromatographic Methods for the Analysis of Non-halal Meats in Food Products
	5.1 Introduction
	5.2 Meat’s Authentication Using Spectroscopic and Chromatographic-Based Methods
	5.3 Analysis of Non-halal Meats in Fresh and Meat-Based Food Products
		5.3.1 Analysis of Pork in Fresh and Meat-Based Food
		5.3.2 Analysis of Wild Boar Meat as Meat Adulterants
	References
6 Identification of Potential Biomarkers of Porcine Gelatin
	6.1 Introduction
	6.2 Principle
		6.2.1 Objective of Experiment
	6.3 Materials
	6.4 Methodology
		6.4.1 Sample Preparation
		6.4.2 SDS-PAGE
		6.4.3 Peptide Mass Fingerprinting (PMF)
	6.5 Results and Discussion
		6.5.1 Protein Identification of Targeted Polypeptide Band
		6.5.2 Potential Marker Peptides as Immunogens
	6.6 Conclusion
	References
7 Gamma Ray Mutagenesis on Bacteria Isolated from Shrimp Farm Mud for Microbial Fuel Cell Enhancement and Degradation of Organic Waste
	7.1 Introduction
		7.1.1 Principle: Microbial Fuel Cell (MFC)
		7.1.2 Principle: Mutation
		7.1.3 Objective of Experiment
	7.2 Materials
	7.3 Methodology
		7.3.1 Bacterial Radiation
		7.3.2 Bacterial Growth
		7.3.3 MFC Power Measurement
		7.3.4 Total Chemical Oxygen Demand (COD)
	7.4 Results and Discussion
		7.4.1 Suitable Gamma Rays Dose by OFAT
		7.4.2 Formation of Biofilm in the Anolyte
		7.4.3 Electricity Performance
		7.4.4 The Effect of COD Removal and Strain Relations
	7.5 Conclusion
	References
8 Effect of Temperature on Antibacterial Activity and Fatty Acid Methyl Esters of Carica Papaya Seed Extract
	8.1 Introduction
	8.2 Methodology
		8.2.1 Plant Material
		8.2.2 Extraction of Phytochemicals
		8.2.3 Effect of Temperature on the Extract
		8.2.4 Percentage of Growth Inhibition
		8.2.5 Effect of Temperature on Fatty Acid Profile in Carica Papaya Seed Extract
		8.2.6 Profiling of Fatty Acid Methyl Esters by GC/MS Analysis
		8.2.7 Statistical Analysis
	8.3 Discussion
		8.3.1 Effect of Temperature on Antibacterial Activity and Fatty Acids Profile of Carica Papaya Seed Extract
		8.3.2 Effect of Temperature on Fatty Acid Methyl Esters Profile of Carica Papaya Seed Extract
		8.3.3 Profile of Cis and Trans Fatty Acid Methyl Esters as Affected by Temperature
	8.4 Conclusion
	References
9 Solid-State Fermentation of Agro-Industrial Waste Using Heterofermentative Lactic Acid Bacteria
	9.1 Introduction
	9.2 Objective
	9.3 Methodology
		9.3.1 Preparation of Medium for Activation of Bacteria
		9.3.2 Bacterial Growth Curve
		9.3.3 Heterofermentative Lactic Acid Bacteria
	9.4 Materials
	9.5 Methods
		9.5.1 Preparation of MRS Broth Medium (MRSB)
		9.5.2 Preparation of MRS Broth Medium Mixed with Solid Substrate (MRSB-S)
		9.5.3 Activation of Bacteria and Growth Curve Analysis
		9.5.4 Solid-State Fermentation Using Bacterial Inoculum
		9.5.5 Measurement of Bacterial Concentration Using Serial Dillution and Standard Plate Count Method
	9.6 Results and Discussion
		9.6.1 Activation of Bacteria from an Agar Media to a Broth Media
		9.6.2 Growth Curve of Bacteria Cultivated Using MRS Broth Mixed with Coffee Pulp
		9.6.3 Growth of Bacteria During Solid-State Fermentation
	9.7 Conclusions
	References
10 Synthesis of Chitosan-Folic Acid Nanoparticles as a Drug Delivery System for Propolis Compounds
	10.1 Introduction
	10.2 Principle
	10.3 Materials and Methods
		10.3.1 Materials
		10.3.2 Extraction of Propolis
		10.3.3 Conjugation of Folic Acid with Chitosan
		10.3.4 Preparation of Chitosan-Folic Acid Nanoparticles
		10.3.5 Characterization of Nanoparticles
		10.3.6 In vitro Study of the Release of Propolis from Nanoparticles
	10.4 Result and Discussion
		10.4.1 Folic Acid-Chitosan Conjugate
		10.4.2 Encapsulation Efficiency of Propolis in Nanoparticles
		10.4.3 FTIR Analysis
		10.4.4 In vitro Study of Propolis Released from NP-KF-P
	10.5 Conclusion
	References