Chaudhari Sandeep A, Singhal Rekha S
Food Engineering and Technology Department, Institute of Chemical Technology, Matunga, Mumbai-400019, India.
Food Engineering and Technology Department, Institute of Chemical Technology, Matunga, Mumbai-400019, India.
Int J Biol Macromol. 2017 May;98:610-621. doi: 10.1016/j.ijbiomac.2017.02.033. Epub 2017 Feb 10.
The major hurdles in commercial exploitation of cutinase (having both esterolytic and lipolytic activities) with potent industrial applications are its high production cost, operational instability and reusability. Although commercially available in immobilized form, its immobilization process (synthesis of support/carrier) makes it expensive. Herein we tried to address multiple issues of production cost, stability, and reusability, associated with cutinase. Waste watermelon rinds, an agroindustrial waste was considered as a cheap support for solid state fermentation (SSF) for cutinase production by newly isolated Fusarium sp. ICT SAC1. Subsequently, carrier free cross-linked enzyme aggregates of cutinase (cut-CLEA) directly from the SSF crude broth were developed. All the process variables affecting CLEA formation along with the different additives were evaluated. It was found that 50% (w/v) of ammonium sulphate, 125μmol of glutaraldehyde, cross-linking for 1h at 30°C and broth pH of 7.0, yielded 58.12% activity recovery. All other additives (hexane, butyric acid, sodium dodecyl sulphate, Trition-X 100, Tween-20, BSA) evaluated presented negative results to our hypothesis. Kinetics and morphology studies confirmed the diffusive nature of cut-CLEA and BSA cut-CLEA. Developed CLEA showed better thermal, solvent, detergent and storage stability, making it more elegant and efficient for industrial biocatalytic process.
角质酶（具有酯解和脂解活性）在工业应用中具有巨大潜力，但其商业开发面临着诸多障碍，包括生产成本高、操作稳定性差以及可重复使用性低等问题。尽管角质酶有固定化形式的商业产品，但其固定化过程（载体的合成）使其价格昂贵。在此，我们试图解决与角质酶相关的生产成本、稳定性和可重复使用性等多个问题。废弃西瓜皮，一种农业工业废弃物，被视为用于新分离的镰刀菌属ICT SAC1固态发酵（SSF）生产角质酶的廉价载体。随后，直接从SSF粗发酵液中制备了无载体的角质酶交联酶聚集体（cut-CLEA）。评估了所有影响CLEA形成的工艺变量以及不同添加剂。结果发现，50%（w/v）的硫酸铵、125μmol戊二醛、在30°C下交联1小时且发酵液pH为7.0时，活性回收率为58.12%。评估的所有其他添加剂（己烷、丁酸、十二烷基硫酸钠、曲拉通X-100、吐温-20、牛血清白蛋白）均与我们的假设结果不符。动力学和形态学研究证实了cut-CLEA和BSA cut-CLEA的扩散性质。所制备的CLEA表现出更好的热稳定性、溶剂稳定性、洗涤剂稳定性和储存稳定性，使其在工业生物催化过程中更加出色和高效。
