2018 International Conference on Energy, Power and Materials Engineering (EPME 2018) ISBN: 978-1-60595-102-7

An Overview and Application of Amylase

Chun-hai ZHAO*, Ning-bo HUO, Hui FENG and Zhen-qing LI Bin Zhou Polytechnic, Key Laboratory of Biotechnology of Binzhou, 256603 China *Corresponding author

Keywords: Amylase, Classification, Application.

Abstract. Amylase is an enzyme that catalyses the hydrolysis of starch into sugars, amylases are glycoside hydrolases and act on α-1,4-glycosidic bonds. It is classified three amylases: α-amylase, β-amylase, γ-amylase. Amylases are important to brewing beer and liquor and other fermentation food made from sugars derived from starch. Research shows that it has new application, such as dishwasher detergents, flour additive. With full use of depth research and bio-engineering technology, amylase application prospects will be broader.

Introduction Amylases are α-1,4- glycosidic bond hydrolysis enzyme, which typically hydrolyze soluble starch, amylose, glycogen and act on α-1,4-glycosidic bonds [1]. α- amylase Widely existing in animals, plants and microorganisms, it's random act on intramolecular α-1,4- glucoside linkage of starch, glycogen, oligosaccharides or polysaccharide They cut α-1.4-glucoside bond randomly, produce lower dextrin and oligosaccharides[2]. α-amylase as an enzyme is currently the most widely used of industrial production, it has been widely used in food, textile, paper, feed and other industries, shortening the production cycle, improving product yield and raw material utilization, improving product quality and food resources conservation, etc., it has an extremely important role in industrial processes [3], α- amylase has become an essential key enzyme in the entire enzyme market, the share of the entire enzyme market is about 25%. Although α- amylase has been widely used in many ways in food industry, but there are many problems of its use, there is still a low activity, poor stability and adaptability, and the activity retention time is short, the difficulty of regeneration and reuse and many other problems. All these problems affect the production efficiency of α- amylase largely and indirectly raise production costs. Therefore, in order to solve these problems, people are constantly exploring a modified method of α-amylase. β-amylase β- amylase begins successively cutting a maltose molecule from the non-reducing end of the starch molecule, hydrolysis α-1,4 glycosidic bond,. which mainly seen in higher plants (barley, wheat, sweet potato, soybean, etc.), but there is also the presence of bacteria, mold in reports. β- amylase can only hydrolyze α-1,4 glycosidic bonds, but it can not hydrolyze α-1,6 glycosidic bonds, and it can not across α-1,6 glycosidic bond hydrolysis and stop hydrolyzing. When β- amylase hydrolysis of amylose, so that amylose molecules shorten gradually, maltose generates slower. While it hydrolyzes amylopectin, amylopectin has a large of branches and non-reducing end, so the formation rate of maltose from hydrolysis of amylopectin is faster. But β- amylase can not hydrolyze α-1,6 glycosidic bonds neither across, so it can only act on the outside part of the branch points, producing the equivalent of 50% -60% of the total amount of maltose from branch starch, while it can not hydrolyze within the part of a branch point[4]. γ-amylase γ-Amylase is also named glucoamylas, it begins cutting the starch molecule successively from the non-reducing end of the starch molecule, producing a lot of glucose. It is not only capable of hydrolyzing α-1,4 glycosidic bond, but also capable of hydrolyzing α-1,6 and α-1,3 glycosidic bonds

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glycosidic bond, but the latter rate of hydrolysis of much slower than that of both the former, therefore, when glucoamylase acts on the amylose and amylopectin, all of them can be hydrolyzed to glucose. When the temperature is between 40-60℃, even at the same temperature within the range of pH3.5-5.0 glucoamylase activity difference is not obvious, as the pH rises, the enzyme activity decreased gradually in some content[5].

Amylase Distribution The source amylase is very extensive, including animals, plants and microorganisms, microorganisms produce a rich source of amylase, which is ease of industrial production to meet the needs of a variety of industrial applications, there are a variety of microorganisms can produce α- amylase, including filamentous fungi, yeasts, bacteria and actinomycetes. The extraction process of amylase from microorganisms is simple, low cost, high yield, stable, mild conditions, microbial amylase hydrolysis method has been completely replaced traditional chemical hydrolysis. Therefore, it has been widely in the industry. But the major industrial amylase derived from microorganisms, it is because: ①Microbial growth is rapid, microorganisms are suitable for mass; ② The medium for microorganisms is low cost; ③Many microorganisms are improved by genetic manipulation of available means; ④ Microbial production of extracellular amylase is mostly easy to extract. However, we know about the amylase, it can not meet the needs of the industrial age. In the prerequisite of respect for biodiversity, a new generation of amylase of microorganisms should be continuing to explore.

Cold Temperature-amylase Low amylase (Cold temperature-amylase) is a relative concept, the study found that a large number of cold temperature-amylase has the following characteristics [6] ①The optimum temperature of cold temperature-amylase is lower 0℃-30℃ comparison with mesophilic enzyme; ②Cold temperature-amylases are characterized by higher Kcat and physiological efficiency (Kcat/Km) and by a lower and rather constant Km at temperatures from 0 to 30℃. ③The stability of cold temperature-amylase is poor. Low amylase is mainly produced by cold-adapted-microorganisms. There is a large number of low-temperature environments on the Earth's surface, so that cold-adapted-microorganisms play an important role in the ecosystem, study the mechanism of cold-adapted microorganisms and the low enzyme has important biological significance; at the same time, the low temperature enzymes can play an effective catalytic characteristic at low temperatures, so that in the field of bio-engineering it has a broad application prospects. Therefore, the study of low-temperature enzymes both in theory and in practice has important significance. In recent years, research on low-temperature enzymes have increased, I believe that with full use of the continuous, in-depth research and bio-engineering technology, low amylase industry prospects will be broader[7].

Research Development of Amylase

The Use in Environmental Protection Amylase is an important enzyme, which has great values. With the world's energy crisis, the new road of industrialization of energy-saving and environmental protection has become an inevitable trend of global industrial development. From the present analysis on the application of amylase, the cold temperature-amylase is found to solve these problems that the temperature of starch treatment process is high by high temperature amylase and mesophilic enzyme. The other problem is

424 deterioration of raw materials in the starch treatment process, the use of cold-adapted-microorganisms will greatly ease the energy crisis. The cold temperature-amylase is more in line with the special nature of the new industrial production. The use of cold temperature-amylase has some advantages in industrial production applications: low temperature fermentation to produce a lot of the flavor of food, energy conservation, reduction pollution and preventing microbial contamination from mesophilic bacteria (especially the in continuous fermentation system) with the range of 0-20℃ (in this case homologous mesophilic enzyme inactive). Cold temperature microorganisms have a high growth rate, high enzyme activity and high catalytic efficiency, which can greatly shorten the processing time and save expensive heating system, so there is considerable progress in energy efficiency; cold temperature-amylase isolated from marine microorganisms can be used in low-temperature medicine, food and so on, which does not affect the quality of the product and so on[8]. The Use in Food Industry In baking process, cold temperature-amylase can shorten the fermentation time to improve the quality of the dough and bread crumbs, while still can maintain a certain flavor and moisture. The cold temperature-amylase can act on starch, bran and flour hemicellulosem, the advantages of the cold temperature-amylase is not only in terms of its highly specific activity, but also in itself is easy inactivated. The enzyme reaction is prevented to continuing by easy inactivation, thus avoiding changing crumb structure, and therefore does not cause that the bread becomes too soft or too sticky. The Use of Detergent Additives The cold temperature-amylase is a good substitute of mesophilic amylase, a cold temperature-amylase is a well-known and the important application is as detergent additives. The most significant advantage is that it saves energy and reduces the clothing chance of wear and tear. The commercial application of cold temperature-amylase is a recombinant enzyme, which can maintain high catalytic efficiency at low temperature and also improve the stability of the enzyme, at the same time to maintain its biological activity and greatly save energy. Studies have been carried out, the cold temperature-amylase is applied in the detergent industry. Applied Microbiology to reduce environmental pollution is an important part of biotechnology. The idea is use of microorganisms to environmental pollution of soil and water; it is a reasonable alternative to physical methods. In suitable areas, the special low-temperature growth of microorganisms in mixed media, which produce cold temperature-amylase, it helps to improve the refractory chemicals biodegradability[9]. The Use in Detergent Textile Because these organisms have high catalytic activity and their specificity at a lower temperature, microbial amylase is an ideal tool for bioremediation. In the textile production process, there is often a variety of fibrous tissue protruding from the outer end of the main fiber cotton fiber, which will affect clothing smoothness and overall appearance, and this clothing will be washed with successive, and it becomes worse. Under appropriate conditions, the pre-treatment low amylase and cellulase not only can reduce the formation of flocks, but also can increase the flexibility and durability of clothing fibrous tissues. Now the processing method is that cold resistant amylase and cellulase have been used in the process, the two enzymes can reduce the temperature and the enzyme concentration[10].

New Application of Glucoamylase Glucoamylase is used very extensively in industrial production, which can be applied into liquor, wine, beer and other brewing, cheese production, the production of fermented food and animal feeding. In the alcohol industry, glucoamylase can replace homemade bran, simplifying the production process, improving the efficiency of production. In starch, sugar industry, In starch, sugar industry, highly specific glucoamylase hydrolyzes to control sugar component and improve the

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product purity, avoiding glycoside bond randomness by inorganic acid, reducing the corrosion of the production equipment from inorganic acid hydrolysis. in dry beer brewing process glucoamylase can increase fermentable sugar content; in the production of wine and liquor glucoamylase can improve alcohol yield and reduce food consumption, improving the taste and quality of wine, the process of glutamic acid and citric acid fermentation, glucoamylase transforms material starch into low molecular sugars, weight into, then by so glucoamylase has wide application value in light industry, food, pharmaceutical, and fermentation industries, more and more scholars pay attention to glucoamylase. With the in-depth study of its enzyme structure, mechanism, and other issues related to the structure and function, it has applications and wider range [11]

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Acknowledgement This research is supported by Grant BS2012SW001 from National Natural Science Foundation of Shan Dong Province and the J14LE58 and J17KA122 Project of Shandong Province Higher Educational Science and Technology Program from Shandong Provincial Education Department and 2014ZC0301 from Bin zhou Science and Technology Bureau, and 2017YJKT01, 2017YJKT0103, 2017YJKT0104 from binzhou polytechnic.

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426 [8] María Laura Foresti, María del Pilar Williams, Ricardo Martínez-García, Analía Vázquez Analysis of a preferential action of α-amylase from B. licheniformis towards amorphous regions of waxy maize starch Carbohydrate Polymers, 2014, 102: 80-87. [9] Marzieh Ghollasi, Maryam Ghanbari-Safari, Khosro Khajeh Improvement of thermal stability of a mutagenised α-amylase by manipulation of the calcium-binding site Enzyme and Microbial Technology, 2013, 53 (6-7): 406-413. [10] Roohi, Mohammed Kuddus Bio-statistical approach for optimization of cold-active α-amylase production by novel psychrotolerant M. foliorum GA2 in solid state fermentation Biocatalysis and Agricultural Biotechnology, 2014, 3 (2): 175-181. [11] Shamsi Emtenani, Ahmad Asoodeh, Shirin Emtenani Gene cloning and characterization of a thermostable organic-tolerant α-amylase from Bacillus subtilis DR8806 International Journal of Biological Macromolecules, 2015, 72 290-298.

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