Partial Purification of Extracellular Amylase From Halotolerant Actinomycetes Streptomyces brasiliensis MML2028

Abstract

Amylase is considered as an industrially important enzyme as it occupies the most important function in the food, paper, and pharmaceutical industries. The present study is concerned with the optimization, production and partial purification of halotolerant amylase from newly isolated Streptomyces brasiliensis MML2028, from Kelambakkam salt pan, Tamil Nadu, India. The primary screening was carried out by well diffusion assay to find the zone of lysis. The assay was observed for each media optimization by measuring the release of reducing sugar (RS) by the 3,5 dinitro salicylic acid (DNS) method and expressed in the international unit (UI). Ammonium sulphate precipitation was used to partially purify the enzyme and then lyophilized. SDS-PAGE was performed to identify the molecular weight. The production medium was optimized with 1% of the starch substrate, 3% of NaCl at 24˚C and pH 9, and incubation of 9 days. The total activity of the partially purified α-amylase was observed to be 1806.9U/mL. The partially purified enzyme was more active with 3% NaCl, pH 8, and 24˚C which is known to be a halotolerant alkaline α-amylase. The enzyme showed tolerance towards magnesium, manganese ions, Triton x-100, and urea. De-inking of α-amylase showed good results proving that the enzyme activity is more efficient. Hence, the alkaliphilic amylase from Halotolerant actinomycetes S. Brasiliensis MML2028 could be a better microbial source that can be used in many industries, especially in paper and textiles.