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Thermo-alkali-stable lipase from a novel Aspergillus niger: statistical optimization, enzyme purification, immobilization and its application in biodiesel production
Preparative Biochemistry & Biotechnology ( IF 2.9 ) Pub Date : 2020-08-18 , DOI: 10.1080/10826068.2020.1805759
Dina H El-Ghonemy 1 , Thanaa H Ali 1 , Naziha M Hassanein 2 , Eman M Abdellah 1 , Mohamed Fadel 1 , Ghada E A Awad 3 , Dalia A M Abdou 2
Affiliation  

Abstract

The influences of nutritional components affecting lipase production from the new Aspergillus niger using wheat bran as substrate were studied by employing Plackett–Burman and central composite statistical designs. Out of the 11 medium components tested, sucrose, KH2PO4 and MgSO4 at final concentrations of 3.0, 1.0 and 0.5 g/L, respectively, were reported to contribute positively to enzyme production (20.09 ± 0.98 U/g ds). The enzyme was purified through ammonium sulfate precipitation followed by Sephadex G-100 gel filtration. Molecular mass of the purified lipase was 57 kDa as evident on SDS-PAGE. Different methods of immobilization were studied and the highest immobilization yield of 81.7 ± 2.18% was reported with agarose (2%) and the optimum temperature was raised from 45 to 50 °C. Immobilized lipase could retain 80% of its original activity at 60 °C after 1 hr of incubation, and was stable at pH values between neutral and alkaline pH. Lipase-catalyzed transesterification process of fungal oil resulted in a fatty acid methyl ester yield consisting of a high percentage of polyunsaturated fatty acids (83.6%), making it appropriate to be used as winter-grade biodiesel. The operational stability studies revealed that the immobilized lipase could keep 70% of its total activity after 5 cycles of the transesterification process.



中文翻译:

一种新型黑曲霉的热碱稳定脂肪酶:统计优化,酶纯化,固定化及其在生物柴油生产中的应用

抽象的

采用Plackett-Burman和中心综合统计设计,研究了营养成分对以小麦麸为底物的新型黑曲霉产生脂肪酶的影响。在测试的11种培养基成分中,蔗糖,KH 2 PO 4和MgSO 4据报道,最终浓度分别为3.0、1.0和0.5 g / L对酶的产生有积极作用(20.09±0.98 U / g ds)。通过硫酸铵沉淀,然后用Sephadex G-100凝胶过滤纯化酶。如在SDS-PAGE上明显的,纯化的脂肪酶的分子量为57kDa。研究了不同的固定方法,使用琼脂糖(2%)报道的最高固定产率为81.7±2.18%,最佳温度从45°C升高至50°C。固定的脂肪酶在孵育1小时后,在60°C下可以保留其80%的原始活性,并且在中性和碱性pH值之间保持稳定。脂肪酶催化的真菌油酯交换过程导致脂肪酸甲酯的收率较高,多百分比的多不饱和脂肪酸占83.6%,使其适合用作冬季级生物柴油。操作稳定性研究表明,固定化脂肪酶在酯交换过程的5个循环后可以保持其总活性的70%。

更新日期:2020-08-18
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