Isolation and Screening of Aspergillus niger and Bacillus coagulans as Potential Candidates for Amylase and Glucose Isomerase Production

Musa, B., Zangina, B. U., Ado, S. A., Hussaini, I. M., Madika, A., Aliyu, M. S.
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Abstract

There is an increasing demand for high fructose corn syrup as an alternative to glucose especially for use by people with diabetes due to its low glycemic index. The high cost of this product coupled with its high demand has attracted the attention of many researchers to search for an alternative and sustainable production route. Therefore, the focus of this work was isolating and screening Aspergillus niger and Bacillus coagulans as possible producers of amylase and glucose isomerase respectively, which are needed to produce high fructose corn syrup. Nine (9) samples of loamy soil were taken from three (3) separate locations (BG = Botanical Garden, RD = Refuse Dumpsite, FB = Flower Bed) to isolate the bacterium and fungus. Based on the lacto-phenol cotton blue stain preparation under a microscope, colonies that were thought to be Aspergillus niger colonies were observed and further identified. Cultural, microscopic, and biochemical characteristics were used to confirm the identification of colonies that were thought to be Bacillus coagulans. The probable isolates of Aspergillus niger and Bacillus coagulans were then screened for amylase and glucose isomerase production respectively. Out of the total of nine (9) soil samples analysed, 5 (55.6 %) were positive for Aspergillus niger. A higher isolation rate of Aspergillus niger, 2 (66.7 %) was recorded in soil samples from botanical garden (BG) and flower bed (FB). The lowest isolation rate, 1 (33.3 %) was observed in soil obtained from the refuse dump. Out of all the isolates screened for their potential to produce amylase, the isolate from the botanical garden showed the highest zone of starch hydrolysis (28 mm), and the isolate from the flower bed showed the least zone of hydrolysis of starch (13 mm). On the other hand, out of the nine (9) soil samples analysed, 6 (66.7 %) were positive for Bacillus coagulans and the highest occurrence of Bacillus coagulans, 3 (100 %) was recorded with soil from botanical garden (BG1), while the least occurrence, 1 (33.3 %) was observed in soil from flower bed (FB2). The probable isolates of Bacillus coagulans screened for glucose isomerase production revealed that all the six (6) isolates produced glucose isomerase, with isolate from refuse dump (RD2) producing the highest concentration of glucose isomerase (4.7014 g/L).
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作为淀粉酶和葡萄糖异构酶潜在候选菌的黑曲霉和凝固芽孢杆菌的分离和筛选
由于高果糖玉米糖浆的血糖指数低,人们对其作为葡萄糖替代品的需求越来越大,尤其是糖尿病患者。该产品的高成本和高需求引起了许多研究人员的注意,寻找替代和可持续的生产路线。因此,本研究的重点是分离和筛选生产高果糖玉米糖浆所需的淀粉酶和葡萄糖异构酶的黑曲霉和凝固芽孢杆菌。从3个不同地点(BG =植物园,RD =垃圾场,FB =花坛)采集9份壤土样品分离细菌和真菌。基于在显微镜下制备的乳酚棉蓝染色,观察并进一步鉴定了被认为是黑曲霉菌落的菌落。培养、显微镜和生化特征被用来确认被认为是凝固芽孢杆菌的菌落的鉴定。然后对可能分离的黑曲霉和凝固芽孢杆菌分别进行淀粉酶和葡萄糖异构酶的筛选。在分析的9份土壤样本中,5份(55.6%)呈黑曲霉阳性。植物园(BG)和花坛(FB)土壤样品中黑曲霉2的分离率较高(66.7%)。垃圾填埋场土壤的隔离率最低,为1(33.3%)。在所有具有淀粉酶潜力的分离株中,来自植物园的分离株淀粉水解区最高(28 mm),而来自花坛的分离株淀粉水解区最低(13 mm)。在9份土壤样品中,6份(66.7%)为凝固芽孢杆菌阳性,其中植物园(BG1)土壤中凝固芽孢杆菌的检出率最高,为3份(100%),花圃(FB2)土壤中最少,为1份(33.3%)。对6株凝固性芽孢杆菌进行葡萄糖异构酶的筛选,结果表明,6株凝固性芽孢杆菌均能产生葡萄糖异构酶,其中垃圾填埋场(RD2)的凝固性芽孢杆菌产生葡萄糖异构酶的浓度最高,为4.7014 g/L。
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