Screening and partial purification of amylase from <i>Aspergillus Niger</i> isolated from deteriorated tomato (<i>Lycopersicon Esculentum</i> mill.) fruits

Abstract

Amylases (EC 3.2.1.1) are cellwall degrading enzymes associated with the pathogenicity of microorganisms in the spoilage of tomato fruits. The use of amylase in many industries has made it very important to optimize production process to achieve maximum yields. Screening and partial purification of Amylase from Aspergillus niger isolated from tomato (Lycopersicon esculentum Mill.) fruits was studied. Amylase producing fungi were isolated from fresh tomatoes kept at ambient temperature (28±1˚C). Isolates were characterized on the basis of their morphological and cultural techniques. Partial purification of amylase was carried out by ammonium sulphate precipitation. The enzyme activity was determined and optimum conditions were obtained. The molecular weights of the crude and partially purified Amylase were determined by SDS PAGE method. A total of five isolates were obtained using basic screening technique for amylase activity, one of the isolates (Isolate code F2) exhibited maximum amylase activity. The fungi isolate code F2 was identified as Aspergillus niger. Optimum conditions for Amylase AMY F2 were ascertained at pH 6.0; temperature 30°C; substrate concentration of 0.3mg/ml, and time of heating of less than 10min. The molecular weights of the crude and partially purified Amylase AMY F2 were found to be 55kDa and 35kDa respectively by SDS PAGE method. Microorganisms had been an encouraging means of economical production of enzymes in large scale for the food and drug industry.

Detergent-stable amylase production by Paenibacillus lactis strain OPSA3 isolated from soil; optimization by response surface methodology

“…These results are higher than the 1.86 purification fold, 9.33 U/mg specific activity and 63.00% yield recorded by Obafemi et al. [ 93 ] using amylase from Aspergillus niger . Almost similarly, Bhaskara Rao et al.…”

Isolation of Polygalacturonase-Producing Bacterial Strain from Tomatoes (Lycopersicon esculentum Mill.)

“…Over a long period of time, there has been an urgent need to isolate and identify microorganisms that are found to often cause spoilage of fruits, as well as to characterize the extracellular enzymes produced by the microorganisms in order to find a way to preserve the tomato fruits all year round and produce enzymes for industrial use [11]. Many pathogenic bacteria and fungi are known to produce some extracellular enzymes which can degrade the cell wall of the tomato fruits [12, 13]. The enzymes are involved in tissue breakdown of the fruits, and this play a major role in helping the microorganism to penetrate the cell wall of the plant [1, 14].…”

Biological control of fungal pathogens of tomato (Lycopersicon esculentum) by chitinolytic bacterial strains

“…After surface sterilization, plant parts were rinsed with sterilized distilled water to remove chemicals. It was then placed onto potato dextrose agar (PDA) plates containing 80% PDA [14] and then incubated at 28°C for 2-3 days. The fungal isolates were morphologically identified based on their infected plant parts, colony growth, and spore morphology under a microscope (http://vegetablemdonline.ppath.cornell.edu).…”