Study with the several resources on Docsity
Earn points by helping other students or get them with a premium plan
Prepare for your exams
Study with the several resources on Docsity
Earn points to download
Earn points by helping other students or get them with a premium plan
Community
Ask the community for help and clear up your study doubts
Discover the best universities in your country according to Docsity users
Free resources
Download our free guides on studying techniques, anxiety management strategies, and thesis advice from Docsity tutors
Basically a thesis in extraction of amylase bacteria from soil
Typology: Schemes and Mind Maps
1 / 59
This page cannot be seen from the preview
Don't miss anything!
i
iii
iv
vi
Certain biochemical reactions are regulated by biological catalysts called enzymes. Extracellular enzymatic activity in a range of bacteria has drawn attention in recent years due to the possibility of using microorganisms as biotechnological sources of industrially relevant enzymes. Proteases and amylases are regarded as the most well-known industrially significant enzymes due to their extensive use in the food, detergent, and brewing industries [Doss, A. & Anand, SP. 2012].The enzyme amylase can hydrolyze starch molecules into polymers that include glucose units. Amylase is used by a number of industries, including the food, fermentation, and pharmaceutical sectors, for its possible use in a variety of industrial processes. Living things, such as plants, animals, and microbes, can provide amylases (Souza, 2010). Amylase is a vital digestive enzyme that plays a central role in breaking down carbohydrates in the body. Produced primarily by the pancreas and salivary glands, it initiates the chemical process of digestion by converting complex starches into simpler sugars. Amylase functions as an endoglycosidase enzyme that specifically targets α-1,4-glycosidic bonds within carbohydrate molecules. TYPES OF AMYLASE Amylases can be broadly classified into three categories (Castro et al., 2018). α- Amylase Alpha-amylase is the most widely distributed form, found in humans, animals, plants, and microbes, functioning as a calcium metalloenzyme that breaks random α-1,4 glycosidic bonds in starch molecules .[ Henrissat, B & Bairoch, A. 1993.] β-Amylase Beta-amylase, primarily occurring in plants and microbes, works differently by cleaving the second α-1,4 glycosidic bond, releasing two maltose molecules at a time.0A wide range of microorganisms contain β-amylases and glucoamylases.[ Pandey, A; Nigam, P; Soccol, CR; Soccol, VT; Singh, D & Mohan, R. 2000.] γ-Amylase γ-amylase breaks down α 1-6 glycosidic bonds, including those at the non-reducing end of
vii amylose and amylopectin, resulting in the production of glucose. γ-amylase, unlike other amylases, performs best in acidic settings with an optimal pH of 3. Table 1.1: Types of Amylase and their properties MARKET POTENTIAL The global enzymes market is poised for substantial growth, projected to reach USD 20. billion by 2029, with a compound annual growth rate (CAGR) of 7.8% from 2024. This expansion is driven by diverse applications across multiple industries and a rising demand for sustainable solutions. The market is categorized into industrial enzymes—used in sectors such as food and beverage processing, bioethanol production, textiles and leather, detergents, and paper and pulp—and specialty enzymes, which serve the pharmaceutical, research and biotechnology, diagnostics, and biocatalyst industries. Within this industry, the amylase market exhibits strong growth potential. Amylase, particularly sourced from barley and wheat, plays a vital role in the brewing sector by breaking down starches into fermentable sugars, enhancing production efficiency and quality. Moreover, amylase enzymes are increasingly recognized as eco-friendly alternatives to conventional synthetic chemicals, as they function under mild conditions and contribute to lower energy consumption. SOURCES OF AMYLASE Plant sources Type Source Function Optimum pH Products α-amylase Animals, plants, and microbes Random cleavage of starch 6.7-7.0 Maltose, dextrins β-amylase Plants, microbes End-wise cleavage 5.4-5.5 Maltose γ-amylase Animals, microbes Complete hydrolysis 4.0-4.5 Glucose
ix natural availability of starch. Soil samples from these sites regularly show higher enzyme activity than other agricultural soils. Amylase Production Amylase can be produced through submerged fermentation (SmF) or solid-state fermentation (SSF), depending on physicochemical factors. SmF is preferred for commercial enzyme production due to its controlled regulation of temperature, pH, aeration, and moisture (Gangadharan et al., 2008). SSF, which mimics natural microbial growth conditions, is favored for value-added products due to its high yield, specificity, and low moisture content, reducing bacterial contamination (Souza, 2010). While SSF is generally suited for fungal amylase production, studies suggest that bacterial cultures can also be adapted for this method (Pandey, 2003). Table 1.2: Optimal conditions for different microorganisms Characteristics/properties of Amylase Physical and chemical factors play a crucial role in microbial growth for amylase production, with bacterial and fungal requirements extensively studied (Gupta et al., 2003; Shalini, 2014). These parameters are essential for optimizing fermentation processes, impacting both economic feasibility and practicality (Saini et al., 2017). Key factors influencing amylase production include pH, temperature, metal ions, carbon and nitrogen sources, surfactants, phosphate levels, and agitation. Applications of α- Amylase Microorganism pH Optimal/Range Temperature Optimal/Range Bacillus subtilis 7.0 37 °C Bacillus amyloliquefaciens 7.0 33 °C Bacillus sp. 4.5 70 °C Aspergillus niger 5.5 70 °C
x Starch is a primary storage component in many key crops like wheat, rice, maize, tapioca, and potatoes, supporting a growing starch processing industry. Over time, starch hydrolysis has shifted from acid-based methods to enzyme-driven processes, with starch-converting enzymes now accounting for 30% of global enzyme production. These enzymes are widely used in producing maltodextrin, modified starches, and glucose/fructose syrup, as well as in industrial applications like detergents and baking. In the food industry, amylases are essential for bread making, brewing, and sweetener production by converting starches into fermentable sugars. In textiles, they remove excess starch from fabrics, while in detergents, they help dissolve starch-based stains. The paper industry uses amylases to enhance sizing and reduce starch consumption. In pharmaceuticals, amylases aid in drug formulation, pancreatic enzyme therapy, and diagnostic testing. They are also expanding into biofuel production, breaking down starch into fermentable sugars for ethanol conversion. OBJECTIVES OF THIS PROJECT
xii
1811 - Kirchhoff discovers starch-degrading enzyme in wheat 1833 - Anselme Payen isolates and names "diastase" (early amylase) 1894 - First industrial fungal amylase used for digestion treatment 1914 - Takamine pioneers fungal amylase production via Solid-State Fermentation (SSF) 1917 - Boidin & Effront commercialize bacterial α-amylase production (Bacillus species) 1925 - Kuhn names "α-amylase" based on hydrolysis product configuration 1930 - Ohlsson identifies and names "β-amylase" 1990s - 3D structures of amylase enzymes analysed and compared Present - Amylases widely used in industrial biotechnology and therapeutics Types of Amylases Amylases, including endoamylases and exoamylases, are enzymes capable of hydrolyzing starch. They are classified based on how they attack glycosidic bonds. Starch-degrading enzymes belong to several glycoside hydrolase families, with the most notable being the GH13 family.[ Coutinho, P & Henrissat, B. 1999.][. Henrissat, B & Bairoch, A. 1993]. Amylase enzymes exist in three main categories, each with distinct characteristics and functions. α-Amylases (EC 3.2.1.1) Endoamylases break α-1,4 glycosidic linkages found in amylose or amylopectin chains. A well-known example is α-amylase, found in a wide spectrum of microbes, including Archaea and bacteria.[Pandey, A; Nigam, P; Soccol, CR; Soccol, VT; Singh, D, & Mohan, R. 2000]. The enzyme generates branched oligosaccharides with an α-configuration and α-limit dextrins. α-Amylases are classified according on their level of substrate hydrolysis: Saccharifying α-amylases break down 50-60% of glycosidic bonds. Liquefying α-amylases break down 30-40% of the connections. α-amylases are metalloenzymes that require calcium for action. They degrade long-chain carbohydrates at random sites throughout the starch molecule, producing maltotriose and
xiii maltose from amylose and maltose, glucose, and limited dextrin from amylopectin. Because of their random activity, α-amylases act faster than β-amylases.[Fukumoto, J, and Okada, S. 1963.] Salivary and pancreatic amylases are part of the α-amylase family. Plants, fungi, and bacteria (including Bacillus species) all contain the enzyme. Exoamylases, the second type of amylase, work by cleaving starch molecules from the outer glucose residues. They are further grouped according to their specificity.
xv Substrate Characteristics and Production Patterns
xvi fungal, and viral amylases has gained prominence due to their ease of large-scale production and low downstream processing costs, as they are extracellular in nature (Ashis et al., 2009). Amylases, namely α-amylases, have been identified from fungi, yeasts, bacteria, and actinomycetes. However, enzymes of fungal and bacterial origin are more commonly utilized in industrial applications and dominate the market. Notably, Aspergillus and Rhizopus species generate amylase with high efficiency (Pandey et al., McVey, 2002). The consistency, cost-effectiveness, reduced time and space requirements for production, and ease of process optimization and modification make bacterial and fungal amylases the preferred choice for industrial applications (Ellaiah et al., 2002). Plant Sources of Amylase Amylases are found in many different organisms, including mammals, fungi, plants, unicellular eukaryotes, bacteria, and archaea. While plants and animals produce amylases, microbial enzymes are most commonly used in commercial applications because to their high productivity, thermostability, and simplicity of cultivation (Reddy, R; Reddy, G & Seenayya, G. 1999). A key advantage of enzymatic production is its high selectivity, efficiency, and specificity, ensuring high yields of the desired product (Kim, S & Dale, BE. 2004.). Bacillus species are the primary bacterial sources used in commercial enzyme production (Olafimihan, C & Akinyanju, J. 1999), while other bacteria such as Escherichia, Pseudomonas, Proteus, Serratia, and Rhizobium also produce significant amounts of amylase (Oliviera, A; Oliviera, L; Andrade, J & Chagas, A. 2007). Fungal species such as Aspergillus, Rhizopus, Mucor, Neurospora, Penicillium, and Candida are also known to produce commercially significant extracellular amylases (Gupta, R; Gigras, P; Mohapatra, H; Goswami, VK & Chauhan, B. 2003). Despite their presence, plant-derived amylases have not been widely explored as enzyme sources (Azad, MAK; Bae, JH; Kim, JS; Lim, JK; Song, KS; Shin, BS & Kim, HR. 2009). However, agricultural waste materials offer a sustainable alternative by reducing pollution while serving as substrates for solid and liquid fermentation, lowering production costs. These carbon- and nitrogen-rich agricultural byproducts include pearl millet starch, orange
