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ROHEENA ABDULLAH, KINZA NISAR, AFSHAN KALEEM & MEHWISH IQTEDAR

Biologia (Lahore), Vol.70, Iss. 1, Pages 20-27

Abstract

The use of alpha amylase as a substitute for chemical processes(Process Optimization for the Production of Alpha Amylase by Aspergillus niger KBT-3 Using Submerged Fermentation) has gained significant momentum in the field of biotechnology. As a result, there is a growing need to explore untapped reserves of fungi with high potential for efficient alpha amylase production. This has led to the isolation of novel fungal strains capable of producing the targeted product(Process Optimization for the Production of Alpha Amylase by Aspergillus niger KBT-3 Using Submerged Fermentation). The aim of this study was to isolate a novel fungal strain and optimization of process parameters in order to enhance the alpha amylase productivity. The production of enzyme indigenously lessen reliance on external sources and promoting self-sufficiency In this study, 15 amylolytic fungal strains were isolated and screened via submerged fermentation. The strain exhibiting the highest alpha amylase activity was identified using conventional methods, including morphology and microscopic features, and confirmed by 18S rDNA gene sequencing using specific ITS primers. The selected strain was then subjected to sequencing and phylogenetic analysis to further characterize its properties. The result indicates the selected strain was found to be A. (Process Optimization for the Production of Alpha Amylase by Aspergillus niger KBT-3 Using Submerged Fermentation)niger and this strain was given code A. niger KBT-3. The Five fermentation media were also screened. The Medium 2 gave higher titer of alpha amylase activity and found to be the best medium. Different parameters including time and temperature of incubation, pH, Inoculum size, volume, carbon and nitrogen sources were also tested. Optimal enzyme production was obtained at 72 h, 30°C, pH 5.5, 1ml inoculum, 50ml volume and 1.5% lactose and 1% yeast extract.

INTRODUCTION

Alpha amylase, which is also referred to as a-1,4 glucan-glucanohydrolase (EC 3.2.1.1), is a widely distributed enzyme that breaks down starch in a variety of natural sources. The random hydrolysis of a-1,4 glucosidic links inside the starch molecule is catalyzed by this extracellular enzyme, which yields(Process Optimization for the Production of Alpha Amylase by Aspergillus niger KBT-3 Using Submerged Fermentation) both monosaccharides and oligosaccharides, such as alpha limit dextrin, glucose, and maltose (Saha et al., 2014; Soy et al., 2021). Alpha amylases are among the most important and frequently used enzymes, having a wide range of uses in pharmaceutical, clinical and analytical chemistry.
They are widely used in many different industries, including culinary(Process Optimization for the Production of Alpha Amylase by Aspergillus niger KBT-3 Using Submerged Fermentation), baking, brewing, detergent, textile, and paper manufacture, in addition to their function in starch saccharification. The need to develop quick and efficient ways in
order to produce alpha amylase is growing due to upsurge in its consumption; the production of enzyme indigenously lessen reliance on external sources and promoting self-sufficiency (Gupta et al., 2008; Balakrishnan et al., 2021; Singh et al., 2022). Numerous creatures, including plants, animals, and microbes, are sources of alpha amylase. However, production from plants and animals is constrained due to several factors. Due to the low concentration of alpha amylase obtained from plants processing(Process Optimization for the Production of Alpha Amylase by Aspergillus niger KBT-3 Using Submerged Fermentation) significant amounts of plant material is necessary. However, the availability of animal-derived enzyme is limited because it is a byproduct of the meat industry. Microbial sources, on the other hand, such as bacteria and fungi, provide a good substitute for large-scale production meeting market demands numerous bacterial and fungal strains have been widely used to produce

alpha amylase, providing a more scalable and effective alternative.
Filamentous fungi are renowned for their natural ability to secrete enzymes that break down starch and cellulose. (Process Optimization for the Production of Alpha Amylase by Aspergillus niger KBT-3 Using Submerged Fermentation)The ability to produce substantial amounts of extracellular proteins renders them a perfect option for industrial enzyme synthesis. Several species of fungi have been commonly utilized for this purpose, including Trichoderma, Thermomyces lanuginosus,
Penicillium griseoroseum, Fusarium moniliformis, Actinomycetes, and Alternaria. These fungi have been exploited for their ability to produce a broad range of enzymes (Elyasi et al., 2020; Balakrishnan et al., 2021).
One of the most important things in developing a biotechnological process is choosing an appropriate strain. (Process Optimization for the Production of Alpha Amylase by Aspergillus niger KBT-3 Using Submerged Fermentation)The ability of the strain to continuously produce high yields, its physiological stability, and incubation time needed for ideal production was consider very crucial for choosing an appropriate strain. The strain’s resistance to several environmental stresses, like temperature, aeration, and shear stress, is also an important factor (Laluce et al., 1991).
The culture process has a major influence on the amount of enzymes produced. Both submerged and solid-state fermentation methods can be used to produce alpha amylase. The submerged fermentation offers more agitation, greater aeration, and easier enzyme separation, (Process Optimization for the Production of Alpha Amylase by Aspergillus niger KBT-3 Using Submerged Fermentation)it is frequently chosen over solid-state fermentation, even if it has its own benefits. Submerged fermentation can result in increased enzyme yields and activity because the liquid culture facilitates more effective oxygen transfer and mixing. It is also easier to separate the enzyme, which makes it a more feasible option for large-scale enzyme production (Lokeswari, 2010; Fatima and Ali, 2012) Optimizing cultural conditions is crucial for maximizing enzyme production. (Process Optimization for the Production of Alpha Amylase by Aspergillus niger KBT-3 Using Submerged Fermentation)This involves carefully selecting the carbon and nitrogen sources, pH, and incubation temperature of the medium, as well as the inoculum size. Finding the optimal temperature and duration is especially crucial for getting the maximum amount of alpha amylase. Enzyme output can be greatly increased by adjusting these variables, which will increase productivity and efficiency across a range of industrial applications(Process Optimization for the Production of Alpha Amylase by Aspergillus niger KBT-3 Using Submerged Fermentation). The aim of this study was to isolate a novel fungal strain and optimization of process parameters in order to enhance the alpha amylase productivity.