Saeed Saba, Shakir Hafiz Abdullah, Qazi Javed Iqbal
Institute of Zoology, University of the Punjab, 54590, Lahore, Punjab, Pakistan.
J Microbiol Methods. 2025 Aug 7;237:107208. doi: 10.1016/j.mimet.2025.107208.
The current study aimed to optimize the amylase producing strain using agro-industrial waste under submerged fermentation. Of six bacterial strains, five gave positive results for amylase production but Bacillus amyloliquefaciens was selected due to highest enzyme production. The medium components were selected using Plackett-Burman design (PBD) while concentration of medium components was determined using the central composite design (CCD) of response surface methodology (RSM). The maximum enzyme production was obtained from mango peels (1.25 %), sodium chloride (0.15 %), potassium nitrate (0.06 %), and magnesium sulfate (0.03 %). It was observed that B. amyloliquefaciens produce maximum amylase at 45 °C, pH 5, and 2 % inoculum size over 24-h incubation period. Furthermore, partial characterization revealed best enzyme activity at 9 pH, 50 °C, 1 % substrate concentration, and 30 min of incubation. Besides, amylase was found more stable at 6 pH and 60 ᴼC. In the presence of metal ions, the enzyme activity decreased by Cu (5 mM) and found maximum in Ca (1 mM). With the increase in concentration of inhibitors, the amylase activity of ethylenediaminetetraacetic acid (EDTA) and 1, 10-phenanthroline decreases. Among organic solvents, surfactants, and oxidizing agents maximum activity was observed for sodium dodecylsulphate (SDS) and hydrogen peroxide (HO). These findings highlight the potential of B. amyloliquefaciens as a promising candidate for sustainable and cost-effective industrial amylase production using agro-industrial waste and can be further explored for scale-up studies.
本研究旨在利用农工业废料在深层发酵条件下优化淀粉酶产生菌株。在六种细菌菌株中,有五种菌株产生淀粉酶呈阳性结果,但由于淀粉酶产量最高,选择了解淀粉芽孢杆菌。使用Plackett-Burman设计(PBD)选择培养基成分,同时使用响应面法(RSM)的中心复合设计(CCD)确定培养基成分的浓度。从芒果皮(1.25%)、氯化钠(0.15%)、硝酸钾(0.06%)和硫酸镁(0.03%)中获得了最大酶产量。观察到解淀粉芽孢杆菌在45℃、pH 5和2%接种量下,经过24小时培养产生的淀粉酶最多。此外,部分特性分析表明,在pH 9、50℃、1%底物浓度和30分钟培养时间下酶活性最佳。此外,发现淀粉酶在pH 6和60℃时更稳定。在金属离子存在的情况下,酶活性因铜(5 mM)而降低,在钙(1 mM)存在时酶活性最高。随着抑制剂浓度的增加,乙二胺四乙酸(EDTA)和1,10-菲咯啉的淀粉酶活性降低。在有机溶剂、表面活性剂和氧化剂中,观察到十二烷基硫酸钠(SDS)和过氧化氢(HO)的活性最高。这些发现突出了解淀粉芽孢杆菌作为利用农工业废料进行可持续且具有成本效益的工业淀粉酶生产的有前景候选菌株的潜力,并且可以进一步进行放大研究。