Out of thirty isolates, eleven of them showed positive for Iodine test on solid media. Among these, the isolate number 7 of A. Ac-7 which gave maximum activity was selected for the optimization of amylase activity in liquid media.
After sterilization, the flasks were cooled to room temperature and 0. An uninoculated flask served as control. The culture broth was filtered using Whatman filter paper No. Amylase activity was determined at room temperature in a reaction mixture containing 1mL of 1mol L -1 sodium acetate buffer pH 6.
After 20 minutes of incubation, the liberated maltose was estimated by dinitrosalicylic acid DNS method The denatured culture filtrate served as control. Out of thirty endophytic isolates screened for the amylolytic activity on solid media, eleven of them showed positive results.
Among these the isolate Ac-7 Cylindrocephalum sp. Mean results are represented in Fig. Optimization of various parameters and manipulation of media are one of the most important techniques used for the production of enzymes in large quantities to meet industrial demands Production of amylase enzyme in fungi is known to depend on both morphological and metabolic state of the culture. Growth of mycelium is crucial for extracellular enzymes 8.
Various physical and chemical factors have been known to affect the production of amylase such as temperature, pH, carbon sources acting as inducers, and nitrogen sources respectively. Interactions of these parameters are reported to have a significant influence on the production of the enzyme The influence of temperature on amylase production is related to the growth of the organism.
Hence, the optimum temperature for enzyme production depends on whether the culture is mesophilic or thermophilic. The pH is one of the important factors that determine the growth and morphology of microorganisms as they are sensitive to the concentration of hydrogen ions present in the medium.
The optimum pH for amylase production was 7. The biomass yield was found to be higher in case of pH 5. Aspergillus oryzae , A. The fungus was able to grow on all the tested carbon sources. There were significant differences in the yield of the biomass and amylase production. Among the various substrates screened for amylase production, maltose gave the highest enzyme activity followed by starch.
There was high increase in biomass yield in case of wheat bran powder and rice bran powder, but the enzyme activity was low in contrast to the findings of Ellaiah et al. This indicates that the nature and amount of carbon source in culture media is important for the biomass growth and production of extracellular amylase. Among chemical parameters, carbon source of the growth medium plays a very important role in inducing enzyme secretion.
Amylase is generally induced in the presence of carbon sources such as starch and its hydrolytic products. Similarly, such findings were reported by Kathiresan and Manivannan 20 as maltose as best carbon source to enhance the amylase activity in P.
Compared to defined carbon sources, the biomass yield was higher in undefined carbon sources similar to the findings of Oliveira et al. Earlier reports show that among various inorganic nitrogen sources tested, ammonium sulphate, ammonium chloride and ammonium hydrogen phosphate favored growth and enzyme secretion Similar observations were recorded by Chandra et al.
However, in our studies, there was no significant increase in enzyme yield in the case of the supplementation with either inorganic or organic nitrogen sources Fig. A marginal increase in amylase activity was noted with the addition of Sodium nitrate. A minimum activity was observed when Ammonium nitrate was used as nitrogen source. There was no significant difference in amylase activity with peptone, tryptone and beef extract indicating that any of these sources can be alternatively used.
In the present study, an attempt has been made to offer an endophytic fungus as source of enzymes for industrial requirements. The growth parameter of Cylindrocepahalum sp. Though, the cheaper agricultural byproducts are feasible for the production of amylase enzyme, but are not significant to the maltose when used as substrate. However, more detailed investigation is required to characterize this enzyme, which may be used in the large-scale production for commercial purpose in future.
The authors are thankful to Dr. Siddamallaya for authenticating the plant sample and maintaining the herbarium in National Ayurveda Dietetics Research Institute. Submitted: August 19, ; Returned to authors for corrections: October 25, ; Approved: June 07, Abrir menu Brasil.
Brazilian Journal of Microbiology. Abrir menu. C Srinivas. About the authors. Alpinia calcarata Roscoe; Endophytic fungi; Cylindrocephalum sp. Roscoe Sunitha. Screening of endophytic fungi for amylase production on solid media Endophytic fungal culture grown on PDA was cut into 5 mm discs with the help of borer.
Optimization of culture conditions for amylase production The procedure adopted for optimization of various parameters influencing amylase production was to evaluate the effect of each parameter independently keeping others as constant.
Enzyme assay The culture broth was filtered using Whatman filter paper No. Akher, M. Optimal conditions of the production of bacterial amylase. Hyg , , Babu, K. Process Biochem , 30, Barnett, H. Illustrated genera of imperfect fungi , fourth edition.
As the ground warms, the amylase in the seed becomes more active. Since the amylase in plants acts as a food storage reserve as the plant grows, temperatures that are too high will typically cause the enzymes will stop functioning.
For this reason, seeds that are exposed to high temperatures will not be able to sprout. Gibberellins are also involved in the process of germination and early plant development. A tetra cyclic diterpene acid, gibberellin is believed to signal the start of the amylase process in the cells for starch-to-sugar conversion.
The gibberellin becomes a chemical messenger for the plant embryo. There are several basic forms of amylase, including alpha amylase, beta amylase, and glucoamylase. Alpha amylase heps to process basic substances like glycogen and starch, and can be found in the tissues and organs of many mammals, including humans. It works by breaking random bonds in the viscosity of starches, and produces glucose. Beta amylase is abundant is seeds. Plant foods rich in amylase include soybeans, sweet potatoes, barley, potatoes, rice, tomatoes and cereal grains.
Because many of the foods rich in amylase are valuable crop plants, amylases have been studied in those plants. Researchers still attempt to elucidate the functions of beta-amylases. Evidence suggests the enzymes play a crucial role in leaf starch turnover in some of these plants. If the genes for beta-amylases are overexpressed, it leads to stunted plants and other problems.
Sources of beta-amylases in food include grains essential to brewing. Beta-amylase affects the malting quality of brewing grains. The malting process requires beta-amylose to produce maltose in the mashing phase, which in turn affects the amount of alcohol yeast can produce.
Scientists face challenges studying these grains because their genetic makeup is redundant and complex, with polyploidy of their genomes being a common feature. Researchers will continue to learn more about how beta-amylase in food works.
She spent nine years working in laboratory and clinical research. A lifelong writer, Dianne is also a content manager and science fiction and fantasy novelist. Dianne features science as well as writing topics on her website, jdiannedotson. What Is Lipase? The Effect of Alcohol on Plants.
Industrial Uses of Pepsin.
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