Amylase is an enzyme which catalyzes the hydrolysis of α (1, 4)-glycosidic linkages in amylose (a linear form of starch), amylopectin (a branched form of starch) and glycogen into simpler carbohydrate molecules such as oligosaccharides or disaccharides. Alpha-amylase is the major form of amylase found in human, most prominently in pancreatic juice and saliva. The salivary amylase is an amylolytic enzyme, which can acts on cooked or boiled starch and converts it in to maltose. So it became interesting to study the behaviour of salivary amylase, when it is secreted as result of different stimuli. And thus began to study the effect of five different stimulatory temperatures, and also the effect of four tastes on the behaviour of salivary amylase.
For the study of stimulatory effect of temperature on salivary amylase, five different temperatures are selected (4, 27, 37, 55 and 75°C). And likewise four tastes also selected (sweet, sour, salt and bitter). The DNS method was done in the both tests to obtain the absorbance at 520 nm. The samples were collected from three people, of same age. The saliva was collected at same time, after one and a half hour of their breakfast in order to maintain a controlled condition for this study. In each cases the incubation temperature also kept as variable (4, 27, 37, 55 and 75°C). This study was also aimed to determine the behaviour of salivary amylase with reference to the kinetic parameters like Km and Vmax of salivary alpha amylase by incubating the enzyme (stimulated by different stimulatory conditions of temperature and taste) with varying concentration of substrate. The study revealed the consistency in kinetic parameters like Km and Vmax of salivary alpha amylase secreted in response to various stimuli.
Table of Contents
1. Introduction
1.1 Industrial use of alpha amylase
1.2 Clinical chemistry and importance
1.3 Objectives
2. Review of literature
2.1 Salivary alpha amylase
2.2 Alpha amylase assay
3. Hypothesis
4. Materials and Methods
4.1 Study area
4.2 Sample collection
4.3 Reagents
4.4 Assay procedure
4.5 Statistical analysis
5. Results and discussion
6. Conclusions
Research Objectives and Key Topics
The primary objective of this study is to investigate how various stimulatory conditions, specifically different temperatures and tastes, influence the enzymatic activity of salivary alpha-amylase and to determine the resulting kinetic parameters, Km and Vmax, under these conditions.
- Enzymatic hydrolysis of starch by salivary alpha-amylase
- Impact of temperature stimuli on amylase activity
- Influence of taste stimuli on amylase secretion and kinetic performance
- Determination of Km and Vmax parameters using the DNS method
- Consistency of kinetic behavior in response to environmental stimuli
Excerpt from the Book
1. Introduction
Amylase - one of a group of amylolytic enzyme that catalyzes the hydrolysis of starch into simpler carbohydrate molecules. There are three forms of amylase enzyme. The α-amylase EC 3.2.1.1 occur in saliva, pancreatic juice, malt, and certain bacteria. It is also present in seeds containing starch as a food reserve, and is secreted by many fungi. It catalyzes the hydrolysis of alpha bonds of large, alpha-linked polysaccharides, such as starch and glycogen, yielding glucose and maltose. The β amylases EC 3.2.1.1 occur in grains, vegetables, malt, and bacteria, is involved in the hydrolysis of starch to maltose. However all amylases are glycoside hydrolases and act on α-1, 4 -glycosidic bonds.
Alpha-amylase is the major form of amylase found in humans and other mammals. Although found in many tissues, amylase is most prominent in pancreatic juice and saliva. Each of which has its own isoform of human α-amylase. They behave differently on isoelectric focusing, and can also be separated in testing by using specific monoclonal antibodies. In humans, all amylase isoforms link to chromosome1p21 (Mandel et al., 2010) Alpha-amylase is one of the major protein components of saliva. Among other proteins, alpha-amylase is synthesized and secreted by acinar cells, after neurotransmitter stimulation (Baum, 1993). Acinar cells are innervated by sympathetic and parasympathetic branches of the autonomic nervous system (Emmelin et al., 1981).
Summary of Chapters
1. Introduction: Provides an overview of amylase enzyme types, their biological functions, industrial applications, and clinical significance.
2. Review of literature: Examines existing research regarding salivary alpha-amylase, its structure, digestive role, and established assay methods.
3. Hypothesis: Defines the core assumptions that environmental stimuli affect salivary amylase activity and kinetic parameters.
4. Materials and Methods: Details the study area, sample collection protocol, chemical reagents, and the specific procedures for DNS assay and kinetic determination.
5. Results and discussion: Analyzes the observed kinetic data, comparing Vmax and Km values under different temperature and taste stimulation conditions.
6. Conclusions: Summarizes the study findings, highlighting the consistent behavior of salivary amylase kinetics despite various environmental stimuli.
Keywords
Amylase, Vmax, Km, 3, 5-dinitrosalicylic acid method, Environment, Salivary alpha-amylase, Enzyme kinetics, Starch hydrolysis, Stimulation, Biomarker, Saliva, DNS assay, Substrate affinity, Isoforms, Digestive enzymes.
Frequently Asked Questions
What is the primary focus of this research?
The research investigates the behavior of salivary alpha-amylase under various reaction environments, specifically focusing on how different temperatures and tastes influence its kinetic parameters, Km and Vmax.
What are the central themes discussed in the work?
The work covers enzyme kinetics, the physiological impact of stimuli (temperature and taste) on saliva secretion, and the biochemical characterization of alpha-amylase.
What is the main research objective?
The goal is to determine if environmental stimuli correlate with changes in the enzymatic activity and to measure the enzyme's affinity and reaction velocity through Km and Vmax calculations.
Which scientific methods are employed?
The study utilizes the DNS (3, 5-dinitrosalicylic acid) method for measuring reducing sugars, spectrophotometric absorbance at 520 nm, and statistical analysis using SPSS and Sigma Plot software.
What does the main body of the text cover?
It covers the classification of amylases, their industrial and clinical roles, detailed assay protocols for maltose standards and enzyme activity, and the evaluation of kinetic data.
How can the work be characterized by keywords?
The work is characterized by terms such as Amylase, Vmax, Km, DNS method, enzyme kinetics, and salivary biomarkers.
How does temperature affect salivary amylase?
The study indicates that salivary amylase stimulated at 37°C shows a comparatively higher substrate affinity compared to other tested temperatures, even though Vmax variations remain relatively narrow.
Does taste stimulation significantly alter amylase performance?
Yes, the study observed that sweet taste stimulation results in a relatively high Vmax and lower Km compared to other tastes, suggesting an influence of stimulus type on the enzyme's kinetic efficiency.
- Quote paper
- Dr. Prem Jose Vazhacharickal (Author), Sajeshkumar N.K (Author), Jiby John Mathew (Author), Twinkle Jose (Author), 2016, Behaviour of Salivary Amylase in Various Reaction Environments with Reference to Km and Vmax. An Overview, Munich, GRIN Verlag, https://www.grin.com/document/367145
