利用部分纯化的真菌木质素酶对玉米棒进行预处理和糖化

IF 3.2 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biofuels Bioproducts & Biorefining-Biofpr Pub Date : 2024-07-02 DOI:10.1002/bbb.2661
Kandukuri Thanuja Reddy, Gurvinder Singh Kocher, Alla Singh
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引用次数: 0

摘要

玉米棒主要由结晶状态的木质纤维素材料组成,包括半纤维素、纤维素和木质素,对微生物的糖化具有抗性。利用玉米芯生产生物乙醇的尝试很少,尤其是使用化学预处理方法。本研究涉及真菌(Phanerochaete chrysosporium MTCC 787 和 Pleurotus florida PAU 22-01)胞外木质素分解酶--木质素过氧化物酶(LiP)、锰过氧化物酶(MnP)和漆酶(Lac)--的生产和纯化,然后利用它们对玉米棒进行生物预处理,并使用商业纤维素酶进行糖化。粗 LiP、MnP 和 Lac 的比活度分别为 2.23、2.1 和 2.63 U/mg.采用二乙基氨基乙基(DEAE)纤维素离子交换色谱一步法纯化粗酶,LiP、MnP 和 Lac 活性分别纯化了 11.3、10.1 和 8.62 倍,部分纯化的木质素酶的比活性分别为 25.1 U/mg(LiP)、21.2 U/mg(MnP)和 22.7 U/mg(Lac)。使用后者,在含有约 200 个单位 Lac、Lip 和 MnP 酶(在 pH 值为 6 的磷酸盐缓冲液中)的 30 毫升反应体积中对 2.5 克玉米棒进行生物预处理,结果是,使用商业纤维素酶,木质素脱除率最高达 78.4%,糖化效率为 97.1%。
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Pretreatment and saccharification of corn cobs using partially purified fungal ligninozymes

Corn cobs consist primarily of a lignocellulosic material comprising hemicellulose, cellulose, and lignin in a crystalline state, which is resistant to microbial saccharification. Bioethanol production from corn cobs has rarely been attempted, especially using chemical pretreatment methods.

The present study deals with the production and purification of fungal (Phanerochaete chrysosporium MTCC 787 and Pleurotus florida PAU 22-01) extracellular ligninolytic enzymes – lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase (Lac) – followed by their utilization for the biological pretreatment of corn cobs along with saccharification using commercial cellulase. Crude LiP, MnP, and Lac demonstrated specific activity of 2.23, 2.1, and 2.63 U/mg, respectively.

The one-step purification of crude enzyme using diethyl amino ethyl (DEAE) cellulose ion exchange chromatography resulted in 11.3, 10.1 and 8.62-fold purification of LiP, MnP and Lac activity, respectively, with corresponding specific activity of 25.1 U/mg (LiP), 21.2 U/mg (MnP) and 22.7 U/mg (Lac) in the partially purified ligninozymes. Using the latter, biological pretreatment of 2.5 g corn cobs in a reaction volume of 30 mL containing approximately 200 units of Lac, Lip and MnP enzymes (in phosphate buffer, pH 6) resulted in a maximum of 78.4% delignification with a saccharification efficiency of 97.1% using commercial cellulases.

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来源期刊
CiteScore
7.80
自引率
5.10%
发文量
122
审稿时长
4.5 months
期刊介绍: Biofuels, Bioproducts and Biorefining is a vital source of information on sustainable products, fuels and energy. Examining the spectrum of international scientific research and industrial development along the entire supply chain, The journal publishes a balanced mixture of peer-reviewed critical reviews, commentary, business news highlights, policy updates and patent intelligence. Biofuels, Bioproducts and Biorefining is dedicated to fostering growth in the biorenewables sector and serving its growing interdisciplinary community by providing a unique, systems-based insight into technologies in these fields as well as their industrial development.
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