Abstract
New isolate obtained from leaf surfaces of Spinacia olevacea, Brassica oleracea and Coriandrum sativum was identified as Aureobasidium pullulans on the basis of cultural, morphological, biochemical and molecular characterization. The factors such as inoculums size, incubation time, sources of starch, and concentration of starch, sources of carbon and nitrogen, effect of initial pH and effect of temperature affecting production of amylase were optimized in present study. The optimum pH, temperature and incubation period for enzyme production were 5, 25OC and 5 days respectively. Among different sources of starch tested potato starch was shown to be the best. Potato starch at 1% was recorded to be the best concentration of starch for enzyme production. Sodium nitrate was ideal nitrogen source. At optimum conditions Aureobasidium pullulans Cau 19 has shown amylase activity 800U/L, which is twofold higher than before optimization. For the improvement of Aureobasidium pullulans Cau 19, the parental strain, after optimization of the cultural conditions, was subjected to UV irradiation for 8 min. Total 286 mutants were checked, out of which 9 mutant strains showing comparatively greater hydrolysis zone were selected for further study. Aureobasidium pullulans UVm 276 mutant has shown amylase activity which is 2.78 fold that of the wild type strain. Thus these findings have more impact on enzyme economy for biotechnological applications of microbial amylases.
Highlights
Amylases are among the most important enzymes used in several industries such as paper industry, detergent industry, food and pharmaceutical industries in various processes such as starch liquefaction and saccharification, textile desizing, manufacture of high fructose containing syrups, treatment of starch processing waste water (Gupta et al, 2003)
New isolate obtained from leaf surfaces of Spinacia olevacea, Brassica oleracea and Coriandrum sativum was identified as Aureobasidium pullulans on the basis of cultural, morphological, biochemical and molecular characterization
We have recently reported the preparation of bioconjugate using biologically synthesized gold nanoparticles and purified amylase from Aureobasidium pullulans. (Mulay and Deopurkar, 2017).The amylases are produced by plants, animals, and microorganisms (Pandey et al, 2000), largely microbial amylases are commercially available, due to ease of cultivation of microorganisms and processing to isolate and purify enzyme
Summary
Amylases are among the most important enzymes used in several industries such as paper industry, detergent industry, food and pharmaceutical industries in various processes such as starch liquefaction and saccharification, textile desizing, manufacture of high fructose containing syrups, treatment of starch processing waste water (Gupta et al, 2003). We have recently reported the preparation of bioconjugate using biologically synthesized gold nanoparticles and purified amylase from Aureobasidium pullulans. (Mulay and Deopurkar, 2017).The amylases are produced by plants, animals, and microorganisms (Pandey et al, 2000), largely microbial amylases are commercially available, due to ease of cultivation of microorganisms and processing to isolate and purify enzyme. Aureobasidium pullulans is yeast like fungus that has been used for industrial production of wide variety of enzymes used in biotechnology in process involving starch hydrolysis. The search for newer yeast is biotechnologically important for the development of efficient, economical and environmentally safe amylolytic hydrolysis of starch. We report the isolation and characterization of amylase producing yeast from phyllosphere of different plants. These isolates were mutagenized to enhance the production of amylase
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