Starch is the commonest storage carbohydrate in plants. There exist two types of amylases, alpha-amylase (a-1,4 glucan glucohydrolase) hydrolyses a-1,4 linkages in a random manner to by-pass 1,6 branch points and beta-amylase (a-1,4 glucan maltohydrolase EC 22.214.171.124) hydrolyses alternate bonds from the non-reducing end of the substrate to produce maltose which results in 55% conversion of starch into maltose as well as large limit dextrin. Germinating seeds/ pulses are known to contain starch degrading-enzymes, a-and b-amylases. Alpha-amylase appears little in developing seeds and is synthesized de novo in germinating seeds. In contrast to alpha-amylase, beta-amylase are synthesized during seed development and stored in dry seeds. In germinating seeds, the enzyme occurs in two forms: activated and latent. The activated enzyme can be extracted with saline solution, whereas the latent one is extracted in the presence of reducing agent. Furthermore, beta-amylase known to occur in isozymic forms such as alpha-amylase. Amylases are turned on during seed sprouting to mobilize the stored resources of the seeds. The amylases were extracted by centrifugation of the cell, salt and buffer mixture. Therefore the protein so obtained was a soluble fraction (supernatant) which contains some soluble periplasmic and cytoplasmic proteins. It therefore became important to purify the enzyme by commonly used conventional purification methods (ammonium sulphate precipitation, dialysis and chromatography). Therefore Aqueous Two Phase extraction System was used to achieve a purified amylase an many polymers are used in the system have stabilizing effects on the biological activity and structure of proteins and enzymes. This technique is also straightforward and requires relatively simple equipments which are easy to operate. The most general method to monitor the purification process is by running a SDS-PAGE of the different steps which gives the measure of the amounts of different proteins/enzymes in the mixture. Amylase is extensively used in textile industry for desizing of fabric. The process of weaving cloth tends to break the threads making up the wrap. In order to give tensile strength to the yarn the woven cloth is treated with gelatinized starch. The starch must be removed from the fabric. Amylase is used to solubilise starch. Process parameters that affect the activity of the enzyme are the ones that affect the desizing of starch. The crude and purified enzyme has been studied as desizer of fabric.
About Author / Additional Info:
Dr. Kirti Rani Sharma,
Assistant Professor (II),
Amity Institute of Biotechnology,
Amity University Uttar Pradesh, Noida
Sec-125, Gautam Buddha Nagar, Noida-201303 (UP), India.
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