Purpose: Amylase being one of the most important industrial enzymes requires large-scale production. When producing an enzyme, high productivity, high purity and low production costs need to be considered. This study focuses on comparing various agro-industrial waste substrates, for production of alpha-amylase using Bacillus amyloliquefaciens. Moreover, it studies the stability and activity of amylase– gold nanoparticles composite. Methods: This study is divided into two parts, in the first part various agro-industrial waste substrates, such as wheat bran, rice bran and potato peel were used to produce alpha-amylase using solid-state fermentation (SSF). The production of the enzyme was quantified and compared in specific enzyme activity units. In the second part, change in the stability and activity of amylase in enzyme– gold nanoparticles (AuNPs) composite has been discussed. Results: Highest enzyme production was observed in wheat bran and potato peel substrate with specific enzyme activity of almost 1. 2 U/ug and 1. 1 U/ug. Among combination substrates, wheat bran with potato peel showed a high enzyme production of 1. 3 U/ug. On the other hand, the optimum temperature for amylase activity shifted to 55 ° C in the composite compared to 37 ° C for free enzyme. Conclusions: Comparison of specific enzyme activity of extracts from various substrates showed that wheat bran alone, and in combination with potato peel, produces active and pure amylases. To stress on various catalytic activities of alphaamylase, the capability of the enzyme to synthesize gold nanoparticles and the effect of conjugation of the nanoparticle on its optimum catalytic activity are also discussed in this paper.

Alpha-amylase is one of the most widely used enzymes in the starch industry. However, industrial application of soluble alpha-amylase is hampered by changes in pH and temperature (adverse effects on enzyme stability) and activity loss, leading to higher costs. Immobilization of alphaamylase is an efficient strategy to reduce the enzyme losing and subsequently reduces costs in this regard. Alpha-amylases are immobilized by adsorption, entrapment, covalent attachment, and cross-linking. A barrier in alpha-amylase immobilization is the large size of its substrate, namely amylose and amylopectin. Most of these immobilization methods decrease the affinity of the enzyme for its substrate as well as the maximum rate of reaction (Vmax). This review aims to study different aspects of alpha-amylase including enzyme activity, applications, structure, starch, immobilization methods, and immobilization’, s obstacles to improve alpha-amylase efficiency in the industry and also lowering the costs related to providing this enzyme.

Digestive enzyme inhibitors are proteinacious or nonproteinacious compounds which reduce an enzyme activity through attaching to its active site and/or its substrate. Nowadays, plant enzyme inhibitors are of great importance because 1) these have considerable effects on insect digestive enzymes and as a result on their development; and 2) the transgenic plants expressing them are safe. In this study, the effects of NaCl, ethylenediamine tetraacetate disodium dehydrate (EDTA), Tris, sodium dodecyl sulfate (SDS) and an alphaamylase inhibitor derived from wheat kernels (WAAI) on alpha-amylase activity of adults of the stripped bug Graphosoma lineatum (L.) during 60 minutes incubation was studied. Distilled water was considered as the control. The results showed that the effects of the inhibitor type on adult alpha-amylase activity was significant (P<0.01). In general, enzyme activities in NaCl, EDTA and Tris were the highest (nearly 54% of the control) and there was not significant differences between them. Enzyme activities in SDS (nearly 31% of the control) and in WAAI (nearly 12.5% of the control) showed significant differences to each other and also to 3 previously mentioned compounds. At low concentrations of NaCl, EDTA, Tris and SDS (1 and 2 mM) and at the beginning of incubation period, female alpha-amylase activity was greater than that of the males which shows that the female enzyme has had more resistance against the inhibitors.

Rapid diagnosis in patients with acute pancreatitis is essential to optimal therapeutic outcomes. Upon clinical suspicion, various methods could be used to confirm the diagnosis based on the symptoms and examinations. Laboratory tests are an important diagnostic method in this regard based on the increased serum amylase/lipase. In this study, we described the case of a 31-year-old male patient presenting with the clinical signs of pancreatitis with normal amylase and elevated triglyceride in the follow-up. In case of strong clinical suspicion, other tests and diagnostic methods are recommended to confirm acute pancreatitis.

Background and Objectives: Enzymes are selective proteins that catalyze all metabolic reactions found in living organisms. Millions of enzymes are produced by prokaryotes and eukaryotes. Marine sponges have developed a enormous quantity of diverse microorganisms such as bacteria, fungi, viruses, protozoa, and single-celled algae and the nature of the sponge-microbe interaction is diverse. Marine sponge's symbiotic bacteria can comprise as much as 40% of sponge tissue volume. At least 32 bacterial phyla and candidate phyla were described from marine sponges by both cultivation-dependent and cultivation-independent techniques. Symbiotic microorganisms in sponges can be sources of various natural products, because metabolites previously ascribed to sponges have recently been demonstrated to be biosynthesized by symbionts. If symbiotic microorganisms from which some natural products are derived can be cultured, the microorganism could be used in a mass production of the bioactive comopounds. The goal of this research is isolation of Amylolytic bacteria from marine sponges and optimization of bacterial growth and amylase production in most amylolytic isolate. Symbiot bacteria were isolated using serial dilution method and differential analyses done for isolated bacteria. Most effective isolate was selected by using qualitative starch hydrolysis method. Methods: Morphological and biochemical characteristics were studied using Bergey’s manual of systematic bacteriology. Differential analyses were performed by catalase and Gram test TSI, H2S, VP, MR tests. Preliminary screening and quantification of amylase activities were analysed in selected isolates by starch agar plate and dinitrosalicylic acid (DNS) method respectively. In order to optimize the enzyme production, the fermentation process was carried out under M9 medium using various sodium chloride concentrations (2–14%). To evaluate the effect of pH on the production of Amylase, the best amylase producer isolate was cultured at M9 medium with different pH rang 4-8. After incubation time it was subjected to amylase activity assay. Isolate SS1 was grown in M1 medium at 28°C for 7 days. DNA extraction was done using Cinnapure DNA Kit for isolation of DNA from gram positive bacteria. PCR amplification of 16S rRNA gene was performed, the specific actinomycete primers, F27 and R1492 were used to amplify 16S rDNA. The resultant 16S rRNA gene sequences were aligned manually with corresponding almost complete sequences. The 16S rRNA gene sequence search was performed using the BLAST program available from the National Centre for Biotechnology Information (NCBI) and the isolate was identified to generic level Using the CLUSTAL-X Multiple Sequence Alignment Program (Strasburg, France), The Phylogenetic tree was constructed by Neighbor-Joining method using MEGA 6. 06. The reliability of the phylogenetic tree was tested by bootstrap analysis using 1, 000 replicates. Findings: Biochemical analyses showed that all tests were catalase and Gram positive none of them produced H2S and TSI, MR, VP tests were negative. According to 16S rRNA sequences result the most effective isolate was belong to Bacillus genus and Optimum growth condition was found at 12% NaCl concentration, 35˚C temperature and pH 12. Maximum amylase activity in optimum condition was 109 Unit (one Unit is amount of enzyme that release one microgram of glucose in a minute). Conclusion: The results showed that enzyme production is related to growth of bacteria conditions and the amylase gene should amplify from selected isolate and clone in suitable host for more studies.

Alpha-amylase reputes for starch modification by breaking of 1-4 glycosidic bands and is widely applied in different industrial sectors. Microorganisms express unique alpha-amylases with thermostable and halotolerant characteristics dependent on the microorganism’ s intrinsic features. Likewise, genetic engineering methods are applied to produce enzymes with higher stability in contrast to wild types. As there are widespread application of α-amylase in industry, optimization methods like RSM are used to improve the production of the enzyme ex vivo. This study aimed to review the latest researches on the production improvement and stability of α-amylase.

a-Amylase inhibitory activities of ethanol extracts from six selected Iranian Salvia species (S. hydrangea DC., S. hypoleuca Benth., S. officinalis L., S. reuterana Boiss., S. verticillata L. and S. virgata Jacq.) were examined, using in vitro model. In addition, antioxidant capacities (DPPH radical scavenging) and total flavonoid cotents of the extracts were also determined. The results showed that the extracts of S. verticillata and S. virgata, significantly and concentration dependently, inhibited a-amylase activity. All the extracts exhibited antioxidant activities, among which S. verticillata, S. virgata, S. officinalis, S. hypoleuca and S. hydrangea were found to be the most active in terms of assay. S. hypoleuca and S. reuterana extracts had the highest total flavonoid contents. However, a favorable correlation was not found between a-amylase inhibitory effects or antioxidant activities and the total flavonoid contents of the extracts.

The starch degrading enzyme α-amylase (EC 3.2.1.1) is widely distributed in nature. This extracellular enzyme hydrolyses a-1, 4 glycosidic linkages randomly throughout the starch molecule in an endo-fashion, producing oligosaccharides and monosaccharides, including maltose, glucose and alpha limit dextrin. a-amylases are widely used in industrial processing of starch-containing materials. The Bacillus α-amylases have found widespread use in industrial processes, and much attention has been devoted to optimizing these enzymes for the very harsh conditions. The majority of a-amylase reactions are conducted at high temperatures, resulting in high demand for the thermostable enzyme. In this study some amino acids from Bacillus amyloliquefaciens a-amylase were substituted with other amino acids, using in silico approaches to finally achieve mutations that can improve the thermostability of the enzyme. The I-Mutant2.0 web server results showed that two of selected mutations improve the thermostability of the enzyme (N190F and N265Y). Therefore, by the Swiss-PdbViewer software and I-Tasser, cph model and Swiss model web servers the mutations were exerted on the 3D structure of the enzyme and mutant 3D models from enzyme structure were achieved. The models were refined by 3Drefine, chiron and KoBaMIN web servers. The models quality assessment was performed by ModFOLD4 server. The best models were selected and then docked with partial amylose polymer to found the effect of mutations on the binding of enzyme with it substrate. Docking technique was performed by AutoDock software. The results showed that after exerting the mutations, the binding of enzyme and its substrate was not negatively affected. Thus, we suggest these two mutations for achieving α-amylases with high thermostability.

Alpha-amylases are starch-degrading enzymes which catalyze the hydrolysis of internal a-1, 4-O-glycosidic bonds in polysaccharides. These enzymes have many applications in food, paper, textile and pharmaceutical industries. About 30 % of world enzyme production market is provided by this enzyme. Alpha amylases are produced by plants, animals and microorganisms. However, microbial sources are the most preferred one for large scale production. Among microorganisms, alpha-amylase is mainly derived from Bacillus genus for commercial applications. Therefore, in this study, 13 different strains of Bacillus genus were tested for their ability to hydrolyze and consume starch. Comparative studies were performed on the strains which were capable to consume starch. The curve of starch consumption by applying logul's solution was depicted for strains. The enzymes activity was assayed by using DNS (3, 5-dinitrosalicylic acid) at various times and temperatures. The best strain according to the enzymatic features was selected. Enzymatic extract was achieved by centrifugation. As alpha-amylase was precipitated by ammonium sulphate, desalting process was performed by dialysis. Then zymography and determination of molecular weight for alpha-amylase were performed. The best strain regard to enzymatic features was Bacillus amyloliquefaciens. Maximum production of this enzyme was after 36 h. The enzyme activity was at the highest level at 70oC. Molecular weight of the enzyme was about 55KD. In conclusion, the selected strain can be an appropriate candidate for industrial production of alpha-amylase.