Department of Biochemistry, College of Basic Sciences and Humanities, CCS Haryana Agricultural University, Hisar, Haryana, 125004, India.
Department of Biotechnology, School of Interdisciplinary and Applied Sciences, Central University of Haryana, Mahendergarh, Haryana, 123031, India.
Bioprocess Biosyst Eng. 2024 Jan;47(1):39-55. doi: 10.1007/s00449-023-02943-9. Epub 2023 Nov 14.
Phytase enzyme found in plants, animals, and microorganisms is mainly involved in catalyzing the systematic removal of a phosphate group from phytic acid. Enzyme immobilization is one of the cost-effective methods for the wide usage of enzymes in the industrial sector. This paper reports the covalent immobilization of phytase on glutaraldehyde-activated aluminum oxide beads. The immobilization yield, efficiency, and activation energy were found to be 47.8%, 71.5%, and 15.78 J/mol, respectively. The bound enzyme displayed a shift in pH optima from 5.5 to 4.5, which is more beneficial to increase digestibility in comparison with the free enzyme. Immobilized phytase retained 42.60% of its activity after 1.0 h incubation at 80 °C, whereas free enzyme retained only 4.20% of its activity. Thermodynami increase in half-lives, D-values, enthalpy and free energy change after covalent immobilization could be credited to the enhanced stability. Immobilized phytase could be reused for five consecutive cycles retaining 51% of its initial activity with sodium phytate. The immobilized phytase was also found effective to hydrolyze the soybean meal, thus increasing the digestibility of poultry feed. The hydrolyzing reaction of soybean meal was carried out for six consecutive cycles and immobilized phytase retained nearly 50% of activity till the fifth cycle. The amount of phosphorus released after treatment with immobilized phytase was far higher than that from free phytase. Immobilization on this support is significant, as this support can sustain high mechanical resistance at high pH and temperature. This considerable stability and reusability of the bound enzyme may be advantageous for its industrial application.
植酸酶存在于植物、动物和微生物中,主要参与催化系统地从植酸中去除磷酸基团。酶固定化是在工业领域广泛应用酶的一种具有成本效益的方法。本文报道了植酸酶通过戊二醛活化的氧化铝珠的共价固定化。固定化产率、效率和活化能分别为 47.8%、71.5%和 15.78 J/mol。结合酶的 pH 最适值从 5.5 转移到 4.5,与游离酶相比,这更有利于提高消化率。固定化植酸酶在 80°C 孵育 1.0 小时后保留了其初始活性的 42.60%,而游离酶仅保留了其初始活性的 4.20%。固定化后半衰期、D 值、焓和自由能变化的热力学增加可以归因于稳定性的提高。固定化植酸酶可以在连续五个循环中重复使用,用植酸钠保留其初始活性的 51%。固定化植酸酶也被发现可以有效地水解豆粕,从而提高家禽饲料的消化率。豆粕的水解反应连续进行了六个循环,固定化植酸酶在第五个循环中保留了近 50%的活性。用固定化植酸酶处理后释放的磷量远远高于游离植酸酶。在这种支持物上进行固定化是有意义的,因为这种支持物可以在高 pH 值和高温下承受高机械阻力。结合酶的这种相当大的稳定性和可重复使用性可能有利于其工业应用。