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利用椰子纤维废料(CPW)在固态发酵中优化和表征 Hexagonia hirta MSF2 产生的漆酶(LccH)。

Optimization and characterization of laccase (LccH) produced by Hexagonia hirta MSF2 in solid-state fermentation using coir pith wastes (CPW).

机构信息

Biocatalysts Laboratory, Department of Agricultural Microbiology, Tamil Nadu Agricultural University (TNAU), Coimbatore, 641003, Tamil Nadu, India.

Biocatalysts Laboratory, Department of Agricultural Microbiology, Tamil Nadu Agricultural University (TNAU), Coimbatore, 641003, Tamil Nadu, India.

出版信息

J Environ Manage. 2024 Apr;356:120625. doi: 10.1016/j.jenvman.2024.120625. Epub 2024 Mar 18.

Abstract

The accumulation of coir pith waste, a byproduct of coconut husk processing, poses environmental and logistical challenges. An innovative and sustainable solution involves using coir pith as a substrate for solid-state fermentation (SSF). In SSF, coir pith can be converted into valuable products, such as enzymes, organic acids, and bioactive compounds. The present study aimed to evaluate laccase production by Hexagonia hirta MSF2 through SSF using the coir pith waste as substrate. Physico-chemical parameters like moisture, pH, temperature, C source, N source, and CuSO concentrations were pre-optimized, and optimized through RSM. Laccase activity of 1585.24 U g of dry substrate was recorded by H. hirta MSF2 on coir pith containing 1 % C source, 0.5 % N source, 0.25 mM of CuSO concentration, moisture content of 75 % at pH 4.6 and temperature 28 °C. Subsequently, the enzyme extraction parameters including, extraction buffer, mode of extraction, and temperature were optimized. The molecular weight of laccase was 66 kDa as observed by SDS-PAGE and native-PAGE. The optimum activity of partially purified laccase was achieved at 40 °C, and pH 4.0. Increasing salt concentration and use of different inhibitors affected the laccase activity. Organic solvents like dimethyl sulphoxide (DMSO) and methanol, and metal ions like BaCl, CaCl, CuSO and MnCl stimulated the laccase activity. Hence, coir pith used in SSF offers a dual benefit of waste management and enzyme synthesis through an eco-friendly and cost-effective approach.

摘要

椰壳纤维废料的积累是椰子壳加工的副产品,对环境和物流构成挑战。一种创新和可持续的解决方案是将椰壳纤维废料用作固态发酵(SSF)的基质。在 SSF 中,椰壳纤维废料可以转化为有价值的产品,如酶、有机酸和生物活性化合物。本研究旨在通过 SSF 使用椰壳纤维废料作为基质来评估 Hexagonia hirta MSF2 产生漆酶的情况。对水分、pH 值、温度、C 源、N 源和 CuSO4 浓度等理化参数进行了预优化,并通过响应面法进行了优化。H. hirta MSF2 在含有 1% C 源、0.5% N 源、0.25mM CuSO4 浓度、75%水分、pH 值 4.6 和温度 28°C 的椰壳纤维废料上记录到 1585.24 U g 干基质的漆酶活性。随后,优化了酶提取参数,包括提取缓冲液、提取方式和温度。SDS-PAGE 和 native-PAGE 观察到漆酶的分子量为 66 kDa。部分纯化漆酶的最佳活性在 40°C 和 pH 值 4.0 时达到。增加盐浓度和使用不同的抑制剂会影响漆酶的活性。有机溶剂如二甲基亚砜(DMSO)和甲醇,以及金属离子如 BaCl、CaCl、CuSO4 和 MnCl 会刺激漆酶的活性。因此,在 SSF 中使用椰壳纤维废料提供了废物管理和酶合成的双重好处,采用了环保且经济有效的方法。

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